Discovery and Synthetic Applications of a NAD(P)H-Dependent Reductive Aminase from .

Reductive amination is one of the most synthetically direct routes to access chiral amines. Several Imine Reductases (IREDs) have been discovered to catalyze reductive amination (Reductive Aminases or RedAms), yet they are dependent on the expensive phosphorylated nicotinamide adenine dinucleotide cofactor NADPH and usually more active at basic pH. Here, we describe the discovery and synthetic potential of an IRED from (RedAm) that catalyzes reductive amination between a series of medium to large carbonyl and amine compounds with conversions of up to >99% and 99% enantiomeric excess at neutral pH. RedAm catalyzes the formation of a substituted γ-lactam and -methyl-1-phenylethanamine with stereochemistry opposite to that of fungal RedAms, giving the ()-enantiomer. This enzyme remarkably uses both NADPH and NADH cofactors with values of 15 and 247 μM and turnover numbers of 3.6 and 9.0 s, respectively, for the reductive amination of hexanal with allylamine. The crystal structure obtained provides insights into the flexibility to also accept NADH, with residues R35 and I69 diverging from that of other IREDs/RedAms in the otherwise conserved Rossmann fold. RedAm thus represents a subfamily of enzymes that enable synthetic applications using NADH-dependent reductive amination to access complementary chiral amine products.