Discovery and Rational Mutagenesis of Methionine Sulfoxide Reductase Biocatalysts To Expand the Substrate Scope of the Kinetic Resolution of Chiral Sulfoxides

Silvia Anselmi; Alexandra T P Carvalho; Angela Serrano-Sanchez; Jose L Ortega-Roldan; Jill Caswell; Iman Omar; Gustavo Perez-Ortiz; Sarah M Barry; Thomas S Moody; Daniele Castagnolo

doi:10.1021/acscatal.3c00372

Discovery and Rational Mutagenesis of Methionine Sulfoxide Reductase Biocatalysts To Expand the Substrate Scope of the Kinetic Resolution of Chiral Sulfoxides

ACS Catal. 2023 Mar 23;13(7):4742-4751. doi: 10.1021/acscatal.3c00372. eCollection 2023 Apr 7.

Authors

Affiliations

¹ Department of Chemistry, University College London, 20 Gordon Street, WC1H 0AJ London, U. K.
² Department of Biocatalysis and Isotope Chemistry, Almac, 20 Seagoe Industrial Estate, Craigavon BT63 5QD, U. K.
³ School of Biosciences, University of Kent, Canterbury CT2 7NJ, U. K.
⁴ Faculty of Natural, Mathematical and Engineering Sciences, Department of Chemistry, King's College London, 7 Trinity Street, SE1 1DB London, U. K.
⁵ Arran Chemical Company Limited, Unit 1 Monksland Industrial Estate, Athlone, Co., Roscommon N37 DN24, Ireland.

Abstract

Methionine sulfoxide reductase A (MsrA) enzymes have recently found applications as nonoxidative biocatalysts in the enantioselective kinetic resolution of racemic sulfoxides. This work describes the identification of selective and robust MsrA biocatalysts able to catalyze the enantioselective reduction of a variety of aromatic and aliphatic chiral sulfoxides at 8-64 mM concentration with high yields and excellent ees (up to 99%). Moreover, with the aim to expand the substrate scope of MsrA biocatalysts, a library of mutant enzymes has been designed via rational mutagenesis utilizing in silico docking, molecular dynamics, and structural nuclear magnetic resonance (NMR) studies. The mutant enzyme MsrA33 was found to catalyze the kinetic resolution of bulky sulfoxide substrates bearing non-methyl substituents on the sulfur atom with ees up to 99%, overcoming a significant limitation of the currently available MsrA biocatalysts.

Abstract

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