Characterization of a novel cytochrome P450 from Amycolatopsis sp. CGMCC1149 for hydroxylation of lovastatin.
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.
The Key Laboratory of Industrial Biotechnology, Ministry of Education and the Research Centre of Industrial Microbiology, Jiangnan University, Wuxi, People's Republic of China.
Hong
H
Zong
Bin
B
Zhuge
Xinyao
X
Lu
Xiaoyu
X
Huo
Huiying
H
Fang
Jian
J
Song
Jin
J
Sun
D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D055162:Biocatalysis; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D006900:Hydroxylation; D008148:Lovastatin / Q000378:metabolism*; D008969:Molecular Sequence Data
D003577:Cytochrome P-450 Enzyme System; D008148:Lovastatin
3-hydroxymethyl lovastatin (product I); Amycolatopsis sp. CGMCC1149; cytochrome P450; hydroxylase; lovastatin