In-silico and In-vitro based studies of Streptomyces peucetius CYP107N3 for oleic acid epoxidation

被引：6

作者：

Bhattarai, Saurabh ^{[1
]}

Niraula, Narayan Prasad ^{[1
]}

Sohng, Jae Kyung ^{[1
]}

Oh, Tae-Jin ^{[1
]}

机构：

[1] SunMoon Univ, Dept Pharmaceut Engn, IBR, Asan 336708, South Korea

来源：

BMB REPORTS | 2012年 / 45卷 / 12期

基金：

新加坡国家研究基金会;

关键词：

Cytochrome P450; Epoxidation; Homology Modeling/docking; In silico; Streptomyces peucetius; FATTY-ACIDS; HETEROLOGOUS EXPRESSION; EPOXYGENASE METABOLITES; POLYKETIDE SYNTHASE; ARACHIDONIC-ACID; BIOSYNTHESIS; OXYLIPINS; MECHANISM; OXIDATION; CLONING;

D O I：

10.5483/BMBRep.2012.45.12.080

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Certain members of the cytochromes P450 superfamily metabolize polyunsaturated long-chain fatty acids to several classes of oxygenated metabolites. An approach based on in silico analysis predicted that Streptomyces peucetius CYP107N3 might be a fatty acid-metabolizing enzyme, showing high homology with epoxidase enzymes. Homology modeling and docking studies of CYP107N3 showed that oleic acid can fit directly into the active site pocket of the double bond of oleic acid within optimum distance of 4.6 angstrom from the Fe. In order to confirm the epoxidation activity proposed by in silico analysis, a gene coding CYP107N3 was expressed in Escherichia coli. The purified CYP107N3 was shown to catalyze C-9-C-10 epoxidation of oleic acid in vitro to 9,10-epoxy stearic acid confirmed by ESI-MS, HPLC-MS and GC-MS spectral analysis. [BMB Reports 2012; 45(12): 736-741]

引用

页码：736 / 741

页数：6

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