Kang, Suk Woo Antoney, James Lupton, David W. Speight, Robert Scott, Colin Jackson, Colin J. 2024-01-19T10:00:10Z 2024-01-19T10:00:10Z 2023-04-06 2023-04 1439-4227 https://pubs.kist.re.kr/handle/201004/113871 Asymmetric reduction by ene-reductases has received considerable attention in recent decades. While several enzyme families possess ene-reductase activity, the Old Yellow Enzyme (OYE) family has received the most scientific and industrial attention. However, there is a limited substrate range and few stereocomplementary pairs of current ene-reductases, necessitating the development of a complementary class. Flavin/deazaflavin oxidoreductases (FDORs) that use the uncommon cofactor F-420 have recently gained attention as ene-reductases for use in biocatalysis due to their stereocomplementarity with OYEs. Although the enzymes of the FDOR-As sub-group have been characterized in this context and reported to catalyse ene-reductions enantioselectively, enzymes from the similarly large, but more diverse, FDOR-B sub-group have not been investigated in this context. In this study, we investigated the activity of eight FDOR-B enzymes distributed across this sub-group, evaluating their specific activity, kinetic properties, and stereoselectivity against alpha,beta-unsaturated compounds. The stereochemical outcomes of the FDOR-Bs are compared with enzymes of the FDOR-A sub-group and OYE family. Computational modelling and induced-fit docking are used to rationalize the observed catalytic behaviour and proposed a catalytic mechanism. English John Wiley & Sons Ltd. Asymmetric Ene-Reduction by F420-Dependent Oxidoreductases B (FDOR-B) from Mycobacterium smegmatis Article 10.1002/cbic.202200797 1 ChemBioChem, v.24, no.8 ChemBioChem 24 8 Y scie scopus 000945793100001 2-s2.0-85149513706 Biochemistry & Molecular Biology Chemistry, Medicinal Biochemistry & Molecular Biology Pharmacology & Pharmacy Article; Early Access F420H2-DEPENDENT REDUCTASES COFACTOR COENZYME FAMILY F-420 GLUCOSE-6-PHOSPHATE-DEHYDROGENASE IDENTIFICATION COVERAGE ENZYMES biocatalysis deazaflavin F-420 ene-reductases Mycobacterium smegmatis