{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T18:40:48Z","timestamp":1792521648511,"version":"4.5.0"},"reference-count":36,"publisher":"Wiley","issue":"20","license":[{"start":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T00:00:00Z","timestamp":1792454400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T00:00:00Z","timestamp":1792454400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22378120"],"award-info":[{"award-number":["22378120"]}],"id":[{"id":"10.13039\/501100001809","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":[[2026,10,28]]},"abstract":"<jats:p>\n                    Indoxacarb is an oxadiazine insecticide bearing a single stereogenic center, of which only the (\n                    <jats:italic>S<\/jats:italic>\n                    )\u2010enantiomer exhibits insecticidal activity. Therefore, the synthesis of the key intermediate (\n                    <jats:italic>S<\/jats:italic>\n                    )\u20105\u2010chloro\u20102\u2010methoxycarbonyl\u20102\u2010hydroxy\u20101\u2010indanone (\n                    <jats:bold>2<\/jats:bold>\n                    ) is crucial for the enantioselective total synthesis of Indoxacarb. In this study, a flavin\u2010containing monooxygenase from\n                    <jats:italic>Bradyrhizobium<\/jats:italic>\n                    sp. 200 (\n                    <jats:italic>Bj<\/jats:italic>\n                    FMO) was employed for the asymmetric synthesis of this chiral intermediate, offering a biocatalytic approach as a potential complement to conventional chemical routes. Based on molecular docking and sequence alignment, several amino acid residues located in the FAD and NADPH\u2010binding domains were selected for site\u2010saturation mutagenesis. In combination with rational design, additional positions potentially affecting catalytic activity and stability were subjected to site\u2010directed mutagenesis. Beneficial single mutations were iteratively combined to generate the final variant,\n                    <jats:italic>Bj<\/jats:italic>\n                    FMO\n                    <jats:sub>M9<\/jats:sub>\n                    . Under optimal conditions,\n                    <jats:italic>Bj<\/jats:italic>\n                    FMO\n                    <jats:sub>M9<\/jats:sub>\n                    displayed a twofold increase in specific activity compared with the wild type. Its half\u2010life at 40\u2009\u00b0C was extended from 1.3 to 3.0\u2009h, and its tolerance toward isopropanol, ethanol, and acetone was markedly enhanced. In a preparative synthesis,\n                    <jats:italic>Bj<\/jats:italic>\n                    FMO\n                    <jats:sub>M9<\/jats:sub>\n                    consistently outperformed the wild type, achieving &gt;80% conversion within 10\u2009h and &gt;90% ee for the (\n                    <jats:italic>S<\/jats:italic>\n                    )\u2010product.\n                  <\/jats:p>","DOI":"10.1002\/cbic.70575","type":"journal-article","created":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T18:34:05Z","timestamp":1792521245000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Discovery and Engineering of a Flavin\u2010Containing Monooxygenase From\n                    <i>Bradyrhizobium<\/i>\n                    sp. 200 for the Asymmetric Synthesis of an Indoxacarb Intermediate"],"prefix":"10.1002","volume":"27","author":[{"given":"Yifei","family":"Hao","sequence":"first","affiliation":[{"name":"Shanghai Collaborative Innovation Center for Biomanufacturing East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Jiang","sequence":"additional","affiliation":[{"name":"Shanghai Collaborative Innovation Center for Biomanufacturing East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Na","family":"Liu","sequence":"additional","affiliation":[{"name":"Shanghai Collaborative Innovation Center for Biomanufacturing East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2498-8012","authenticated-orcid":false,"given":"Xiaoyan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Collaborative Innovation Center for Biomanufacturing East China University of Science and Technology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6973-581X","authenticated-orcid":false,"given":"Yunpeng","family":"Bai","sequence":"additional","affiliation":[{"name":"Shanghai Collaborative Innovation Center for Biomanufacturing East China University of Science and Technology  Shanghai China"},{"name":"Shaanxi R&amp;D Centre of Biomaterials and Fermentation Engineering School of Chemical Engineering Northwest University  Xi\u2019an Shaanxi 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