[Cloning and application of a novel hydroxylase in lovastatin conversion].
Wuxistatin, a novel and potent statin, is converted from lovastatin by Amycolatopsis sp. CGMCC1149. In the bioconversion, lovastatin is firstly hydroxylated by a hydroxylase. To obtain the critical hydroxylase, a novel hydroxylase gene was isolated from Amycolatopsis sp. CGMCC1149 by Degenerate PCR and Self-Formed Adaptor PCR and expressed in Escherichia coli. BLAST sequence analysis revealed that the gene belonged to cytochrome P450 gene superfamily and could encode a 403-amino-acid protein with a molecular weight of 44.8 kDa. The secondary structure prediction result showed that this protein contained many typical functional regions of P450, such as oxygen binding site, ion-pair region and heme binding region. Meanwhile, a catalytic function verification system was constructed by NADH, ferredoxin and ferredoxin reductase which could catalyze lovastatin hydroxylation into the target product. These would be helpful for further studies in large-scale production of wuxistatin.
Xiaoyu
X
Huo
Bin
B
Zhuge
Huiying
H
Fang
Hong
H
Zong
Jian
J
Song
Jian
J
Zhuge
D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D002087:Butyrates / Q000378:metabolism*; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D006900:Hydroxylation; D007218:Industrial Microbiology; D008148:Lovastatin / Q000378:metabolism*; D008969:Molecular Sequence Data
D002087:Butyrates; C533002:wuxistatin; D003577:Cytochrome P-450 Enzyme System; D008148:Lovastatin