Characterization of a novel cytochrome P450 from Amycolatopsis sp. CGMCC1149 for hydroxylation of lovastatin

Abstract

Wuxistatin, a novel and potent statin, is converted from lovastatin by Amycolatopsis sp. CGMCC1149. 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 Amycolatopsis sp. CGMCC1149 by degenerate PCR and self-formed adaptor PCR and expressed in Escherichia 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 P450 (CYP) 105 family, CYP105A44. 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 CYP105A44 in a final volume of 200 µL Tris HCl buffer (50 mM, pH 7.4). These would be helpful for further studies on the hydroxylation of statins.

Bioconversion of lovastatin to simvastatin by Streptomyces carpaticus toward the inhibition of HMG‐CoA activity

“…Simvastatin production is mostly facilitated by the biotransformation, when the microbes are supplemented with lovastatin. A plethora of studies documented the microbial biotransformation of simvastatin from lovastatin by various microbes 48 . In this study, we have screened 40 Actinobacteria strains from soil samples to reveal the ability of individual strain in biotransforming lovastatin to other statins.…”

“…A plethora of studies documented the microbial biotransformation of simvastatin from lovastatin by various microbes. 48 In this study, we have screened 40 Actinobacteria strains from soil samples to reveal the ability of individual strain in biotransforming lovastatin to other statins. In order to characterize the biotransformed statins, we employed TLC, through which, we identified three different strains that produced simvastatin.…”

Detection of 4a,5-dihydropravastatin as Impurity in the Cholesterol Lowering Drug Pravastatin

“…Subsequently, pravastatin is produced by enzymatic hydroxylation of compactin by a bacterial P450 [ 14 ], which most likely at the same time also will hydroxylate the 4a,5-dihydrocompactin present in the fermentation broth [ 15 ] into 4a,5-dihydropravastatin. Promiscuity of statin P450 enzymes with respect to the absence or presence of both double bonds in the decalin core or the methylbutyrate side chain was demonstrated by C6 hydroxylation of dihydromonacolin L [ 22 ] and lovastatin [ 24 ], making it very likely that 4a,5-dihydropravastatin is derived from hydroxylating 4a,5-dihydrocompactin by the P450 monooxygenase.…”