{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T13:27:07Z","timestamp":1786541227401,"version":"build-2736575974"},"reference-count":44,"publisher":"American Chemical Society (ACS)","issue":"31","license":[{"start":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T00:00:00Z","timestamp":1785542400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T00:00:00Z","timestamp":1785542400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T00:00:00Z","timestamp":1785542400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100018532","name":"Major Scientific and Technological Innovation Project of Shandong Province","doi-asserted-by":"publisher","award":["2022CXGC020414"],"award-info":[{"award-number":["2022CXGC020414"]}],"id":[{"id":"10.13039\/501100018532","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Marine Aquaculture Innovation and Entrepreneurship Community Project of Shandong Province","award":["YZ2024004"],"award-info":[{"award-number":["YZ2024004"]}]},{"name":"Key R&D Program of Shandong Province","award":["2022TZXD005"],"award-info":[{"award-number":["2022TZXD005"]}]},{"name":"Key R&D Program of Shandong Province","award":["2024TZXD003"],"award-info":[{"award-number":["2024TZXD003"]}]},{"name":"Key R&D Program of Shandong Province","award":["2024TZXD046"],"award-info":[{"award-number":["2024TZXD046"]}]},{"name":"Taishan Scholars Program","award":["tstp20250716"],"award-info":[{"award-number":["tstp20250716"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Rational sequence optimization is essential for transforming food-derived peptides into more potent angiotensin-converting enzyme (ACE) inhibitors. LYPVK and its sequence-designed derivative YWLKP exhibit potent activity; however, their binding mechanisms, transport routes, and in vivo antihypertensive effects remain unclear. Here, they were systematically investigated by biotechnological preparation, biolayer interferometry (BLI), molecular dynamics (MD) simulation, Caco-2 transport assays, and studies on spontaneously hypertensive rats (SHRs). BLI revealed that YWLKP bound to ACE with a subnanomolar affinity constant KD of (1.08 \u00b1 0.40) \u00d7 10\u20139 M, consistent with MD results showing a more stable peptide\u2013ACE complex. Caco-2 monolayer transport showed that LYPVK mainly permeated via the paracellular pathway, whereas YWLKP exhibited PepT1-mediated transport. In SHRs, both peptides significantly reduced blood pressure after single and long-term administration, accompanied by renin\u2013angiotensin system modulation, reduced renal and cardiac impairment, and improved endothelial dysfunction. Collectively, these findings support YWLKP as a promising antihypertensive candidate for functional food applications.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c08999","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T14:33:38Z","timestamp":1785854018000},"page":"24457-24476","source":"Crossref","is-referenced-by-count":0,"title":["Heterologous Expression,\nAntihypertensive Effect,\nand Underlying Mechanism of Sequence-Designed ACE Inhibitory Peptide\nYWLKP"],"prefix":"10.1021","volume":"74","author":[{"given":"Qingping","family":"Liang","sequence":"first","affiliation":[{"name":"Ocean University of China , , ,","place":["Qingdao, China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nana","family":"Sun","sequence":"additional","affiliation":[{"name":"Ocean University of China , , ,","place":["Qingdao, China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changliang","family":"Zhu","sequence":"additional","affiliation":[{"name":"Ocean University of China , , ,","place":["Qingdao, China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dongyu","family":"Li","sequence":"additional","affiliation":[{"name":"Ocean University of China , , ,","place":["Qingdao, China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhemin","family":"Liu","sequence":"additional","affiliation":[{"name":"Ocean University of China , , ,","place":["Qingdao, China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3667-1562","authenticated-orcid":true,"given":"Haijin","family":"Mou","sequence":"additional","affiliation":[{"name":"Ocean University of China , , ,","place":["Qingdao, China, 266404"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,1]]},"reference":[{"issue":"3","key":"2026081208091254800_cit1","doi-asserted-by":"publisher","first-page":"1772","DOI":"10.3390\/ijms23031772","article-title":"Immunomodulatory activity of the\nmost commonly used antihypertensive drugs\u2013Angiotensin converting\nenzyme inhibitors and angiotensin II receptor blockers","volume":"23","author":"Bryniarski","year":"2022","journal-title":"Int. J. Mol. Sci"},{"key":"2026081208091254800_cit2","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1016\/j.ijbiomac.2022.01.114","article-title":"Exploration of interaction between\nangiotensin I-converting enzyme (ACE) and the inhibitory peptide from\nWakame (Undaria pinnatifida)","volume":"204","author":"Feng","year":"2022","journal-title":"Int. J. Biol. Macromol"},{"issue":"5","key":"2026081208091254800_cit3","doi-asserted-by":"publisher","first-page":"408","DOI":"10.2174\/0929866529666220106100225","article-title":"Current trends and\napplications of food-derived antihypertensive peptides for the management\nof cardiovascular disease","volume":"29","author":"Shukla","year":"2022","journal-title":"Protein Peptide Lett"},{"issue":"5","key":"2026081208091254800_cit4","doi-asserted-by":"publisher","first-page":"2376","DOI":"10.1002\/fft2.70076","article-title":"Improving activity of ACE inhibitory\npeptide by sequence-based rational design: Properties, function on\nendothelial cells, and potential antihypertensive mechanism","volume":"6","author":"Liang","year":"2025","journal-title":"Food Front"},{"issue":"3","key":"2026081208091254800_cit5","doi-asserted-by":"publisher","first-page":"759","DOI":"10.1021\/acs.jafc.9b05618","article-title":"Antihypertensive\neffects in vitro and in vivo of\nnovel angiotensin-converting enzyme inhibitory peptides from bovine\nbone gelatin hydrolysate","volume":"68","author":"Cao","year":"2020","journal-title":"J. Agric. Food Chem"},{"issue":"4","key":"2026081208091254800_cit6","doi-asserted-by":"publisher","first-page":"3645","DOI":"10.1039\/C9FO02687H","article-title":"Antihypertensive effects\nof Trichiurus lepturus myosin hydrolysate in spontaneously\nhypertensive rats","volume":"11","author":"Fu","year":"2020","journal-title":"Food Funct"},{"issue":"1","key":"2026081208091254800_cit7","doi-asserted-by":"publisher","DOI":"10.1111\/jfbc.12571","article-title":"Influence\nof peptide characteristics on their stability,\nintestinal transport, and in vitro bioavailability: A review","volume":"43","author":"Wang","year":"2019","journal-title":"J. Food Biochem"},{"issue":"2","key":"2026081208091254800_cit8","doi-asserted-by":"publisher","first-page":"1150","DOI":"10.1111\/1541-4337.12711","article-title":"Activity and bioavailability of food\nprotein-derived angiotensin-I-converting enzyme\u2013inhibitory\npeptides","volume":"20","author":"Xue","year":"2021","journal-title":"Compr. Rev. Food. Sci. Food Saf"},{"key":"2026081208091254800_cit9","doi-asserted-by":"publisher","first-page":"140274","DOI":"10.1016\/j.ijbiomac.2025.140274","article-title":"Heterologous expression of a recombinant ACE inhibitory peptide LYPVK\nand its potential antihypertensive action mechanism","volume":"300","author":"Liang","year":"2025","journal-title":"Int. J. Biol. Macromol"},{"issue":"20","key":"2026081208091254800_cit10","doi-asserted-by":"publisher","first-page":"6207","DOI":"10.3390\/molecules26206207","article-title":"Construction of\na tandem repeat peptide sequence with pepsin cutting sites to produce\nrecombinant \u03b1-melanocyte-stimulating hormone","volume":"26","author":"Jiang","year":"2021","journal-title":"Molecules"},{"key":"2026081208091254800_cit11","doi-asserted-by":"publisher","first-page":"104443","DOI":"10.1016\/j.tifs.2024.104443","article-title":"Research progress in the preparation and structure-activity relationship\nof bioactive peptides derived from aquatic foods","volume":"147","author":"Mao","year":"2024","journal-title":"Trends Food Sci. Technol"},{"issue":"8","key":"2026081208091254800_cit12","doi-asserted-by":"publisher","first-page":"2435","DOI":"10.1007\/s11947-023-03278-9","article-title":"Identification novel\nsalty-enhancing peptides from sea cucumber collagen:\nAlphaFold2 modeling and molecular simulation","volume":"17","author":"Bu","year":"2024","journal-title":"Food Bioprocess Technol"},{"key":"2026081208091254800_cit13","doi-asserted-by":"publisher","first-page":"142965","DOI":"10.1016\/j.foodchem.2025.142965","article-title":"Transport and action\nof sesame protein-derived ACE inhibitory peptides ITAPHW and IRPNGL","volume":"472","author":"Du","year":"2025","journal-title":"Food Chem"},{"issue":"29","key":"2026081208091254800_cit14","doi-asserted-by":"publisher","first-page":"8149","DOI":"10.1021\/acs.jafc.9b02276","article-title":"Isolation and characterization of three antihypertension\npeptides from the mycelia of Ganoderma lucidum (Agaricomycetes)","volume":"67","author":"Wu","year":"2019","journal-title":"J. Agric. Food Chem"},{"issue":"45","key":"2026081208091254800_cit15","doi-asserted-by":"publisher","first-page":"17420","DOI":"10.1021\/acs.jafc.3c03752","article-title":"Screening, characterization,\nand mechanistic evaluation\nof angiotensin converting enzyme inhibitory peptides derived from\nmilk fermented with Lactobacillus delbrueckii QS306\nwith and without ultrahigh-pressure treatment","volume":"71","author":"Wu","year":"2023","journal-title":"J. Agric. Food Chem"},{"key":"2026081208091254800_cit16","doi-asserted-by":"publisher","first-page":"115155","DOI":"10.1016\/j.foodres.2024.115155","article-title":"Exploring the salty peptides of enzymatically hydrolyzed Volvariella volvacea protein: Structural analysis and taste\nperception insights","volume":"197","author":"Jiang","year":"2024","journal-title":"Food Res. Int"},{"issue":"8","key":"2026081208091254800_cit17","doi-asserted-by":"publisher","first-page":"4155","DOI":"10.1021\/acs.jafc.3c08795","article-title":"Novel multitarget\nACE inhibitory\npeptides from bovine colostrum immunoglobulin G: Cellular transport,\nefficacy in regulating endothelial dysfunction, and network pharmacology\nstudies","volume":"72","author":"Wu","year":"2024","journal-title":"J. Agric. Food Chem"},{"key":"2026081208091254800_cit18","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1007\/s00217-015-2597-5","article-title":"Separation and purification\nof angiotensin-I-converting enzyme (ACE) inhibitory peptides from\nwalnuts (Juglans regia L.) meal","volume":"242","author":"Wang","year":"2016","journal-title":"Eur. Food Res. Technol"},{"issue":"7","key":"2026081208091254800_cit19","doi-asserted-by":"publisher","first-page":"2149","DOI":"10.1021\/acs.jafc.0c07048","article-title":"Transport, in vivo antihypertensive effect, and pharmacokinetics of\nan Angiotensin-Converting Enzyme (ACE) inhibitory peptide LVLPGE","volume":"69","author":"Pei","year":"2021","journal-title":"J. Agric. Food Chem"},{"issue":"28","key":"2026081208091254800_cit20","doi-asserted-by":"publisher","first-page":"10638","DOI":"10.1021\/acs.jafc.3c01149","article-title":"Angiotensin\nI-converting enzyme inhibitory activity of two peptides derived from\nin vitro digestion products of pork sausage with partial substitution\nof NaCl by KCl","volume":"71","author":"Zou","year":"2023","journal-title":"J. Agric. Food Chem"},{"issue":"34","key":"2026081208091254800_cit21","doi-asserted-by":"publisher","first-page":"7406","DOI":"10.1021\/acs.jafc.7b02176","article-title":"Transport study of\negg-derived antihypertensive peptides (LKP and IQW) using Caco-2 and\nHT29 coculture monolayers","volume":"65","author":"Xu","year":"2017","journal-title":"J. Agric. Food Chem"},{"key":"2026081208091254800_cit22","doi-asserted-by":"publisher","first-page":"127196","DOI":"10.1016\/j.ijbiomac.2023.127196","article-title":"Identification of novel angiotensin\nconverting enzyme (ACE) inhibitory peptides from Pacific saury: In vivo antihypertensive effect and transport route","volume":"254","author":"Wang","year":"2024","journal-title":"Int. J. Biol. Macromol"},{"issue":"37","key":"2026081208091254800_cit23","doi-asserted-by":"publisher","first-page":"8143","DOI":"10.1021\/acs.jafc.5b01824","article-title":"Transport of antihypertensive\npeptide RVPSL, ovotransferrin 328\u2013332, in human intestinal\nCaco-2 cell monolayers","volume":"63","author":"Ding","year":"2015","journal-title":"J. Agric. Food Chem"},{"key":"2026081208091254800_cit24","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1016\/j.foodres.2016.08.037","article-title":"Angiotensin\nI\u2013converting enzyme\u2013inhibitory peptides from bovine\ncollagen: Insights into inhibitory mechanism and transepithelial transport","volume":"89","author":"Fu","year":"2016","journal-title":"Food Res. Int"},{"key":"2026081208091254800_cit25","doi-asserted-by":"publisher","first-page":"137537","DOI":"10.1016\/j.foodchem.2023.137537","article-title":"Two novel\nangiotensin\nI-converting enzyme inhibitory peptides from garlic protein: In silico screening, stability, antihypertensive effects in vivo and underlying mechanisms","volume":"435","author":"Xiang","year":"2024","journal-title":"Food Chem"},{"issue":"47","key":"2026081208091254800_cit26","doi-asserted-by":"publisher","first-page":"30458","DOI":"10.1021\/acs.jafc.5c06316","article-title":"Antihypertension\nand improvement effects of hexapeptide APPLRP on exosomes-mediated\nvascular endothelial inflammation in spontaneously hypertensive rats","volume":"73","author":"Cai","year":"2025","journal-title":"J. Agric. Food Chem"},{"key":"2026081208091254800_cit27","doi-asserted-by":"publisher","first-page":"103395","DOI":"10.1016\/j.jff.2019.05.051","article-title":"Identification of angiotensin converting\nenzyme 2 (ACE2) up-regulating peptides from pea protein hydrolysate","volume":"60","author":"Liao","year":"2019","journal-title":"J. Funct. Foods"},{"issue":"4","key":"2026081208091254800_cit28","doi-asserted-by":"publisher","first-page":"912","DOI":"10.1021\/jf405189y","article-title":"Antihypertensive effect\nof angiotensin-converting enzyme inhibitory peptide RVPSL on spontaneously\nhypertensive rats by regulating gene expression of the renin\u2013angiotensin\nsystem","volume":"62","author":"Yu","year":"2014","journal-title":"J. Agric. Food Chem"},{"key":"2026081208091254800_cit29","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1016\/j.phrs.2019.01.019","article-title":"The\ninterplay among gut microbiota, hypertension and kidney diseases:\nThe role of short-chain fatty acids","volume":"141","author":"Felizardo","year":"2019","journal-title":"Pharmacol.\nRes"},{"issue":"9","key":"2026081208091254800_cit30","doi-asserted-by":"publisher","DOI":"10.1161\/CIR.0000000000000757","article-title":"Heart\nDisease and Stroke Statistics\u25002020 Update: A Report From the\nAmerican Heart Association","volume":"141","author":"Virani","year":"2020","journal-title":"Circulation"},{"issue":"1","key":"2026081208091254800_cit31","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.tcm.2019.01.007","article-title":"Endothelin-1\nlevels and cardiovascular events","volume":"30","author":"Jankowich","year":"2020","journal-title":"Trends Cardiovasc.\nMed"},{"issue":"2","key":"2026081208091254800_cit32","doi-asserted-by":"publisher","first-page":"626","DOI":"10.3390\/jcm14020626","article-title":"The balance between\nthe natriuretic\npeptides and the renin-angiotensin-aldosterone system in the preservation\nof ideal cardiovascular health","volume":"14","author":"Rubattu","year":"2025","journal-title":"J. Clin. Med"},{"issue":"9","key":"2026081208091254800_cit33","doi-asserted-by":"publisher","first-page":"1194","DOI":"10.1038\/s41440-021-00692-4","article-title":"Dairy-originated\ndigestion-resistant and bioactive peptides increase the risk of hypertension:\nTehran lipid and glucose study","volume":"44","author":"Barati","year":"2021","journal-title":"Hypertens Res"},{"key":"2026081208091254800_cit34","doi-asserted-by":"publisher","first-page":"304","DOI":"10.1186\/s13075-019-2068-7","article-title":"Significant association between clinical\ncharacteristics and changes in peripheral immuno-phenotype in large\nvessel vasculitis","volume":"21","author":"Matsumoto","year":"2019","journal-title":"Arthritis Res. Ther"},{"issue":"6","key":"2026081208091254800_cit35","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1002\/iub.1390","article-title":"Neopterin:\nAn immune biomarker of coronary artery disease and its association\nwith other CAD markers","volume":"67","author":"Firoz","year":"2015","journal-title":"IUBMB Life"},{"key":"2026081208091254800_cit36","doi-asserted-by":"publisher","first-page":"1081119","DOI":"10.3389\/fphys.2022.1081119","article-title":"Paradigms of endothelial\nstiffening in cardiovascular disease and vascular aging","volume":"13","author":"Aguilar","year":"2023","journal-title":"Front. Physiol"},{"issue":"2","key":"2026081208091254800_cit37","doi-asserted-by":"publisher","first-page":"72","DOI":"10.31083\/j.rcm2502072","article-title":"Molecular mechanisms\nunderlying vascular remodeling in hypertension","volume":"25","author":"Zeng","year":"2024","journal-title":"Rev. Cardiovasc. Med"},{"issue":"4","key":"2026081208091254800_cit38","doi-asserted-by":"publisher","first-page":"620","DOI":"10.1161\/CIRCRESAHA.115.306301","article-title":"Endothelial\ncell dysfunction and the pathobiology of atherosclerosis","volume":"118","author":"Gimbrone","year":"2016","journal-title":"Circ. Res"},{"key":"2026081208091254800_cit39","doi-asserted-by":"publisher","first-page":"1437","DOI":"10.1021\/acs.jafc.8b06341","article-title":"Vasorelaxant and Antihypertensive\nEffects That Are Dependent on the Endothelial NO System Exhibited\nby Rice Bran-Derived Tripeptide","volume":"67","author":"Shobako","year":"2019","journal-title":"J. Agric. Food\nChem"},{"issue":"10","key":"2026081208091254800_cit40","doi-asserted-by":"publisher","first-page":"1437","DOI":"10.1080\/10408398.2021.1964433","article-title":"Advancement and prospects\nof production, transport, functional activity and structure-activity\nrelationship of food-derived angiotensin converting enzyme (ACE) inhibitory\npeptides","volume":"63","author":"Xiang","year":"2023","journal-title":"Crit. Rev. Food Sci. Nutr"},{"issue":"6","key":"2026081208091254800_cit41","doi-asserted-by":"publisher","first-page":"1623","DOI":"10.1016\/j.fshw.2022.06.021","article-title":"Structural\nrequirements and interaction mechanisms of ACE inhibitory peptides:\nMolecular simulation and thermodynamics studies on LAPYK and its modified\npeptides","volume":"11","author":"Zhang","year":"2022","journal-title":"Food Sci. Hum. Wellness"},{"issue":"3","key":"2026081208091254800_cit42","doi-asserted-by":"publisher","first-page":"215","DOI":"10.1080\/08927022.2018.1557327","article-title":"Insight into the binding of ACE-inhibitory peptides to angiotensin-converting\nenzyme: A molecular simulation","volume":"45","author":"Jiang","year":"2019","journal-title":"Mol. Simul"},{"key":"2026081208091254800_cit43","doi-asserted-by":"publisher","first-page":"24463","DOI":"10.1021\/acs.jafc.4c05518","article-title":"Research on the synergistic inhibition of angiotensin-converting\nenzyme (ACE) by the gastrointestinal digestion products of the ACE\ninhibitory peptide FPPDVA","volume":"72","author":"Liang","year":"2024","journal-title":"J. Agric. Food Chem"},{"key":"2026081208091254800_cit44","doi-asserted-by":"publisher","first-page":"384","DOI":"10.1016\/j.jff.2013.11.005","article-title":"Structural\nand functional characterization of hemp seed (Cannabis sativa L.)\nprotein-derived antioxidant and antihypertensive peptides","volume":"6","author":"Girgih","year":"2014","journal-title":"J. Funct. Foods"}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c08999","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/31\/24457\/66353560\/acs.jafc.6c08999.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/31\/24457\/66353560\/acs.jafc.6c08999.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T12:27:30Z","timestamp":1786537650000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/31\/24457\/5238256\/Heterologous-Expression-Antihypertensive-Effect"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,1]]},"references-count":44,"journal-issue":{"issue":"31","published-online":{"date-parts":[[2026,8,1]]},"published-print":{"date-parts":[[2026,8,12]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c08999","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,1]]}}}