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This study evaluated the bioactive potential and peptidomic profile of kefirs from both species before and after in vitro digestion. Peptides from undigested, gastric, and intestinal phases were characterized by LC-MS\/MS. In silico tools identified sequences homologous to known bioactive peptides, with functions such as antihypertensive, immunomodulatory, and anticancer. \u03b2-casein was identified as the main precursor protein, with the highest number of peptides identified during the gastric phase. Undigested kefir showed antioxidant activity in ABTS and FRAP assays, and peptide extracts from both kefirs exhibited activity in all antioxidant assays across digestion phases. Antimicrobial activity against E. coli and S. aureus was observed before digestion for both kefir peptides; however, after digestion, only goat kefir peptides remained active against E. coli. This data indicates that kefir peptides are promising bioactive compounds for food and pharmaceutical applications.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c08235","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T18:01:37Z","timestamp":1786471297000},"page":"26625-26639","source":"Crossref","is-referenced-by-count":0,"title":["Peptidomic Characterization\nand Functional Properties\nof Goat and Cow Milk Kefir before and after\n                    <i>In Vitro<\/i>\n                    Digestion"],"prefix":"10.1021","volume":"74","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1428-0413","authenticated-orcid":true,"given":"Rayssa Cruz","family":"Lima","sequence":"first","affiliation":[{"name":"Fluminense Federal University (UFF) , , Vital Brazil Filho , , ,","place":["Nitero\u0301i, Rio de Janeiro, Brazil, 24230-340"]},{"name":"Federal University of Rio de Janeiro (UFRJ), Cidade Universita\u0301ria , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-598"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9747-4111","authenticated-orcid":true,"given":"Bum Jin","family":"Kim","sequence":"additional","affiliation":[{"name":"Oregon State University , , , ,","place":["Corvallis, Oregon, United States, 97331"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carini Aparecida","family":"Lelis","sequence":"additional","affiliation":[{"name":"Federal University of Rio de Janeiro (UFRJ), Cidade Universita\u0301ria , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-598"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brianne","family":"Wai","sequence":"additional","affiliation":[{"name":"Oregon State University , , , ,","place":["Corvallis, Oregon, United States, 97331"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rohit","family":"Kumar","sequence":"additional","affiliation":[{"name":"Oregon State University , , , ,","place":["Corvallis, Oregon, United States, 97331"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4886-302X","authenticated-orcid":true,"given":"Jelmir Craveiro","family":"de Andrade","sequence":"additional","affiliation":[{"name":"Federal University of Rio de Janeiro (UFRJ), Cidade Universita\u0301ria , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-598"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4415-184X","authenticated-orcid":true,"given":"Marie Biondi","family":"Ryan","sequence":"additional","affiliation":[{"name":"Oregon State University , , , ,","place":["Corvallis, Oregon, United States, 97331"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9696-0967","authenticated-orcid":true,"given":"David C.","family":"Dallas","sequence":"additional","affiliation":[{"name":"Oregon State University , , , ,","place":["Corvallis, Oregon, United States, 97331"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6133-5080","authenticated-orcid":true,"given":"Carlos Adam","family":"Conte-Junior","sequence":"additional","affiliation":[{"name":"Fluminense Federal University (UFF) , , Vital Brazil Filho , , ,","place":["Nitero\u0301i, Rio de Janeiro, Brazil, 24230-340"]},{"name":"Federal University of Rio de Janeiro (UFRJ), Cidade Universita\u0301ria , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-598"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"key":"2026090104465153600_cit1","doi-asserted-by":"publisher","first-page":"8908","DOI":"10.3168\/jds.2024-24998","article-title":"Label-free-based\nproteomics analysis reveals differential proteins\nof sheep, goat, and cow milk","volume":"107","author":"Zhu","year":"2024","journal-title":"J. Dairy Sci."},{"key":"2026090104465153600_cit2","doi-asserted-by":"publisher","first-page":"456","DOI":"10.1016\/j.foodchem.2019.05.178","article-title":"Proteomic\nanalysis of goat milk kefir: Profiling the fermentation-time\ndependent protein digestion and identification of potential peptides\nwith biological activity","volume":"295","author":"Izquierdo-Gonza\u0301lez","year":"2019","journal-title":"Food Chem."},{"key":"2026090104465153600_cit3","doi-asserted-by":"publisher","first-page":"1517","DOI":"10.3168\/jds.2014-8755","article-title":"Manufacture and characterization\nof kefir made from\ncow and buffalo milk, using kefir grain and starter culture","volume":"98","author":"Gul","year":"2015","journal-title":"J. Dairy Sci."},{"key":"2026090104465153600_cit4","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1016\/j.foodchem.2015.10.116","article-title":"Peptidomic analysis reveals proteolytic activity of kefir microorganisms\non bovine milk proteins","volume":"197","author":"Dallas","year":"2016","journal-title":"Food Chem."},{"key":"2026090104465153600_cit5","doi-asserted-by":"publisher","first-page":"27","DOI":"10.3168\/jds.2009-2501","article-title":"Short\ncommunication: Effect of kefir grains on proteolysis\nof major milk proteins","volume":"93","author":"Ferreira","year":"2010","journal-title":"J. Dairy Sci."},{"key":"2026090104465153600_cit6","doi-asserted-by":"publisher","first-page":"1263","DOI":"10.1007\/s13197-021-05091-8","article-title":"Functional dairy products\nas a source of bioactive peptides and probiotics:\ncurrent trends and future prospectives","volume":"59","author":"Ali","year":"2022","journal-title":"J. Food\nSci. 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Macromol."},{"key":"2026090104465153600_cit20","doi-asserted-by":"publisher","first-page":"105970","DOI":"10.1016\/j.idairyj.2024.105970","article-title":"Dynamics of\npeptides released from cow milk fermented by kefir microorganisms\nduring fermentation and storage periods","volume":"155","author":"Aksoy","year":"2024","journal-title":"Int.\nDairy J."},{"key":"2026090104465153600_cit21","doi-asserted-by":"publisher","first-page":"2295","DOI":"10.1007\/s13197-021-05243-w","article-title":"Peptidomic\nprofiling of fermented\ngoat milk: considering the fermentation-time dependent proteolysis\nby Lactobacillus and characterization of novel peptides with Antioxidative\nactivity","volume":"59","author":"Panchal","year":"2022","journal-title":"J. Food Sci. Technol."},{"issue":"1","key":"2026090104465153600_cit22","doi-asserted-by":"publisher","first-page":"11065","DOI":"10.1038\/s41598-022-15297-1","article-title":"Identification of bioactive peptides from a\nBrazilian kefir sample, and their anti-Alzheimer potential in Drosophila\nmelanogaster","volume":"12","author":"Malta","year":"2022","journal-title":"Sci. Rep."},{"key":"2026090104465153600_cit23","doi-asserted-by":"publisher","first-page":"1208970","DOI":"10.3389\/fsufs.2023.1208970","article-title":"Peptidomics\napproaches to the discovery and ACE inhibitory effect of casein peptides\nderived from fermented bovine milk by kefir grains","volume":"7","author":"Wang","year":"2023","journal-title":"Front. Sustain. Food Syst."},{"key":"2026090104465153600_cit24","doi-asserted-by":"publisher","first-page":"8047","DOI":"10.1007\/s00216-022-04338-3","article-title":"Validated\nmultiplex-competitive ELISA using gluten-incurred yogurt calibrant\nfor the quantitation of wheat gluten in fermented dairy products","volume":"414","author":"Panda","year":"2022","journal-title":"Anal. Bioanal. Chem."},{"key":"2026090104465153600_cit25","doi-asserted-by":"publisher","first-page":"991","DOI":"10.1038\/s41596-018-0119-1","article-title":"INFOGEST static in vitro simulation\nof gastrointestinal\nfood digestion","volume":"14","author":"Brodkorb","year":"2019","journal-title":"Nat. Protoc."},{"key":"2026090104465153600_cit26","doi-asserted-by":"publisher","first-page":"11510","DOI":"10.1080\/10408398.2023.2240396","article-title":"Bioactive milk peptides:\nan updated comprehensive overview\nand database","volume":"64","author":"Nielsen","year":"2024","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"2026090104465153600_cit27","doi-asserted-by":"publisher","first-page":"673","DOI":"10.1016\/j.foodchem.2017.04.056","article-title":"Milk bioactive peptide\ndatabase: A comprehensive database of milk protein-derived bioactive\npeptides and novel visualization","volume":"232","author":"Nielsen","year":"2017","journal-title":"Food Chem."},{"key":"2026090104465153600_cit28","doi-asserted-by":"publisher","first-page":"4259","DOI":"10.3168\/jds.2023-24364","article-title":"Evaluation\nof kefir grain microbiota, grain viability, and bioactivity from fermenting\ndairy processing by-products","volume":"107","author":"McGovern","year":"2024","journal-title":"J. Dairy Sci."},{"key":"2026090104465153600_cit29","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1006\/abio.1996.0292","article-title":"The Ferric Reducing\nAbility of Plasma (FRAP) as a measure\nof \u201cAntioxidant Power\u201d: The FRAP assay","volume":"239","author":"Benzie","year":"1996","journal-title":"Anal. Biochem."},{"key":"2026090104465153600_cit30","doi-asserted-by":"publisher","first-page":"100331","DOI":"10.1016\/j.afres.2023.100331","article-title":"Green ultrasound-assisted extraction of bioactive compounds of babassu\n(Attalea speciosa) mesocarp: Effects of solid-liquid ratio extraction,\nantioxidant capacity, and antimicrobial activity","volume":"3","author":"Lima","year":"2023","journal-title":"Appl. Food Res."},{"key":"2026090104465153600_cit31","doi-asserted-by":"publisher","first-page":"101","DOI":"10.5614\/crbb.2020.2.1\/HPMQ5042","article-title":"The effect of temperature, incubation\nand storage time on lactic acid content, pH, and viscosity of goat\nmilk kefir","volume":"2","author":"Putri","year":"2020","journal-title":"Curr. Res. Biosci. Biotechnol."},{"key":"2026090104465153600_cit32","doi-asserted-by":"publisher","first-page":"406","DOI":"10.1007\/s11694-020-00645-4","article-title":"An in-vitro\nassessment of antifungal\nand antibacterial activity of cow, camel, ewe, and goat milk kefir\nand probiotic yogurt","volume":"15","author":"Azizkhani","year":"2021","journal-title":"J. Food Meas. Charact."},{"key":"2026090104465153600_cit33","doi-asserted-by":"publisher","first-page":"1648","DOI":"10.1039\/D2FO03248A","article-title":"Raw milk kefir: microbiota,\nbioactive peptides, and immune modulation","volume":"14","author":"Baars","year":"2023","journal-title":"Food\nFunct."},{"key":"2026090104465153600_cit34","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1007\/s00253-006-0427-1","article-title":"Proteolytic\nsystems of lactic acid\nbacteria","volume":"71","author":"Savijoki","year":"2006","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"2026090104465153600_cit35","doi-asserted-by":"publisher","first-page":"239","DOI":"10.15567\/mljekarstvo.2018.0401","article-title":"Goat\u2019s\nmilk-derived bioactive components - a\nreview","volume":"68","author":"Biada\u0142a","year":"2018","journal-title":"Mljekarstvo"},{"key":"2026090104465153600_cit36","doi-asserted-by":"publisher","first-page":"126419","DOI":"10.1016\/j.foodchem.2020.126419","article-title":"Differences\nin peptide generation following in vitro gastrointestinal\ndigestion of yogurt and milk from cow, sheep and goat","volume":"317","author":"Nguyen","year":"2020","journal-title":"Food Chem."},{"key":"2026090104465153600_cit37","doi-asserted-by":"publisher","first-page":"1865","DOI":"10.1021\/acs.jafc.6b05385","article-title":"Identification\nand Relative Quantification of Bioactive\nPeptides Sequentially Released during Simulated Gastrointestinal Digestion\nof Commercial Kefir","volume":"65","author":"Liu","year":"2017","journal-title":"J. Agric. Food Chem."},{"key":"2026090104465153600_cit38","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/j.jprot.2015.01.005","article-title":"Peptide profiling of\nbovine kefir reveals 236 unique peptides released\nfrom caseins during its production by starter culture or kefir grains","volume":"117","author":"Ebner","year":"2015","journal-title":"J. Proteomics"},{"key":"2026090104465153600_cit39","doi-asserted-by":"publisher","DOI":"10.1111\/1541-4337.70164","article-title":"Comprehensive\nreview of enzymes (protease, lipase)\nin milk: Impact on storage quality, detection methods, and control\nstrategies","volume":"24","author":"Khan","year":"2025","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"2026090104465153600_cit40","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1002\/med.21986","article-title":"Antioxidant peptides, the guardian of life from oxidative\nstress","volume":"44","author":"Zhu","year":"2024","journal-title":"Med. Res. 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