title: In-silico and In-vitro based studies of Streptomyces peucetius CYP107N3 for oleic acid epoxidation.
abstract: 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 Å 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.
authors.affiliation: Institute of Biomolecule Reconstruction (IBR), Department of Pharmaceutical Engineering, SunMoon University, Asan 336-708, Korea.
authors.forename: Saurabh
authors.initials: S
authors.lastname: Bhattarai
authors.forename: Narayan Prasad
authors.initials: NP
authors.lastname: Niraula
authors.forename: Jae Kyung
authors.initials: JK
authors.lastname: Sohng
authors.forename: Tae-Jin
authors.initials: TJ
authors.lastname: Oh
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D020134:Catalytic Domain; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004852:Epoxy Compounds / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D019301:Oleic Acid / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D021241:Spectrometry, Mass, Electrospray Ionization; D013229:Stearic Acids / Q000378:metabolism; D013302:Streptomyces / Q000378:metabolism*
chemical_list: D004852:Epoxy Compounds; D011994:Recombinant Proteins; D013229:Stearic Acids; C013519:9,10-epoxystearic acid; D019301:Oleic Acid; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; C023111:epoxidase