{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T17:24:44Z","timestamp":1790270684736,"version":"4.1.0"},"reference-count":52,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T00:00:00Z","timestamp":1690243200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NORTH-WEST EUROPE INTERREG","award":["NWE 639"],"award-info":[{"award-number":["NWE 639"]}]},{"name":"VITO (Leen Bastiaens and Queenie Simons)","award":["NWE 639"],"award-info":[{"award-number":["NWE 639"]}]},{"name":"Thomas More University","award":["NWE 639"],"award-info":[{"award-number":["NWE 639"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>Inflammation, hypertension, and negative heart health outcomes including cardiovascular disease are closely linked but the mechanisms by which inflammation can cause high blood pressure are not yet fully elucidated. Cyclooxygenase (COX) enzymes play a role in pain, inflammation, and hypertension development, and inhibition of these enzymes is currently of great interest to researchers and pharmaceutical companies. Non-steroidal anti-inflammatory drugs are the drug of choice in terms of COX inhibition but can have negative side effects for consumers. Functional food ingredients containing cyclooxygenase inhibitors offer a strategy to inhibit cyclooxygenases without negative side effects. Several COX inhibitors have been discovered, to date, from marine and other resources. We describe here, for the first time, the generation and characterization of a bioactive hydrolysate generated using Viscozyme\u00ae and Alcalase from the red microalga Porphyridium sp. The hydrolysate demonstrates in vitro COX-1 inhibitory activity and antihypertensive activity in vivo, assessed using spontaneously hypertensive rats (SHRs). Peptides were identified and sequenced using MS and assessed using an in silico computational approach for potential bioactivities. The peptides predicted to be bioactive, including GVDYVRFF, AIPAAPAAPAGPKLY, and LIHADPPGVGL were chemically synthesized and cyclooxygenase inhibition was confirmed. Peptides AIPAAPAAPAGPKLY and LIHADPPGVGL had COX-1 IC50 values of 0.2349 mg\/mL (0.16 \u00b5M) and 0.2193 mg\/mL (0.2 \u00b5M), respectively. The hydrolysate was included in a food carrier (jelly candies) and an antihypertensive effect was observed in SHRs.<\/jats:p>","DOI":"10.3390\/md21080422","type":"journal-article","created":{"date-parts":[[2023,7,26]],"date-time":"2023-07-26T01:09:01Z","timestamp":1690333741000},"page":"422","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["Generation of Bioactive Peptides from Porphyridium sp. and Assessment of Their Potential for Use in the Prevention of Hypertension, Inflammation and Pain"],"prefix":"10.3390","volume":"21","author":[{"given":"Maria","family":"Hayes","sequence":"first","affiliation":[{"name":"Department of Food BioSciences, Teagasc Food Research Centre, Ashtown, D15 DY05 Dublin, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9025-2513","authenticated-orcid":false,"given":"Rotimi E.","family":"Aluko","sequence":"additional","affiliation":[{"name":"Department of Food and Human Nutritional Sciences, University of Manitoba, Winnipeg, MB R3T 2N2, Canada"},{"name":"Richardson Centre for Food Technology and Research, University of Manitoba, Winnipeg, MB R3T 2N2, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Aurino","sequence":"additional","affiliation":[{"name":"Department of Food BioSciences, Teagasc Food Research Centre, Ashtown, D15 DY05 Dublin, Ireland"},{"name":"Department of Chemical, Materials and Industrial Production Engineering, University of Naples Federico II, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1571-5781","authenticated-orcid":false,"given":"Leticia","family":"Mora","sequence":"additional","affiliation":[{"name":"Instituto de Agroqu\u00edmica y Tecnolog\u00eda de Alimentos (CSIC), Avenue Agust\u00edn Escardino 7, Valencia, 46980 Paterna, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"7204","DOI":"10.18632\/oncotarget.23208","article-title":"Inflammatory responses and inflammation-associated diseases in organs","volume":"9","author":"Chen","year":"2017","journal-title":"Oncotarget"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"eabj9954","DOI":"10.1126\/scitranslmed.abj9954","article-title":"Acute inflammatory response via neutrophil activation protects against the development of chronic pain","volume":"14","author":"Parisien","year":"2022","journal-title":"Sci. Transl. Med."},{"key":"ref_3","first-page":"247","article-title":"Anti-inflammatory and side effects of cyclooxygenase inhibitors","volume":"59","author":"Demircan","year":"2007","journal-title":"Pharmacol. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1111\/bcp.13048","article-title":"Cardiovascular effects of cyclooxygenase-2 inhibitors: A mechanistic and clinical perspective","volume":"82","author":"Patrono","year":"2016","journal-title":"Br. J. Clin. Pharmacol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"976","DOI":"10.3389\/fphar.2018.00976","article-title":"Raging the War Against Inflammation With Natural Products","volume":"9","author":"Attiq","year":"2018","journal-title":"Front. Pharmacol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.ijbiomac.2009.03.013","article-title":"Preparation of different molecular weight polysaccharides from Porphyridium cruentum and their antioxidant activities","volume":"45","author":"Sun","year":"2009","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_7","unstructured":"Dillon, H., Sumanchi, A., and Rao, K. (2007). Methods and Compositions for Reducing Inflammation and Preventing Oxidative Damage, Current Holdrs TerraVia Holdings Inc."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1007\/s10811-007-9188-1","article-title":"Phycobiliproteins as a commodity: Trends in applied research, patents and commercialization","volume":"20","author":"Sekar","year":"2008","journal-title":"J. Appl. Phycol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Risjani, Y., Mutmainnah, N., Manurung, P., Wulan, S.N. (2021). Exopolysaccharide from Porphyridium cruentum (purpureum) is Not Toxic and Stimulates Immune Response against Vibriosis: The Assessment Using Zebrafish and White Shrimp Litopenaeus vannamei. Mar. Drugs., 19.","DOI":"10.20944\/preprints202102.0422.v1"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1007\/s10529-018-2621-5","article-title":"Atheroprotective effect of novel peptides from Porphyridium purpureum in RAW 264.7 macrophage cell line and its molecular docking study","volume":"41","author":"Kavitha","year":"2019","journal-title":"Biotechnol. Lett."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Mora, L., Gallego, M., and Toldr\u00e1, F. (2018). ACEI-Inhibitory Peptides Naturally Generated in Meat and Meat Products and Their Health Relevance. Nutrients, 10.","DOI":"10.3390\/nu10091259"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.meatsci.2011.07.008","article-title":"Assessment of the angiotensin-I-converting enzyme (ACE-I) inhibitory and antioxidant activities of hydrolysates of bovine brisket sarcoplasmic proteins produced by papain and characterisation of associated bioactive peptidic fractions","volume":"90","author":"Mullen","year":"2012","journal-title":"Meat Sci."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Hayes, M., Mora, L., and Lucakova, S. (2022). Identification of Bioactive Peptides from Nannochloropsis oculata Using a Combination of Enzymatic Treatment, in Silico Analysis and Chemical Synthesis. Biomolecules, 12.","DOI":"10.3390\/biom12121806"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Bleakley, S., Hayes, M., O\u2019 Shea, N., Gallagher, E., and Lafarga, T. (2017). Predicted Release and Analysis of Novel ACE-I, Renin, and DPP-IV Inhibitory Peptides from Common Oat (Avena sativa) Protein Hydrolysates Using in Silico Analysis. Foods, 6.","DOI":"10.3390\/foods6120108"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1007\/s10811-022-02900-1","article-title":"In vitro enzyme inhibitory effects of green and brown Australian seaweeds and potential impact on metabolic syndrome","volume":"35","author":"Shannon","year":"2023","journal-title":"J. Appl. Phycol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1128\/AEM.00096-07","article-title":"Casein fermentate of Lactobacillus animalis DPC6134 contains a range of novel propeptide Angiotensin-Converting Enzyme inhibitors","volume":"73","author":"Hayes","year":"2007","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1515\/opag-2021-0029","article-title":"Development and characterization of healthy gummy jellies containing natural fruits","volume":"6","author":"Almeida","year":"2021","journal-title":"Open Agric."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Mooney, C., Haslam, N., Pollastri, G., and Shields, D.C. (2012). Towards the improved discovery and design of functional peptides: Common features of diverse classes permit generalised prediction of bioactivity. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0045012"},{"key":"ref_19","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_20","doi-asserted-by":"crossref","unstructured":"Walker, J.M. (2005). Protein Identification and Analysis Tools on the Expasy Server, Full text\u2014Copyright Humana Press. The Proteomics Protocols Handbook, Humana Press.","DOI":"10.1385\/1592598900"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4202","DOI":"10.1002\/elps.200700324","article-title":"Identification of novel angiotensin-converting enzyme inhibitory peptides from ovine milk proteins by CE-MS and chromatographic techniques","volume":"28","author":"Ramos","year":"2007","journal-title":"Electrophoresis"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1909","DOI":"10.1016\/j.jff.2013.09.012","article-title":"Inhibition of dipeptidyl peptidase IV (DPP-IV) by proline containing casein derived peptides","volume":"5","author":"Nongonierma","year":"2013","journal-title":"J. Funct. Foods"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.foodchem.2014.11.131","article-title":"Analyzing a dipeptide library to identify human dipeptidyl peptidase IV inhibitor","volume":"175","author":"Lan","year":"2015","journal-title":"Food Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1002\/cbf.1448","article-title":"Hydrolysis of various bioactive peptides by goat brain dipeptidyl peptidase III","volume":"23","author":"Dhanda","year":"2008","journal-title":"Cell Biochem. Funct."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Hatakenaka, T., Kato, T., and Okamoto, K. (2023). In Vitro and In Silico Studies on Angiotensin I-Converting Enzyme Inhibitory Peptides Found in Hydrophobic Domains of Porcine Elastin. Molecules, 28.","DOI":"10.3390\/molecules28083337"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Naseri, A., Jacobsen, C., Sejberg, J.J.P., Pedersen, T.E., Larsen, J., Hansen, K.M., and Holdt, S.L. (2020). Multi-Extraction and Quality of Protein and Carrageenan from Commercial Spinosum (Eucheuma denticulatum). Foods, 9.","DOI":"10.3390\/foods9081072"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Blanco-Llamero, C., Garc\u00eda-Garc\u00eda, P., and Se\u00f1or\u00e1ns, F.J. (2021). Combination of Synergic Enzymes and Ultrasounds as an Effective Pretreatment Process to Break Microalgal Cell Wall and Enhance Algal Oil Extraction. Foods, 10.","DOI":"10.3390\/foods10081928"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.biortech.2017.05.012","article-title":"Energy consumption and water-soluble protein release by cell wall disruption of Nannochloropsis gaditana","volume":"239","author":"Safi","year":"2017","journal-title":"Bioresour. Technol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Toldr\u00e1, F., and Mora, L. (2021). Proteins and Bioactive Peptides in High Protein Content Foods. Foods, 10.","DOI":"10.3390\/foods10061186"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Purcell, D., Packer, M.A., and Hayes, M. (2022). Angiotensin-I-Converting Enzyme Inhibitory Activity of Protein Hydrolysates Generated from the Macroalga Laminaria digitata (Hudson) JV Lamouroux 1813. Foods, 11.","DOI":"10.3390\/foods11121792"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"7421","DOI":"10.1021\/jf301361c","article-title":"Isolation and Characteri zation of Bioactive Pro-Peptides with in Vitro Renin Inhibitory Activities from the Macroalga Palmaria palmata","volume":"60","author":"Fitzgerald","year":"2012","journal-title":"J. Agric. Food Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1111\/1750-3841.14011","article-title":"Bioactive peptides derived from seaweed protein and their health benefits: Antihypertensive, antioxidant and antidiabetic properties","volume":"83","author":"Admassu","year":"2018","journal-title":"J. Food Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1007\/s11130-022-00963-6","article-title":"In Vitro Anti-Inflammatory Activity of Three Peptides Derived from the Byproduct of Rice Processing","volume":"77","author":"Qu","year":"2022","journal-title":"Plant Foods Hum Nutr."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"109002","DOI":"10.1016\/j.foodres.2020.109002","article-title":"A biotechnological approach for the production of branched chain amino acid containing bioactive peptides to improve human health: A review","volume":"131","author":"Dullius","year":"2020","journal-title":"Food Res. Int."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"383","DOI":"10.21010\/ajtcam.v14i2.39","article-title":"Structural characteristics and antihypertensive effects of angiotensin-1-converting enzyme inhibitory peptides in the Renin-Angiotensin and Kallikrein Kinin systems","volume":"14","author":"Manoharan","year":"2017","journal-title":"Afr. J. Tradit. Complement. Altern. Med."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"He, R., Aluko, R.E., and Ju, X.-R. (2014). Evaluating Molecular Mechanism of Hypotensive Peptides Interactions with Renin and Angiotensin Converting Enzyme. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0091051"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1021\/jf051263l","article-title":"Structural requirements of Angiotensin-I-converting enzyme inhibitory activity\u2014Structure activity relationship study of di- and tripeptides","volume":"54","author":"Wu","year":"2006","journal-title":"J. Agric. Food Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"77","DOI":"10.12944\/CRNFSJ.4.Special-Issue-October.10","article-title":"Effect of in vitro gastrointestinal digestion on bioactivity of poultry protein hydrolysate","volume":"4","author":"Torkova","year":"2016","journal-title":"Curr. Res. Nutr. Food Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jff.2014.10.027","article-title":"Isolation and characterization of three antioxidant peptides from protein hydrolysate of bluefin leatherjacket (Navodon septentrionalis) heads","volume":"12","author":"Chi","year":"2015","journal-title":"J. Funct. Foods"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"He, J., Guo, H., Zhang, M., Wang, M., Sun, L., and Zhuang, Y. (2022). Purification and characterization of a novel calcium-binding heptapeptide from the hydrolysate of tilapia bone with its osteogenic activity. Foods, 11.","DOI":"10.3390\/foods11030468"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2251","DOI":"10.1021\/jm0402059","article-title":"Adverse cardiovascular effects of the coxibs","volume":"48","author":"Dogne","year":"2005","journal-title":"J. Med. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.drudis.2014.10.003","article-title":"Peptide therapeutics: Current status and future directions","volume":"20","author":"Fosgerau","year":"2015","journal-title":"Drug Discov. Today"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Z\u0142otek, U., Jakubczyk, A., Rybczy\u0144ska-Tkaczyk, K., \u0106wiek, P., Baraniak, B., and Lewicki, S. (2020). Characteristics of New Peptides GQLGEHGGAGMG, GEHGGAGMGGGQFQPV, EQGFLPGPEESGR, RLARAGLAQ, YGNPVGGVGH, and GNPVGGVGHGTTGT as Inhibitors of Enzymes Involved in Metabolic Syndrome and Antimicrobial Potential. Molecules, 25.","DOI":"10.3390\/molecules25112492"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1271\/bbb.70.517","article-title":"Blood pressure-depressing activity of a peptide derived from silkworm fibroin in spontaneously hypertensive rats","volume":"70","author":"Igarashi","year":"2006","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1016\/S0196-9781(03)00174-8","article-title":"New antihypertensive peptides isolated from rapeseed","volume":"24","author":"Marczak","year":"2003","journal-title":"Peptides"},{"key":"ref_46","first-page":"57","article-title":"Antihypertensive activities of royal jelly protein hydrolysate and its fractions in spontaneously hypertensive rats","volume":"63","author":"Hashimoto","year":"2009","journal-title":"Acta Med. Okayama."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"9877","DOI":"10.1021\/acs.jafc.2c02982","article-title":"Bovine Bone Gelatin-Derived Peptides: Food Processing Characteristics and Evaluation of Antihypertensive and Antihyperlipidemic Activities","volume":"70","author":"Cao","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1111\/j.1365-2621.1994.tb06901.x","article-title":"Quality of fish protein hydrolysates from herring (Clupea harengus)","volume":"59","author":"Hoyle","year":"1994","journal-title":"J. Food Sci."},{"key":"ref_49","unstructured":"Association of Official Analytical Chemists (1998). Official Methods of Analysis of AOAC International, AOAC International."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Willenberg, I., Meschede, A.K., Gueler, F., Jang, M.-S., Shushakova, N., and Schebb, N.H. (2015). Food Polyphenols Fail to Cause a Biologically Relevant Reduction of COX-2 Activity. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0139147"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"129","DOI":"10.3389\/fgene.2019.00129","article-title":"PreAIP: Computational prediction of anti-inflammatory peptides by integrating multiple complementary features","volume":"10","author":"Khatun","year":"2019","journal-title":"Front. Genet."},{"key":"ref_52","first-page":"25","article-title":"Formulation of jelly from candy Betel (Piper betle L.)\u2014Suji (Pleomele angustofolia) leaf extract","volume":"2","author":"Purwaningtyas","year":"2017","journal-title":"J. Teknol. Dan Ind. Pangan"}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/21\/8\/422\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:18:31Z","timestamp":1760127511000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/21\/8\/422"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,25]]},"references-count":52,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2023,8]]}},"alternative-id":["md21080422"],"URL":"https:\/\/doi.org\/10.3390\/md21080422","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,25]]}}}