{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T07:49:22Z","timestamp":1791964162459,"version":"4.3.4"},"reference-count":39,"publisher":"Wiley","issue":"17","license":[{"start":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T00:00:00Z","timestamp":1789344000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T00:00:00Z","timestamp":1789344000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Basic Research Program of China","doi-asserted-by":"publisher","award":["2024YFA0917800"],"award-info":[{"award-number":["2024YFA0917800"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2026,9,14]]},"abstract":"<jats:p>\n                    Efficient enzymaticsynthesis of L\u2010carnosine (L\u2010Car) is often limited by competing hydrolysis and side reactions that reduce substrate utilization and product yield. In this study, a \u03b2\u2010aminopeptidase (\n                    <jats:italic>Cb<\/jats:italic>\n                    AP) from\n                    <jats:italic>Caulobacter<\/jats:italic>\n                    sp. was systematically engineered to enhance catalytic selectivity toward L\u2010Car using \u03b2\u2010alaninamide and L\u2010histidine as substrates. Analysis of \u03b2\u2010alaninamide\u2010derived byproducts revealed two major competing pathways, namely \u03b2\u2010alaninamide hydrolysis and self\u2010ligation into short oligopeptides. To evaluate the balance between L\u2010Car synthesis and competing reactions, a composite apparent selectivity index (\n                    <jats:italic>R<\/jats:italic>\n                    <jats:sub>S<\/jats:sub>\n                    ) was introduced. Structure\u2010guided mutagenesis targeting residues within 4\u20136\u2009\u00c5 of the active site identified several improved variants. Iterative combination of beneficial mutations yielded the optimal mutant\n                    <jats:italic>Cb<\/jats:italic>\n                    AP\n                    <jats:sub>Q138FF96Y<\/jats:sub>\n                    (\n                    <jats:italic>Cb<\/jats:italic>\n                    AP\n                    <jats:sub>M2<\/jats:sub>\n                    ), exhibiting an approximately 2.3\u2010fold increase in\n                    <jats:italic>R<\/jats:italic>\n                    <jats:sub>S<\/jats:sub>\n                    relative to Q138F and an approximately 3.1\u2010fold increase relative to the wild\u2010type enzyme, with substantial suppression of undesired side reactions. Molecular dynamics simulations indicated that these improvements arise from reduced substrate binding stability and disrupted hydrogen\u2010bonding interactions with \u03b2\u2010alaninamide and L\u2010Car. Under optimized 10\u2009mL\u2010scale conditions,\n                    <jats:italic>Cb<\/jats:italic>\n                    AP\n                    <jats:sub>M2<\/jats:sub>\n                    produced 37.3\u2009mM L\u2010Car within 1\u2009h, corresponding to a 74.6% yield based on \u03b2\u2010alanine methyl ester, with an STY of 8.44\u2009g\u2009L\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>1<\/jats:sup>\n                    \u2009h\n                    <jats:sup>\u22121<\/jats:sup>\n                    .\n                  <\/jats:p>","DOI":"10.1002\/cbic.70530","type":"journal-article","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T08:17:18Z","timestamp":1789373838000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An Engineered \u03b2\u2010Aminopeptidase With Better Product Selectivity for L\u2010Carnosine Production"],"prefix":"10.1002","volume":"27","author":[{"given":"Bi\u2010Han","family":"Guan","sequence":"first","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering School of Biotechnology East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wu\u2010Tong","family":"Yin","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering School of Biotechnology East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8871-542X","authenticated-orcid":false,"given":"Chen","family":"Zhao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering School of Biotechnology East China University of Science and Technology  Shanghai 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