Nutrient chapter

Nattokinase / subtilisin NAT

The purified mature 275-residue bacterial subtilisin-family serine protease of Bacillus subtilis var. natto, sequence-derived mass 27,728 Da, EC 3.4.21.62. This entity is the purified enzyme only. Natto the food, the crude fermentation powder, the NSK-SD product and a generic supplement are separate entities in this chapter because they differ in composition in ways that change the answer to a clinical question.

130 recorded mechanisms · 4 availability situations · 2 preserved sources. Draft and verified records are labeled separately.

Read recorded mechanisms Open recursive mind map

The mechanisms

What the sources say this nutrient does, one relationship at a time. Plain wording comes first; the technical statement follows.

A strong fibrinolytic activity averaging about 40 CU, in plasmin units, per gram wet weight was demonstrated in natto, and the enzyme responsible was extracted with saline and named nattokinase.
Natto, fermented soybean food → Nattokinase / subtilisin NAT source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Natto, fermented soybean food
exposure
Not applicable
limitations
The discovery report. Its estimates of molecular weight, about 20,000, and isoelectric point, 8.6, predate the sequence and are superseded by the 275-residue chain of 27,728 Da. Activity is given in CU, plasmin units, which is a different unit system from the FU used by every product in this chapter and is not convertible to it. Diisopropyl fluorophosphate strongly inhibited the activity, consistent with the serine-protease assignment made later.
organism
Natto, fermented soybean food
plain_language
A strong fibrinolytic activity averaging about 40 CU, in plasmin units, per gram wet weight was demonstrated in natto, and the enzyme responsible was extracted with saline and named nattokinase.
primary_references
A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet. (1987) https://pubmed.ncbi.nlm.nih.gov/3478223/ DOI: 10.1007/BF01956052
route
In vitro
tissue
Saline extract of the food
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 26–26
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Natto, fermented soybean food · source_derived_draft · unverified_draft
A strong fibrinolytic activity averaging about 40 CU, in plasmin units, per gram wet weight was demonstrated in natto, and the enzyme responsible was extracted with saline and named nattokinase.
Complete structured claim and evidence
The aprN gene of Bacillus subtilis (natto) encodes the subtilisin NAT preproenzyme.
Bacillus subtilis natto aprN gene → Subtilisin NAT preproenzyme source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Bacillus subtilis var. natto
exposure
Not applicable
limitations
PubMed carries no abstract for this record, so only the nucleotide-sequence result named in the title is taken from it. The commonly quoted 29-residue signal peptide and 77-residue propeptide lengths are not sourced by anything read during this curation and are recorded as an open question instead.
organism
Bacillus subtilis var. natto
plain_language
The aprN gene of Bacillus subtilis (natto) encodes the subtilisin NAT preproenzyme.
primary_references
Nucleotide sequence of the subtilisin NAT gene, aprN, of Bacillus subtilis (natto). (1992) https://pubmed.ncbi.nlm.nih.gov/1369081/ DOI: 10.1271/bbb.56.1869
route
In vitro
tissue
Bacterial gene expression
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 37–37
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Bacillus subtilis var. natto · source_derived_draft · unverified_draft
The aprN gene of Bacillus subtilis (natto) encodes the subtilisin NAT preproenzyme.
Complete structured claim and evidence
The mature enzyme purified from natto is a single 275-residue polypeptide of 27,728 Da that is highly homologous to the subtilisins.
Subtilisin NAT preproenzyme → Nattokinase / subtilisin NAT source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Bacillus subtilis var. natto
exposure
Not applicable
limitations
Sequence determined by Edman degradation of the uncleaved molecule and an enzymatically derived peptide. The 1987 discovery report estimated the mass at about 20,000 by a different method; the sequence-derived value supersedes it.
organism
Bacillus subtilis var. natto
plain_language
The mature enzyme purified from natto is a single 275-residue polypeptide of 27,728 Da that is highly homologous to the subtilisins.
primary_references
Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan. (1993) https://pubmed.ncbi.nlm.nih.gov/8280151/ DOI: 10.1006/bbrc.1993.2624
route
In vitro
tissue
Purified protein
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 48–48
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Bacillus subtilis var. natto · source_derived_draft · unverified_draft
The mature enzyme purified from natto is a single 275-residue polypeptide of 27,728 Da that is highly homologous to the subtilisins.
Complete structured claim and evidence
Phenylmethylsulfonyl fluoride inhibited both the fibrinolytic and the amidolytic activity of the purified enzyme, placing it in the serine-protease class.
Phenylmethylsulfonyl fluoride → Nattokinase / subtilisin NAT source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified nattokinase
exposure
PMSF
limitations
The most sensitive synthetic substrate was succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, a subtilisin substrate. Inhibitor sensitivity identifies the catalytic class, not the physiological substrate.
organism
Purified nattokinase
plain_language
Phenylmethylsulfonyl fluoride inhibited both the fibrinolytic and the amidolytic activity of the purified enzyme, placing it in the serine-protease class.
primary_references
Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan. (1993) https://pubmed.ncbi.nlm.nih.gov/8280151/ DOI: 10.1006/bbrc.1993.2624
route
In vitro
tissue
Enzyme assay
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 59–59
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified nattokinase · source_derived_draft · unverified_draft
Phenylmethylsulfonyl fluoride inhibited both the fibrinolytic and the amidolytic activity of the purified enzyme, placing it in the serine-protease class.
Complete structured claim and evidence
Alanine substitution at Ser33, Asp60, Ser62 or Thr220 raised the free energy of the transition state and lowered kcat.
Nattokinase Ser33Ala, Asp60Ala, Ser62Ala and Thr220Ala mutants → Nattokinase catalytic turnover on a model peptide substrate source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Recombinant subtilisin NK mutants
exposure
Ser33Ala, Asp60Ala, Ser62Ala and Thr220Ala single mutants
limitations
Measured against succinyl-Ala-Ala-Pro-Phe-p-nitroanilide. The exact kinetic constants were not extracted for this ledger, and an effect size on a chromogenic model substrate does not transfer to fibrin, PAI-1 or von Willebrand factor.
organism
Recombinant subtilisin NK mutants
plain_language
Alanine substitution at Ser33, Asp60, Ser62 or Thr220 raised the free energy of the transition state and lowered kcat.
primary_references
Probing the importance of hydrogen bonds in the active site of the subtilisin nattokinase by site-directed mutagenesis and molecular dynamics simulation. (2006) https://pubmed.ncbi.nlm.nih.gov/16411898/ DOI: 10.1042/BJ20050772
route
In vitro
tissue
Enzyme kinetics with molecular dynamics and free-energy perturbation
trigger_kind
machinery_impairment Imported condition classification; unverified.
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 70–70
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Recombinant subtilisin NK mutants · source_derived_draft · unverified_draft
Alanine substitution at Ser33, Asp60, Ser62 or Thr220 raised the free energy of the transition state and lowered kcat.
Complete structured claim and evidence
The same four substitutions left Km essentially unchanged, so the bonds support catalysis rather than ground-state substrate binding.
Nattokinase Ser33Ala, Asp60Ala, Ser62Ala and Thr220Ala mutants → Nattokinase ground-state substrate binding source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Recombinant subtilisin NK mutants
exposure
Ser33Ala, Asp60Ala, Ser62Ala and Thr220Ala single mutants
limitations
A measured null on one model substrate. Simulation partially released Asp32, His64 and Asn155 in the mutants, which is the proposed explanation rather than a separate measurement.
organism
Recombinant subtilisin NK mutants
plain_language
The same four substitutions left Km essentially unchanged, so the bonds support catalysis rather than ground-state substrate binding.
primary_references
Probing the importance of hydrogen bonds in the active site of the subtilisin nattokinase by site-directed mutagenesis and molecular dynamics simulation. (2006) https://pubmed.ncbi.nlm.nih.gov/16411898/ DOI: 10.1042/BJ20050772
route
In vitro
tissue
Enzyme kinetics with molecular dynamics and free-energy perturbation
trigger_kind
machinery_impairment Imported condition classification; unverified.
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 81–81
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Recombinant subtilisin NK mutants · source_derived_draft · unverified_draft
The same four substitutions left Km essentially unchanged, so the bonds support catalysis rather than ground-state substrate binding.
Complete structured claim and evidence
The deposited 1.36 A subtilisin NAT structure models four calcium ions across its two chains, and the 1.74 A native structure is a monomeric assembly of one 275-residue chain.
Nattokinase / subtilisin NAT → Calcium ion source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
RCSB Protein Data Bank entry metadata reviewed. No functional assay is attached to a structure record.
experimental_model
Bacillus subtilis var. natto
exposure
Not applicable
limitations
A deposited crystal structure is not a solution stoichiometry. Calcium occupancy here is what was modelled into density, and no functional calcium requirement is measured by these entries.
organism
Bacillus subtilis var. natto
plain_language
The deposited 1.36 A subtilisin NAT structure models four calcium ions across its two chains, and the 1.74 A native structure is a monomeric assembly of one 275-residue chain.
primary_references
[rcsb-3vyv] RCSB PDB entry 3VYV: crystal structure of subtilisin NAT at 1.36 A. (2013) https://www.rcsb.org/structure/3VYV Not indexed in PubMed.
route
In vitro
tissue
X-ray crystallography
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 92–92
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Bacillus subtilis var. natto · source_derived_draft · unverified_draft
The deposited 1.36 A subtilisin NAT structure models four calcium ions across its two chains, and the 1.74 A native structure is a monomeric assembly of one 275-residue chain.
Complete structured claim and evidence
Purified nattokinase cleaves cross-linked fibrin directly in a clot-lysis assay.
Nattokinase / subtilisin NAT → Cross-linked fibrin source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Bibliographic record and the EFSA opinion's account of this study reviewed. The primary article itself was not retrieved and is not indexed in PubMed.
experimental_model
Human protein substrates
exposure
Purified nattokinase
limitations
The primary report is in a journal PubMed does not index for this article and was not retrieved; the result is corroborated by the EFSA opinion, which cites the same work for the cross-linked fibrin finding. No validated preference map across fibrin alpha, beta, gamma and gamma-gamma chains under clinically relevant conditions was available in the sources read here.
organism
Human protein substrates
plain_language
Purified nattokinase cleaves cross-linked fibrin directly in a clot-lysis assay.
primary_references
[fujita-fibrin-1995] Characterization of nattokinase-degraded products from human fibrinogen or cross-linked fibrin. (1995) Not indexed in PubMed.
route
In vitro
tissue
Fibrin clot
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 103–103
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human protein substrates · source_derived_draft · unverified_draft
Purified nattokinase cleaves cross-linked fibrin directly in a clot-lysis assay.
Complete structured claim and evidence
Purified nattokinase proteolyzes human fibrinogen.
Nattokinase / subtilisin NAT → Human fibrinogen source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Bibliographic record and the EFSA opinion's account of this study reviewed. The primary article itself was not retrieved and is not indexed in PubMed.
experimental_model
Human protein substrates
exposure
Purified nattokinase
limitations
Same access limitation as the fibrin record. The 1993 purification paper independently reports that the enzyme digests fibrin.
organism
Human protein substrates
plain_language
Purified nattokinase proteolyzes human fibrinogen.
primary_references
[fujita-fibrin-1995] Characterization of nattokinase-degraded products from human fibrinogen or cross-linked fibrin. (1995) Not indexed in PubMed.
route
In vitro
tissue
Purified fibrinogen
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 114–114
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human protein substrates · source_derived_draft · unverified_draft
Purified nattokinase proteolyzes human fibrinogen.
Complete structured claim and evidence
The enhancement of clot lysis seen without added PAI-1 is attributed to direct fibrin dissolution by the enzyme rather than to plasminogen activation.
Nattokinase / subtilisin NAT → Human plasminogen source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
In vitro fibrin clot system
exposure
Subtilisin NAT, 0.06-1 nM
limitations
This is the authors' attribution of a partition between two arms of the same assay, not a plasminogen-depleted control. A plasminogen-free system was not reported.
organism
In vitro fibrin clot system
plain_language
The enhancement of clot lysis seen without added PAI-1 is attributed to direct fibrin dissolution by the enzyme rather than to plasminogen activation.
primary_references
The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis Cleaves and inactivates plasminogen activator inhibitor type 1. (2001) https://pubmed.ncbi.nlm.nih.gov/11325965/ DOI: 10.1074/jbc.M101751200
route
In vitro
tissue
tPA-induced clot lysis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 125–125
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · In vitro fibrin clot system · source_derived_draft · unverified_draft
The enhancement of clot lysis seen without added PAI-1 is attributed to direct fibrin dissolution by the enzyme rather than to plasminogen activation.
Complete structured claim and evidence
Subtilisin NAT cleaved active recombinant PAI-1 at its reactive-site Arg346-Met347 bond and PAI-1 lost specific activity dose-dependently over 0.02-1.0 nM, with a half-maximal effect near 0.1 nM.
Nattokinase / subtilisin NAT → Human SERPINE1 / plasminogen activator inhibitor 1 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Recombinant human PAI-1
exposure
Subtilisin NAT, 0.02-1.0 nM
limitations
The substrate was active recombinant prokaryotic PAI-1. Whether latent or vitronectin-bound PAI-1 is cleaved was not established by this work or by anything else read here.
organism
Recombinant human PAI-1
plain_language
Subtilisin NAT cleaved active recombinant PAI-1 at its reactive-site Arg346-Met347 bond and PAI-1 lost specific activity dose-dependently over 0.02-1.0 nM, with a half-maximal effect near 0.1 nM.
primary_references
The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis Cleaves and inactivates plasminogen activator inhibitor type 1. (2001) https://pubmed.ncbi.nlm.nih.gov/11325965/ DOI: 10.1074/jbc.M101751200
route
In vitro
tissue
Purified-protein proteolysis with MALDI-TOF and peptide sequencing
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 136–136
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Recombinant human PAI-1 · source_derived_draft · unverified_draft
Subtilisin NAT cleaved active recombinant PAI-1 at its reactive-site Arg346-Met347 bond and PAI-1 lost specific activity dose-dependently over 0.02-1.0 nM, with a half-maximal effect near 0.1 nM.
Complete structured claim and evidence
At 1 nM and without added PAI-1, subtilisin NAT enhanced tPA-induced fibrin clot lysis by 48 +/- 1.4%.
Nattokinase / subtilisin NAT → Tissue-plasminogen-activator-induced fibrin clot lysis in vitro source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
In vitro fibrin clot system
exposure
Subtilisin NAT, 1 nM
limitations
Paired with the PAI-1-present arm below. The difference between the two is consistent with an added PAI-1 arm but is not a formal component-additivity model.
organism
In vitro fibrin clot system
plain_language
At 1 nM and without added PAI-1, subtilisin NAT enhanced tPA-induced fibrin clot lysis by 48 +/- 1.4%.
primary_references
The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis Cleaves and inactivates plasminogen activator inhibitor type 1. (2001) https://pubmed.ncbi.nlm.nih.gov/11325965/ DOI: 10.1074/jbc.M101751200
route
In vitro
tissue
tPA-induced clot lysis, PAI-1 absent
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 147–147
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · In vitro fibrin clot system · source_derived_draft · unverified_draft
At 1 nM and without added PAI-1, subtilisin NAT enhanced tPA-induced fibrin clot lysis by 48 +/- 1.4%.
Complete structured claim and evidence
At the same 1 nM and with active PAI-1 present, the same enzyme enhanced tPA-induced clot lysis by 78 +/- 2.0%.
Nattokinase / subtilisin NAT → Tissue-plasminogen-activator-induced fibrin clot lysis in vitro source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
In vitro fibrin clot system
exposure
Subtilisin NAT, 1 nM
limitations
The stronger enhancement when PAI-1 is present is the evidence for a second, inhibitor-directed arm. It is one assay in one laboratory.
organism
In vitro fibrin clot system
plain_language
At the same 1 nM and with active PAI-1 present, the same enzyme enhanced tPA-induced clot lysis by 78 +/- 2.0%.
primary_references
The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis Cleaves and inactivates plasminogen activator inhibitor type 1. (2001) https://pubmed.ncbi.nlm.nih.gov/11325965/ DOI: 10.1074/jbc.M101751200
route
In vitro
tissue
tPA-induced clot lysis, active recombinant PAI-1 present
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 158–158
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · In vitro fibrin clot system · source_derived_draft · unverified_draft
At the same 1 nM and with active PAI-1 present, the same enzyme enhanced tPA-induced clot lysis by 78 +/- 2.0%.
Complete structured claim and evidence
Nattokinase directly cleaved the folded von Willebrand factor A2 domain and the unstructured vWF73 peptide under static conditions, with Thr1608-Gly1609 as the primary site.
Nattokinase / subtilisin NAT → Human von Willebrand factor / VWF source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified human von Willebrand factor constructs
exposure
Purified nattokinase
limitations
A purified-substrate system. The cleaved bond sits three residues downstream of the ADAMTS13 site Tyr1605-Met1606. No multimer endpoint and no human oral exposure were measured.
organism
Purified human von Willebrand factor constructs
plain_language
Nattokinase directly cleaved the folded von Willebrand factor A2 domain and the unstructured vWF73 peptide under static conditions, with Thr1608-Gly1609 as the primary site.
primary_references
Degradation mechanism of the von Willebrand factor A2 domain by nattokinase. (2026) https://pubmed.ncbi.nlm.nih.gov/42163571/ DOI: 10.1002/1873-3468.70366
route
In vitro
tissue
In vitro enzymology with mass spectrometry
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 169–169
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified human von Willebrand factor constructs · source_derived_draft · unverified_draft
Nattokinase directly cleaved the folded von Willebrand factor A2 domain and the unstructured vWF73 peptide under static conditions, with Thr1608-Gly1609 as the primary site.
Complete structured claim and evidence
Nattokinase showed a catalytic efficiency on the tested vWF A2 substrate about 200-fold that of ADAMTS13.
Nattokinase / subtilisin NAT → Human ADAMTS13 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified human von Willebrand factor constructs
exposure
Purified nattokinase compared with ADAMTS13
limitations
A kcat/Km ratio on one substrate under one set of conditions. It is not a statement about systemic potency after oral dosing, where the enzyme concentration reaching plasma is unmeasured.
organism
Purified human von Willebrand factor constructs
plain_language
Nattokinase showed a catalytic efficiency on the tested vWF A2 substrate about 200-fold that of ADAMTS13.
primary_references
Degradation mechanism of the von Willebrand factor A2 domain by nattokinase. (2026) https://pubmed.ncbi.nlm.nih.gov/42163571/ DOI: 10.1002/1873-3468.70366
route
In vitro
tissue
In vitro kinetics
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 180–180
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified human von Willebrand factor constructs · source_derived_draft · unverified_draft
Nattokinase showed a catalytic efficiency on the tested vWF A2 substrate about 200-fold that of ADAMTS13.
Complete structured claim and evidence
Unlike ADAMTS13, nattokinase cleaved the A2 domain without the shear-dependent substrate unfolding that exposes the ADAMTS13 scissile bond.
Nattokinase / subtilisin NAT → Shear-induced unfolding of the von Willebrand factor A2 domain source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified human von Willebrand factor constructs
exposure
Purified nattokinase
limitations
Establishes independence from unfolding in this assay. It does not establish what happens to circulating multimers, where local shear and platelet binding also apply.
organism
Purified human von Willebrand factor constructs
plain_language
Unlike ADAMTS13, nattokinase cleaved the A2 domain without the shear-dependent substrate unfolding that exposes the ADAMTS13 scissile bond.
primary_references
Degradation mechanism of the von Willebrand factor A2 domain by nattokinase. (2026) https://pubmed.ncbi.nlm.nih.gov/42163571/ DOI: 10.1002/1873-3468.70366
route
In vitro
tissue
Static in vitro cleavage
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 191–191
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified human von Willebrand factor constructs · source_derived_draft · unverified_draft
Unlike ADAMTS13, nattokinase cleaved the A2 domain without the shear-dependent substrate unfolding that exposes the ADAMTS13 scissile bond.
Complete structured claim and evidence
Nattokinase degraded amyloid fibrils in vitro, and the study determined the conditions under which it does so.
Nattokinase / subtilisin NAT → Amyloid-beta fibrils, sequence specified by study source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified fibril preparations
exposure
Nattokinase
limitations
In vitro proteolysis of a purified fibril preparation. It says nothing about oral dosing, brain entry or clinical benefit in any amyloid disease.
organism
Purified fibril preparations
plain_language
Nattokinase degraded amyloid fibrils in vitro, and the study determined the conditions under which it does so.
primary_references
Amyloid-degrading ability of nattokinase from Bacillus subtilis natto. (2009) https://pubmed.ncbi.nlm.nih.gov/19117402/ DOI: 10.1021/jf803072r
route
In vitro
tissue
In vitro fibril proteolysis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 202–202
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified fibril preparations · source_derived_draft · unverified_draft
Nattokinase degraded amyloid fibrils in vitro, and the study determined the conditions under which it does so.
Complete structured claim and evidence
Subtilisin Carlsberg degraded the same amyloid fibrils.
Subtilisin Carlsberg → Amyloid-beta fibrils, sequence specified by study source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified fibril preparations
exposure
Subtilisin Carlsberg
limitations
Recorded because it removes the uniqueness of the nattokinase result, not because subtilisin Carlsberg is of interest here in itself.
organism
Purified fibril preparations
plain_language
Subtilisin Carlsberg degraded the same amyloid fibrils.
primary_references
Amyloid-degrading ability of nattokinase from Bacillus subtilis natto. (2009) https://pubmed.ncbi.nlm.nih.gov/19117402/ DOI: 10.1021/jf803072r
route
In vitro
tissue
In vitro fibril proteolysis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 213–213
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified fibril preparations · source_derived_draft · unverified_draft
Subtilisin Carlsberg degraded the same amyloid fibrils.
Complete structured claim and evidence
Proteinase K degraded the same amyloid fibrils.
Proteinase K → Amyloid-beta fibrils, sequence specified by study source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified fibril preparations
exposure
Proteinase K
limitations
As above.
organism
Purified fibril preparations
plain_language
Proteinase K degraded the same amyloid fibrils.
primary_references
Amyloid-degrading ability of nattokinase from Bacillus subtilis natto. (2009) https://pubmed.ncbi.nlm.nih.gov/19117402/ DOI: 10.1021/jf803072r
route
In vitro
tissue
In vitro fibril proteolysis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 224–224
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified fibril preparations · source_derived_draft · unverified_draft
Proteinase K degraded the same amyloid fibrils.
Complete structured claim and evidence
Plasmin did not degrade the fibrils that nattokinase, proteinase K and subtilisin Carlsberg degraded.
Plasmin → Amyloid-beta fibrils, sequence specified by study source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified fibril preparations
exposure
Plasmin
limitations
This measured null is the informative half of the comparison: the activity tracks the subtilisin family rather than proteolysis in general.
organism
Purified fibril preparations
plain_language
Plasmin did not degrade the fibrils that nattokinase, proteinase K and subtilisin Carlsberg degraded.
primary_references
Amyloid-degrading ability of nattokinase from Bacillus subtilis natto. (2009) https://pubmed.ncbi.nlm.nih.gov/19117402/ DOI: 10.1021/jf803072r
route
In vitro
tissue
In vitro fibril proteolysis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 235–235
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified fibril preparations · source_derived_draft · unverified_draft
Plasmin did not degrade the fibrils that nattokinase, proteinase K and subtilisin Carlsberg degraded.
Complete structured claim and evidence
Trypsin likewise did not degrade the fibrils.
Trypsin → Amyloid-beta fibrils, sequence specified by study source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified fibril preparations
exposure
Trypsin
limitations
The second measured null in the same comparison.
organism
Purified fibril preparations
plain_language
Trypsin likewise did not degrade the fibrils.
primary_references
Amyloid-degrading ability of nattokinase from Bacillus subtilis natto. (2009) https://pubmed.ncbi.nlm.nih.gov/19117402/ DOI: 10.1021/jf803072r
route
In vitro
tissue
In vitro fibril proteolysis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 246–246
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified fibril preparations · source_derived_draft · unverified_draft
Trypsin likewise did not degrade the fibrils.
Complete structured claim and evidence
Nattokinase degraded SARS-CoV-2 spike protein in transfected cell lysates in a dose- and time-dependent way, and degraded surface spike when added to the culture medium.
Nattokinase / subtilisin NAT → SARS-CoV-2 spike protein source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Transfected HEK293 cells and their lysates
exposure
Nattokinase, dose range not extracted here
limitations
No live virus, no infectivity endpoint, no animal or human data, and no link between the concentrations used and any plasma concentration achievable by mouth.
organism
Transfected HEK293 cells and their lysates
plain_language
Nattokinase degraded SARS-CoV-2 spike protein in transfected cell lysates in a dose- and time-dependent way, and degraded surface spike when added to the culture medium.
primary_references
Degradative Effect of Nattokinase on Spike Protein of SARS-CoV-2. (2022) https://pubmed.ncbi.nlm.nih.gov/36080170/ DOI: 10.3390/molecules27175405
route
In vitro
tissue
Cell lysate and cell-surface protein degradation
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 257–257
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Transfected HEK293 cells and their lysates · source_derived_draft · unverified_draft
Nattokinase degraded SARS-CoV-2 spike protein in transfected cell lysates in a dose- and time-dependent way, and degraded surface spike when added to the culture medium.
Complete structured claim and evidence
On a molar basis, nattokinase restored 62.0 +/- 5.3% of arterial blood flow in a chemically induced rat carotid thrombosis model.
Nattokinase / subtilisin NAT → Arterial blood flow recovery in the rat carotid thrombosis model source_derived_draftungraded
Experimental context and source evidence
duration
60 min monitoring
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase, equimolar with the comparators
limitations
A composite in vivo thrombolysis endpoint, not an enzyme specific-activity measurement. The occlusive thrombus was platelet-rich by CD61 staining. Administration was parenteral, so this says nothing about what an oral dose does.
organism
Rat
plain_language
On a molar basis, nattokinase restored 62.0 +/- 5.3% of arterial blood flow in a chemically induced rat carotid thrombosis model.
primary_references
Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat. (1995) https://pubmed.ncbi.nlm.nih.gov/8593442/ DOI: 10.1248/bpb.18.1387
route
In vivo, intravenous
tissue
Common carotid artery, acetic-acid endothelial injury
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 268–268
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
On a molar basis, nattokinase restored 62.0 +/- 5.3% of arterial blood flow in a chemically induced rat carotid thrombosis model.
Complete structured claim and evidence
Plasmin restored 15.8 +/- 0.7% of blood flow in the same model on the same molar basis.
Plasmin → Arterial blood flow recovery in the rat carotid thrombosis model source_derived_draftungraded
Experimental context and source evidence
duration
60 min monitoring
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Plasmin, equimolar with nattokinase
limitations
The ratio of these two numbers, about 3.9, is the origin of the widely repeated claim that nattokinase is four times more fibrinolytic than plasmin. It is a blood-flow recovery ratio in one rat model, not a purified-enzyme specific-activity comparison.
organism
Rat
plain_language
Plasmin restored 15.8 +/- 0.7% of blood flow in the same model on the same molar basis.
primary_references
Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat. (1995) https://pubmed.ncbi.nlm.nih.gov/8593442/ DOI: 10.1248/bpb.18.1387
route
In vivo, intravenous
tissue
Common carotid artery, acetic-acid endothelial injury
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 279–279
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Plasmin restored 15.8 +/- 0.7% of blood flow in the same model on the same molar basis.
Complete structured claim and evidence
Elastase restored 0% of blood flow in the same model.
Elastase, as used in the 1995 rat thrombolysis comparison → Arterial blood flow recovery in the rat carotid thrombosis model source_derived_draftungraded
Experimental context and source evidence
duration
60 min monitoring
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Elastase, equimolar with nattokinase
limitations
A measured zero, which is what makes the comparison informative: a fibrino(geno)lytic enzyme is not automatically thrombolytic in this model.
organism
Rat
plain_language
Elastase restored 0% of blood flow in the same model.
primary_references
Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat. (1995) https://pubmed.ncbi.nlm.nih.gov/8593442/ DOI: 10.1248/bpb.18.1387
route
In vivo, intravenous
tissue
Common carotid artery, acetic-acid endothelial injury
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 290–290
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Elastase restored 0% of blood flow in the same model.
Complete structured claim and evidence
Oral nattokinase for one week delayed ferric-chloride-induced carotid occlusion dose-dependently, doubling occlusion time at 160 mg/kg and fully preventing occlusion at 500 mg/kg.
Nattokinase / subtilisin NAT → Time to arterial occlusion after ferric-chloride injury in rat carotid artery source_derived_draftungraded
Experimental context and source evidence
duration
1 week of pre-treatment
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase 160 or 500 mg/kg
limitations
An oral in vivo effect, which makes it one of the few records here that clears the absorption question by outcome rather than by measurement. It does not identify what crossed the gut. The test article is described only as nattokinase extracted from fermented soybean.
organism
Rat
plain_language
Oral nattokinase for one week delayed ferric-chloride-induced carotid occlusion dose-dependently, doubling occlusion time at 160 mg/kg and fully preventing occlusion at 500 mg/kg.
primary_references
Nattokinase improves blood flow by inhibiting platelet aggregation and thrombus formation. (2013) https://pubmed.ncbi.nlm.nih.gov/24396387/ DOI: 10.5625/lar.2013.29.4.221
route
In vivo, oral
tissue
Carotid artery, 35% FeCl3 injury, laser Doppler flow
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 301–301
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Oral nattokinase for one week delayed ferric-chloride-induced carotid occlusion dose-dependently, doubling occlusion time at 160 mg/kg and fully preventing occlusion at 500 mg/kg.
Complete structured claim and evidence
Aspirin at 30 mg/kg fully prevented occlusion in the same model, as the 500 mg/kg nattokinase dose did.
Aspirin / acetylsalicylic acid → Time to arterial occlusion after ferric-chloride injury in rat carotid artery source_derived_draftungraded
Experimental context and source evidence
duration
1 week of pre-treatment
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Aspirin 30 mg/kg
limitations
The comparator arm. Equivalence of endpoint at these two doses is not equivalence of mechanism, of potency per milligram, or of bleeding risk.
organism
Rat
plain_language
Aspirin at 30 mg/kg fully prevented occlusion in the same model, as the 500 mg/kg nattokinase dose did.
primary_references
Nattokinase improves blood flow by inhibiting platelet aggregation and thrombus formation. (2013) https://pubmed.ncbi.nlm.nih.gov/24396387/ DOI: 10.5625/lar.2013.29.4.221
route
In vivo, oral
tissue
Carotid artery, 35% FeCl3 injury, laser Doppler flow
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 312–312
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Aspirin at 30 mg/kg fully prevented occlusion in the same model, as the 500 mg/kg nattokinase dose did.
Complete structured claim and evidence
Supplementing nattokinase at 4,000, 8,000 or 12,000 FU/kg reduced carrageenan-induced thrombosis in mice.
Nattokinase / subtilisin NAT → Carrageenan-induced tail thrombosis in mice source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse
exposure
Nattokinase 4,000-12,000 FU/kg
limitations
Reported as the preventive arm of a study whose main subject is intestinal transport. Group sizes were six per dose.
organism
Mouse
plain_language
Supplementing nattokinase at 4,000, 8,000 or 12,000 FU/kg reduced carrageenan-induced thrombosis in mice.
primary_references
Study on the transport and internalisation mechanism of dietary supplement nattokinase in the small intestine using animal and Caco-2 cell monolayer models. (2023) https://pubmed.ncbi.nlm.nih.gov/37971898/ DOI: 10.1080/00498254.2023.2284249
route
In vivo, oral
tissue
Tail thrombosis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 323–323
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse · source_derived_draft · unverified_draft
Supplementing nattokinase at 4,000, 8,000 or 12,000 FU/kg reduced carrageenan-induced thrombosis in mice.
Complete structured claim and evidence
Nattokinase inhibited both collagen-induced and thrombin-induced aggregation of rabbit platelets.
Nattokinase / subtilisin NAT → Rabbit platelet aggregation, agonist specified source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rabbit platelet-rich plasma
exposure
Nattokinase, concentration range not stated in the abstract
limitations
Rabbit platelets, so a route crossing into the human platelet node is inferred through orthology. No proximal platelet target was identified: no GPVI, P2Y12, integrin alphaIIbbeta3, COX-1 or thromboxane-synthase step is measured anywhere in this chapter.
organism
Rabbit platelet-rich plasma
plain_language
Nattokinase inhibited both collagen-induced and thrombin-induced aggregation of rabbit platelets.
primary_references
Nattokinase improves blood flow by inhibiting platelet aggregation and thrombus formation. (2013) https://pubmed.ncbi.nlm.nih.gov/24396387/ DOI: 10.5625/lar.2013.29.4.221
route
In vitro
tissue
Platelet aggregometry
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 334–334
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rabbit platelet-rich plasma · source_derived_draft · unverified_draft
Nattokinase inhibited both collagen-induced and thrombin-induced aggregation of rabbit platelets.
Complete structured claim and evidence
Nattokinase reduced thromboxane B2 formation from collagen-activated platelets in a concentration-dependent manner.
Nattokinase / subtilisin NAT → Thromboxane B2 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rabbit washed platelets
exposure
Nattokinase
limitations
The authors read this as blockade of thromboxane formation upstream of aggregation, but the enzyme or pathway step that is blocked is not shown. Concentrations used in vitro are high relative to anything measured in plasma after oral dosing.
organism
Rabbit washed platelets
plain_language
Nattokinase reduced thromboxane B2 formation from collagen-activated platelets in a concentration-dependent manner.
primary_references
Nattokinase improves blood flow by inhibiting platelet aggregation and thrombus formation. (2013) https://pubmed.ncbi.nlm.nih.gov/24396387/ DOI: 10.5625/lar.2013.29.4.221
route
In vitro
tissue
Thromboxane B2 assay
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 345–345
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rabbit washed platelets · source_derived_draft · unverified_draft
Nattokinase reduced thromboxane B2 formation from collagen-activated platelets in a concentration-dependent manner.
Complete structured claim and evidence
Nattokinase markedly inhibited the rise in intracellular calcium in thrombin-stimulated human platelets.
Nattokinase / subtilisin NAT → Platelet intracellular calcium concentration source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human platelets
exposure
Nattokinase
limitations
Attributed by the authors to attenuated release from internal calcium stores rather than to influx. The receptor-proximal step is not identified.
organism
Human platelets
plain_language
Nattokinase markedly inhibited the rise in intracellular calcium in thrombin-stimulated human platelets.
primary_references
Mechanisms of Nattokinase in protection of cerebral ischemia. (2014) https://pubmed.ncbi.nlm.nih.gov/25446567/ DOI: 10.1016/j.ejphar.2014.10.024
route
In vitro
tissue
Platelet calcium mobilisation
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 356–356
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human platelets · source_derived_draft · unverified_draft
Nattokinase markedly inhibited the rise in intracellular calcium in thrombin-stimulated human platelets.
Complete structured claim and evidence
Nattokinase significantly increased cyclic AMP levels in the cerebral ischemia model.
Nattokinase / subtilisin NAT → Cyclic adenosine monophosphate source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase
limitations
Reported together with activation of JAK1/STAT1 signalling in the injured region. Neither the adenylate cyclase step nor the receptor upstream of it is identified, and the route of administration for this arm is not stated in the abstract.
organism
Rat
plain_language
Nattokinase significantly increased cyclic AMP levels in the cerebral ischemia model.
primary_references
Mechanisms of Nattokinase in protection of cerebral ischemia. (2014) https://pubmed.ncbi.nlm.nih.gov/25446567/ DOI: 10.1016/j.ejphar.2014.10.024
route
Not stated here
tissue
Injured brain tissue
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 367–367
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Nattokinase significantly increased cyclic AMP levels in the cerebral ischemia model.
Complete structured claim and evidence
Incubating blood with nattokinase produced a significant dose-dependent decrease in red blood cell aggregation.
Nattokinase / subtilisin NAT → Red blood cell aggregation source_derived_draftungraded
Experimental context and source evidence
duration
30 min at 37 C
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human blood ex vivo
exposure
Nattokinase at 15.6, 31.3, 62.5 and 125 units/mL
limitations
The units here are the study's own and are not FU, CU or IU. They cannot be mapped onto an oral dose or a plasma concentration, so this result does not bridge to the human trials.
organism
Human blood ex vivo
plain_language
Incubating blood with nattokinase produced a significant dose-dependent decrease in red blood cell aggregation.
primary_references
Effects of nattokinase, a pro-fibrinolytic enzyme, on red blood cell aggregation and whole blood viscosity. (2006) https://pubmed.ncbi.nlm.nih.gov/16899918/
route
In vitro
tissue
Myrenne aggregometry
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 378–378
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human blood ex vivo · source_derived_draft · unverified_draft
Incubating blood with nattokinase produced a significant dose-dependent decrease in red blood cell aggregation.
Complete structured claim and evidence
The same incubation produced a significant dose-dependent decrease in low-shear whole-blood viscosity.
Nattokinase / subtilisin NAT → Low-shear whole-blood viscosity source_derived_draftungraded
Experimental context and source evidence
duration
30 min at 37 C
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human blood ex vivo
exposure
Nattokinase at 15.6, 31.3, 62.5 and 125 units/mL
limitations
Same unit problem. The authors note the concentrations are similar to those reached in earlier animal work, which is an argument by analogy rather than a measurement.
organism
Human blood ex vivo
plain_language
The same incubation produced a significant dose-dependent decrease in low-shear whole-blood viscosity.
primary_references
Effects of nattokinase, a pro-fibrinolytic enzyme, on red blood cell aggregation and whole blood viscosity. (2006) https://pubmed.ncbi.nlm.nih.gov/16899918/
route
In vitro
tissue
Scanning capillary rheometry over 1-1000 per second
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 389–389
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human blood ex vivo · source_derived_draft · unverified_draft
The same incubation produced a significant dose-dependent decrease in low-shear whole-blood viscosity.
Complete structured claim and evidence
Nattokinase relaxed rat thoracic aortic rings dose-dependently, and the relaxation required an intact endothelium.
Nattokinase / subtilisin NAT → Rat thoracic aortic ring tension source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase
limitations
The proteolytic target that starts this response is not identified. Endothelium dependence narrows where it begins without naming the step.
organism
Rat
plain_language
Nattokinase relaxed rat thoracic aortic rings dose-dependently, and the relaxation required an intact endothelium.
primary_references
Mechanisms of Nattokinase in protection of cerebral ischemia. (2014) https://pubmed.ncbi.nlm.nih.gov/25446567/ DOI: 10.1016/j.ejphar.2014.10.024
route
Ex vivo
tissue
Thoracic aortic ring, contracted with KCl or norepinephrine
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 400–400
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Nattokinase relaxed rat thoracic aortic rings dose-dependently, and the relaxation required an intact endothelium.
Complete structured claim and evidence
N-nitro-L-arginine methyl ester attenuated the relaxation, implicating nitric-oxide synthase in it.
NOS-dependent nitric oxide production, model specified → Rat thoracic aortic ring tension source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase with L-NAME
limitations
Recorded under a blocked pathway: the finding is what happens when NOS is inhibited. The isoform is not identified, and indomethacin was tested in the same design without its result being separable from the abstract.
organism
Rat
plain_language
N-nitro-L-arginine methyl ester attenuated the relaxation, implicating nitric-oxide synthase in it.
primary_references
Mechanisms of Nattokinase in protection of cerebral ischemia. (2014) https://pubmed.ncbi.nlm.nih.gov/25446567/ DOI: 10.1016/j.ejphar.2014.10.024
route
Ex vivo
tissue
Thoracic aortic ring
trigger_kind
machinery_impairment Imported condition classification; unverified.
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 411–411
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
N-nitro-L-arginine methyl ester attenuated the relaxation, implicating nitric-oxide synthase in it.
Complete structured claim and evidence
Nattokinase reduced receptor-operated calcium influx in vascular smooth muscle.
Nattokinase / subtilisin NAT → Receptor-operated calcium influx in vascular smooth muscle source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase
limitations
Recorded by the authors as one of three contributions to relaxation, alongside promotion of nitric-oxide synthesis and release. The channel involved is not named.
organism
Rat
plain_language
Nattokinase reduced receptor-operated calcium influx in vascular smooth muscle.
primary_references
Mechanisms of Nattokinase in protection of cerebral ischemia. (2014) https://pubmed.ncbi.nlm.nih.gov/25446567/ DOI: 10.1016/j.ejphar.2014.10.024
route
Ex vivo
tissue
Vascular smooth muscle
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 422–422
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Nattokinase reduced receptor-operated calcium influx in vascular smooth muscle.
Complete structured claim and evidence
Culture solutions of Bacillus subtilis natto and market nattokinase, deactivated by heating at 121 C for 15 min, raised tissue-plasminogen-activator activity in HeLa cells about 20-fold and 24-fold and in human umbilical vein endothelial cells about 5-fold.
Heat-inactivated nattokinase → Human tissue-type plasminogen activator / tPA / PLAT source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human cells
exposure
Heat-deactivated nattokinase and heat-deactivated B. subtilis natto culture solution
limitations
The subject here is the heat-inactivated material, not the active protease: the same report shows that heating to 70 C or above destroys the fibrinolytic and amidolytic activity. Cell counts did not change, so the effect is not a viability artefact. This is a regulated cellular response, not proteolytic generation of tPA, and it conflicts in direction with the endothelial result recorded next.
organism
Human cells
plain_language
Culture solutions of Bacillus subtilis natto and market nattokinase, deactivated by heating at 121 C for 15 min, raised tissue-plasminogen-activator activity in HeLa cells about 20-fold and 24-fold and in human umbilical vein endothelial cells about 5-fold.
primary_references
Nattokinase-promoted tissue plasminogen activator release from human cells. (2008) https://pubmed.ncbi.nlm.nih.gov/19996631/ DOI: 10.1159/000252817
route
In vitro
tissue
HeLa and human umbilical vein endothelial cells
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 433–433
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human cells · source_derived_draft · unverified_draft
Culture solutions of Bacillus subtilis natto and market nattokinase, deactivated by heating at 121 C for 15 min, raised tissue-plasminogen-activator activity in HeLa cells about 20-fold and 24-fold and in human umbilical vein endothelial cells about 5-fold.
Complete structured claim and evidence
In human umbilical vein endothelial cells, nattokinase reduced adrenaline-stimulated secretion of tissue-type plasminogen activator.
Nattokinase / subtilisin NAT → Human tissue-type plasminogen activator / tPA / PLAT source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human umbilical vein endothelial cells
exposure
Nattokinase
limitations
Opposite in direction to the heat-inactivated result in the same cell type. The two differ in the state of the enzyme and in whether the cells were stimulated, and this chapter records the disagreement rather than choosing between them.
organism
Human umbilical vein endothelial cells
plain_language
In human umbilical vein endothelial cells, nattokinase reduced adrenaline-stimulated secretion of tissue-type plasminogen activator.
primary_references
Mechanisms of Nattokinase in protection of cerebral ischemia. (2014) https://pubmed.ncbi.nlm.nih.gov/25446567/ DOI: 10.1016/j.ejphar.2014.10.024
route
In vitro
tissue
Cultured endothelium, adrenaline-stimulated
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 444–444
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human umbilical vein endothelial cells · source_derived_draft · unverified_draft
In human umbilical vein endothelial cells, nattokinase reduced adrenaline-stimulated secretion of tissue-type plasminogen activator.
Complete structured claim and evidence
In the same cells, nattokinase reduced adrenaline-stimulated secretion of plasminogen activator inhibitor 1.
Nattokinase / subtilisin NAT → Human SERPINE1 / plasminogen activator inhibitor 1 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human umbilical vein endothelial cells
exposure
Nattokinase
limitations
A second route to lower PAI-1 that is not proteolysis of the inhibitor. Whether it needs catalytic activity was not tested.
organism
Human umbilical vein endothelial cells
plain_language
In the same cells, nattokinase reduced adrenaline-stimulated secretion of plasminogen activator inhibitor 1.
primary_references
Mechanisms of Nattokinase in protection of cerebral ischemia. (2014) https://pubmed.ncbi.nlm.nih.gov/25446567/ DOI: 10.1016/j.ejphar.2014.10.024
route
In vitro
tissue
Cultured endothelium, adrenaline-stimulated
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 455–455
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human umbilical vein endothelial cells · source_derived_draft · unverified_draft
In the same cells, nattokinase reduced adrenaline-stimulated secretion of plasminogen activator inhibitor 1.
Complete structured claim and evidence
Nattokinase suppressed lipopolysaccharide-induced TLR4 activation in RAW264.7 macrophages.
Nattokinase / subtilisin NAT → Mouse Toll-like receptor 4 / Tlr4 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse RAW264.7 macrophages
exposure
Nattokinase
limitations
No proximal receptor, protease-activated-receptor cleavage or peptide mediator is identified that would connect an extracellular protease to this intracellular pathway. Whether catalytic activity is required was not tested with an inactivated control.
organism
Mouse RAW264.7 macrophages
plain_language
Nattokinase suppressed lipopolysaccharide-induced TLR4 activation in RAW264.7 macrophages.
primary_references
Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress. (2020) https://pubmed.ncbi.nlm.nih.gov/32193146/ DOI: 10.1016/j.redox.2020.101500
route
In vitro
tissue
Cultured macrophages, LPS-stimulated
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 466–466
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse RAW264.7 macrophages · source_derived_draft · unverified_draft
Nattokinase suppressed lipopolysaccharide-induced TLR4 activation in RAW264.7 macrophages.
Complete structured claim and evidence
Nattokinase suppressed lipopolysaccharide-induced NOX2 activation in the same macrophages.
Nattokinase / subtilisin NAT → Mouse CYBB / gp91phox source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse RAW264.7 macrophages
exposure
Nattokinase
limitations
As above.
organism
Mouse RAW264.7 macrophages
plain_language
Nattokinase suppressed lipopolysaccharide-induced NOX2 activation in the same macrophages.
primary_references
Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress. (2020) https://pubmed.ncbi.nlm.nih.gov/32193146/ DOI: 10.1016/j.redox.2020.101500
route
In vitro
tissue
Cultured macrophages, LPS-stimulated
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 477–477
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse RAW264.7 macrophages · source_derived_draft · unverified_draft
Nattokinase suppressed lipopolysaccharide-induced NOX2 activation in the same macrophages.
Complete structured claim and evidence
Suppressing TLR4 and NOX2 activation repressed the corresponding reactive-oxygen-species production.
Nattokinase / subtilisin NAT → Reactive oxygen species generation in mouse RAW 264.7 macrophages source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse RAW264.7 macrophages
exposure
Nattokinase
limitations
Reported as a consequence of the two steps above rather than as an independent measurement.
organism
Mouse RAW264.7 macrophages
plain_language
Suppressing TLR4 and NOX2 activation repressed the corresponding reactive-oxygen-species production.
primary_references
Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress. (2020) https://pubmed.ncbi.nlm.nih.gov/32193146/ DOI: 10.1016/j.redox.2020.101500
route
In vitro
tissue
Cultured macrophages, LPS-stimulated
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 488–488
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse RAW264.7 macrophages · source_derived_draft · unverified_draft
Suppressing TLR4 and NOX2 activation repressed the corresponding reactive-oxygen-species production.
Complete structured claim and evidence
Nattokinase repressed translocation of NF-kappa-B from the cytoplasm to the nucleus in stimulated macrophages, along with MAPK activation.
Nattokinase / subtilisin NAT → NF-kappa-B signaling source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse RAW264.7 macrophages
exposure
Nattokinase
limitations
Nuclear translocation is the measured step; downstream mediator output (TNF-alpha, IL-6, nitric oxide and PAI-1) is reported as following from it.
organism
Mouse RAW264.7 macrophages
plain_language
Nattokinase repressed translocation of NF-kappa-B from the cytoplasm to the nucleus in stimulated macrophages, along with MAPK activation.
primary_references
Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress. (2020) https://pubmed.ncbi.nlm.nih.gov/32193146/ DOI: 10.1016/j.redox.2020.101500
route
In vitro
tissue
Cultured macrophages, LPS-stimulated
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 499–499
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse RAW264.7 macrophages · source_derived_draft · unverified_draft
Nattokinase repressed translocation of NF-kappa-B from the cytoplasm to the nucleus in stimulated macrophages, along with MAPK activation.
Complete structured claim and evidence
MAPK activation was repressed in the same stimulated macrophages.
Nattokinase / subtilisin NAT → Mouse p38 MAP kinase family, isoform unresolved source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse RAW264.7 macrophages
exposure
Nattokinase
limitations
The abstract names the family, not the isoform, so this is recorded against the mouse p38 family node the ledger already uses for isoform-unresolved reports.
organism
Mouse RAW264.7 macrophages
plain_language
MAPK activation was repressed in the same stimulated macrophages.
primary_references
Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress. (2020) https://pubmed.ncbi.nlm.nih.gov/32193146/ DOI: 10.1016/j.redox.2020.101500
route
In vitro
tissue
Cultured macrophages, LPS-stimulated
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 510–510
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse RAW264.7 macrophages · source_derived_draft · unverified_draft
MAPK activation was repressed in the same stimulated macrophages.
Complete structured claim and evidence
Nattokinase markedly inhibited the rise in serum PAI-1 induced by lipopolysaccharide in mice.
Nattokinase / subtilisin NAT → Mouse SERPINE1 / plasminogen activator inhibitor 1 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse
exposure
Nattokinase
limitations
This is a third and separate route from nattokinase to PAI-1, after direct proteolysis of the protein and reduced secretion from endothelium. Here the mechanism proposed is suppressed transcription downstream of NF-kappa-B, in a different species. The three are not assumed to be the same mechanism.
organism
Mouse
plain_language
Nattokinase markedly inhibited the rise in serum PAI-1 induced by lipopolysaccharide in mice.
primary_references
Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress. (2020) https://pubmed.ncbi.nlm.nih.gov/32193146/ DOI: 10.1016/j.redox.2020.101500
route
Not stated here
tissue
Serum, LPS-induced acute kidney injury model
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 521–521
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse · source_derived_draft · unverified_draft
Nattokinase markedly inhibited the rise in serum PAI-1 induced by lipopolysaccharide in mice.
Complete structured claim and evidence
Lower serum PAI-1 was accompanied by blocked fibrin deposition in the glomeruli of endotoxin-treated animals.
Nattokinase / subtilisin NAT → Fibrin deposition in glomeruli of endotoxin-treated mice source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Mouse
exposure
Nattokinase
limitations
An in vivo consequence in an endotoxin model. Whether it runs through PAI-1, through direct fibrin proteolysis or through both is not apportioned.
organism
Mouse
plain_language
Lower serum PAI-1 was accompanied by blocked fibrin deposition in the glomeruli of endotoxin-treated animals.
primary_references
Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress. (2020) https://pubmed.ncbi.nlm.nih.gov/32193146/ DOI: 10.1016/j.redox.2020.101500
route
Not stated here
tissue
Renal glomeruli, LPS-induced acute kidney injury model
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 532–532
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Mouse · source_derived_draft · unverified_draft
Lower serum PAI-1 was accompanied by blocked fibrin deposition in the glomeruli of endotoxin-treated animals.
Complete structured claim and evidence
Nattokinase pretreatment enhanced Nrf2 activation in particulate-matter-exposed human gingival fibroblasts, confirmed by antioxidant-response-element luciferase assay.
Nattokinase / subtilisin NAT → Human Nrf2 / NFE2L2 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human gingival fibroblasts (HGF-1)
exposure
Nattokinase pretreatment
limitations
A cell model of periodontal inflammation, not a vascular one. The proximal target by which an extracellular protease reaches Nrf2 is not established.
organism
Human gingival fibroblasts (HGF-1)
plain_language
Nattokinase pretreatment enhanced Nrf2 activation in particulate-matter-exposed human gingival fibroblasts, confirmed by antioxidant-response-element luciferase assay.
primary_references
Nuclear factor erythroid 2-related factor 2/heme oxygenase-1 activation by nattokinase reduces pro-inflammatory and matrix-degrading mediators in human gingival fibroblasts. (2026) https://pubmed.ncbi.nlm.nih.gov/41585176/ DOI: 10.1016/j.jds.2025.10.022
route
In vitro
tissue
Cell culture, particulate-matter exposure
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 543–543
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human gingival fibroblasts (HGF-1) · source_derived_draft · unverified_draft
Nattokinase pretreatment enhanced Nrf2 activation in particulate-matter-exposed human gingival fibroblasts, confirmed by antioxidant-response-element luciferase assay.
Complete structured claim and evidence
Nattokinase raised heme oxygenase 1 expression in the same cells.
Nattokinase / subtilisin NAT → Human heme oxygenase 1 / HMOX1 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human gingival fibroblasts (HGF-1)
exposure
Nattokinase pretreatment
limitations
Reported as the effector arm of the Nrf2 response.
organism
Human gingival fibroblasts (HGF-1)
plain_language
Nattokinase raised heme oxygenase 1 expression in the same cells.
primary_references
Nuclear factor erythroid 2-related factor 2/heme oxygenase-1 activation by nattokinase reduces pro-inflammatory and matrix-degrading mediators in human gingival fibroblasts. (2026) https://pubmed.ncbi.nlm.nih.gov/41585176/ DOI: 10.1016/j.jds.2025.10.022
route
In vitro
tissue
Cell culture, particulate-matter exposure
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 554–554
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human gingival fibroblasts (HGF-1) · source_derived_draft · unverified_draft
Nattokinase raised heme oxygenase 1 expression in the same cells.
Complete structured claim and evidence
Inhibition of Nrf2 or of heme oxygenase 1 abolished the protective effect of nattokinase on COX-2, prostaglandin E2 and MMP-1 induction.
Human Nrf2 / NFE2L2 → Particulate-matter-induced COX-2, prostaglandin E2 and MMP-1 induction in gingival fibroblasts source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human gingival fibroblasts (HGF-1)
exposure
Nattokinase with Nrf2 or HO-1 inhibitors
limitations
A necessity test under a blocked pathway. Pharmacological inhibitors carry off-target risk that a genetic control would not, and no inactivated-enzyme arm was run.
organism
Human gingival fibroblasts (HGF-1)
plain_language
Inhibition of Nrf2 or of heme oxygenase 1 abolished the protective effect of nattokinase on COX-2, prostaglandin E2 and MMP-1 induction.
primary_references
Nuclear factor erythroid 2-related factor 2/heme oxygenase-1 activation by nattokinase reduces pro-inflammatory and matrix-degrading mediators in human gingival fibroblasts. (2026) https://pubmed.ncbi.nlm.nih.gov/41585176/ DOI: 10.1016/j.jds.2025.10.022
route
In vitro
tissue
Cell culture with pharmacological pathway inhibition
trigger_kind
machinery_impairment Imported condition classification; unverified.
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 565–565
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human gingival fibroblasts (HGF-1) · source_derived_draft · unverified_draft
Inhibition of Nrf2 or of heme oxygenase 1 abolished the protective effect of nattokinase on COX-2, prostaglandin E2 and MMP-1 induction.
Complete structured claim and evidence
Nattokinase suppressed NADPH-oxidase-derived reactive-oxygen-species generation in particulate-exposed gingival fibroblasts.
Nattokinase / subtilisin NAT → Reactive oxygen species source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human gingival fibroblasts (HGF-1)
exposure
Nattokinase pretreatment
limitations
The reported pathway has reciprocal regulation between ROS, PI3K/Akt and MAPK, so a single arrow into ROS understates a loop the authors describe as bidirectional.
organism
Human gingival fibroblasts (HGF-1)
plain_language
Nattokinase suppressed NADPH-oxidase-derived reactive-oxygen-species generation in particulate-exposed gingival fibroblasts.
primary_references
Nuclear factor erythroid 2-related factor 2/heme oxygenase-1 activation by nattokinase reduces pro-inflammatory and matrix-degrading mediators in human gingival fibroblasts. (2026) https://pubmed.ncbi.nlm.nih.gov/41585176/ DOI: 10.1016/j.jds.2025.10.022
route
In vitro
tissue
Cell culture, particulate-matter exposure
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 576–576
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human gingival fibroblasts (HGF-1) · source_derived_draft · unverified_draft
Nattokinase suppressed NADPH-oxidase-derived reactive-oxygen-species generation in particulate-exposed gingival fibroblasts.
Complete structured claim and evidence
RNA sequencing and Q-PCR showed nattokinase upregulated serum response factor in endothelial cells.
Nattokinase / subtilisin NAT → Human serum response factor / SRF source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase
limitations
A transcript-level measurement. The step by which an extracellular protease changes transcription is not identified.
organism
Endothelial cells
plain_language
RNA sequencing and Q-PCR showed nattokinase upregulated serum response factor in endothelial cells.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium, transcriptional profiling
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 587–587
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
RNA sequencing and Q-PCR showed nattokinase upregulated serum response factor in endothelial cells.
Complete structured claim and evidence
Nattokinase upregulated thrombospondin 1 in the same cells.
Nattokinase / subtilisin NAT → Human thrombospondin-1 / THBS1 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase
limitations
As above.
organism
Endothelial cells
plain_language
Nattokinase upregulated thrombospondin 1 in the same cells.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium, transcriptional profiling
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 598–598
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
Nattokinase upregulated thrombospondin 1 in the same cells.
Complete structured claim and evidence
SREBF1 messenger RNA appeared in the sequencing screen but was unaffected on Q-PCR confirmation.
Nattokinase / subtilisin NAT → Sterol regulatory element-binding protein 1 / SREBP-1 source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase
limitations
A measured null from the confirmation step of a screen, recorded because it bounds which transcripts the screen actually supports.
organism
Endothelial cells
plain_language
SREBF1 messenger RNA appeared in the sequencing screen but was unaffected on Q-PCR confirmation.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium, transcriptional profiling
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 609–609
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
SREBF1 messenger RNA appeared in the sequencing screen but was unaffected on Q-PCR confirmation.
Complete structured claim and evidence
Nattokinase treatment induced autophagy in endothelial cells.
Nattokinase / subtilisin NAT → Autophagy source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase
limitations
Reported alongside a mouse arm in which nattokinase reduced vascular inflammation by the same route.
organism
Endothelial cells
plain_language
Nattokinase treatment induced autophagy in endothelial cells.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 620–620
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
Nattokinase treatment induced autophagy in endothelial cells.
Complete structured claim and evidence
Nattokinase inhibited NLRP3 inflammasome formation in endothelial cells.
Nattokinase / subtilisin NAT → NLRP3 inflammasome source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase
limitations
Placed downstream of autophagy induction by the necessity experiments rather than measured as an independent arm.
organism
Endothelial cells
plain_language
Nattokinase inhibited NLRP3 inflammasome formation in endothelial cells.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 631–631
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
Nattokinase inhibited NLRP3 inflammasome formation in endothelial cells.
Complete structured claim and evidence
Nattokinase inhibited necroptosis in endothelial cells.
Nattokinase / subtilisin NAT → Necroptosis source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase
limitations
As above.
organism
Endothelial cells
plain_language
Nattokinase inhibited necroptosis in endothelial cells.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 642–642
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
Nattokinase inhibited necroptosis in endothelial cells.
Complete structured claim and evidence
Small interfering RNA against SRF suppressed the autophagy induction and restored NLRP3 inflammasome formation and necroptosis.
Human serum response factor / SRF → Autophagy source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase with SRF siRNA
limitations
A necessity test recorded under a broken pathway. It places SRF between nattokinase and autophagy without showing how nattokinase raises SRF.
organism
Endothelial cells
plain_language
Small interfering RNA against SRF suppressed the autophagy induction and restored NLRP3 inflammasome formation and necroptosis.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium with siRNA knockdown
trigger_kind
machinery_impairment Imported condition classification; unverified.
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 653–653
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
Small interfering RNA against SRF suppressed the autophagy induction and restored NLRP3 inflammasome formation and necroptosis.
Complete structured claim and evidence
Small interfering RNA against THBS1 had the same effect: autophagy was suppressed and NLRP3 and necroptosis signals returned.
Human thrombospondin-1 / THBS1 → Autophagy source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Endothelial cells
exposure
Nattokinase with THBS1 siRNA
limitations
As above. Whether SRF and THBS1 act in series or in parallel is not resolved by these two experiments.
organism
Endothelial cells
plain_language
Small interfering RNA against THBS1 had the same effect: autophagy was suppressed and NLRP3 and necroptosis signals returned.
primary_references
Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1. (2024) https://pubmed.ncbi.nlm.nih.gov/38679250/ DOI: 10.1016/j.ijbiomac.2024.131779
route
In vitro
tissue
Cultured endothelium with siRNA knockdown
trigger_kind
machinery_impairment Imported condition classification; unverified.
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 664–664
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Endothelial cells · source_derived_draft · unverified_draft
Small interfering RNA against THBS1 had the same effect: autophagy was suppressed and NLRP3 and necroptosis signals returned.
Complete structured claim and evidence
Heat treatment at 120 C for 15 min markedly increased the ACE-inhibitory activity of subtilisin NAT while inactivating it as a protease and fragmenting it into small peptides.
Heat-inactivated nattokinase → Human angiotensin-converting enzyme / ACE source_derived_draftungraded
Experimental context and source evidence
duration
15 min heating
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified subtilisin NAT
exposure
Subtilisin NAT, heat-treated at 120 C for 15 min
limitations
This is the clearest evidence in the chapter that destroying the enzyme creates a different pharmacological species rather than nothing. The authors state the inhibition is not due to an enzymatic action of the protease. Inhibition was of mixed type: Vmax fell and Km rose.
organism
Purified subtilisin NAT
plain_language
Heat treatment at 120 C for 15 min markedly increased the ACE-inhibitory activity of subtilisin NAT while inactivating it as a protease and fragmenting it into small peptides.
primary_references
Inhibition of angiotensin I converting enzyme by subtilisin NAT (nattokinase) in natto, a Japanese traditional fermented food. (2012) https://pubmed.ncbi.nlm.nih.gov/22453301/ DOI: 10.1039/c2fo10245e
route
In vitro
tissue
ACE inhibition assay
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 675–675
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified subtilisin NAT · source_derived_draft · unverified_draft
Heat treatment at 120 C for 15 min markedly increased the ACE-inhibitory activity of subtilisin NAT while inactivating it as a protease and fragmenting it into small peptides.
Complete structured claim and evidence
Heat fragmentation and simulated gastrointestinal digestion both release ACE-inhibitory peptides from the nattokinase sequence.
Nattokinase / subtilisin NAT → Nattokinase-derived digestion peptide pool source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified nattokinase
exposure
Nattokinase
limitations
In vitro generation. Whether these peptides are generated in a human gut, absorbed, or reach concentrations that inhibit ACE in vivo has not been measured.
organism
Purified nattokinase
plain_language
Heat fragmentation and simulated gastrointestinal digestion both release ACE-inhibitory peptides from the nattokinase sequence.
primary_references
Identification and mechanistic characterization of novel ACE-inhibitory peptides derived from Nattokinase. (2026) https://pubmed.ncbi.nlm.nih.gov/41844106/ DOI: 10.1016/j.foodchem.2026.148875
route
In vitro
tissue
Simulated digestion with peptide identification by HPLC-MS
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 686–686
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified nattokinase · source_derived_draft · unverified_draft
Heat fragmentation and simulated gastrointestinal digestion both release ACE-inhibitory peptides from the nattokinase sequence.
Complete structured claim and evidence
The Leu-Tyr dipeptide, liberated by fragmentation of subtilisin NAT, inhibited angiotensin-converting enzyme.
Leu-Tyr dipeptide → Human angiotensin-converting enzyme / ACE source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified enzyme assay
exposure
LY dipeptide
limitations
Its presence in the heat-inactivated material was confirmed by HPLC-MS. No human exposure measurement exists for it.
organism
Purified enzyme assay
plain_language
The Leu-Tyr dipeptide, liberated by fragmentation of subtilisin NAT, inhibited angiotensin-converting enzyme.
primary_references
Inhibition of angiotensin I converting enzyme by subtilisin NAT (nattokinase) in natto, a Japanese traditional fermented food. (2012) https://pubmed.ncbi.nlm.nih.gov/22453301/ DOI: 10.1039/c2fo10245e
route
In vitro
tissue
ACE inhibition kinetics
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 697–697
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified enzyme assay · source_derived_draft · unverified_draft
The Leu-Tyr dipeptide, liberated by fragmentation of subtilisin NAT, inhibited angiotensin-converting enzyme.
Complete structured claim and evidence
The Phe-Tyr dipeptide, liberated in the same way, inhibited angiotensin-converting enzyme.
Phe-Tyr dipeptide → Human angiotensin-converting enzyme / ACE source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified enzyme assay
exposure
FY dipeptide
limitations
As above.
organism
Purified enzyme assay
plain_language
The Phe-Tyr dipeptide, liberated in the same way, inhibited angiotensin-converting enzyme.
primary_references
Inhibition of angiotensin I converting enzyme by subtilisin NAT (nattokinase) in natto, a Japanese traditional fermented food. (2012) https://pubmed.ncbi.nlm.nih.gov/22453301/ DOI: 10.1039/c2fo10245e
route
In vitro
tissue
ACE inhibition kinetics
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 708–708
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified enzyme assay · source_derived_draft · unverified_draft
The Phe-Tyr dipeptide, liberated in the same way, inhibited angiotensin-converting enzyme.
Complete structured claim and evidence
The Leu-Gly-Gly peptide identified from simulated digestion inhibited ACE competitively with an IC50 of 10.78 micromolar.
Leu-Gly-Gly peptide → Human angiotensin-converting enzyme / ACE source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
In vitro simulated digestion
exposure
LGG peptide
limitations
An IC50 against purified ACE. Docking and network-pharmacology analyses in the same report are modelling, not measurement.
organism
In vitro simulated digestion
plain_language
The Leu-Gly-Gly peptide identified from simulated digestion inhibited ACE competitively with an IC50 of 10.78 micromolar.
primary_references
Identification and mechanistic characterization of novel ACE-inhibitory peptides derived from Nattokinase. (2026) https://pubmed.ncbi.nlm.nih.gov/41844106/ DOI: 10.1016/j.foodchem.2026.148875
route
In vitro
tissue
ACE inhibition kinetics with molecular docking
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 719–719
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · In vitro simulated digestion · source_derived_draft · unverified_draft
The Leu-Gly-Gly peptide identified from simulated digestion inhibited ACE competitively with an IC50 of 10.78 micromolar.
Complete structured claim and evidence
The Thr-Trp peptide from the same digestion acted as a noncompetitive ACE inhibitor.
Thr-Trp dipeptide → Human angiotensin-converting enzyme / ACE source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
In vitro simulated digestion
exposure
TW peptide
limitations
As above. A different inhibition mode from LGG in the same peptide pool.
organism
In vitro simulated digestion
plain_language
The Thr-Trp peptide from the same digestion acted as a noncompetitive ACE inhibitor.
primary_references
Identification and mechanistic characterization of novel ACE-inhibitory peptides derived from Nattokinase. (2026) https://pubmed.ncbi.nlm.nih.gov/41844106/ DOI: 10.1016/j.foodchem.2026.148875
route
In vitro
tissue
ACE inhibition kinetics with molecular docking
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 730–730
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · In vitro simulated digestion · source_derived_draft · unverified_draft
The Thr-Trp peptide from the same digestion acted as a noncompetitive ACE inhibitor.
Complete structured claim and evidence
Catalytically active intact nattokinase is not required for the ACE-inhibitory activity of the material, because heat inactivation raises rather than removes it.
Nattokinase / subtilisin NAT → Human angiotensin-converting enzyme / ACE source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Purified subtilisin NAT
exposure
Active compared with heat-inactivated subtilisin NAT
limitations
This is a statement about one activity only. Every direct-proteolysis mechanism in this chapter does require the intact active enzyme.
organism
Purified subtilisin NAT
plain_language
Catalytically active intact nattokinase is not required for the ACE-inhibitory activity of the material, because heat inactivation raises rather than removes it.
primary_references
Inhibition of angiotensin I converting enzyme by subtilisin NAT (nattokinase) in natto, a Japanese traditional fermented food. (2012) https://pubmed.ncbi.nlm.nih.gov/22453301/ DOI: 10.1039/c2fo10245e
route
In vitro
tissue
ACE inhibition assay
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 741–741
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Purified subtilisin NAT · source_derived_draft · unverified_draft
Catalytically active intact nattokinase is not required for the ACE-inhibitory activity of the material, because heat inactivation raises rather than removes it.
Complete structured claim and evidence
Nattokinase injected into a tied intestinal loop at 80 mg/kg was followed by fibrinogen degradation products in plasma in all twenty rats, with a 270 kDa fragment appearing within 0.5 to 5 h.
Nattokinase / subtilisin NAT → Rat fibrinogen source_derived_draftungraded
Experimental context and source evidence
duration
Sampling to 5 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (male Wistar, n = 20)
exposure
Nattokinase 80 mg/kg
limitations
The stomach was bypassed, so this cannot establish gastric survival after ordinary oral dosing, and there was no control group receiving the same surgery with placebo. Breakdown patterns differed between animals: fourteen of twenty showed a 270 kDa fragment at 3 to 5 h with no further degradation, while six showed it at 0.5 h followed by degradation to a 105 kDa fragment via a 200 kDa intermediate. The abstract narrates only the six-rat pattern; the fourteen-rat pattern is recorded in the EFSA account of the same study.
organism
Rat (male Wistar, n = 20)
plain_language
Nattokinase injected into a tied intestinal loop at 80 mg/kg was followed by fibrinogen degradation products in plasma in all twenty rats, with a 270 kDa fragment appearing within 0.5 to 5 h.
primary_references
Transport of nattokinase across the rat intestinal tract. (1995) https://pubmed.ncbi.nlm.nih.gov/8845803/ DOI: 10.1248/bpb.18.1194
route
In vivo, intraduodenal into a ligated loop
tissue
Ligated intestinal loop including duodenum and jejunum, femoral-vein plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 752–752
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (male Wistar, n = 20) · source_derived_draft · unverified_draft
Nattokinase injected into a tied intestinal loop at 80 mg/kg was followed by fibrinogen degradation products in plasma in all twenty rats, with a 270 kDa fragment appearing within 0.5 to 5 h.
Complete structured claim and evidence
In parallel with the fibrinogen degradation, plasma recalcification time was prolonged.
Nattokinase / subtilisin NAT → Plasma recalcification time source_derived_draftungraded
Experimental context and source evidence
duration
Sampling to 5 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (male Wistar, n = 20)
exposure
Nattokinase 80 mg/kg
limitations
Uncontrolled, and a functional consequence of the same uncontrolled experiment rather than independent support for it.
organism
Rat (male Wistar, n = 20)
plain_language
In parallel with the fibrinogen degradation, plasma recalcification time was prolonged.
primary_references
Transport of nattokinase across the rat intestinal tract. (1995) https://pubmed.ncbi.nlm.nih.gov/8845803/ DOI: 10.1248/bpb.18.1194
route
In vivo, intraduodenal into a ligated loop
tissue
Femoral-vein plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 763–763
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (male Wistar, n = 20) · source_derived_draft · unverified_draft
In parallel with the fibrinogen degradation, plasma recalcification time was prolonged.
Complete structured claim and evidence
Nattokinase is a moderately absorbed biomolecule whose transport through enterocytes is energy- and time-dependent.
Nattokinase / subtilisin NAT → Intestinal epithelium source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Caco-2 cell monolayer and rat
exposure
Nattokinase
limitations
Energy dependence rules out passive paracellular leak as the whole story. Crossing an epithelium in a monolayer or an everted sac is not the same as arriving intact and active in human plasma, which is the step nothing in this chapter measures.
organism
Caco-2 cell monolayer and rat
plain_language
Nattokinase is a moderately absorbed biomolecule whose transport through enterocytes is energy- and time-dependent.
primary_references
Study on the transport and internalisation mechanism of dietary supplement nattokinase in the small intestine using animal and Caco-2 cell monolayer models. (2023) https://pubmed.ncbi.nlm.nih.gov/37971898/ DOI: 10.1080/00498254.2023.2284249
route
In vitro
tissue
Intestinal epithelium, everted gut sac and ligated loop models
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 774–774
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Caco-2 cell monolayer and rat · source_derived_draft · unverified_draft
Nattokinase is a moderately absorbed biomolecule whose transport through enterocytes is energy- and time-dependent.
Complete structured claim and evidence
Chlorpromazine inhibited nattokinase uptake in Caco-2 monolayers, implicating clathrin-mediated endocytosis.
Nattokinase / subtilisin NAT → Clathrin-mediated endocytosis source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Caco-2 cell monolayer and rat
exposure
Nattokinase with endocytosis inhibitors
limitations
Transport through enterocytes was energy- and time-dependent and the authors call the molecule moderately absorbed. Cellular uptake is not the same as intact active enzyme reaching human plasma.
organism
Caco-2 cell monolayer and rat
plain_language
Chlorpromazine inhibited nattokinase uptake in Caco-2 monolayers, implicating clathrin-mediated endocytosis.
primary_references
Study on the transport and internalisation mechanism of dietary supplement nattokinase in the small intestine using animal and Caco-2 cell monolayer models. (2023) https://pubmed.ncbi.nlm.nih.gov/37971898/ DOI: 10.1080/00498254.2023.2284249
route
In vitro
tissue
Intestinal epithelium, everted gut sac and ligated loop models
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 785–785
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Caco-2 cell monolayer and rat · source_derived_draft · unverified_draft
Chlorpromazine inhibited nattokinase uptake in Caco-2 monolayers, implicating clathrin-mediated endocytosis.
Complete structured claim and evidence
Nystatin inhibited uptake in the same system, implicating a caveolae-mediated pathway.
Nattokinase / subtilisin NAT → Caveolae/lipid-raft-associated endocytosis source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Caco-2 cell monolayer and rat
exposure
Nattokinase with nystatin
limitations
As above. Inhibitor selectivity is the basis for each pathway assignment.
organism
Caco-2 cell monolayer and rat
plain_language
Nystatin inhibited uptake in the same system, implicating a caveolae-mediated pathway.
primary_references
Study on the transport and internalisation mechanism of dietary supplement nattokinase in the small intestine using animal and Caco-2 cell monolayer models. (2023) https://pubmed.ncbi.nlm.nih.gov/37971898/ DOI: 10.1080/00498254.2023.2284249
route
In vitro
tissue
Intestinal epithelium
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 796–796
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Caco-2 cell monolayer and rat · source_derived_draft · unverified_draft
Nystatin inhibited uptake in the same system, implicating a caveolae-mediated pathway.
Complete structured claim and evidence
EIPA inhibited uptake in the same system, implicating macropinocytosis.
Nattokinase / subtilisin NAT → Macropinocytosis source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Caco-2 cell monolayer and rat
exposure
Nattokinase with EIPA
limitations
As above. Three pathways were implicated to varying degrees; the report does not apportion them.
organism
Caco-2 cell monolayer and rat
plain_language
EIPA inhibited uptake in the same system, implicating macropinocytosis.
primary_references
Study on the transport and internalisation mechanism of dietary supplement nattokinase in the small intestine using animal and Caco-2 cell monolayer models. (2023) https://pubmed.ncbi.nlm.nih.gov/37971898/ DOI: 10.1080/00498254.2023.2284249
route
In vitro
tissue
Intestinal epithelium
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 807–807
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Caco-2 cell monolayer and rat · source_derived_draft · unverified_draft
EIPA inhibited uptake in the same system, implicating macropinocytosis.
Complete structured claim and evidence
A single 2,000 FU oral NSK-SD dose produced anti-nattokinase immunoreactivity in serum that rose significantly from baseline between 2 and 24 h and peaked at about 13.3 +/- 2.5 h.
NSK-SD fermented soybean extract → Plasma nattokinase-immunoreactive material source_derived_draftungraded
Experimental context and source evidence
duration
Single dose, sampling to 48 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (11 healthy adults, 5 male and 6 female, ages 21-65)
exposure
NSK-SD softgel, 2,000 FU, about 100 mg
limitations
No control group and a non-validated polyclonal rabbit anti-nattokinase capture ELISA. EFSA reviewed this study and noted the same limitations, adding that it did not assess the biological activity of nattokinase in blood. The authors themselves recommend that future work look separately for intact enzyme and for bioactive peptides.
organism
Human (11 healthy adults, 5 male and 6 female, ages 21-65)
plain_language
A single 2,000 FU oral NSK-SD dose produced anti-nattokinase immunoreactivity in serum that rose significantly from baseline between 2 and 24 h and peaked at about 13.3 +/- 2.5 h.
primary_references
A pilot study on the serum pharmacokinetics of nattokinase in humans following a single, oral, daily dose. (2013) https://pubmed.ncbi.nlm.nih.gov/23709455/
route
In vivo, oral
tissue
Serum
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 818–818
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (11 healthy adults, 5 male and 6 female, ages 21-65) · source_derived_draft · unverified_draft
A single 2,000 FU oral NSK-SD dose produced anti-nattokinase immunoreactivity in serum that rose significantly from baseline between 2 and 24 h and peaked at about 13.3 +/- 2.5 h.
Complete structured claim and evidence
A polyclonal ELISA signal does not by itself distinguish intact catalytically active nattokinase from fragments, complexes or cross-reactive material.
Plasma nattokinase-immunoreactive material → Systemic exposure to intact catalytically active nattokinase after oral dosing source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human
exposure
Anti-nattokinase polyclonal capture ELISA
limitations
Recorded as an assay-scope limit, not as a negative finding. No study in the retrieved evidence immunocaptured the material and then measured catalytic activity on a substrate.
organism
Human
plain_language
A polyclonal ELISA signal does not by itself distinguish intact catalytically active nattokinase from fragments, complexes or cross-reactive material.
primary_references
A pilot study on the serum pharmacokinetics of nattokinase in humans following a single, oral, daily dose. (2013) https://pubmed.ncbi.nlm.nih.gov/23709455/
route
In vivo, oral
tissue
Serum
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 829–829
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human · source_derived_draft · unverified_draft
A polyclonal ELISA signal does not by itself distinguish intact catalytically active nattokinase from fragments, complexes or cross-reactive material.
Complete structured claim and evidence
EFSA concluded that the information provided on absorption, distribution, metabolism and excretion did not allow conclusions to be drawn on the absorption of active nattokinase or any functional metabolites derived from it.
NSK-SD fermented soybean extract → Systemic exposure to intact catalytically active nattokinase after oral dosing source_derived_draftungraded
Experimental context and source evidence
duration
Dossier review
evidence_access
EFSA Journal opinion retrieved and read in full during this curation. Quoted figures are from the opinion text and its specification tables.
experimental_model
Regulatory assessment
exposure
NSK-SD, up to 100 mg/day
limitations
The Panel noted in the same passage that nattokinase has in vitro fibrinolytic activity and in vivo thrombolytic activity in animals when administered parenterally, which is the distinction this chapter turns on. An applicant assay of the softgel product at pH 2.0 mimicking gastric fluid was submitted; the opinion records that the results were provided but does not state what they showed.
organism
Regulatory assessment
plain_language
EFSA concluded that the information provided on absorption, distribution, metabolism and excretion did not allow conclusions to be drawn on the absorption of active nattokinase or any functional metabolites derived from it.
primary_references
Safety of fermented soybean extract NSK-SD as a novel food. (2016) DOI: 10.2903/j.efsa.2016.4541 Not indexed in PubMed.
route
In vivo, oral
tissue
Whole-body disposition
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 840–840
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Regulatory assessment · source_derived_draft · unverified_draft
EFSA concluded that the information provided on absorption, distribution, metabolism and excretion did not allow conclusions to be drawn on the absorption of active nattokinase or any functional metabolites derived from it.
Complete structured claim and evidence
D-dimer concentrations rose significantly 6 and 8 h after a single 2,000 FU oral dose.
NSK-SD fermented soybean extract → Plasma D-dimer concentration source_derived_draftungraded
Experimental context and source evidence
duration
Single dose, sampling to 8 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (12 healthy young males)
exposure
NSK-SD, 2,000 FU single dose
limitations
Double-blind placebo-controlled crossover, but very small, with multiple time-point comparisons and no measurement of what was absorbed. All changes stayed within the normal reference range.
organism
Human (12 healthy young males)
plain_language
D-dimer concentrations rose significantly 6 and 8 h after a single 2,000 FU oral dose.
primary_references
A single-dose of oral nattokinase potentiates thrombolysis and anti-coagulation profiles. (2015) https://pubmed.ncbi.nlm.nih.gov/26109079/ DOI: 10.1038/srep11601
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 851–851
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (12 healthy young males) · source_derived_draft · unverified_draft
D-dimer concentrations rose significantly 6 and 8 h after a single 2,000 FU oral dose.
Complete structured claim and evidence
Fibrin and fibrinogen degradation products rose significantly 4 h after the same single dose.
NSK-SD fermented soybean extract → Blood fibrin/fibrinogen degradation products source_derived_draftungraded
Experimental context and source evidence
duration
Single dose, sampling to 8 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (12 healthy young males)
exposure
NSK-SD, 2,000 FU single dose
limitations
As above. Within the normal range.
organism
Human (12 healthy young males)
plain_language
Fibrin and fibrinogen degradation products rose significantly 4 h after the same single dose.
primary_references
A single-dose of oral nattokinase potentiates thrombolysis and anti-coagulation profiles. (2015) https://pubmed.ncbi.nlm.nih.gov/26109079/ DOI: 10.1038/srep11601
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 862–862
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (12 healthy young males) · source_derived_draft · unverified_draft
Fibrin and fibrinogen degradation products rose significantly 4 h after the same single dose.
Complete structured claim and evidence
Factor VIII activity declined significantly at 4 and 6 h after the same single dose.
NSK-SD fermented soybean extract → Human coagulation factor VIII / F8 source_derived_draftungraded
Experimental context and source evidence
duration
Single dose, sampling to 8 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (12 healthy young males)
exposure
NSK-SD, 2,000 FU single dose
limitations
Within the normal range. No direct proteolytic cleavage of factor VIII by purified nattokinase has been shown anywhere in the retrieved evidence, so the step producing this is unidentified.
organism
Human (12 healthy young males)
plain_language
Factor VIII activity declined significantly at 4 and 6 h after the same single dose.
primary_references
A single-dose of oral nattokinase potentiates thrombolysis and anti-coagulation profiles. (2015) https://pubmed.ncbi.nlm.nih.gov/26109079/ DOI: 10.1038/srep11601
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 873–873
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (12 healthy young males) · source_derived_draft · unverified_draft
Factor VIII activity declined significantly at 4 and 6 h after the same single dose.
Complete structured claim and evidence
Blood antithrombin concentration was higher at 2 and 4 h after the same single dose.
NSK-SD fermented soybean extract → Human antithrombin / SERPINC1 source_derived_draftungraded
Experimental context and source evidence
duration
Single dose, sampling to 8 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (12 healthy young males)
exposure
NSK-SD, 2,000 FU single dose
limitations
Within the normal range. A rise in an inhibitor is harder to explain by proteolysis than a fall would be.
organism
Human (12 healthy young males)
plain_language
Blood antithrombin concentration was higher at 2 and 4 h after the same single dose.
primary_references
A single-dose of oral nattokinase potentiates thrombolysis and anti-coagulation profiles. (2015) https://pubmed.ncbi.nlm.nih.gov/26109079/ DOI: 10.1038/srep11601
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 884–884
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (12 healthy young males) · source_derived_draft · unverified_draft
Blood antithrombin concentration was higher at 2 and 4 h after the same single dose.
Complete structured claim and evidence
Activated partial thromboplastin time was prolonged significantly at 2 and 4 h after the same single dose.
NSK-SD fermented soybean extract → Activated partial thromboplastin time source_derived_draftungraded
Experimental context and source evidence
duration
Single dose, sampling to 8 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (12 healthy young males)
exposure
NSK-SD, 2,000 FU single dose
limitations
Within the normal range. This is one of the two findings that make an additive effect with an antithrombotic drug biologically plausible.
organism
Human (12 healthy young males)
plain_language
Activated partial thromboplastin time was prolonged significantly at 2 and 4 h after the same single dose.
primary_references
A single-dose of oral nattokinase potentiates thrombolysis and anti-coagulation profiles. (2015) https://pubmed.ncbi.nlm.nih.gov/26109079/ DOI: 10.1038/srep11601
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 895–895
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (12 healthy young males) · source_derived_draft · unverified_draft
Activated partial thromboplastin time was prolonged significantly at 2 and 4 h after the same single dose.
Complete structured claim and evidence
Prothrombin time and fibrinogen showed no detected difference in the same study.
NSK-SD fermented soybean extract → Prothrombin time source_derived_draftungraded
Experimental context and source evidence
duration
Single dose, sampling to 8 h
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (12 healthy young males)
exposure
NSK-SD, 2,000 FU single dose
limitations
A measured null in a twelve-person crossover, so absence of a detected effect rather than evidence of none. It contrasts with the eight-week study, where prothrombin time did rise within group.
organism
Human (12 healthy young males)
plain_language
Prothrombin time and fibrinogen showed no detected difference in the same study.
primary_references
A single-dose of oral nattokinase potentiates thrombolysis and anti-coagulation profiles. (2015) https://pubmed.ncbi.nlm.nih.gov/26109079/ DOI: 10.1038/srep11601
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 906–906
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (12 healthy young males) · source_derived_draft · unverified_draft
Prothrombin time and fibrinogen showed no detected difference in the same study.
Complete structured claim and evidence
Plasma fibrinogen declined by 7 to 10% across three subject groups over two months of 4,000 FU per day.
Commercial nattokinase supplement, composition study-specific → Human fibrinogen source_derived_draftungraded
Experimental context and source evidence
duration
2 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (45 subjects: healthy, cardiovascular-risk and dialysis groups)
exposure
Nattokinase, two 2,000 FU capsules daily
limitations
Open-label and self-controlled with no placebo arm. A significant time effect but no group effect, so the decline was similar across strata. The mechanism producing lower factor levels is not established.
organism
Human (45 subjects: healthy, cardiovascular-risk and dialysis groups)
plain_language
Plasma fibrinogen declined by 7 to 10% across three subject groups over two months of 4,000 FU per day.
primary_references
Nattokinase decreases plasma levels of fibrinogen, factor VII, and factor VIII in human subjects. (2009) https://pubmed.ncbi.nlm.nih.gov/19358933/ DOI: 10.1016/j.nutres.2009.01.009
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 917–917
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (45 subjects: healthy, cardiovascular-risk and dialysis groups) · source_derived_draft · unverified_draft
Plasma fibrinogen declined by 7 to 10% across three subject groups over two months of 4,000 FU per day.
Complete structured claim and evidence
Factor VII declined by 7 to 14% over the same two months.
Commercial nattokinase supplement, composition study-specific → Human coagulation factor VII / F7 source_derived_draftungraded
Experimental context and source evidence
duration
2 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (45 subjects across three groups)
exposure
Nattokinase, 4,000 FU per day
limitations
As above. Direct proteolytic cleavage of factor VII by purified nattokinase has not been shown, so this association has no identified molecular step behind it.
organism
Human (45 subjects across three groups)
plain_language
Factor VII declined by 7 to 14% over the same two months.
primary_references
Nattokinase decreases plasma levels of fibrinogen, factor VII, and factor VIII in human subjects. (2009) https://pubmed.ncbi.nlm.nih.gov/19358933/ DOI: 10.1016/j.nutres.2009.01.009
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 928–928
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (45 subjects across three groups) · source_derived_draft · unverified_draft
Factor VII declined by 7 to 14% over the same two months.
Complete structured claim and evidence
Factor VIII declined by 17 to 19% over the same two months.
Commercial nattokinase supplement, composition study-specific → Human coagulation factor VIII / F8 source_derived_draftungraded
Experimental context and source evidence
duration
2 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (45 subjects across three groups)
exposure
Nattokinase, 4,000 FU per day
limitations
As above, and the largest of the three declines. It agrees in direction with the single-dose crossover result, which is a different design in a different population.
organism
Human (45 subjects across three groups)
plain_language
Factor VIII declined by 17 to 19% over the same two months.
primary_references
Nattokinase decreases plasma levels of fibrinogen, factor VII, and factor VIII in human subjects. (2009) https://pubmed.ncbi.nlm.nih.gov/19358933/ DOI: 10.1016/j.nutres.2009.01.009
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 939–939
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (45 subjects across three groups) · source_derived_draft · unverified_draft
Factor VIII declined by 17 to 19% over the same two months.
Complete structured claim and evidence
Blood lipids were unaffected over the same two months.
Commercial nattokinase supplement, composition study-specific → Blood total cholesterol concentration source_derived_draftungraded
Experimental context and source evidence
duration
2 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (45 subjects across three groups)
exposure
Nattokinase, 4,000 FU per day
limitations
A measured null in an uncontrolled study, recorded because the lipid question is contested elsewhere in this chapter.
organism
Human (45 subjects across three groups)
plain_language
Blood lipids were unaffected over the same two months.
primary_references
Nattokinase decreases plasma levels of fibrinogen, factor VII, and factor VIII in human subjects. (2009) https://pubmed.ncbi.nlm.nih.gov/19358933/ DOI: 10.1016/j.nutres.2009.01.009
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 950–950
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (45 subjects across three groups) · source_derived_draft · unverified_draft
Blood lipids were unaffected over the same two months.
Complete structured claim and evidence
Collagen-epinephrine closure time rose significantly more with nattokinase than with placebo over eight weeks in hypercholesterolemic subjects.
Commercial nattokinase supplement, composition study-specific → Collagen-epinephrine closure time source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (100 nondiabetic hypercholesterolemic subjects, 50 per arm)
exposure
Nattokinase, dose not stated in the abstract
limitations
Randomized and placebo-controlled. Closure time is a platelet-function surrogate, not a clinical event endpoint. The dose is unavailable from the abstract, which blocks any comparison with the trials that do state FU per day. PubMed links an erratum to this article.
organism
Human (100 nondiabetic hypercholesterolemic subjects, 50 per arm)
plain_language
Collagen-epinephrine closure time rose significantly more with nattokinase than with placebo over eight weeks in hypercholesterolemic subjects.
primary_references
The effects of nattokinase supplementation on collagen-epinephrine closure time, prothrombin time and activated partial thromboplastin time in nondiabetic and hypercholesterolemic subjects. (2019) https://pubmed.ncbi.nlm.nih.gov/31070609/ DOI: 10.1039/c8fo02324g Correction on record: Erratum in: Food Funct. 2019 Jul 17;10(7):4454. doi: 10.1039/c9fo90033k. PMID 31287455.
route
In vivo, oral
tissue
Whole blood
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 961–961
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (100 nondiabetic hypercholesterolemic subjects, 50 per arm) · source_derived_draft · unverified_draft
Collagen-epinephrine closure time rose significantly more with nattokinase than with placebo over eight weeks in hypercholesterolemic subjects.
Complete structured claim and evidence
Activated partial thromboplastin time also rose significantly more with nattokinase than with placebo over the same eight weeks.
Commercial nattokinase supplement, composition study-specific → Activated partial thromboplastin time source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (100 nondiabetic hypercholesterolemic subjects)
exposure
Nattokinase, dose not stated in the abstract
limitations
Agrees in direction with the single-dose crossover finding at a different dose and duration. Same erratum note as above.
organism
Human (100 nondiabetic hypercholesterolemic subjects)
plain_language
Activated partial thromboplastin time also rose significantly more with nattokinase than with placebo over the same eight weeks.
primary_references
The effects of nattokinase supplementation on collagen-epinephrine closure time, prothrombin time and activated partial thromboplastin time in nondiabetic and hypercholesterolemic subjects. (2019) https://pubmed.ncbi.nlm.nih.gov/31070609/ DOI: 10.1039/c8fo02324g Correction on record: Erratum in: Food Funct. 2019 Jul 17;10(7):4454. doi: 10.1039/c9fo90033k. PMID 31287455.
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 972–972
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (100 nondiabetic hypercholesterolemic subjects) · source_derived_draft · unverified_draft
Activated partial thromboplastin time also rose significantly more with nattokinase than with placebo over the same eight weeks.
Complete structured claim and evidence
Prothrombin time rose significantly within the nattokinase group over eight weeks, but the between-group comparison against placebo was significant only for closure time and aPTT.
Commercial nattokinase supplement, composition study-specific → Prothrombin time source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (100 nondiabetic hypercholesterolemic subjects)
exposure
Nattokinase, dose not stated in the abstract
limitations
Recorded with the within-group and between-group results kept apart, because they differ here. The single-dose crossover found no prothrombin-time change at all.
organism
Human (100 nondiabetic hypercholesterolemic subjects)
plain_language
Prothrombin time rose significantly within the nattokinase group over eight weeks, but the between-group comparison against placebo was significant only for closure time and aPTT.
primary_references
The effects of nattokinase supplementation on collagen-epinephrine closure time, prothrombin time and activated partial thromboplastin time in nondiabetic and hypercholesterolemic subjects. (2019) https://pubmed.ncbi.nlm.nih.gov/31070609/ DOI: 10.1039/c8fo02324g Correction on record: Erratum in: Food Funct. 2019 Jul 17;10(7):4454. doi: 10.1039/c9fo90033k. PMID 31287455.
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 983–983
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (100 nondiabetic hypercholesterolemic subjects) · source_derived_draft · unverified_draft
Prothrombin time rose significantly within the nattokinase group over eight weeks, but the between-group comparison against placebo was significant only for closure time and aPTT.
Complete structured claim and evidence
Net changes of -5.55 mmHg systolic and -2.84 mmHg diastolic against placebo were reported after eight weeks at 2,000 FU per day.
NSK-SD fermented soybean extract → Arterial blood pressure source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (86 randomized, 73 completed, untreated systolic 130-159 mmHg, ages 20-80)
exposure
Nattokinase, 2,000 FU per capsule
limitations
Randomized, double-blind and placebo-controlled, but small and short, with industry affiliations. The confidence intervals reach close to zero (-10.5 to -0.57 and -5.33 to -0.33). No mechanistic mediator beyond plasma renin activity was measured.
organism
Human (86 randomized, 73 completed, untreated systolic 130-159 mmHg, ages 20-80)
plain_language
Net changes of -5.55 mmHg systolic and -2.84 mmHg diastolic against placebo were reported after eight weeks at 2,000 FU per day.
primary_references
Effects of nattokinase on blood pressure: a randomized, controlled trial. (2008) https://pubmed.ncbi.nlm.nih.gov/18971533/ DOI: 10.1291/hypres.31.1583
route
In vivo, oral
tissue
Arterial blood pressure
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 994–994
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (86 randomized, 73 completed, untreated systolic 130-159 mmHg, ages 20-80) · source_derived_draft · unverified_draft
Net changes of -5.55 mmHg systolic and -2.84 mmHg diastolic against placebo were reported after eight weeks at 2,000 FU per day.
Complete structured claim and evidence
The net change in plasma renin activity was -1.17 ng/mL/h against placebo in the same trial.
NSK-SD fermented soybean extract → Plasma renin activity source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (73 completers with elevated blood pressure)
exposure
Nattokinase, 2,000 FU per day
limitations
The only mediator measured in that trial. Its direction is not reproduced by the North American trial, where an exploratory low-renin subgroup moved the other way.
organism
Human (73 completers with elevated blood pressure)
plain_language
The net change in plasma renin activity was -1.17 ng/mL/h against placebo in the same trial.
primary_references
Effects of nattokinase on blood pressure: a randomized, controlled trial. (2008) https://pubmed.ncbi.nlm.nih.gov/18971533/ DOI: 10.1291/hypres.31.1583
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1005–1005
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (73 completers with elevated blood pressure) · source_derived_draft · unverified_draft
The net change in plasma renin activity was -1.17 ng/mL/h against placebo in the same trial.
Complete structured claim and evidence
Average diastolic blood pressure fell from 87 to 84 mmHg with nattokinase against no change on placebo over eight weeks, reaching 86 to 81 mmHg in males.
NSK-SD fermented soybean extract → Arterial blood pressure source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (79 enrolled, 74 completed, systolic 130 or above or diastolic 90 or above)
exposure
NSK-SD, 100 mg per day
limitations
Randomized, double-blind, placebo-controlled and multicentre, with industry sponsorship and several exploratory subgroup analyses. The dose is stated in milligrams, not FU, so it is not directly comparable with the 2,000 FU trials without the product specification.
organism
Human (79 enrolled, 74 completed, systolic 130 or above or diastolic 90 or above)
plain_language
Average diastolic blood pressure fell from 87 to 84 mmHg with nattokinase against no change on placebo over eight weeks, reaching 86 to 81 mmHg in males.
primary_references
Consumption of nattokinase is associated with reduced blood pressure and von Willebrand factor, a cardiovascular risk marker: results from a randomized, double-blind, placebo-controlled, multicenter North American clinical trial. (2016) https://pubmed.ncbi.nlm.nih.gov/27785095/ DOI: 10.2147/IBPC.S99553
route
In vivo, oral
tissue
Arterial blood pressure
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1016–1016
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (79 enrolled, 74 completed, systolic 130 or above or diastolic 90 or above) · source_derived_draft · unverified_draft
Average diastolic blood pressure fell from 87 to 84 mmHg with nattokinase against no change on placebo over eight weeks, reaching 86 to 81 mmHg in males.
Complete structured claim and evidence
A decrease in von Willebrand factor was seen in the female participants taking nattokinase, at p < 0.1.
NSK-SD fermented soybean extract → Human von Willebrand factor / VWF source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (74 completers with elevated blood pressure)
exposure
NSK-SD, 100 mg per day
limitations
Exploratory, sex-restricted and above the conventional significance threshold. It is the only human measurement in this chapter that touches the von Willebrand factor mechanism, which makes it worth recording and makes over-reading it easy.
organism
Human (74 completers with elevated blood pressure)
plain_language
A decrease in von Willebrand factor was seen in the female participants taking nattokinase, at p < 0.1.
primary_references
Consumption of nattokinase is associated with reduced blood pressure and von Willebrand factor, a cardiovascular risk marker: results from a randomized, double-blind, placebo-controlled, multicenter North American clinical trial. (2016) https://pubmed.ncbi.nlm.nih.gov/27785095/ DOI: 10.2147/IBPC.S99553
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1027–1027
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (74 completers with elevated blood pressure) · source_derived_draft · unverified_draft
A decrease in von Willebrand factor was seen in the female participants taking nattokinase, at p < 0.1.
Complete structured claim and evidence
Among participants with baseline plasma renin activity below 0.29 ng/mL/h, 66% showed an increase after eight weeks against 8% on placebo, at p < 0.1.
NSK-SD fermented soybean extract → Plasma renin activity source_derived_draftungraded
Experimental context and source evidence
duration
8 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human, exploratory low-renin subgroup
exposure
NSK-SD, 100 mg per day
limitations
A post-hoc subgroup at p < 0.1, in the opposite direction to the whole-group fall reported by the Korean trial. Recorded as a conflict rather than resolved.
organism
Human, exploratory low-renin subgroup
plain_language
Among participants with baseline plasma renin activity below 0.29 ng/mL/h, 66% showed an increase after eight weeks against 8% on placebo, at p < 0.1.
primary_references
Consumption of nattokinase is associated with reduced blood pressure and von Willebrand factor, a cardiovascular risk marker: results from a randomized, double-blind, placebo-controlled, multicenter North American clinical trial. (2016) https://pubmed.ncbi.nlm.nih.gov/27785095/ DOI: 10.2147/IBPC.S99553
route
In vivo, oral
tissue
Plasma
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1038–1038
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human, exploratory low-renin subgroup · source_derived_draft · unverified_draft
Among participants with baseline plasma renin activity below 0.29 ng/mL/h, 66% showed an increase after eight weeks against 8% on placebo, at p < 0.1.
Complete structured claim and evidence
Across six randomized trials with 546 participants, pooled systolic blood pressure fell by 3.45 mmHg and diastolic by 2.32 mmHg against placebo.
Commercial nattokinase supplement, composition study-specific → Arterial blood pressure source_derived_draftungraded
Experimental context and source evidence
duration
Trial durations differ
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human, systematic review and meta-analysis of randomized controlled trials
exposure
Nattokinase supplementation, doses and products differing between trials
limitations
Few small trials with different populations, doses and products, and predominantly surrogate outcomes. The pooled estimate inherits the industry affiliation of several component trials and does not include the three-year randomized trial that found no blood-pressure effect.
organism
Human, systematic review and meta-analysis of randomized controlled trials
plain_language
Across six randomized trials with 546 participants, pooled systolic blood pressure fell by 3.45 mmHg and diastolic by 2.32 mmHg against placebo.
primary_references
Nattokinase Supplementation and Cardiovascular Risk Factors: A Systematic Review and Meta-Analysis of Randomized Controlled Trials. (2023) https://pubmed.ncbi.nlm.nih.gov/39076715/ DOI: 10.31083/j.rcm2408234
route
In vivo, oral
tissue
Arterial blood pressure
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1049–1049
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human, systematic review and meta-analysis of randomized controlled trials · source_derived_draft · unverified_draft
Across six randomized trials with 546 participants, pooled systolic blood pressure fell by 3.45 mmHg and diastolic by 2.32 mmHg against placebo.
Complete structured claim and evidence
At relatively low total dosage the pooled mean difference in total cholesterol was +5.27 against control, and it remained +3.18 at relatively high dosage.
Commercial nattokinase supplement, composition study-specific → Blood total cholesterol concentration source_derived_draftungraded
Experimental context and source evidence
duration
Trial durations differ
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human, meta-analysis of randomized controlled trials
exposure
Nattokinase supplementation
limitations
The direction is unfavourable, which is the opposite of what the uncontrolled cohort studies in this chapter report. Units are as given in the source and the analysis pools heterogeneous assays.
organism
Human, meta-analysis of randomized controlled trials
plain_language
At relatively low total dosage the pooled mean difference in total cholesterol was +5.27 against control, and it remained +3.18 at relatively high dosage.
primary_references
Nattokinase Supplementation and Cardiovascular Risk Factors: A Systematic Review and Meta-Analysis of Randomized Controlled Trials. (2023) https://pubmed.ncbi.nlm.nih.gov/39076715/ DOI: 10.31083/j.rcm2408234
route
In vivo, oral
tissue
Blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1060–1060
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human, meta-analysis of randomized controlled trials · source_derived_draft · unverified_draft
At relatively low total dosage the pooled mean difference in total cholesterol was +5.27 against control, and it remained +3.18 at relatively high dosage.
Complete structured claim and evidence
At relatively low total dosage the pooled mean difference in high-density lipoprotein cholesterol was -2.76 against control.
Commercial nattokinase supplement, composition study-specific → Blood high-density lipoprotein cholesterol concentration source_derived_draftungraded
Experimental context and source evidence
duration
Trial durations differ
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human, meta-analysis of randomized controlled trials
exposure
Nattokinase supplementation
limitations
Also unfavourable in direction, and directly contradicted by the open active-comparator study that reported a rise. No significant difference was found at high dosage.
organism
Human, meta-analysis of randomized controlled trials
plain_language
At relatively low total dosage the pooled mean difference in high-density lipoprotein cholesterol was -2.76 against control.
primary_references
Nattokinase Supplementation and Cardiovascular Risk Factors: A Systematic Review and Meta-Analysis of Randomized Controlled Trials. (2023) https://pubmed.ncbi.nlm.nih.gov/39076715/ DOI: 10.31083/j.rcm2408234
route
In vivo, oral
tissue
Blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1071–1071
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human, meta-analysis of randomized controlled trials · source_derived_draft · unverified_draft
At relatively low total dosage the pooled mean difference in high-density lipoprotein cholesterol was -2.76 against control.
Complete structured claim and evidence
At relatively low total dosage the pooled mean difference in low-density lipoprotein cholesterol was +6.49 against control.
Commercial nattokinase supplement, composition study-specific → Blood low-density lipoprotein cholesterol concentration source_derived_draftungraded
Experimental context and source evidence
duration
Trial durations differ
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human, meta-analysis of randomized controlled trials
exposure
Nattokinase supplementation
limitations
Wide interval (0.83 to 12.15). No significant difference at high dosage.
organism
Human, meta-analysis of randomized controlled trials
plain_language
At relatively low total dosage the pooled mean difference in low-density lipoprotein cholesterol was +6.49 against control.
primary_references
Nattokinase Supplementation and Cardiovascular Risk Factors: A Systematic Review and Meta-Analysis of Randomized Controlled Trials. (2023) https://pubmed.ncbi.nlm.nih.gov/39076715/ DOI: 10.31083/j.rcm2408234
route
In vivo, oral
tissue
Blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1082–1082
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human, meta-analysis of randomized controlled trials · source_derived_draft · unverified_draft
At relatively low total dosage the pooled mean difference in low-density lipoprotein cholesterol was +6.49 against control.
Complete structured claim and evidence
Pooled blood glucose rose by 0.40 against placebo across the same randomized trials.
Commercial nattokinase supplement, composition study-specific → Blood glucose concentration source_derived_draftungraded
Experimental context and source evidence
duration
Trial durations differ
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human, meta-analysis of randomized controlled trials
exposure
Nattokinase supplementation
limitations
Small and of unstated clinical meaning, but signed and significant in the pooled analysis. It is absent from every narrative summary of nattokinase read during this curation.
organism
Human, meta-analysis of randomized controlled trials
plain_language
Pooled blood glucose rose by 0.40 against placebo across the same randomized trials.
primary_references
Nattokinase Supplementation and Cardiovascular Risk Factors: A Systematic Review and Meta-Analysis of Randomized Controlled Trials. (2023) https://pubmed.ncbi.nlm.nih.gov/39076715/ DOI: 10.31083/j.rcm2408234
route
In vivo, oral
tissue
Blood glucose
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1093–1093
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human, meta-analysis of randomized controlled trials · source_derived_draft · unverified_draft
Pooled blood glucose rose by 0.40 against placebo across the same randomized trials.
Complete structured claim and evidence
Over a median three years at 2,000 FU per day, the annualized rate of change in carotid intima-media thickness did not differ from placebo.
NSK-SD fermented soybean extract → Carotid artery intima-media thickness source_derived_draftungraded
Experimental context and source evidence
duration
Median 3 years
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (265 randomized, median age 65.3, no clinical cardiovascular disease)
exposure
Nattokinase, 2,000 fibrinolytic units per day
limitations
Double-blinded and randomized, and by far the longest trial here. The null applies to this dose, this formulation, this low-risk population and these endpoints; it is not evidence that every product or dose is inert. It is also not an active-enzyme pharmacokinetic study.
organism
Human (265 randomized, median age 65.3, no clinical cardiovascular disease)
plain_language
Over a median three years at 2,000 FU per day, the annualized rate of change in carotid intima-media thickness did not differ from placebo.
primary_references
Nattokinase atherothrombotic prevention study: A randomized controlled trial. (2021) https://pubmed.ncbi.nlm.nih.gov/33843667/ DOI: 10.3233/CH-211147
route
In vivo, oral
tissue
Carotid artery ultrasound
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1104–1104
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (265 randomized, median age 65.3, no clinical cardiovascular disease) · source_derived_draft · unverified_draft
Over a median three years at 2,000 FU per day, the annualized rate of change in carotid intima-media thickness did not differ from placebo.
Complete structured claim and evidence
Carotid arterial stiffness likewise did not differ from placebo over the same period.
NSK-SD fermented soybean extract → Carotid arterial stiffness source_derived_draftungraded
Experimental context and source evidence
duration
Median 3 years
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (265 randomized, low cardiovascular risk)
exposure
Nattokinase, 2,000 FU per day
limitations
The co-primary endpoint of the same trial.
organism
Human (265 randomized, low cardiovascular risk)
plain_language
Carotid arterial stiffness likewise did not differ from placebo over the same period.
primary_references
Nattokinase atherothrombotic prevention study: A randomized controlled trial. (2021) https://pubmed.ncbi.nlm.nih.gov/33843667/ DOI: 10.3233/CH-211147
route
In vivo, oral
tissue
Carotid artery ultrasound
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1115–1115
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (265 randomized, low cardiovascular risk) · source_derived_draft · unverified_draft
Carotid arterial stiffness likewise did not differ from placebo over the same period.
Complete structured claim and evidence
There was no significant effect on blood pressure over the same three years.
NSK-SD fermented soybean extract → Arterial blood pressure source_derived_draftungraded
Experimental context and source evidence
duration
Median 3 years
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (265 randomized, low cardiovascular risk)
exposure
Nattokinase, 2,000 FU per day
limitations
This is the direct contradiction of the eight-week blood-pressure trials and of the pooled estimate. The populations differ: these participants were low-risk and not selected for elevated blood pressure.
organism
Human (265 randomized, low cardiovascular risk)
plain_language
There was no significant effect on blood pressure over the same three years.
primary_references
Nattokinase atherothrombotic prevention study: A randomized controlled trial. (2021) https://pubmed.ncbi.nlm.nih.gov/33843667/ DOI: 10.3233/CH-211147
route
In vivo, oral
tissue
Arterial blood pressure
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1126–1126
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (265 randomized, low cardiovascular risk) · source_derived_draft · unverified_draft
There was no significant effect on blood pressure over the same three years.
Complete structured claim and evidence
There was no significant effect on any laboratory determination, covering metabolic factors, blood rheology, coagulation and fibrinolysis factors, inflammatory markers and monocyte/macrophage activation markers.
NSK-SD fermented soybean extract → Plasma D-dimer concentration source_derived_draftungraded
Experimental context and source evidence
duration
Median 3 years
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (265 randomized, low cardiovascular risk)
exposure
Nattokinase, 2,000 FU per day
limitations
The broadest measured null in the chapter, and it covers the same classes of marker that the short trials moved. Assay panels differ between studies, and a null over three years does not exclude a transient change hours after a dose.
organism
Human (265 randomized, low cardiovascular risk)
plain_language
There was no significant effect on any laboratory determination, covering metabolic factors, blood rheology, coagulation and fibrinolysis factors, inflammatory markers and monocyte/macrophage activation markers.
primary_references
Nattokinase atherothrombotic prevention study: A randomized controlled trial. (2021) https://pubmed.ncbi.nlm.nih.gov/33843667/ DOI: 10.3233/CH-211147
route
In vivo, oral
tissue
Blood laboratory panel
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1137–1137
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (265 randomized, low cardiovascular risk) · source_derived_draft · unverified_draft
There was no significant effect on any laboratory determination, covering metabolic factors, blood rheology, coagulation and fibrinolysis factors, inflammatory markers and monocyte/macrophage activation markers.
Complete structured claim and evidence
In a retrospective cohort of 1,062 participants, 10,800 FU per day for 12 months was followed by reduced carotid intima-media thickness and carotid plaque size, with improvement rates of 66.5 to 95.4%.
Commercial nattokinase supplement, composition study-specific → Carotid plaque area source_derived_draftungraded
Experimental context and source evidence
duration
12 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (1,062 outpatients)
exposure
Nattokinase, 10,800 FU per day
limitations
Uncontrolled and retrospective, with selection and regression-to-the-mean risk, self-reported lifestyle data and manufacturer-employed authors. Causal inference from it is low certainty. The effect sizes are far larger than the randomized trials at lower doses found, which is the pattern a dose threshold would produce and also the pattern uncontrolled design produces.
organism
Human (1,062 outpatients)
plain_language
In a retrospective cohort of 1,062 participants, 10,800 FU per day for 12 months was followed by reduced carotid intima-media thickness and carotid plaque size, with improvement rates of 66.5 to 95.4%.
primary_references
Effective management of atherosclerosis progress and hyperlipidemia with nattokinase: A clinical study with 1,062 participants. (2022) https://pubmed.ncbi.nlm.nih.gov/36072877/ DOI: 10.3389/fcvm.2022.964977 Correction on record: Erratum in: Front Cardiovasc Med. 2022 Dec 05;9:1076420. doi: 10.3389/fcvm.2022.1076420. PMID 36545015.
route
In vivo, oral
tissue
Carotid artery imaging and blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1148–1148
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (1,062 outpatients) · source_derived_draft · unverified_draft
In a retrospective cohort of 1,062 participants, 10,800 FU per day for 12 months was followed by reduced carotid intima-media thickness and carotid plaque size, with improvement rates of 66.5 to 95.4%.
Complete structured claim and evidence
The same cohort found nattokinase ineffective for lipids and atherosclerosis progression at 3,600 FU per day.
Commercial nattokinase supplement, composition study-specific → Carotid artery intima-media thickness source_derived_draftungraded
Experimental context and source evidence
duration
12 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (1,062 outpatients)
exposure
Nattokinase, 3,600 FU per day
limitations
Same design limitations. Recorded because it is the only dose-comparison in the chapter and because it sits directly against the three-year randomized null at 2,000 FU per day. PubMed links an erratum to this article.
organism
Human (1,062 outpatients)
plain_language
The same cohort found nattokinase ineffective for lipids and atherosclerosis progression at 3,600 FU per day.
primary_references
Effective management of atherosclerosis progress and hyperlipidemia with nattokinase: A clinical study with 1,062 participants. (2022) https://pubmed.ncbi.nlm.nih.gov/36072877/ DOI: 10.3389/fcvm.2022.964977 Correction on record: Erratum in: Front Cardiovasc Med. 2022 Dec 05;9:1076420. doi: 10.3389/fcvm.2022.1076420. PMID 36545015.
route
In vivo, oral
tissue
Carotid artery imaging and blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1159–1159
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (1,062 outpatients) · source_derived_draft · unverified_draft
The same cohort found nattokinase ineffective for lipids and atherosclerosis progression at 3,600 FU per day.
Complete structured claim and evidence
Co-administration of vitamin K2 and aspirin with nattokinase produced what the authors described as a synergistic effect on the same outcomes.
Aspirin / acetylsalicylic acid → Commercial nattokinase supplement, composition study-specific source_derived_draftungraded
Experimental context and source evidence
duration
12 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (1,062 outpatients)
exposure
Nattokinase with vitamin K2 and aspirin
limitations
A subgroup comparison inside an uncontrolled retrospective study by manufacturer-employed authors, with no formal interaction design. It points the opposite way from the case report in which nattokinase with aspirin preceded a cerebellar hemorrhage, and neither can settle the other.
organism
Human (1,062 outpatients)
plain_language
Co-administration of vitamin K2 and aspirin with nattokinase produced what the authors described as a synergistic effect on the same outcomes.
primary_references
Effective management of atherosclerosis progress and hyperlipidemia with nattokinase: A clinical study with 1,062 participants. (2022) https://pubmed.ncbi.nlm.nih.gov/36072877/ DOI: 10.3389/fcvm.2022.964977 Correction on record: Erratum in: Front Cardiovasc Med. 2022 Dec 05;9:1076420. doi: 10.3389/fcvm.2022.1076420. PMID 36545015.
route
In vivo, oral
tissue
Carotid artery imaging and blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1170–1170
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (1,062 outpatients) · source_derived_draft · unverified_draft
Co-administration of vitamin K2 and aspirin with nattokinase produced what the authors described as a synergistic effect on the same outcomes.
Complete structured claim and evidence
At 6,000 FU per day for 26 weeks, carotid plaque area fell 36.6% against 11.5% in the simvastatin arm, with plaque area falling from 0.25 to 0.16 square centimetres and intima-media thickness from 1.13 to 1.01 mm.
Commercial nattokinase supplement, composition study-specific → Carotid plaque area source_derived_draftungraded
Experimental context and source evidence
duration
26 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (82 enrolled, 76 completed: 39 nattokinase, 37 statin)
exposure
Nattokinase 6,000 FU per day against simvastatin 20 mg per day
limitations
Randomized in allocation but open-label with no placebo, single-centre, and reported in Chinese. An active comparator that also works makes a between-arm difference hard to read, and the imaging endpoints are surrogates.
organism
Human (82 enrolled, 76 completed: 39 nattokinase, 37 statin)
plain_language
At 6,000 FU per day for 26 weeks, carotid plaque area fell 36.6% against 11.5% in the simvastatin arm, with plaque area falling from 0.25 to 0.16 square centimetres and intima-media thickness from 1.13 to 1.01 mm.
primary_references
[A clinical study on the effect of nattokinase on carotid artery atherosclerosis and hyperlipidaemia]. (2017) https://pubmed.ncbi.nlm.nih.gov/28763875/ DOI: 10.3760/cma.j.issn.0376-2491.2017.26.005
route
In vivo, oral
tissue
Common carotid artery ultrasound
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1181–1181
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (82 enrolled, 76 completed: 39 nattokinase, 37 statin) · source_derived_draft · unverified_draft
At 6,000 FU per day for 26 weeks, carotid plaque area fell 36.6% against 11.5% in the simvastatin arm, with plaque area falling from 0.25 to 0.16 square centimetres and intima-media thickness from 1.13 to 1.01 mm.
Complete structured claim and evidence
Nattokinase significantly raised high-density lipoprotein cholesterol, while the statin arm showed no change in it.
Commercial nattokinase supplement, composition study-specific → Blood high-density lipoprotein cholesterol concentration source_derived_draftungraded
Experimental context and source evidence
duration
26 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (76 completers)
exposure
Nattokinase 6,000 FU per day
limitations
Open-label. It is the direct opposite in direction to the pooled randomized estimate, which found HDL lower at low dose.
organism
Human (76 completers)
plain_language
Nattokinase significantly raised high-density lipoprotein cholesterol, while the statin arm showed no change in it.
primary_references
[A clinical study on the effect of nattokinase on carotid artery atherosclerosis and hyperlipidaemia]. (2017) https://pubmed.ncbi.nlm.nih.gov/28763875/ DOI: 10.3760/cma.j.issn.0376-2491.2017.26.005
route
In vivo, oral
tissue
Blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1192–1192
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (76 completers) · source_derived_draft · unverified_draft
Nattokinase significantly raised high-density lipoprotein cholesterol, while the statin arm showed no change in it.
Complete structured claim and evidence
The lipid-lowering seen in the nattokinase arm did not correlate with the reduction in intima-media thickness or plaque area.
Blood low-density lipoprotein cholesterol concentration → Carotid plaque area source_derived_draftungraded
Experimental context and source evidence
duration
26 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (39 nattokinase-arm completers)
exposure
Nattokinase 6,000 FU per day
limitations
A within-arm correlation of r = 0.35 at p = 0.09, which is an absence of detected correlation in a small sample rather than established independence. The authors read it as showing a mechanism separate from lipid lowering.
organism
Human (39 nattokinase-arm completers)
plain_language
The lipid-lowering seen in the nattokinase arm did not correlate with the reduction in intima-media thickness or plaque area.
primary_references
[A clinical study on the effect of nattokinase on carotid artery atherosclerosis and hyperlipidaemia]. (2017) https://pubmed.ncbi.nlm.nih.gov/28763875/ DOI: 10.3760/cma.j.issn.0376-2491.2017.26.005
route
In vivo, oral
tissue
Carotid imaging against blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1203–1203
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (39 nattokinase-arm completers) · source_derived_draft · unverified_draft
The lipid-lowering seen in the nattokinase arm did not correlate with the reduction in intima-media thickness or plaque area.
Complete structured claim and evidence
A nattokinase-only formula showed no effect on blood lipids through six months, while the same enzyme combined with red yeast rice improved every lipid measured from month one.
Commercial nattokinase supplement, composition study-specific → Blood low-density lipoprotein cholesterol concentration source_derived_draftungraded
Experimental context and source evidence
duration
6 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (47 patients with hyperlipidemia in three arms)
exposure
Nattokinase 50 mg per capsule, two capsules twice daily, alone or with 300 mg red yeast rice extract
limitations
Randomized, double-blind and placebo-controlled but small. The combination arm cannot attribute anything to the nattokinase, since red yeast rice contains monacolin K. The mono-arm null is the informative part.
organism
Human (47 patients with hyperlipidemia in three arms)
plain_language
A nattokinase-only formula showed no effect on blood lipids through six months, while the same enzyme combined with red yeast rice improved every lipid measured from month one.
primary_references
Combined nattokinase with red yeast rice but not nattokinase alone has potent effects on blood lipids in human subjects with hyperlipidemia. (2009) https://pubmed.ncbi.nlm.nih.gov/19786378/
route
In vivo, oral
tissue
Blood lipids
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1214–1214
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (47 patients with hyperlipidemia in three arms) · source_derived_draft · unverified_draft
A nattokinase-only formula showed no effect on blood lipids through six months, while the same enzyme combined with red yeast rice improved every lipid measured from month one.
Complete structured claim and evidence
At 8,000 FU per day for six months there was no between-group difference in Montreal Cognitive Assessment change, with a mean difference of 0.038.
Commercial nattokinase supplement, composition study-specific → Global cognition score, instrument specified by study source_derived_draftungraded
Experimental context and source evidence
duration
6 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (120 randomized, 88 completed, asymptomatic intracranial or carotid stenosis, mean age 58.3)
exposure
Nattokinase, 8,000 FU per day
limitations
Double-blind, placebo-controlled and single-centre, with substantial attrition. The highest dose tested in any randomized trial here. A null on cognition does not test plaque or thrombotic endpoints, which is what the high-dose cohort studies report.
organism
Human (120 randomized, 88 completed, asymptomatic intracranial or carotid stenosis, mean age 58.3)
plain_language
At 8,000 FU per day for six months there was no between-group difference in Montreal Cognitive Assessment change, with a mean difference of 0.038.
primary_references
Nattokinase supplementation for cognitive enhancement in asymptomatic intracranial/carotid stenosis: A randomized controlled trial. (2026) https://pubmed.ncbi.nlm.nih.gov/41325794/ DOI: 10.1016/j.jstrokecerebrovasdis.2025.108511
route
In vivo, oral
tissue
Cognitive assessment with multimodal brain MRI
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1225–1225
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (120 randomized, 88 completed, asymptomatic intracranial or carotid stenosis, mean age 58.3) · source_derived_draft · unverified_draft
At 8,000 FU per day for six months there was no between-group difference in Montreal Cognitive Assessment change, with a mean difference of 0.038.
Complete structured claim and evidence
Exploratory analysis of the same trial found a between-group difference in visuospatial function change of 0.350 and a reduced risk of visuospatial decline.
Commercial nattokinase supplement, composition study-specific → Visuospatial function domain score source_derived_draftungraded
Experimental context and source evidence
duration
6 months
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (88 completers with asymptomatic stenosis)
exposure
Nattokinase, 8,000 FU per day
limitations
Exploratory and one of several domains, in a trial whose primary endpoint was null. Recorded as a domain-level finding, not as a cognitive benefit.
organism
Human (88 completers with asymptomatic stenosis)
plain_language
Exploratory analysis of the same trial found a between-group difference in visuospatial function change of 0.350 and a reduced risk of visuospatial decline.
primary_references
Nattokinase supplementation for cognitive enhancement in asymptomatic intracranial/carotid stenosis: A randomized controlled trial. (2026) https://pubmed.ncbi.nlm.nih.gov/41325794/ DOI: 10.1016/j.jstrokecerebrovasdis.2025.108511
route
In vivo, oral
tissue
Cognitive domain scores
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1236–1236
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (88 completers with asymptomatic stenosis) · source_derived_draft · unverified_draft
Exploratory analysis of the same trial found a between-group difference in visuospatial function change of 0.350 and a reduced risk of visuospatial decline.
Complete structured claim and evidence
Nattokinase increased peripheral blood irisin in rat stroke models, and the authors place this on the path to improved neurogenesis and cognition.
Nattokinase / subtilisin NAT → Irisin source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase
limitations
An animal disease model. Whether nattokinase raises irisin by a direct action or as a consequence of the stroke protection is not separated, and human relevance is unestablished.
organism
Rat
plain_language
Nattokinase increased peripheral blood irisin in rat stroke models, and the authors place this on the path to improved neurogenesis and cognition.
primary_references
Nattokinase Promotes Post-stroke Neurogenesis and Cognition Recovery via Increasing Circulating Irisin. (2023) https://pubmed.ncbi.nlm.nih.gov/37466380/ DOI: 10.1021/acs.jafc.2c08718
route
In vivo, oral
tissue
Peripheral blood and hippocampus, post-stroke
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1247–1247
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Nattokinase increased peripheral blood irisin in rat stroke models, and the authors place this on the path to improved neurogenesis and cognition.
Complete structured claim and evidence
Nattokinase promoted neurogenesis in rat stroke models and improved cognitive function.
Nattokinase / subtilisin NAT → Hippocampal neurogenesis in rat source_derived_draftungraded
Experimental context and source evidence
duration
Not stated here
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase
limitations
Proposed to run through peripheral irisin as a blood-to-brain axis, which is the hypothesis the paper advances rather than a measured causal chain.
organism
Rat
plain_language
Nattokinase promoted neurogenesis in rat stroke models and improved cognitive function.
primary_references
Nattokinase Promotes Post-stroke Neurogenesis and Cognition Recovery via Increasing Circulating Irisin. (2023) https://pubmed.ncbi.nlm.nih.gov/37466380/ DOI: 10.1021/acs.jafc.2c08718
route
In vivo, oral
tissue
Hippocampus, post-stroke
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1258–1258
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Nattokinase promoted neurogenesis in rat stroke models and improved cognitive function.
Complete structured claim and evidence
Oral nattokinase at 65 or 130 mg/kg per day improved aluminium and amyloid plaque accumulation in the brain and enhanced brain volume in a rat model.
Nattokinase / subtilisin NAT → Amyloid plaque and aluminium accumulation in rat brain source_derived_draftungraded
Experimental context and source evidence
duration
10 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (40 Wistar in four groups)
exposure
Nattokinase 65 or 130 mg/kg per day, on AlCl3 and D-galactose
limitations
A chemically induced model rather than a genetic or human disease. Doses are far above any human supplement exposure on a body-weight basis. Morris water maze performance improved only partially.
organism
Rat (40 Wistar in four groups)
plain_language
Oral nattokinase at 65 or 130 mg/kg per day improved aluminium and amyloid plaque accumulation in the brain and enhanced brain volume in a rat model.
primary_references
Effect of Nattokinase in D-galactose- and Aluminum Chloride-induced Alzheimer's Disease Model of Rat. (2024) https://pubmed.ncbi.nlm.nih.gov/39477413/ DOI: 10.21873/invivo.13744
route
In vivo, oral
tissue
Brain histochemistry and micro-computed tomography
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1269–1269
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (40 Wistar in four groups) · source_derived_draft · unverified_draft
Oral nattokinase at 65 or 130 mg/kg per day improved aluminium and amyloid plaque accumulation in the brain and enhanced brain volume in a rat model.
Complete structured claim and evidence
The same treatment increased free-form amyloid-beta in cerebrospinal fluid.
Nattokinase / subtilisin NAT → Free-form amyloid-beta concentration in rat cerebrospinal fluid source_derived_draftungraded
Experimental context and source evidence
duration
10 weeks
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (40 Wistar in four groups)
exposure
Nattokinase 65 or 130 mg/kg per day
limitations
Mechanistically ambiguous and recorded as measured rather than interpreted. A rise in free amyloid-beta is consistent with fibril breakdown releasing monomer and with impaired clearance, and this experiment does not distinguish them.
organism
Rat (40 Wistar in four groups)
plain_language
The same treatment increased free-form amyloid-beta in cerebrospinal fluid.
primary_references
Effect of Nattokinase in D-galactose- and Aluminum Chloride-induced Alzheimer's Disease Model of Rat. (2024) https://pubmed.ncbi.nlm.nih.gov/39477413/ DOI: 10.21873/invivo.13744
route
In vivo, oral
tissue
Cerebrospinal fluid
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1280–1280
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (40 Wistar in four groups) · source_derived_draft · unverified_draft
The same treatment increased free-form amyloid-beta in cerebrospinal fluid.
Complete structured claim and evidence
Nattokinase given by mouth for seven days before middle cerebral artery occlusion dose-dependently reduced infarct volume.
Nattokinase / subtilisin NAT → Cerebral infarct volume after transient middle cerebral artery occlusion in rat source_derived_draftungraded
Experimental context and source evidence
duration
7 days of pre-treatment
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase 65 or 130 mg/kg
limitations
Pre-treatment, not treatment: the enzyme was on board before the insult, which is not how an acute stroke is treated. High doses on a body-weight basis. No human clinical inference follows.
organism
Rat
plain_language
Nattokinase given by mouth for seven days before middle cerebral artery occlusion dose-dependently reduced infarct volume.
primary_references
Nattokinase Attenuates Acute Cerebral Infarction in a Rat Model of Middle Cerebral Artery Occlusion. (2026) https://pubmed.ncbi.nlm.nih.gov/41760340/ DOI: 10.21873/invivo.14243
route
In vivo, oral
tissue
Brain, transient middle cerebral artery occlusion
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1291–1291
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
Nattokinase given by mouth for seven days before middle cerebral artery occlusion dose-dependently reduced infarct volume.
Complete structured claim and evidence
The same oral pre-treatment prolonged clotting time in the model animals.
Nattokinase / subtilisin NAT → Whole-blood clotting time in rat source_derived_draftungraded
Experimental context and source evidence
duration
7 days of pre-treatment
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat
exposure
Nattokinase 65 or 130 mg/kg
limitations
Described by the authors as a mild and transient anticoagulant effect. It is the clearest oral in vivo coagulation readout in the chapter, and it is in rat.
organism
Rat
plain_language
The same oral pre-treatment prolonged clotting time in the model animals.
primary_references
Nattokinase Attenuates Acute Cerebral Infarction in a Rat Model of Middle Cerebral Artery Occlusion. (2026) https://pubmed.ncbi.nlm.nih.gov/41760340/ DOI: 10.21873/invivo.14243
route
In vivo, oral
tissue
Whole blood
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1302–1302
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat · source_derived_draft · unverified_draft
The same oral pre-treatment prolonged clotting time in the model animals.
Complete structured claim and evidence
A high-nattokinase diet inhibited the rise in circulating advanced glycation end products in streptozotocin-diabetic rats.
Nattokinase / subtilisin NAT → Advanced glycation end-product formation source_derived_draftungraded
Experimental context and source evidence
duration
14 days
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (streptozotocin-induced diabetes)
exposure
Diet containing 0.6 mg nattokinase per gram
limitations
The low-dose diet at 0.2 mg/g did nothing, so this is dose-dependent. The authors state explicitly that it is unclear whether the effect is caused by intact nattokinase or by peptides derived from it during digestion, which is the same open question this whole chapter turns on.
organism
Rat (streptozotocin-induced diabetes)
plain_language
A high-nattokinase diet inhibited the rise in circulating advanced glycation end products in streptozotocin-diabetic rats.
primary_references
Effect of nattokinase on the pathological conditions in streptozotocin induced diabetic rats. (2024) https://pubmed.ncbi.nlm.nih.gov/38586318/ DOI: 10.1016/j.heliyon.2024.e28835
route
In vivo, oral
tissue
Circulating AGEs and renal histopathology
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1313–1313
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (streptozotocin-induced diabetes) · source_derived_draft · unverified_draft
A high-nattokinase diet inhibited the rise in circulating advanced glycation end products in streptozotocin-diabetic rats.
Complete structured claim and evidence
The same high-nattokinase diet inhibited glycogen deposition in the renal tubules.
Nattokinase / subtilisin NAT → Glycogen deposition in renal tubules of streptozotocin-diabetic rat source_derived_draftungraded
Experimental context and source evidence
duration
14 days
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (streptozotocin-induced diabetes)
exposure
Diet containing 0.6 mg nattokinase per gram
limitations
Dose-dependent, as above.
organism
Rat (streptozotocin-induced diabetes)
plain_language
The same high-nattokinase diet inhibited glycogen deposition in the renal tubules.
primary_references
Effect of nattokinase on the pathological conditions in streptozotocin induced diabetic rats. (2024) https://pubmed.ncbi.nlm.nih.gov/38586318/ DOI: 10.1016/j.heliyon.2024.e28835
route
In vivo, oral
tissue
Renal tubules
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1324–1324
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (streptozotocin-induced diabetes) · source_derived_draft · unverified_draft
The same high-nattokinase diet inhibited glycogen deposition in the renal tubules.
Complete structured claim and evidence
Fasting blood glucose was not reduced by either nattokinase diet.
Nattokinase / subtilisin NAT → Blood glucose concentration source_derived_draftungraded
Experimental context and source evidence
duration
14 days
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (streptozotocin-induced diabetes)
exposure
Diet containing 0.2 or 0.6 mg nattokinase per gram
limitations
A measured null, and the reason the authors describe the renal and AGE effects as occurring without glycaemic correction. It agrees in sign with the small glucose rise in the human meta-analysis only in the sense that neither shows glucose falling.
organism
Rat (streptozotocin-induced diabetes)
plain_language
Fasting blood glucose was not reduced by either nattokinase diet.
primary_references
Effect of nattokinase on the pathological conditions in streptozotocin induced diabetic rats. (2024) https://pubmed.ncbi.nlm.nih.gov/38586318/ DOI: 10.1016/j.heliyon.2024.e28835
route
In vivo, oral
tissue
Fasting blood glucose
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1335–1335
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (streptozotocin-induced diabetes) · source_derived_draft · unverified_draft
Fasting blood glucose was not reduced by either nattokinase diet.
Complete structured claim and evidence
Natto food contains vitamin K2, and one 50 g pack typically carries 1,400 to 2,000 FU of nattokinase activity with a reported range of 600 to 3,290 FU per pack.
Natto, fermented soybean food → Menaquinone-7 / MK-7 source_derived_draftungraded
Experimental context and source evidence
duration
Not applicable
evidence_access
EFSA Journal opinion retrieved and read in full during this curation. Quoted figures are from the opinion text and its specification tables.
experimental_model
Food composition
exposure
Natto, 50 g portion
limitations
Whole fermented soybeans also carry poly-gamma-glutamate, soy protein, peptides and live fermentation organisms. A typical pack is close in nattokinase activity to a 2,000 FU supplement capsule, which makes the vitamin K2 content the sharpest difference between them.
organism
Food composition
plain_language
Natto food contains vitamin K2, and one 50 g pack typically carries 1,400 to 2,000 FU of nattokinase activity with a reported range of 600 to 3,290 FU per pack.
primary_references
Safety of fermented soybean extract NSK-SD as a novel food. (2016) DOI: 10.2903/j.efsa.2016.4541 Not indexed in PubMed.
route
In vivo, oral
tissue
Fermented soybean food
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1346–1346
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Food composition · source_derived_draft · unverified_draft
Natto food contains vitamin K2, and one 50 g pack typically carries 1,400 to 2,000 FU of nattokinase activity with a reported range of 600 to 3,290 FU per pack.
Complete structured claim and evidence
Vitamin K2 is removed during NSK-SD manufacture, to a specification of 0.1 ppm or less, and was not detected in any of five analysed batches.
NSK-SD fermented soybean extract → Menaquinone-7 / MK-7 source_derived_draftungraded
Experimental context and source evidence
duration
Not applicable
evidence_access
EFSA Journal opinion retrieved and read in full during this curation. Quoted figures are from the opinion text and its specification tables.
experimental_model
Product specification and batch analysis
exposure
NSK-SD, 30% fermented soybean extract and 70% resistant dextrin, 20,000-28,000 FU/g
limitations
This is the specification of one product. It does not describe crude fermentation powders or supplements from other manufacturers, and it is the reason natto and NSK-SD must be modelled separately in any question about vitamin K antagonists.
organism
Product specification and batch analysis
plain_language
Vitamin K2 is removed during NSK-SD manufacture, to a specification of 0.1 ppm or less, and was not detected in any of five analysed batches.
primary_references
Safety of fermented soybean extract NSK-SD as a novel food. (2016) DOI: 10.2903/j.efsa.2016.4541 Not indexed in PubMed.
route
In vivo, oral
tissue
Standardised fermented soybean extract
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1357–1357
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Product specification and batch analysis · source_derived_draft · unverified_draft
Vitamin K2 is removed during NSK-SD manufacture, to a specification of 0.1 ppm or less, and was not detected in any of five analysed batches.
Complete structured claim and evidence
In product testing at 25 C, NSK-SD retained activity for 24 h between pH 5.5 and 10 but was labile below pH 5.0.
Acidity below pH 5 → Nattokinase / subtilisin NAT source_derived_draftungraded
Experimental context and source evidence
duration
24 h
evidence_access
EFSA Journal opinion retrieved and read in full during this curation. Quoted figures are from the opinion text and its specification tables.
experimental_model
Product stability testing
exposure
NSK-SD
limitations
A product stability measurement, not a human gastric-survival measurement: fasting gastric pH is well below 5.0. An applicant assay of the softgel at pH 2.0 mimicking gastric fluid was submitted to EFSA, but the opinion records only that the results were provided, not what they were.
organism
Product stability testing
plain_language
In product testing at 25 C, NSK-SD retained activity for 24 h between pH 5.5 and 10 but was labile below pH 5.0.
primary_references
Safety of fermented soybean extract NSK-SD as a novel food. (2016) DOI: 10.2903/j.efsa.2016.4541 Not indexed in PubMed.
route
In vitro
tissue
25% aqueous product solution at 25 C
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1368–1368
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Product stability testing · source_derived_draft · unverified_draft
In product testing at 25 C, NSK-SD retained activity for 24 h between pH 5.5 and 10 but was labile below pH 5.0.
Complete structured claim and evidence
In a 90-day oral study in Sprague-Dawley rats at 0, 100, 300 and 1,000 mg/kg body weight per day, the no-observed-adverse-effect level was 1,000 mg/kg per day, the highest dose tested.
NSK-SD fermented soybean extract → Adverse findings in a 90-day rat oral toxicology battery source_derived_draftungraded
Experimental context and source evidence
duration
90 days
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Rat (Sprague-Dawley, 12 per sex per group)
exposure
NSK-SD at 21,900 FU/g, up to 1,000 mg/kg body weight per day
limitations
A NOAEL at the highest dose tested is a floor, not a ceiling. The material was non-mutagenic and non-clastogenic in vitro, and a 4-week human tolerance study at 10 mg/kg per day reported no problems. None of this tests interaction with antithrombotic drugs or long-term disease outcomes. This study and the regulatory opinion share the same applicant dossier and are not independent of one another.
organism
Rat (Sprague-Dawley, 12 per sex per group)
plain_language
In a 90-day oral study in Sprague-Dawley rats at 0, 100, 300 and 1,000 mg/kg body weight per day, the no-observed-adverse-effect level was 1,000 mg/kg per day, the highest dose tested.
primary_references
Toxicological assessment of nattokinase derived from Bacillus subtilis var. natto. (2016) https://pubmed.ncbi.nlm.nih.gov/26740078/ DOI: 10.1016/j.fct.2015.12.025
route
In vivo, oral gavage
tissue
Subchronic oral toxicology battery
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1379–1379
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Rat (Sprague-Dawley, 12 per sex per group) · source_derived_draft · unverified_draft
In a 90-day oral study in Sprague-Dawley rats at 0, 100, 300 and 1,000 mg/kg body weight per day, the no-observed-adverse-effect level was 1,000 mg/kg per day, the highest dose tested.
Complete structured claim and evidence
A patient taking aspirin for secondary stroke prevention had an acute cerebellar hemorrhage after seven consecutive days of nattokinase at 400 mg daily, with multiple cerebral microbleeds on MRI.
Commercial nattokinase supplement, composition study-specific → Hemorrhage source_derived_draftungraded
Experimental context and source evidence
duration
7 days
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (single case report)
exposure
Nattokinase 400 mg daily with concurrent aspirin
limitations
One case with strong predispositions, so neither causality nor incidence can be inferred from it. It is recorded because formal pharmacokinetic and pharmacodynamic interaction studies with warfarin, direct oral anticoagulants, aspirin or clopidogrel are absent from everything read here, and because the human aPTT and closure-time findings make an additive effect plausible.
organism
Human (single case report)
plain_language
A patient taking aspirin for secondary stroke prevention had an acute cerebellar hemorrhage after seven consecutive days of nattokinase at 400 mg daily, with multiple cerebral microbleeds on MRI.
primary_references
Cerebellar hemorrhage provoked by combined use of nattokinase and aspirin in a patient with cerebral microbleeds. (2008) https://pubmed.ncbi.nlm.nih.gov/18310985/ DOI: 10.2169/internalmedicine.47.0620
route
In vivo, oral
tissue
Cerebellum, with cerebral microbleeds on MR imaging
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1390–1390
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (single case report) · source_derived_draft · unverified_draft
A patient taking aspirin for secondary stroke prevention had an acute cerebellar hemorrhage after seven consecutive days of nattokinase at 400 mg daily, with multiple cerebral microbleeds on MRI.
Complete structured claim and evidence
A patient who substituted nattokinase for warfarin after aortic valve replacement with a mechanical prosthesis developed thrombus on the valve after nearly a year and required repeat valve replacement.
Commercial nattokinase supplement, composition study-specific → Thrombus formation on a mechanical prosthetic heart valve source_derived_draftungraded
Experimental context and source evidence
duration
Nearly one year
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (single case report)
exposure
Nattokinase replacing warfarin
limitations
A single case, and one of treatment failure rather than of intrinsic toxicity. It records what happened when an in vitro fibrinolytic property was treated as anticoagulation, which is the distinction the rest of this chapter is built around.
organism
Human (single case report)
plain_language
A patient who substituted nattokinase for warfarin after aortic valve replacement with a mechanical prosthesis developed thrombus on the valve after nearly a year and required repeat valve replacement.
primary_references
Consequence of patient substitution of nattokinase for warfarin after aortic valve replacement with a mechanical prosthesis. (2015) https://pubmed.ncbi.nlm.nih.gov/25552810/ DOI: 10.1080/08998280.2015.11929198
route
In vivo, oral
tissue
Mechanical aortic valve prosthesis
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1401–1401
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (single case report) · source_derived_draft · unverified_draft
A patient who substituted nattokinase for warfarin after aortic valve replacement with a mechanical prosthesis developed thrombus on the valve after nearly a year and required repeat valve replacement.
Complete structured claim and evidence
All thirteen patients with natto allergy in an allergy-clinic cohort had positive skin prick and basophil activation tests to poly-gamma-glutamic acid.
Poly-gamma-glutamic acid → Sensitization to poly-gamma-glutamic acid source_derived_draftungraded
Experimental context and source evidence
duration
Cross-sectional questionnaire and testing
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (140 food-allergy outpatients, 13 with natto allergy)
exposure
Natto food
limitations
Poly-gamma-glutamate causes late-onset anaphylaxis to natto. The sensitizing exposure this study identified was marine sports, especially surfing, at an odds ratio of 278, and not natto consumption: there was no significant relation between the amount of natto eaten and sensitization. The finding is about natto the food, and does not establish whether any given purified supplement contains poly-gamma-glutamate, residual soy protein or neither.
organism
Human (140 food-allergy outpatients, 13 with natto allergy)
plain_language
All thirteen patients with natto allergy in an allergy-clinic cohort had positive skin prick and basophil activation tests to poly-gamma-glutamic acid.
primary_references
Surfing as a risk factor for sensitization to poly(γ-glutamic acid) in fermented soybeans, natto, allergy. (2018) https://pubmed.ncbi.nlm.nih.gov/29175280/ DOI: 10.1016/j.alit.2017.11.001
route
In vivo, oral
tissue
Skin prick and basophil activation testing
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1412–1412
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (140 food-allergy outpatients, 13 with natto allergy) · source_derived_draft · unverified_draft
All thirteen patients with natto allergy in an allergy-clinic cohort had positive skin prick and basophil activation tests to poly-gamma-glutamic acid.
Complete structured claim and evidence
Poly-gamma-glutamic acid is a constituent of natto and the allergen responsible for late-onset anaphylaxis to it.
Natto, fermented soybean food → Poly-gamma-glutamic acid source_derived_draftungraded
Experimental context and source evidence
duration
Not applicable
evidence_access
Primary PubMed abstract and indexed metadata reviewed. Full-text method details not stated here remain unresolved.
experimental_model
Human (allergy clinic cohort)
exposure
Natto food
limitations
Recorded so that natto the food and a purified preparation are not treated as the same exposure in an allergy question. Purified products may differ in residual soy protein, poly-gamma-glutamate and enzyme content, and no study read here measures that across products.
organism
Human (allergy clinic cohort)
plain_language
Poly-gamma-glutamic acid is a constituent of natto and the allergen responsible for late-onset anaphylaxis to it.
primary_references
Surfing as a risk factor for sensitization to poly(γ-glutamic acid) in fermented soybeans, natto, allergy. (2018) https://pubmed.ncbi.nlm.nih.gov/29175280/ DOI: 10.1016/j.alit.2017.11.001
route
In vivo, oral
tissue
Fermented soybean food
Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23) · lines 1423–1423
Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · supports · Human (allergy clinic cohort) · source_derived_draft · unverified_draft
Poly-gamma-glutamic acid is a constituent of natto and the allergen responsible for late-onset anaphylaxis to it.
Complete structured claim and evidence
MK-7 at 45 micrograms/day lowered mean INR and uncarboxylated factor II by about 40%; lower doses altered anticoagulation in some participants.
Menaquinone-7 / MK-7 → International normalized ratio / INR source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
evidence_span
{"source_cache": "artifacts/k2-research/23530987.abstract.txt", "locator": "Exact primary indexed abstract; zero-based, end-exclusive Unicode character offsets", "file_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af", "start_char": 0, "end_char": 1740, "text_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af"}
experimental_model
Sequential dose-response intervention during anticoagulation
exposure
Acenocoumarol followed by MK-7 10, 20 and 45 micrograms/day
limitations
Small pharmacological study; no dose here is a recommended safe self-adjustment. Drug was acenocoumarol, not warfarin.
nutrient_topic
Vitamin K2 research collection; topical membership is not evidence of a direct dietary effect. · Vitamin K2 / menaquinone family
organism
18 healthy adults; 15 attained target INR
plain_language
K2 is involved in clotting as well as extrahepatic functions.
primary_references
[k2-p23530987] Effect of low-dose supplements of menaquinone-7 (vitamin K2 ) on the stability of oral anticoagulant treatment: dose-response relationship in healthy volunteers. (2013). https://pubmed.ncbi.nlm.nih.gov/23530987/ DOI: 10.1111/jth.12203
tissue_or_cell_type
Coagulation, osteocalcin and MGP markers
trigger_kind
machinery_impairment Imported condition classification; unverified.
Vitamin K2: menaquinone forms, carboxylation, recycling and nutrient interactions (2026-09-17) · lines 1046–1057
AI-assisted literature curation; primary study URLs and scope retained in the document and extraction. Not publisher full text. · supports · Sequential dose-response intervention during anticoagulation · source_derived_draft · unverified_draft
### k2-mk7-inr MK-7 at 45 micrograms/day lowered mean INR and uncarboxylated factor II by about 40%; lower doses altered anticoagulation in some participants. Condition category: machinery_impairment nutrient_topic: Vitamin K2 research collection; topical membership is not evidence of a direct dietary effect. plain_language: K2 is involved in clotting as well as extrahepatic functions. organism: 18 healthy adults; 15 attained target INR tissue_or_cell_type: Coagulation, osteocalcin and MGP markers experimental_model: Sequential dose-response intervention during anticoagulation limitations: Small pharmacological study; no dose here is a recommended safe self-adjustment. Drug was acenocoumarol, not warfarin. exposure: Acenocoumarol followed by MK-7 10, 20 and 45 micrograms/day evidence_span: {"source_cache": "artifacts/k2-research/23530987.abstract.txt", "locator": "Exact primary indexed abstract; zero-based, end-exclusive Unicode character offsets", "file_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af", "start_char": 0, "end_char": 1740, "text_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af"} [k2-p23530987] Effect of low-dose supplements of menaquinone-7 (vitamin K2 ) on the stability of oral anticoagulant treatment: dose-response relationship in healthy volunteers. (2013). https://pubmed.ncbi.nlm.nih.gov/23530987/ DOI: 10.1111/jth.12203
Complete structured claim and evidence
MK-7 at 10 and 20 micrograms/day increased endogenous thrombin potential by approximately 20% and 30%, respectively; ucOC and dp-ucMGP did not change.
Menaquinone-7 / MK-7 → Endogenous thrombin generation potential source_derived_draftungraded
Experimental context and source evidence
availability_state
machinery_impairment Imported condition classification; unverified.
evidence_span
{"source_cache": "artifacts/k2-research/23530987.abstract.txt", "locator": "Exact primary indexed abstract; zero-based, end-exclusive Unicode character offsets", "file_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af", "start_char": 0, "end_char": 1740, "text_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af"}
experimental_model
Sequential dose-response intervention during anticoagulation
exposure
Acenocoumarol followed by MK-7 10, 20 and 45 micrograms/day
limitations
Small pharmacological study; no dose here is a recommended safe self-adjustment. Drug was acenocoumarol, not warfarin.
nutrient_topic
Vitamin K2 research collection; topical membership is not evidence of a direct dietary effect. · Vitamin K2 / menaquinone family
organism
18 healthy adults; 15 attained target INR
plain_language
A clotting response appeared without the same response in the other measured proteins.
primary_references
[k2-p23530987] Effect of low-dose supplements of menaquinone-7 (vitamin K2 ) on the stability of oral anticoagulant treatment: dose-response relationship in healthy volunteers. (2013). https://pubmed.ncbi.nlm.nih.gov/23530987/ DOI: 10.1111/jth.12203
tissue_or_cell_type
Coagulation, osteocalcin and MGP markers
trigger_kind
machinery_impairment Imported condition classification; unverified.
Vitamin K2: menaquinone forms, carboxylation, recycling and nutrient interactions (2026-09-17) · lines 1059–1070
AI-assisted literature curation; primary study URLs and scope retained in the document and extraction. Not publisher full text. · supports · Sequential dose-response intervention during anticoagulation · source_derived_draft · unverified_draft
### k2-mk7-thrombin MK-7 at 10 and 20 micrograms/day increased endogenous thrombin potential by approximately 20% and 30%, respectively; ucOC and dp-ucMGP did not change. Condition category: machinery_impairment nutrient_topic: Vitamin K2 research collection; topical membership is not evidence of a direct dietary effect. plain_language: A clotting response appeared without the same response in the other measured proteins. organism: 18 healthy adults; 15 attained target INR tissue_or_cell_type: Coagulation, osteocalcin and MGP markers experimental_model: Sequential dose-response intervention during anticoagulation limitations: Small pharmacological study; no dose here is a recommended safe self-adjustment. Drug was acenocoumarol, not warfarin. exposure: Acenocoumarol followed by MK-7 10, 20 and 45 micrograms/day evidence_span: {"source_cache": "artifacts/k2-research/23530987.abstract.txt", "locator": "Exact primary indexed abstract; zero-based, end-exclusive Unicode character offsets", "file_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af", "start_char": 0, "end_char": 1740, "text_sha256": "95455ea6f2fc6de10ed4d46cad9c1587c004ccd445d7b08066108261f161e6af"} [k2-p23530987] Effect of low-dose supplements of menaquinone-7 (vitamin K2 ) on the stability of oral anticoagulant treatment: dose-response relationship in healthy volunteers. (2013). https://pubmed.ncbi.nlm.nih.gov/23530987/ DOI: 10.1111/jth.12203
Complete structured claim and evidence

Availability and dependencies

Each situation shows the normal role first, then what the sources report under a specific condition. A shortfall in the diet, a fault in the machinery, and a low blood reading are kept separate because they are not the same thing.

Knocking down SRF or THBS1 reopens the inflammasome route nattokinase closes

Condition: machinery_impairment · Small interfering RNA against SRF, or against THBS1, applied alongside nattokinase.

Normal role: Nattokinase raises SRF and THBS1 transcript levels, induces autophagy, and lowers NLRP3 inflammasome formation and necroptosis in endothelial cells. SREBF1 appeared in the same sequencing screen and was unchanged on confirmation.

Recorded consequence: Autophagy induction was suppressed and the NLRP3 and necroptosis signals returned, so the anti-inflammatory effect runs through these two transcripts rather than around them.

Scope: One endothelial cell model with a supporting mouse arm, using transcript and pathway readouts.

Blocking Nrf2 or heme oxygenase 1 removes the protection nattokinase gives

Condition: machinery_impairment · Pharmacological inhibition of Nrf2 or of heme oxygenase 1.

Normal role: Nattokinase pretreatment raises Nrf2 activation and heme oxygenase 1 expression and suppresses the COX-2, prostaglandin E2 and MMP-1 induction that particulate matter causes, along with NADPH-oxidase-derived reactive oxygen species.

Recorded consequence: The protective effect was abolished, placing the Nrf2/HO-1 axis on the path rather than beside it.

Scope: One human cell line in a periodontal inflammation model, with antioxidant-response-element luciferase confirmation of the Nrf2 step.

Blocking nitric-oxide synthase removes the relaxation nattokinase produces

Condition: machinery_impairment · N-nitro-L-arginine methyl ester, which inhibits nitric-oxide synthase.

Normal role: Nattokinase relaxes rat thoracic aortic rings dose-dependently, and the relaxation needs an intact endothelium. The authors attribute it to promotion of nitric-oxide synthesis and release together with reduced receptor-operated calcium influx.

Recorded consequence: The relaxation was attenuated, which places nitric-oxide synthase on the path rather than beside it.

Scope: One ex vivo preparation contracted with KCl or norepinephrine, with indomethacin tested in the same design.

Removing transition-state hydrogen bonds costs turnover and not binding

Condition: machinery_impairment · Single alanine substitution at each of the four positions.

Normal role: Four residues around the catalytic triad, Ser33, Asp60, Ser62 and Thr220, form hydrogen bonds that stabilise the transition state of peptide-bond hydrolysis.

Recorded consequence: The free energy of the transition state rose and kcat fell, while Km was essentially unchanged, so the bonds serve catalysis rather than ground-state substrate binding. Simulation showed the mutations partially releasing Asp32, His64 and Asn155.

Scope: Enzyme kinetics on succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, with free-energy perturbation and molecular dynamics.

The sources

Every document behind this chapter is preserved word for word. Open one to read it in full with its recorded conflicts marked in place.

Nattokinase: what the purified enzyme cleaves, what survives being eaten, and the gap between the two (2026-09-23)Original AI-assisted curation built from a supplied entity-first document of 105 entities and 129 claims. Every reference in that document was resolved against live PubMed with its abstract read and its DOI cross-checked on 2026-09-23, and the EFSA novel-food opinion was retrieved and read in full. That check corrected two PMIDs that pointed at unrelated papers, two DOIs, and two papers recorded as carrying no erratum that do carry one; it also reversed three findings the supplied document had stated backwards. Two papers carry a published correction, recorded as such and not as a retraction. Three sources are not indexed in PubMed and are cited by what they have. Laboratory lineages are recorded, so the four papers from one group, the three from another and the two readings of a single applicant dossier cannot be counted as separate lines of support. Study-specific doses, units, populations and limitations retained; activity units are never converted between systems. Not publisher full text. · unverified_draftRead preserved source
Vitamin K2: menaquinone forms, carboxylation, recycling and nutrient interactions (2026-09-17)AI-assisted literature curation; primary study URLs and scope retained in the document and extraction. Not publisher full text. · unverified_draftRead preserved source

Recorded disagreements

Where two sources say different things, both are kept and the difference is explained. You can discuss a disagreement or propose a mechanism that might account for it.

33bef151-bdc7-5892-aff6-0345ab149739:nattokinase-alone-does-not-improve-blood-lipids / 33bef151-bdc7-5892-aff6-0345ab149739:a-retrospective-cohort-reports-plaque-reduction-at-high-doseThe ledger explicitly records that ‘A nattokinase-only formula showed no effect on blood lipids through six months, while the same enzyme combined with red yeast rice improved every lipid measured from month one.’ contradicts ‘In a retrospective cohort of 1,062 participants, 10,800 FU per day for 12 months was followed by reduced carotid intima-media thickness and carotid plaque size, with improvement rates of 66.5 to 95.4%.’. A randomized placebo-controlled mono-arm found no lipid effect over six months where the uncontrolled cohort reports significant improvement.Read the recorded disagreement
Nattokinase raises tPA in one endothelial study and lowers it in anotherYatagai 2008 raised tissue-plasminogen-activator activity about five-fold in human umbilical vein endothelial cells using heat-deactivated material, while Ji 2014 found nattokinase reducing adrenaline-stimulated tPA secretion from the same cell type. The two differ in whether the enzyme was catalytically active and in whether the cells were stimulated, and either difference could explain the sign. Both readings are load-bearing: the first is the only evidence that inactivated material has a cellular effect, and the second is part of the mechanism proposed for cerebral protection.Read the recorded disagreement
Nattokinase with aspirin is reported as synergistic benefit and as a bleeding eventA retrospective cohort by manufacturer-employed authors reports that co-administering aspirin and vitamin K2 with nattokinase improved carotid and lipid outcomes. A case report describes a cerebellar hemorrhage in a patient on aspirin with cerebral microbleeds after seven days of nattokinase. These are not the same endpoint and cannot refute each other, but they are the entire evidence base on this combination, and they point opposite ways.Read the recorded disagreement
Plasma renin activity falls in one blood-pressure trial and rises in a subgroup of anotherThe Korean trial reported a net fall in plasma renin activity of 1.17 ng/mL/h against placebo. The North American trial found renin rising in 66% of an exploratory subgroup with low baseline renin, against 8% on placebo. Both trials reported a blood-pressure fall, so the mediator moves in opposite directions while the outcome moves the same way. That is a reason to doubt renin is the mediator at all.Read the recorded disagreement
Randomized trials pool to worse lipids while uncontrolled studies report better onesThe meta-analysis of six randomized trials found total cholesterol and LDL higher and HDL lower at low dose. The open active-comparator study at 6,000 FU per day found HDL rising, and the retrospective cohort at 10,800 FU per day reported significant lipid improvement. Dose and design differ together, so neither can be separated from the other by the evidence available.Read the recorded disagreement
Short trials find a blood-pressure reduction and the one long trial finds noneTwo eight-week randomized trials and a meta-analysis of six report systolic reductions of roughly 3.5 to 5.5 mmHg. The three-year randomized trial at the same 2,000 FU dose found no effect on blood pressure at all. The populations differ: the short trials enrolled people with elevated blood pressure and the long trial enrolled low-risk adults not selected for it, so regression to the mean and a floor effect are both available explanations alongside a real difference in duration or population.Read the recorded disagreement

Open questions in this collection

Questions the curators could not answer from the sources in front of them, kept here with the reason each one is still open. These are gaps in this collection, not findings or proof that no one has studied them.

Does any intact catalytically active nattokinase reach human plasma after an ordinary oral dose, and at what concentration?Every direct-proteolysis mechanism in this chapter depends on this step and nothing measures it. The one human pharmacokinetic study used an unvalidated polyclonal ELISA with no control group and no activity assay, and EFSA concluded the dossier did not allow a conclusion on absorption of the active enzyme. The decisive experiment is immunocapture followed by a substrate-selective catalytic assay, with inhibitor controls.
What happened in the applicant assay of the softgel product at pH 2.0 mimicking gastric fluid?EFSA records that the assay was performed and that its results were provided to the Panel, and does not state what they showed. The product itself is labile below pH 5.0 and fasting gastric pH is far below that, so this is the single most load-bearing unpublished result in the chapter.
Do the ACE-inhibitory peptides LY, FY, LGG and TW reach relevant concentrations in human plasma after oral nattokinase?Heat inactivation raises ACE inhibition rather than removing it, so destroying the enzyme generates a different active species. That makes the peptides a live alternative explanation for the human blood-pressure signal, and no human study has measured them.
What proximal target connects an extracellular protease to Nrf2, NF-kappa-B and SRF/THBS1 transcription?Four separate cell studies report intracellular transcriptional changes from an extracellular enzyme without identifying a receptor, a protease-activated-receptor cleavage event or a peptide mediator. A protease-activated receptor is the obvious candidate and is untested.
Do any of the signalling effects require catalytic activity, or would heat-inactivated enzyme reproduce them?No study in this chapter ran an inactivated-enzyme arm alongside an active one for the signalling endpoints. The one experiment that did compare them, on ACE inhibition, found the inactivated material more active, which is a reason to expect the answer to be surprising.
Was the test article in the platelet and thromboxane work purified subtilisin NAT or a crude fermentation material?The abstract says only "nattokinase extracted from fermented soybean". The supplied document asserted a spray-dried soy-containing powder and built a correction on that assertion; nothing read here supports it, so the identity is left open rather than decided in either direction.
Is nattokinase absorbed intact enough to explain the oral rat thrombosis results, or do those run through another species?Oral pre-treatment delayed ferric-chloride arterial occlusion in rat, reduced carrageenan thrombosis in mouse, prolonged clotting time in rat and reduced infarct volume in rat. Those are outcomes without an identified circulating agent, and the diabetic-rat study's own authors raise the same question about their result.
Does nattokinase cleave latent or vitronectin-bound PAI-1, or only the active conformer?The cleavage experiment used active recombinant prokaryotic PAI-1. Most circulating PAI-1 is latent or vitronectin-bound, so the answer decides how much of the in vitro potency could matter in plasma.
Does oral nattokinase change von Willebrand factor multimer distribution in humans?The A2 cleavage is the strongest new molecular result in the chapter and the only human measurement touching it is an exploratory sex-restricted decrease at p < 0.1. A site-specific Thr1608-Gly1609 neoepitope assay alongside multimer analysis would settle it.
What is the step between oral nattokinase and lower factor VII and factor VIII?Both fall in human studies, in one case within hours of a single dose. Direct proteolytic cleavage of either by purified nattokinase has not been shown, so an arrow from the supplement to these factors currently has no molecular step behind it.
Is FU the wrong dose currency for the non-protease mechanisms?FU standardises fibrinogen-degrading activity. Peptide yield depends on sequence, formulation and digestion instead, so two products matched on FU need not be matched on peptide exposure. This would show up as poor dose-response coherence, which is what the chapter contains.
Why does a 12-month uncontrolled cohort at 10,800 FU per day report large plaque reduction when a 3-year randomized trial at 2,000 FU per day reports none?A dose threshold and uncontrolled design produce the same pattern. The cohort reports its own null at 3,600 FU per day, which is a dose-response argument; its authors are manufacturer-employed, which is the competing explanation. No randomized trial above 2,000 FU per day has measured a vascular structural endpoint.
How much of the enzyme is in a preparation labelled by activity rather than by mass?The only specific-activity figure available is an applicant calculation of about 480 FU/mg from a purified preparation, used to infer that nattokinase is 89 to 93% of the NSK-SD protein fraction. It is product- and assay-specific and cannot be transferred between preparations.
What are the signal peptide and propeptide lengths of the subtilisin NAT precursor?The commonly repeated 29-residue signal and 77-residue propeptide were not sourced by anything read during this curation: PubMed carries no abstract for the gene-sequence paper. The 275-residue mature chain is well sourced and is what this chapter records.
On what does the conclusion that the product is safe actually rest?EFSA concluded NSK-SD safe under the applicant conditions of up to 100 mg per day, a maximum of 2,800 FU per day, in healthy adults over 35 excluding pregnant and lactating women. But in the same opinion the Panel found the human studies too low-powered and too inconsistent to conclude on safety from them, and rested the conclusion on margins of exposure from animal toxicology instead: 700 on a mass basis and 550 on an activity basis. It is a safety opinion under applicant-defined conditions and assessed no health claim, so it says nothing about efficacy and nothing about co-medication with an antithrombotic drug.
Does nattokinase convert prourokinase to urokinase?A 2010 crystallography paper states in its introduction that nattokinase can activate pro-urokinase and tPA. That is a background assertion with no primary result behind it in anything read here, and EFSA cites the heat-inactivation study for the tPA half. The conversion is recorded as unverified rather than as a mechanism.

Chapters are assembled from supplied drafts and curated literature summaries. Statements remain unverified against the primary studies, and the ledger is not medical advice.

Evidence, AI assistance and curation standards

Nattokinase / subtilisin NAT

The mechanisms

A strong fibrinolytic activity averaging about 40 CU, in plasmin units, per gram wet weight was demonstrated in natto, and the enzyme responsible was extracted with saline and named nattokinase.

The aprN gene of Bacillus subtilis (natto) encodes the subtilisin NAT preproenzyme.

The mature enzyme purified from natto is a single 275-residue polypeptide of 27,728 Da that is highly homologous to the subtilisins.

Phenylmethylsulfonyl fluoride inhibited both the fibrinolytic and the amidolytic activity of the purified enzyme, placing it in the serine-protease class.

Alanine substitution at Ser33, Asp60, Ser62 or Thr220 raised the free energy of the transition state and lowered kcat.

The same four substitutions left Km essentially unchanged, so the bonds support catalysis rather than ground-state substrate binding.

The deposited 1.36 A subtilisin NAT structure models four calcium ions across its two chains, and the 1.74 A native structure is a monomeric assembly of one 275-residue chain.

Purified nattokinase cleaves cross-linked fibrin directly in a clot-lysis assay.

Purified nattokinase proteolyzes human fibrinogen.

The enhancement of clot lysis seen without added PAI-1 is attributed to direct fibrin dissolution by the enzyme rather than to plasminogen activation.

Subtilisin NAT cleaved active recombinant PAI-1 at its reactive-site Arg346-Met347 bond and PAI-1 lost specific activity dose-dependently over 0.02-1.0 nM, with a half-maximal effect near 0.1 nM.

At 1 nM and without added PAI-1, subtilisin NAT enhanced tPA-induced fibrin clot lysis by 48 +/- 1.4%.

At the same 1 nM and with active PAI-1 present, the same enzyme enhanced tPA-induced clot lysis by 78 +/- 2.0%.

Nattokinase directly cleaved the folded von Willebrand factor A2 domain and the unstructured vWF73 peptide under static conditions, with Thr1608-Gly1609 as the primary site.

Nattokinase showed a catalytic efficiency on the tested vWF A2 substrate about 200-fold that of ADAMTS13.

Unlike ADAMTS13, nattokinase cleaved the A2 domain without the shear-dependent substrate unfolding that exposes the ADAMTS13 scissile bond.

Nattokinase degraded amyloid fibrils in vitro, and the study determined the conditions under which it does so.

Subtilisin Carlsberg degraded the same amyloid fibrils.

Proteinase K degraded the same amyloid fibrils.

Plasmin did not degrade the fibrils that nattokinase, proteinase K and subtilisin Carlsberg degraded.

Trypsin likewise did not degrade the fibrils.

Nattokinase degraded SARS-CoV-2 spike protein in transfected cell lysates in a dose- and time-dependent way, and degraded surface spike when added to the culture medium.

On a molar basis, nattokinase restored 62.0 +/- 5.3% of arterial blood flow in a chemically induced rat carotid thrombosis model.

Plasmin restored 15.8 +/- 0.7% of blood flow in the same model on the same molar basis.

Elastase restored 0% of blood flow in the same model.

Oral nattokinase for one week delayed ferric-chloride-induced carotid occlusion dose-dependently, doubling occlusion time at 160 mg/kg and fully preventing occlusion at 500 mg/kg.

Aspirin at 30 mg/kg fully prevented occlusion in the same model, as the 500 mg/kg nattokinase dose did.

Supplementing nattokinase at 4,000, 8,000 or 12,000 FU/kg reduced carrageenan-induced thrombosis in mice.

Nattokinase inhibited both collagen-induced and thrombin-induced aggregation of rabbit platelets.

Nattokinase reduced thromboxane B2 formation from collagen-activated platelets in a concentration-dependent manner.

Nattokinase markedly inhibited the rise in intracellular calcium in thrombin-stimulated human platelets.

Nattokinase significantly increased cyclic AMP levels in the cerebral ischemia model.

Incubating blood with nattokinase produced a significant dose-dependent decrease in red blood cell aggregation.

The same incubation produced a significant dose-dependent decrease in low-shear whole-blood viscosity.

Nattokinase relaxed rat thoracic aortic rings dose-dependently, and the relaxation required an intact endothelium.

N-nitro-L-arginine methyl ester attenuated the relaxation, implicating nitric-oxide synthase in it.

Nattokinase reduced receptor-operated calcium influx in vascular smooth muscle.

Culture solutions of Bacillus subtilis natto and market nattokinase, deactivated by heating at 121 C for 15 min, raised tissue-plasminogen-activator activity in HeLa cells about 20-fold and 24-fold and in human umbilical vein endothelial cells about 5-fold.

In human umbilical vein endothelial cells, nattokinase reduced adrenaline-stimulated secretion of tissue-type plasminogen activator.

In the same cells, nattokinase reduced adrenaline-stimulated secretion of plasminogen activator inhibitor 1.

Nattokinase suppressed lipopolysaccharide-induced TLR4 activation in RAW264.7 macrophages.

Nattokinase suppressed lipopolysaccharide-induced NOX2 activation in the same macrophages.

Suppressing TLR4 and NOX2 activation repressed the corresponding reactive-oxygen-species production.

Nattokinase repressed translocation of NF-kappa-B from the cytoplasm to the nucleus in stimulated macrophages, along with MAPK activation.

MAPK activation was repressed in the same stimulated macrophages.

Nattokinase markedly inhibited the rise in serum PAI-1 induced by lipopolysaccharide in mice.

Lower serum PAI-1 was accompanied by blocked fibrin deposition in the glomeruli of endotoxin-treated animals.

Nattokinase pretreatment enhanced Nrf2 activation in particulate-matter-exposed human gingival fibroblasts, confirmed by antioxidant-response-element luciferase assay.

Nattokinase raised heme oxygenase 1 expression in the same cells.

Inhibition of Nrf2 or of heme oxygenase 1 abolished the protective effect of nattokinase on COX-2, prostaglandin E2 and MMP-1 induction.

Nattokinase suppressed NADPH-oxidase-derived reactive-oxygen-species generation in particulate-exposed gingival fibroblasts.

RNA sequencing and Q-PCR showed nattokinase upregulated serum response factor in endothelial cells.

Nattokinase upregulated thrombospondin 1 in the same cells.

SREBF1 messenger RNA appeared in the sequencing screen but was unaffected on Q-PCR confirmation.

Nattokinase treatment induced autophagy in endothelial cells.

Nattokinase inhibited NLRP3 inflammasome formation in endothelial cells.

Nattokinase inhibited necroptosis in endothelial cells.

Small interfering RNA against SRF suppressed the autophagy induction and restored NLRP3 inflammasome formation and necroptosis.

Small interfering RNA against THBS1 had the same effect: autophagy was suppressed and NLRP3 and necroptosis signals returned.

Heat treatment at 120 C for 15 min markedly increased the ACE-inhibitory activity of subtilisin NAT while inactivating it as a protease and fragmenting it into small peptides.

Heat fragmentation and simulated gastrointestinal digestion both release ACE-inhibitory peptides from the nattokinase sequence.

The Leu-Tyr dipeptide, liberated by fragmentation of subtilisin NAT, inhibited angiotensin-converting enzyme.

The Phe-Tyr dipeptide, liberated in the same way, inhibited angiotensin-converting enzyme.

The Leu-Gly-Gly peptide identified from simulated digestion inhibited ACE competitively with an IC50 of 10.78 micromolar.

The Thr-Trp peptide from the same digestion acted as a noncompetitive ACE inhibitor.

Catalytically active intact nattokinase is not required for the ACE-inhibitory activity of the material, because heat inactivation raises rather than removes it.

Nattokinase injected into a tied intestinal loop at 80 mg/kg was followed by fibrinogen degradation products in plasma in all twenty rats, with a 270 kDa fragment appearing within 0.5 to 5 h.

In parallel with the fibrinogen degradation, plasma recalcification time was prolonged.

Nattokinase is a moderately absorbed biomolecule whose transport through enterocytes is energy- and time-dependent.

Chlorpromazine inhibited nattokinase uptake in Caco-2 monolayers, implicating clathrin-mediated endocytosis.

Nystatin inhibited uptake in the same system, implicating a caveolae-mediated pathway.

EIPA inhibited uptake in the same system, implicating macropinocytosis.

A single 2,000 FU oral NSK-SD dose produced anti-nattokinase immunoreactivity in serum that rose significantly from baseline between 2 and 24 h and peaked at about 13.3 +/- 2.5 h.

A polyclonal ELISA signal does not by itself distinguish intact catalytically active nattokinase from fragments, complexes or cross-reactive material.

EFSA concluded that the information provided on absorption, distribution, metabolism and excretion did not allow conclusions to be drawn on the absorption of active nattokinase or any functional metabolites derived from it.

D-dimer concentrations rose significantly 6 and 8 h after a single 2,000 FU oral dose.

Fibrin and fibrinogen degradation products rose significantly 4 h after the same single dose.

Factor VIII activity declined significantly at 4 and 6 h after the same single dose.