Sequence-Based Discovery, Characterisation, and Biocatalytic Application of a Set of Imine Reductases

Overview

Journal ChemCatChem

Date 2018 Sep 11

PMID 30197686

Citations 17

Authors

Stefan Velikogne

Verena Resch

Carina Dertnig

Joerg H Schrittwieser

Wolfgang Kroutil

Affiliations

Soon will be listed here.

Abstract

Imine reductases (IREDs) have recently become a primary focus of research in biocatalysis, complementing other classes of amine-forming enzymes such as transaminases and amine dehydrogenases. Following in the footsteps of other research groups, we have established a set of IRED biocatalysts by sequence-based enzyme discovery. In this study, we present basic characterisation data for these novel IREDs and explore their activity and stereoselectivity using a panel of structurally diverse cyclic imines as substrates. Specific activities of >1 U/mg and excellent stereoselectivities (>99 %) were observed in many cases, and the enzymes proved surprisingly tolerant towards elevated substrate loadings. Co-expression of the IREDs with an alcohol dehydrogenase for cofactor regeneration led to whole-cell biocatalysts capable of efficiently reducing imines at 100 mM initial concentration with no need for the addition of extracellular nicotinamide cofactor. Preparative biotransformations on gram scale using these 'designer cells' afforded chiral amines in good yield and excellent optical purity.

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