Regiodivergent and Enantioselective Hydroxylation of C−H bonds by Synergistic Use of Protein Engineering and Exogenous Dual‐Functional Small Molecules. Issue 4 (14th December 2022)

• Record Type: Journal Article
• Authors: Chen, Jie; Dong, Sheng; Fang, Wenhan; Jiang, Yiping; Chen, Zhifeng; Qin, Xiangquan; Wang, Cong; Zhou, Haifeng; Jin, Longyi; Feng, Yingang; Wang, Binju; Cong, Zhiqi
• Is Part Of: Angewandte Chemie international edition. Volume 62:Issue 4(2023)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-12-14
• Subjects: Cytochrome P450 Enzymes -- Divergent Hydroxylation -- Enantioselectivity -- Enzyme Catalysis -- Regioselectivity; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.202215088
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: It is a great challenge to optionally access diverse hydroxylation products from a given substrate bearing multiple reaction sites of sp 3 and sp 2 C−H bonds. Herein, we report the highly selective divergent hydroxylation of alkylbenzenes by an engineered P450 peroxygenase driven by a dual‐functional small molecule (DFSM). Using combinations of various P450BM3 variants with DFSMs enabled access to more than half of all possible hydroxylated products from each substrate with excellent regioselectivity (up to >99 %), enantioselectivity (up to >99 % ee ), and high total turnover numbers (up to 80963). Crystal structure analysis, molecular dynamic simulations, and theoretical calculations revealed that synergistic effects between exogenous DFSMs and the protein environment controlled regio‐ and enantioselectivity. This work has implications for exogenous‐molecule‐modulated enzymatic regiodivergent and enantioselective hydroxylation with potential applications in synthetic chemistry. Abstract : An enzymatic approach is reported for multiple‐site highly regio‐ and enantioselective hydroxylation of alkylbenzenes by a combination of P450 variants and exogenous dual‐functional small molecules (DFSMs). Crystallographic analysis and molecular dynamic simulations provided strong evidence for the synergistic effects of engineered P450s and DFSMs, thus establishing a new synthetic toolbox for sustainable divergent biohydroxylation of C−H bonds.