{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T15:11:53Z","timestamp":1786115513112,"version":"build-2736575974"},"reference-count":36,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T00:00:00Z","timestamp":1779235200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T00:00:00Z","timestamp":1779235200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100014103","name":"Key Technology Research and Development Program of Shandong Province","doi-asserted-by":"publisher","award":["2023CXGC010714"],"award-info":[{"award-number":["2023CXGC010714"]}],"id":[{"id":"10.13039\/100014103","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["scijournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J Sci Food Agric"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>BACKGROUND<\/jats:title>\n                    <jats:p>\n                      Allitol is a rare sugar alcohol with potential as a low\u2010calorie sweetener and food ingredient; however, its biocatalytic production from\n                      <jats:sc>d<\/jats:sc>\n                      \u2010allulose is limited by the thermostability and cofactor efficiency of the ribitol dehydrogenases (RDH).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>RESULTS<\/jats:title>\n                    <jats:p>\n                      In this study, the RDH from\n                      <jats:italic>Klebsiella oxytoca<\/jats:italic>\n                      was engineered for enhanced thermostability using consensus\u2010guided mutagenesis targeting non\u2010conserved, non\u2010active\u2010site residues, followed by stepwise combination. The resulting M4 mutant showed markedly improved thermostability, with its melting temperature (T\n                      <jats:sub>m<\/jats:sub>\n                      ) elevated by 19.6\u2009\u00b0C relative to the wild type, accompanied by a 10\u2009\u00b0C increase in the optimum reaction temperature and a substantially broader pH activity range. Molecular dynamics simulations indicated that the four substitutions clustered at subunit interfaces and reinforced hydrophobic packing, stabilizing the tetrameric assembly and contributing to enhanced thermostability. An enzyme complex combining M4 with formate dehydrogenase (FDH) was constructed to enable efficient cofactor regeneration. This increased the conversion by 28% compared with the free enzyme combination and achieved nearly complete substrate conversion under optimized conditions.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>CONCLUSION<\/jats:title>\n                    <jats:p>This study significantly improved the thermostability of RDH using consensus\u2010guided interfacial design and constructed a cofactor\u2010regenerating enzyme complex for the efficient biocatalytic production of allitol. \u00a9 2026 Society of Chemical Industry.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/jsfa.70736","type":"journal-article","created":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T04:51:35Z","timestamp":1779339095000},"page":"7034-7043","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rational engineering of ribitol dehydrogenase for enhanced thermostability and efficient biocatalytic production of allitol"],"prefix":"10.1002","volume":"106","author":[{"given":"Tianwen","family":"Shang","sequence":"first","affiliation":[{"name":"College of Biological Engineering Tianjin University of Science and Technology  Tianjin China"},{"name":"National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology Chinese Academy of Sciences  Tianjin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9463-8862","authenticated-orcid":false,"given":"Jiangang","family":"Yang","sequence":"additional","affiliation":[{"name":"National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology Chinese Academy of Sciences  Tianjin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Men","sequence":"additional","affiliation":[{"name":"National Engineering Laboratory for Industrial Enzymes, Tianjin 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