Gaseous Alkane Hydroxylation by Deceiving Cytochrome P450BM3 Using Decoy Molecules

Abstract

Cytochrome P450BM3 (P450BM3) catalyses the monooxygenation of fatty acids. However, in the case of non-native substrates possessing different structures from fatty acids, P450BM3 remains in an inactive state and catalysis does not proceed. This review will introduce a unique approach, wherein native substrate mimics (decoy molecules) are employed to trick P450BM3 into mistakenly hydroxylating non-native substrates. The decoy molecule system has the advantage that wild-type enzyme can be employed and catalytic activity improved by developing appropriate decoy molecules. With a focus on the hydroxylation of gaseous alkanes, key developments since the first discovery of decoy molecules to the present day will be covered briefly herein. Notably, ethane was efficiently hydroxylated by wild-type P450BM3 with assistance of the systematically evolved N-substituted dipeptidic decoy molecule N-enanthoyl-L-pipecolyl-L-phenylalanine (C7AMPipPhe), achieving a TOF of 82.7 min⁻¹ P450⁻¹, representing the highest TOF among reported values catalysed by P450s including engineered P450s.