Semi-artificial hydroxylating enzymes created by flavins binding to cytochrome P450 2B4 and by bleomycin binding to NADPH-cytochrome P450 reductase

Abstract

To create a semi-artificial monomolecular oxygenase system, FAD or FMN were covalently bound to cytochrome P450 2B4 as electron donor centers and bleomycin to NADPH-cytochrome P450 reductase as a generator of active oxygen species. The most catalytically active was the conjugate of cytochrome P450 with FMN, able to initiate the reactions of dimethylaniline and aminopyrine demethylation along with the reaction of aniline p-hydroxylation. The conjugate of cytochrome P450 with FAD oxidized these substrates at a much slower rate. The bleomycin-reductase complex was capable of demethylating dimethylaniline and aminopyrine but failed to oxidize aniline.