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Compared to cytochrome P450 enzymes (P450s), the catalytic potential of UPOs has been less investigated, largely due to their limited natural diversity and the challenges associated with their optimization through enzyme engineering. In this study, we engineered an UPO from\n                    <jats:italic>Aspergillus niger<\/jats:italic>\n                    (\n                    <jats:italic>Ani<\/jats:italic>\n                    UPO) to catalyze the enantioselective \u03b1\u2010hydroxylation of \u03b2\u2010ketoesters, a valuable transformation yet to be realized in biocatalysis. Through enzyme engineering, two\n                    <jats:italic>Ani<\/jats:italic>\n                    UPO variants,\n                    <jats:italic>Ani<\/jats:italic>\n                    UPO\u2010M3 and\n                    <jats:italic>Ani<\/jats:italic>\n                    UPO\u2010M6, were developed to produce a wide range of enantioenriched \u03b1\u2010hydroxy\u2010\u03b2\u2010ketoesters, achieving up to 97% yield, 4140 total turnover number (TTN), and &gt;99:1 enantiomeric ratio (er). The biocatalytic process operates under mild conditions and is scalable for preparative applications. This study broadens the catalytic repertoire of UPOs and enhances their potential for industrial applications.\n                  <\/jats:p>","DOI":"10.1002\/anie.202509359","type":"journal-article","created":{"date-parts":[[2025,6,6]],"date-time":"2025-06-06T04:58:58Z","timestamp":1749185938000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Engineering Unspecific Peroxygenases for Enantioselective \u03b1\u2010Hydroxylation of \u03b2\u2010Ketoesters"],"prefix":"10.1002","volume":"64","author":[{"given":"Zhen\u2010Yu","family":"Wang","sequence":"first","affiliation":[{"name":"Key Laboratory of Drug\u2010Targeting and Drug Delivery System of the Education Ministry and Sichuan Province, Sichuan Engineering Laboratory for Plant\u2010Sourced Drug and Sichuan Research Center for Drug Precision Industrial Technology, Department of Biopharmaceutics, West China School of Pharmacy Sichuan University  Chengdu 610041 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Drug\u2010Targeting and Drug Delivery System of the Education Ministry and Sichuan Province, Sichuan Engineering Laboratory for Plant\u2010Sourced Drug and Sichuan Research Center for Drug Precision Industrial Technology, Department of Biopharmaceutics, West China School of Pharmacy Sichuan University  Chengdu 610041 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-7249-8180","authenticated-orcid":false,"given":"Peng","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Birth Defects and Related Disease of Women and Children, Children's Medicine Key Laboratory of Sichuan Province, Department of Pharmacy\/Evidence\u2010Based Pharmacy Center, West China Second University Hospital Sichuan University  Chengdu 610041 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongxiang","family":"Zheng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Drug\u2010Targeting and Drug 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