{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T18:20:30Z","timestamp":1790619630364,"version":"4.1.0"},"reference-count":37,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Self-Deployed Project of the Institute of Oceanology, Chinese Academy of Sciences","award":["IOCASZZZX107"],"award-info":[{"award-number":["IOCASZZZX107"]}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"crossref","award":["2025M772964"],"award-info":[{"award-number":["2025M772964"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Science and Technology Project of Fujian Province","award":["2024T3057"],"award-info":[{"award-number":["2024T3057"]}]},{"name":"Presidential Fund Initiative for Universities in Qingdao West Coast New Area"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Deep-sea mollusks represent untapped resources for searching novel biologically active peptides effectual against many chronic diseases. Here we presented the identification of four novel angiotensin I-converting enzyme (ACE) inhibitory peptides from the deep-sea mollusk Gigantidas platifrons by using a combined approach of peptidomics and virtual screening. Fifteen protein hydrolysates from five deep-sea macroorganisms were prepared using three different proteases and were determined for their ACE inhibitory activities. Pepsin hydrolysate of G. platifrons protein (GPp) demonstrated the highest inhibition rate against ACE at 400 \u03bcg\/mL. Then, targeted investigation was conducted on the GPp with peptidomic profiling; more than 3000 peptides were de novo identified, which were then subject to virtual screening using the docking software Smina. Subsequently, 29 peptides were selected and synthesized based on the affinity threshold and the interactions with ACE active sites. More than 58% peptides were biologically active, showing more than 50% inhibition to ACE at 400 \u03bcM. Four peptides, LAAHFAR, YAAPYR, NGAGPYGRP, and FTTFGK, exhibited low micromolar inhibition. The most potent peptide, LAAHFAR with an IC50 of 6.01 \u00b1 1.06 \u03bcM, was subject to molecular dynamics simulations for revealing atomistic interaction analysis. LAAHFAR forms comprehensively stable hydrogen bonds with the classic active site of ACE, and its N terminal arginine residue is anchored by additional hydrogen bonding to Cys370, Asp377, and Thr372. This study highlights deep-sea mollusks as an important source of novel ACE inhibitory peptides, contributing to the development of new therapeutic ingredients or functional food agents against hypertension.<\/jats:p>","DOI":"10.3390\/molecules31050757","type":"journal-article","created":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T13:53:20Z","timestamp":1771941200000},"page":"757","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["From Multi-Species Screening to Targeted Investigation: Discovery of ACE Inhibitory Peptides in Gigantidas platifrons via Peptidomics, Virtual Screening, and Molecular Dynamics Simulations"],"prefix":"10.3390","volume":"31","author":[{"given":"Haorui","family":"Zhang","sequence":"first","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 88 Haijun Road, Qingdao 266000, China"},{"name":"College of Life Sciences, Nanjing Agricultural University, Nanjing 210014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhong","family":"Ouyang","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 88 Haijun Road, Qingdao 266000, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qishan","family":"Suo","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 88 Haijun Road, Qingdao 266000, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Chen","sequence":"additional","affiliation":[{"name":"Center of Deep Sea Research, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1344-1206","authenticated-orcid":false,"given":"Jie","family":"Cui","sequence":"additional","affiliation":[{"name":"College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7736-0405","authenticated-orcid":false,"given":"Yang","family":"Yue","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 88 Haijun Road, Qingdao 266000, China"},{"name":"Laboratory for Marine Biology and Biotechnology, Qingdao Marine Science and Technology Center, No. 1 Wenhai Road, Qingdao 266237, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1016\/S0140-6736(21)01330-1","article-title":"Worldwide trends in hypertension prevalence and progress in treatment and control from 1990 to 2019: A pooled analysis of 1201 population-representative studies with 104 million participants","volume":"398","author":"Zhou","year":"2021","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1038\/s41581-019-0244-2","article-title":"The global epidemiology of hypertension","volume":"16","author":"Mills","year":"2020","journal-title":"Nat. Rev. Nephrol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1038\/nrendo.2015.6","article-title":"Novel RAAS agonists and antagonists: Clinical applications and controversies","volume":"11","author":"Romero","year":"2015","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1038\/nature01370","article-title":"Crystal structure of the human angiotensin-converting enzyme-lisinopril complex","volume":"421","author":"Natesh","year":"2003","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"10141","DOI":"10.1021\/acs.jmedchem.8b01309","article-title":"Molecular basis for multiple omapatrilat binding sites within the ACE C-domain: Implications for drug design","volume":"61","author":"Cozier","year":"2018","journal-title":"J. Med. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"146183","DOI":"10.1016\/j.foodchem.2025.146183","article-title":"Potentials of food-derived peptides as novel antihypertensive agents and their acting mechanisms","volume":"494","author":"Guo","year":"2025","journal-title":"Food Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1016\/j.tifs.2021.08.017","article-title":"Bioactive peptides derived from marine sources: Biological and functional properties","volume":"119","author":"Cunha","year":"2022","journal-title":"Trends Food Sci. Technol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jff.2010.01.003","article-title":"Development and biological activities of marine-derived bioactive peptides: A review","volume":"2","author":"Kim","year":"2010","journal-title":"J. Funct. Foods"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1007\/s10989-023-10495-5","article-title":"In vitro structure-activity relationship study of a novel octapeptide angiotensin-I converting enzyme (ACE) inhibitor from the freshwater mussel Lamellidens marginalis","volume":"29","author":"Das","year":"2023","journal-title":"Int. J. Pept. Res. Ther."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1624","DOI":"10.1016\/j.biortech.2005.01.001","article-title":"Angiotensin I converting enzyme (ACE) inhibitory peptide derived from the sauce of fermented blue mussel. Mytilus edulis","volume":"96","author":"Je","year":"2005","journal-title":"Bioresour. Technol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Jo, D.-M., Khan, F., Park, S.-K., Ko, S.-C., Kim, K.W., Yang, D., Kim, J.-Y., Oh, G.-W., Choi, G., and Lee, D.-S. (2024). From sea to lab: Angiotensin I-converting enzyme inhibition by marine peptides\u2014Mechanisms and applications. Mar. Drugs, 22.","DOI":"10.3390\/md22100449"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"175185","DOI":"10.1016\/j.scitotenv.2024.175185","article-title":"Transcriptomic responses and evolutionary insights of deep-sea and shallow-water mussels under high hydrostatic pressure condition","volume":"949","author":"Zhong","year":"2024","journal-title":"Sci. Total Environ."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Heo, S.Y., Kang, N., Kim, E.A., Kim, J., Lee, S.H., Ahn, G., Oh, J.H., Shin, A.Y., Kim, D., and Heo, S.J. (2023). Purification and molecular docking study on the angiotensin I-converting enzyme (ACE)-inhibitory peptide isolated from hydrolysates of the deep-sea mussel Gigantidas vrijenhoeki. Mar. Drugs, 21.","DOI":"10.3390\/md21080458"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"115308","DOI":"10.1016\/j.foodres.2024.115308","article-title":"Identification and anticoagulant mechanisms of novel factor XIa inhibitory peptides by virtual screening of a in silico generated deep-sea peptide database","volume":"197","author":"Ouyang","year":"2024","journal-title":"Food Res. Int."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"135873","DOI":"10.1016\/j.ijbiomac.2024.135873","article-title":"Discovery of a novel nanomolar angiotensin-I converting enzyme inhibitory peptide with unusual binding mechanisms derived from Chlorella pyrenoidosa","volume":"280","author":"Suo","year":"2024","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1016\/j.foodchem.2017.02.039","article-title":"Antihypertensive peptides from animal products, marine organisms, and plants","volume":"228","author":"Lee","year":"2017","journal-title":"Food Chem."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Pujiastuti, D.Y., Ghoyatul Amin, M.N., Alamsjah, M.A., and Hsu, J.L. (2019). Marine organisms as potential sources of bioactive peptides that inhibit the activity of angiotensin I-converting enzyme: A review. Molecules, 24.","DOI":"10.3390\/molecules24142541"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.procbio.2018.11.017","article-title":"Isolation and identification of an angiotensin I-converting enzyme inhibitory peptide from pearl oyster (Pinctada fucata) shell protein hydrolysate","volume":"77","author":"Sasaki","year":"2019","journal-title":"Process Biochem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"116113","DOI":"10.1016\/j.foodres.2025.116113","article-title":"Clam peptides: Preparation, flavor properties, health benefits, and safety risks","volume":"207","author":"Luo","year":"2025","journal-title":"Food Res. Int."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Sun, X.P., Wang, M., Xu, C.J., Wang, S.L., Li, L., Zou, S.C., Yu, J., and Wei, Y.X. (2022). Positive effect of a pea-clam two-peptide composite on hypertension and organ protection in spontaneously hypertensive rats. Nutrients, 14.","DOI":"10.3390\/nu14194069"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1007\/s12010-012-0024-y","article-title":"Review on the angiotensin-I-converting enzyme (ACE) inhibitor peptides from marine proteins","volume":"169","author":"He","year":"2013","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Chai, T.T., Wong, C.C.C., Sabri, M.Z., and Wong, F.C. (2022). Seafood paramyosins as sources of anti-angiotensin-converting-enzyme and anti-dipeptidyl-peptidase peptides after gastrointestinal digestion: A Cheminformatic Investigation. Molecules, 27.","DOI":"10.3390\/molecules27123864"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"7831","DOI":"10.1039\/D2FO00275B","article-title":"Seventeen novel angiotensin converting enzyme (ACE) inhibitory peptides from the protein hydrolysate of Mytilus edulis: Isolation, identification, molecular docking study, and protective function on HUVECs","volume":"13","author":"Suo","year":"2022","journal-title":"Food Funct."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/j.procbio.2008.02.019","article-title":"ACE-inhibitory peptides identified from the muscle protein hydrolysate of hard clam (Meretrix lusoria)","volume":"43","author":"Tsai","year":"2008","journal-title":"Process Biochem."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Li, Y., Sadiq, F.A., Fu, L., Zhu, H., Zhong, M.H., and Sohail, M. (2016). Identification of angiotensin I-converting enzyme inhibitory peptides derived from enzymatic hydrolysates of razor Clam Sinonovacula constricta. Mar. Drugs, 14.","DOI":"10.3390\/md14060110"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.foodchem.2008.03.059","article-title":"Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats","volume":"111","author":"Wang","year":"2008","journal-title":"Food Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"8718","DOI":"10.1021\/bi049480n","article-title":"Structural details on the binding of antihypertensive drugs captopril and enalaprilat to human testicular angiotensin I-converting enzyme","volume":"43","author":"Natesh","year":"2004","journal-title":"Biochemistry"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"118213","DOI":"10.1016\/j.envres.2024.118213","article-title":"The role of the balance between energy production and ammonia detoxification mediated by key amino acids in divergent hypersaline adaptation among crassostrea oysters","volume":"248","author":"Zhao","year":"2024","journal-title":"Environ. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"9108","DOI":"10.1039\/D2FO01207C","article-title":"Isolation, identification and in vivo antihypertensive effect of novel angiotensin I-converting enzyme (ACE) inhibitory peptides from Spirulina protein hydrolysate","volume":"13","author":"Suo","year":"2022","journal-title":"Food Funct."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3891","DOI":"10.1021\/acs.jcim.1c00203","article-title":"AutoDock Vina 1.2.0: New docking methods, expanded force field, and python bindings","volume":"61","author":"Eberhardt","year":"2021","journal-title":"J. Chem. Inf. Model."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading","volume":"31","author":"Trott","year":"2010","journal-title":"J. Comput. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1893","DOI":"10.1021\/ci300604z","article-title":"Lessons learned in empirical scoring with smina from the CSAR 2011 benchmarking exercise","volume":"53","author":"Koes","year":"2013","journal-title":"J. Chem. Inf. Model."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Minkiewicz, P., Iwaniak, A., and Darewicz, M. (2019). BIOPEP-UWM database of bioactive peptides: Current opportunities. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20235978"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/0010-4655(95)00042-E","article-title":"Gromacs\u2014A message-passing parallel molecular-dynamics implementation","volume":"91","author":"Berendsen","year":"1995","journal-title":"Comput. Phys. Commun."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2135","DOI":"10.1002\/jcc.23354","article-title":"CHARMM36 all-atom additive protein force field: Validation based on comparison to NMR data","volume":"34","author":"Huang","year":"2013","journal-title":"J. Comput. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3144","DOI":"10.1021\/ci300363c","article-title":"Automation of the CHARMM General Force Field (CGenFF) I: Bond perception and atom typing","volume":"52","author":"Vanommeslaeghe","year":"2012","journal-title":"J. Chem. Inf. Model."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"6281","DOI":"10.1021\/acs.jctc.1c00645","article-title":"gmx_MMPBSA: A new tool to perform end-state free energy calculations with GROMACS","volume":"17","author":"Valiente","year":"2021","journal-title":"J. Chem. Theory Comput."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/5\/757\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T05:34:10Z","timestamp":1772084050000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/5\/757"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,24]]},"references-count":37,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["molecules31050757"],"URL":"https:\/\/doi.org\/10.3390\/molecules31050757","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,24]]}}}