Nattokinase / subtilisin NAT

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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%.

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%.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.