Cytochrome P450105D1 (CYP105D1) from Streptomyces griseus: Heterologous expression, activity, and activation effects of multiple xenobiotics

The open reading frame of CYP105D1, a soluble cytochrome P450 from om Strep tomyces griseus, was cloned behind the too promoter of the bacterial expres sion vector pSPg1910L and expressed in Escherichia coli. The recombinant pr otein retained normal spectral characteristics having a Soret peak at 448 n m in the reduced carbon monoxide difference spectrum. CYP105D1 was active, obtaining reducing equivalents from endogenous E. coli ferredoxin and ferre doxin reductase redox partners present in E. coil. lit vitro activity studi es revealed CYP105D1 to catalyse the NADH- and NADPH-dependent oxidation of the xenobiotic substrates benzo[alpha]pyrene, erythromycin, warfarin, and testosterone. Furthermore, this activity could be stimulated in the presenc e of either cu-benzoflavone or beta-benzoflavone in an analogous manner to that reported for mammalian P450 forms including human liver cytochrome P45 03A4 (CYP3A4). The system produces an alternative to whole-cell biotransfor mation of xenobiotic for the production of drug metabolites and an experime ntal system for probing the structural features of a cytochrome P450 with a broad substrate range. (C) 1999 Academic Press.