查看文章

Styrene monooxygenase (SMO) is a two-component flavoprotein catalyzing the selective epoxidation of various CC double bonds. For cell-free catalysis, traditionally a cascade of NAD(P)H:flavin oxidoreductase, nicotinamide cofactor (NADH), and NADH regeneration enzyme is required to supply StyA with the reduced flavin adenine dinucleotide cofactor (FADH₂) required for catalysis. Herein, we present a more direct and efficient FADH₂ regeneration system using the nicotinamide cofactor mimic 1-benzyl-1,4-dihydronicotinamide (BNAH) as the sole reductant. Thus, BNAH replaces two enzymes and the nicotinamide cofactor, resulting in a significantly simplified reaction system.