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Cloning and application of a novel hydroxylase in lovastatin conversion / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1590-1598, 2013.
Article in Chinese | WPRIM | ID: wpr-242434
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.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Industrial Microbiology / Actinomycetales / Butyrates / Lovastatin / Molecular Sequence Data / Amino Acid Sequence / Cloning, Molecular / Cytochrome P-450 Enzyme System / Genetics / Hydroxylation Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2013 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Industrial Microbiology / Actinomycetales / Butyrates / Lovastatin / Molecular Sequence Data / Amino Acid Sequence / Cloning, Molecular / Cytochrome P-450 Enzyme System / Genetics / Hydroxylation Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2013 Type: Article