{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T07:58:07Z","timestamp":1790927887418,"version":"4.1.0"},"reference-count":20,"publisher":"Wiley","issue":"21","license":[{"start":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T00:00:00Z","timestamp":1786320000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T00:00:00Z","timestamp":1786320000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Rapid Comm Mass Spectrometry"],"published-print":{"date-parts":[[2026,11,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Rationale<\/jats:title>\n                    <jats:p>The proteolytic behavior of chymosins during cheese ripening impacts cheese texture, flavor, and quality. Especially, the specific hydrolysis of \u03b1S1\u2010casein to \u03b1S1\u2010I\u2010casein is relevant for early cheese texture development, and this activity varies between chymosin variants. Accurate profiling of \u03b1S1\u2010casein and \u03b1S1\u2010I\u2010casein during cheese ripening is crucial for understanding these differences between chymosins and the impact on cheese quality.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Intact\u2010protein liquid chromatography\u2013high\u2010resolution mass spectrometry (LC\u2010HRMS) was employed to monitor the hydrolysis of \u03b1S1\u2010casein to \u03b1S1\u2010I\u2010casein during mozzarella cheese ripening. Cheese samples, produced with both bovine and camel chymosin variants, were collected at multiple ripening stages. Protein extraction, separation, and quantification were performed using optimized LC\u2010HRMS protocols to assess hydrolysis kinetics and phosphorylation variants.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>LC\u2010HRMS analysis revealed that the tested bovine chymosins displayed significantly higher rates of \u03b1S1\u2010casein hydrolysis to \u03b1S1\u2010I\u2010casein compared to camel chymosins throughout mozzarella cheese ripening. Distinct phosphorylation variants were confidently detected and quantified. Kinetic profiles demonstrated consistent proteolytic activity differences between coagulants.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>LC\u2010HRMS allows tracking of \u03b1S1\u2010casein hydrolysis kinetics throughout mozzarella cheese ripening. This approach not only highlights the pronounced and faster proteolytic activity of bovine chymosins compared to camel chymosins, resulting in more rapid \u03b1S1\u2010I\u2010casein formation, but also facilitates the monitoring of phosphorylation variants present in the cheese. These findings emphasize the enzyme\u2010specific effects on cheese texture and quality, demonstrating the importance of advanced protein analysis for comprehensively understanding cheese production and maturation processes.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/rcm.70153","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T06:55:29Z","timestamp":1786431329000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Profiling Hydrolysis Rates of \u03b1S1\u2010Casein to \u03b1S1\u2010I\u2010Casein During Cheese Ripening by Intact\u2010Protein Liquid Chromatography\u2013High Resolution Mass Spectrometry"],"prefix":"10.1002","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-2940-1668","authenticated-orcid":false,"given":"Sebastiaan","family":"Dolman","sequence":"first","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marjolein","family":"van der Eijk","sequence":"additional","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Loes","family":"Bevers","sequence":"additional","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kees","family":"Muijlwijk","sequence":"additional","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mee Loke","family":"Lim","sequence":"additional","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3387-9253","authenticated-orcid":false,"given":"Margriet","family":"Hendriks","sequence":"additional","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-0953-8077","authenticated-orcid":false,"given":"Nicolas","family":"Abello","sequence":"additional","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rob","family":"van der Hoeven","sequence":"additional","affiliation":[{"name":"dsm\u2010firmenich  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,10]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/9781444323740.ch3"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.3168\/jds.S0022-0302(03)73869-7"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0958-6946(98)00040-5"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444913003260"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0022029900020707"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0022029900027734"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(85)80923-6"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/jf302890h"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.idairyj.2022.105346"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.idairyj.2023.105835"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.3168\/jds.S0022-0302(82)82289-3"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.3168\/jds.S0022-0302(06)72154-3"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2024.142034"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0163471"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.3168\/jds.2019-16273"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.3168\/jds.2024-26179"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.3168\/jds.2016-11250"},{"key":"e_1_2_9_19_1","unstructured":"\u201cJMP [Computer Program] \u201d Version 18.2. 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