Increasing the Activity and Efficiency of Stereoselective Oxidations by using Decoy Molecules in Combination with Rate‐Enhancing Variants of P450Bm3

Abstract The use of rate‐accelerating variants of P450Bm3 coupled with decoy molecules is described, resulting in improved catalytic activity for hydroxylation and epoxidation reactions. Prochiral substrates were investigated to ascertain the effect of the mutant enzymes and the decoy molecules on the regio‐ and enantioselectivity of the oxidations. For the alkyl and alkene substituted benzene substrates tested, large improvements in the product formation activity over the wild‐type enzyme were obtained. The product formation rates for the substrates tested ranged from 660 to 2210 nmol (nmol P450) −1 min −1 for variants containing the R47L and Y51F mutations. Although the regioselectivity was not significantly altered in most of the turnovers, some adjustment in the enantioselectivity was observed with smaller substrates. The addition of decoy molecules often resulted in improved enantioselectivity and counteracted reductions arising from the rate‐accelerating mutants.