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Computational rational design of unspecific peroxygenase for C-H oxidation
Ruichen Gao1, Xiaodi Fu2, Zonglin Li3
1Lab of Applied Biocatalysis, Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, School of Food Science and Engineering, South China University of Technology, Guangdong 510006, China.
None:
Computational rational design has emerged as a transformative approach to engineer enzymes with tailored selectivity and efficiency. In the context of carbon-hydrogen oxidation, a key challenge in synthetic chemistry, unspecific peroxygenases (UPOs) directly oxidize unactivated carbon-hydrogen bonds using hydrogen peroxide, yet their utility is limited by low activity and imperfect selectivity. By computational rational design, this study systematically navigated vast sequence spaces to identify mutations that enhance catalytic performance of UPOs, lastly yielded UPO variants with 13-fold enhanced activity and >99% enantioselectivity, and revealed the dominant role of residue Lys165 in activity and enantioselectivity. This study shows how computational strategies overcome evolutionary constraints to deliver efficient biocatalysts for synthetic chemistry.
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