{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,5]],"date-time":"2026-11-05T01:50:29Z","timestamp":1793843429987,"version":"4.6.1"},"reference-count":29,"publisher":"American Chemical Society (ACS)","issue":"16","license":[{"start":{"date-parts":[[2018,4,13]],"date-time":"2018-04-13T00:00:00Z","timestamp":1523577600000},"content-version":"vor","delay-in-days":9,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_ccbyncnd_termsofuse.html"}],"funder":[{"DOI":"10.13039\/na","name":"FrieslandCampina","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"NanoNextNL","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,4,25]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Based on trypsin specificity (for lysines and arginines), trypsins from different sources are expected to hydrolyze a given protein to the same theoretical maximum degree of hydrolysis (DHmax,theo). This is in contrast with experiments. Using \u03b1-lactalbumin and \u03b2-casein, this study aims to reveal if the differences among experimental DHmax (DHmax,exp) by bovine, porcine, and human trypsins are due to their secondary specificity. Peptide analysis showed that \u223c78% of all the cleavage sites were efficiently hydrolyzed by porcine trypsin, and \u223c47 and \u223c53% were efficiently hydrolyzed by bovine and human trypsins, respectively. These differences were explained by the enzyme secondary specificity, that is, their sensitivities to the amino acids around the cleavage sites. The DHmax predictions based on the secondary specificity were 4 times closer to the DHmax,exp than the predictions based on trypsin specificity alone (DHmax,theo). Proposed preliminary relations between binding sites and trypsin secondary specificity allow DHmax,exp estimations of tryptic hydrolysis of other proteins.<\/jats:p>","DOI":"10.1021\/acs.jafc.8b00679","type":"journal-article","created":{"date-parts":[[2018,4,4]],"date-time":"2018-04-04T10:46:58Z","timestamp":1522838818000},"page":"4219-4232","source":"Crossref","is-referenced-by-count":41,"title":["Comparison of Protein Hydrolysis\nCatalyzed by Bovine, Porcine, and\nHuman Trypsins"],"prefix":"10.1021","volume":"66","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9772-0340","authenticated-orcid":true,"given":"Yuxi","family":"Deng","sequence":"first","affiliation":[{"name":"Wageningen University , , P.O. Bus 17, ,","place":["Wageningen, The Netherlands, 6708 AA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harry","family":"Gruppen","sequence":"additional","affiliation":[{"name":"Wageningen University , , P.O. Bus 17, ,","place":["Wageningen, The Netherlands, 6708 AA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7799-6676","authenticated-orcid":true,"given":"Peter A.","family":"Wierenga","sequence":"additional","affiliation":[{"name":"Wageningen University , , P.O. Bus 17, ,","place":["Wageningen, The Netherlands, 6708 AA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2018,4,4]]},"reference":[{"key":"2026090218050343300_cit1","doi-asserted-by":"publisher","first-page":"210","DOI":"10.1016\/j.foodhyd.2017.02.007","article-title":"Effect of\nMaillard induced glycation on protein hydrolysis by lysine\/arginine\nand non-lysine\/arginine specific proteases","volume":"69","author":"Deng","year":"2017","journal-title":"Food Hydrocolloids"},{"key":"2026090218050343300_cit2","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1016\/j.procbio.2017.11.006","article-title":"Towards\npredicting protein hydrolysis by bovine trypsin","volume":"65","author":"Deng","year":"2018","journal-title":"Process Biochem."},{"issue":"24","key":"2026090218050343300_cit3","doi-asserted-by":"publisher","first-page":"5827","DOI":"10.1007\/s00216-014-8006-2","article-title":"Introducing\nenzyme\nselectivity: A quantitative parameter to describe enzymatic protein\nhydrolysis","volume":"406","author":"Butre\u0301","year":"2014","journal-title":"Anal. Bioanal. Chem."},{"issue":"10","key":"2026090218050343300_cit4","doi-asserted-by":"publisher","first-page":"886","DOI":"10.1136\/gut.20.10.886","article-title":"Trypsinogen variants in pancreatic\njuice of healthy volunteers, chronic alcoholics, and patients with\npancreatitis and cancer of the pancreas","volume":"20","author":"Rinderknecht","year":"1979","journal-title":"Gut"},{"key":"2026090218050343300_cit5","doi-asserted-by":"publisher","first-page":"88","DOI":"10.1016\/j.bej.2012.10.001","article-title":"\u03b2-Lactoglobulin tryptic digestion: A model approach for peptide\nrelease","volume":"70","author":"Ferna\u0301ndez","year":"2013","journal-title":"Biochem. Eng. J."},{"issue":"12","key":"2026090218050343300_cit6","doi-asserted-by":"publisher","first-page":"5666","DOI":"10.1021\/pr400611h","article-title":"Comprehensive\nanalysis of protein digestion using six\ntrypsins reveals the origin of trypsin as a significant source of\nvariability in proteomics","volume":"12","author":"Walmsley","year":"2013","journal-title":"J. Proteome Res."},{"key":"2026090218050343300_cit7","doi-asserted-by":"crossref","DOI":"10.1007\/978-3-642-48380-6","volume-title":"Specificity of proteolysis","author":"Keil","year":"1992"},{"issue":"6","key":"2026090218050343300_cit8","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1002\/mas.21376","article-title":"Getting intimate with trypsin, the\nleading protease in proteomics","volume":"32","author":"Vandermarliere","year":"2013","journal-title":"Mass Spectrom.\nRev."},{"issue":"2","key":"2026090218050343300_cit9","doi-asserted-by":"publisher","first-page":"157","DOI":"10.1016\/S0006-291X(67)80055-X","article-title":"On the size of the active site in proteases. I. Papain","volume":"27","author":"Schechter","year":"1967","journal-title":"Biochem. Biophys. Res. 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I.\n          \n          Introducing enzyme selectivity as a quantitative parameter to describe\nthe effects of substrate concentration on protein hydrolysis. Ph.D. Thesis, Wageningen University & Research, Wageningen, The Netherlands, 2014."},{"issue":"14","key":"2026090218050343300_cit14","doi-asserted-by":"publisher","first-page":"5445","DOI":"10.1021\/jf070337l","article-title":"Prediction of molar\nextinction coefficients of proteins\nand peptides using UV absorption of the constituent amino acids at\n214 nm to enable quantitative reverse phase high-performance liquid\nchromatography-mass spectrometry analysis","volume":"55","author":"Kuipers","year":"2007","journal-title":"J.\nAgric. 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Substrate regulation of peptide bond demasking\nand hydrolysis. Liquid chromatography of hydrolyzates","volume":"30","author":"Vorobe\u0300v","year":"1986","journal-title":"Nahrung"},{"key":"2026090218050343300_cit29","doi-asserted-by":"publisher","first-page":"S426","DOI":"10.1016\/j.molcatb.2017.03.005","article-title":"Demasking\nkinetics of peptide bond cleavage for whey protein isolate hydrolysed\nby Bacillus licheniformis protease","volume":"133","author":"Vorob\u2019ev","year":"2016","journal-title":"J. Mol. Catal. 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