{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:44:45Z","timestamp":1792784685032,"version":"4.5.0"},"reference-count":40,"publisher":"American Chemical Society (ACS)","issue":"36","license":[{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"name":"Xing Dian Plan \u201cYouth Talent Program\u201d of Yunnan Province"},{"name":"National Natural Science Foundation of China (NSFC)","award":["32160217"],"award-info":[{"award-number":["32160217"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Thermostability is critical yet challenging to engineer in industrial enzymes. Here, we present an integrated workflow for enhancing the thermostability of GH11 xylanases through systematic Ser\/Thr optimization. Gibbs free energy calculations on XynCDBFV identified 169 stabilizing single-site mutations involving Ser\/Thr residues, which were combinatorially integrated via rational design and a machine-learning model (ESM2-RF). Both strategies generated multisite variants with superior thermal tolerance. Top performers included RD-4M3 (\u0394Tm = 7.10 \u00b0C) and ER-4M6 (+56.19% residual activity). Molecular dynamics simulations revealed that R148\u2013D57 competition in the WT at high temperature disrupts the R148\u2013E109 interaction, and its elimination in Ser\/Thr variants induces 150-loop rigidification, thereby enhancing thermostability. This finding was validated by transplanting two Ser\/Thr substituted combinations to a second GH11 xylanase, PjxA. This work establishes a systematic Ser\/Thr optimization strategy for engineering thermostable xylanases and underscores the efficiency of multisite combinatorial design.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c06344","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T09:14:41Z","timestamp":1788426881000},"page":"28818-28830","source":"Crossref","is-referenced-by-count":0,"title":["An Integrated Workflow\nfor Improving Thermostability:\nMechanism and Engineering of GH11 Xylanases via Ser\/Thr Substitutions"],"prefix":"10.1021","volume":"74","author":[{"given":"Yuanyuan","family":"Zhe","sequence":"first","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]},{"name":"Southwest United Graduate School , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sisi","family":"Zhang","sequence":"additional","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enheng","family":"Zhu","sequence":"additional","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongyang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Southwest United Graduate School , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huabiao","family":"Miao","sequence":"additional","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]},{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xia","family":"Xiang","sequence":"additional","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]},{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ran","family":"Zhu","sequence":"additional","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dejiao","family":"Li","sequence":"additional","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9789-1596","authenticated-orcid":true,"given":"Nanyu","family":"Han","sequence":"additional","affiliation":[{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]},{"name":"Yunnan Normal University , , ,","place":["Kunming, China, 650500"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,1]]},"reference":[{"issue":"3","key":"2026091609094848800_cit1","doi-asserted-by":"publisher","first-page":"273","DOI":"10.5114\/bta.2024.141806","article-title":"A Review on Xylanase Sources, Classification, Mode\nof Action, Fermentation Processes, and Applications as a Promising\nBiocatalyst","volume":"105","author":"Abena","year":"2024","journal-title":"BioTechnologia"},{"issue":"1","key":"2026091609094848800_cit2","doi-asserted-by":"publisher","first-page":"263","DOI":"10.1146\/annurev-arplant-042809-112315","article-title":"Hemicelluloses","volume":"61","author":"Scheller","year":"2010","journal-title":"Annu. 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Macromol."},{"issue":"1","key":"2026091609094848800_cit5","doi-asserted-by":"publisher","first-page":"127","DOI":"10.3390\/microorganisms14010127","article-title":"Review\nof Xylanases:\nSources, Engineering and Biotechnological Use","volume":"14","author":"Pavlova","year":"2026","journal-title":"Microorganisms"},{"key":"2026091609094848800_cit6","doi-asserted-by":"publisher","first-page":"108148","DOI":"10.1016\/j.biotechadv.2023.108148","article-title":"Endo-1,4-\u03b2-Xylanase-Containing\nGlycoside Hydrolase Families: Characteristics, Singularities and Similarities","volume":"65","author":"Mendonc\u0327a","year":"2023","journal-title":"Biotechnol. 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