Characterization of a novel cytochrome P450 fromAmycolatopsissp. CGMCC1149 for hydroxylation of lovastatin

Abstract Wuxistatin, a novel and potent statin, is converted from lovastatin by A mycolatopsis sp. CGMCC 1149. In the bioconversion, lovastatin is firstly hydroxylated to 3‐hydroxymethyl lovastatin (product I) by a hydroxylase. In the current study, a novel hydroxylase gene p450lov was isolated from A mycolatopsis sp. CGMCC 1149 by degenerate PCR and self‐formed adaptor PCR and expressed in E scherichia coli . The gene encodes a 403‐amino‐acid protein with a molecular weight of 44.8 kDa and was designated as a new member of cytochrome P 450 ( CYP ) 105 family, CYP 105 A 44. Meanwhile, a lovastatin catalytic in vitro system was established, and an optimal hydroxylation reaction system contained 30 µM lovastatin, 600 µM NADH , 120 µM ferredoxin, 0.04 U ferredoxin−nicotinamide adenine dinucleotide phosphate reductase, and 100 µM CYP 105 A 44 in a final volume of 200 µL Tris HC l buffer (50 mM, pH 7.4). These would be helpful for further studies on the hydroxylation of statins.