Issue 18, 2021

Enabling highly (R)-enantioselective epoxidation of styrene by engineering unique non-natural P450 peroxygenases

Abstract

Unlike the excellent (S)-enantioselective epoxidation of styrene performed by natural styrene monooxygenases (ee > 99%), the (R)-enantioselective epoxidation of styrene has not yet achieved a comparable efficiency using natural or engineered oxidative enzymes. This report describes the H2O2-dependent (R)-enantioselective epoxidation of unfunctionalized styrene and its derivatives by site-mutated variants of a unique non-natural P450BM3 peroxygenase, working in tandem with a dual-functional small molecule (DFSM). The observed (R)-enantiomeric excess (ee) of styrene epoxidation is up to 99% with a turnover number (TON) of 918 by the best enantioselective mutant F87A/T268I/L181Q, while the best active mutant F87A/T268I/V78A/A184L (with 98% ee) gave a catalytic TON of 4350, representing the best activity of a P450 peroxygenase towards styrene epoxidation to date. Following this approach, a set of styrene derivatives, such as o-, m-, p-chlorostyrenes and fluorostyrenes, could also be epoxidized with modest to very good TONs (362–3480) and high (R)-enantioselectivities (95–99% ee). The semi-preparative scale synthesis of (R)-styrene oxide performed at 0 °C with high conversion, maintaining enantioselectivity, and moderate isolated yields, further suggests the potential application of the current P450 enzymatic system in styrene epoxidation. This study indicates that the synergistic use of protein engineering and an exogenous DFSM constitutes an efficient strategy to control the enantioselectivity of styrene epoxidation, thus substantially expanding the chemical scope of P450 enzymes as useful bio-oxidative catalysts.

Graphical abstract: Enabling highly (R)-enantioselective epoxidation of styrene by engineering unique non-natural P450 peroxygenases

Supplementary files

Article information

Article type
Edge Article
Submitted
15 1月 2021
Accepted
21 3月 2021
First published
22 3月 2021
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2021,12, 6307-6314

Enabling highly (R)-enantioselective epoxidation of styrene by engineering unique non-natural P450 peroxygenases

P. Zhao, J. Chen, N. Ma, J. Chen, X. Qin, C. Liu, F. Yao, L. Yao, L. Jin and Z. Cong, Chem. Sci., 2021, 12, 6307 DOI: 10.1039/D1SC00317H

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