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Dual-Function Enzyme Catalysis for Enantioselective Carbon–Nitrogen Bond Formation

preprint
submitted on 20.04.2021, 06:07 and posted on 20.04.2021, 13:10 by Zhen Liu, Carla Calvó-Tusell, Andrew Z. Zhou, kai chen, Marc Garcia-Borràs, Frances H. Arnold

Whereas enzymatic asymmetric carbene N–H insertion is a powerful method for preparation of chiral amines in principle, it has suffered from limited enantioselectivity in practice. In this work, we demonstrate that engineered cytochrome P450 enzymes can catalyze this abiological C–N bond-forming reaction with excellent activity and selectivity (up to 32,100 TTN, >99% yield and 98% e.e.) to prepare a series of bioactive α-amino lactones, which have not been accessed previously using a carbene insertion strategy. The enzymes are dual-function catalysts, effecting both carbene transfer and enantioselective proton-transfer catalysis, in a single active site. To gain insight into the mechanism of the enzymatic transformation, especially in the asymmetric protonation step, we performed extensive molecular dynamics simulations and density functional theory (DFT) calculations. Computational studies uncover the important roles of active-site residues that enable high activity and selectivity through interacting with the carbene intermediate and the amine substrate, and directing water molecules for selective proton transfer.

History

Email Address of Submitting Author

zhenliu@caltech.edu

Institution

California Institute of Technology

Country

United States

ORCID For Submitting Author

0000-0002-1119-0693

Declaration of Conflict of Interest

A provisional patent application based on this manuscript has been filed through the California Institute of Technology.

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