Engineering Unspecific Peroxygenases for Enantioselective α‐Hydroxylation of β‐Ketoesters

• Published in: Angewandte Chemie International Edition Vol. 64; no. 34; pp. e202509359 - n/a
• Main Authors: Wang, Zhen‐Yu, Ma, Xin‐Yuan, Wu, Ying, Liu, Yang, Lin, Guang‐Xin, Liu, Xiao‐Qi, Zhang, Chun, Chen, Peng, Zheng, Yongxiang, Jia, Zhi‐Jun
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 18.08.2025
• Edition: International ed. in English
• Subjects: Aspergillus niger - enzymology; Biocatalysis; Biocatalysts; Catalysis; Cytochrome P450; Cytochromes P450; Enantiomers; Enantioselectivity; Enzyme engineering; Hydroxylation; Industrial applications; Ketoesters; Ketones - chemistry; Ketones - metabolism; Mixed Function Oxygenases - chemistry; Mixed Function Oxygenases - genetics; Mixed Function Oxygenases - metabolism; Oxyfunctionalization; Protein Engineering; Stereoisomerism; Unspecific peroxygenase; Oxyfunctionalization; Biocatalysis; Enantioselectivity; Enzyme engineering; Unspecific peroxygenase
• ISSN: 1433-7851; 1521-3773; 1521-3773
• DOI: 10.1002/anie.202509359

Unspecific peroxygenases (UPOs) are promising biocatalysts for selective oxyfunctionalization. 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 Aspergillus niger (AniUPO) to catalyze the enantioselective α‐hydroxylation of β‐ketoesters, a valuable transformation yet to be realized in biocatalysis. Through enzyme engineering, two AniUPO variants, AniUPO‐M3 and AniUPO‐M6, were developed to produce a wide range of enantioenriched α‐hydroxy‐β‐ketoesters, achieving up to 97% yield, 4140 total turnover number (TTN), and >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. An unspecific peroxygenase from Aspergillus niger (AniUPO) has been successfully engineered to catalyze the enantioselective α‐hydroxylation of β‐ketoesters. This biocatalytic process demonstrates high efficiency and selectivity with scalability to preparative levels, expanding the catalytic repertoire and synthetic applicability of UPOs in selective oxyfunctionalization.