Cloning and application of a novel hydroxylase in lovastatin conversion.
- Author:
Xiaoyu HUO
;
Bin ZHUGE
;
Huiying FANG
;
Hong ZONG
;
Jian SONG
;
Jian ZHUGE
- Publication Type:Journal Article
- MeSH:
Actinomycetales;
enzymology;
genetics;
Amino Acid Sequence;
Butyrates;
metabolism;
Cloning, Molecular;
Cytochrome P-450 Enzyme System;
genetics;
metabolism;
Hydroxylation;
Industrial Microbiology;
Lovastatin;
metabolism;
Molecular Sequence Data
- From:
Chinese Journal of Biotechnology
2013;29(11):1590-1598
- CountryChina
- Language:Chinese
-
Abstract:
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.
