title: Screening of bacterial cytochrome P450s responsible for regiospecific hydroxylation of (iso)flavonoids.
abstract: Screening of cytochrome P450 monoxygenases responsible for the regiospecific hydroxylation of flavones, isoflavones and chalcones was attempted using a P450 library constructed from Streptomyces avermitilis MA4680, Bacillus and Nocardia farcinica IFM10152 strains. As electron transfer redox partners with the P450s in Escherichia coli system, putidaredoxin reductase (PdR) and putidaredoxin (Pdx) from Pseudomonas putida were used. Among the 50 soluble P450s in the library screened, three cytochrome P450s, i.e. CYP107Y1, CYP125A2 and CYP107P2 from S. avermitilis MA4680 showed good hydroxylation activities towards flavones and isoflavones. However, low product yields prevented us from identifying complete structure of the products. By using S. avermitilis MA4680 as their expression host, further analysis identified that CYP107Y1(SAV2377), CYP125A2(SAV5841) and CYP107P2(SAV4539) showed good regiospecific hydroxylation activities towards genistein (4',5,7-trihydroxyisoflavone), chrysin (5,7-dihydroxyisoflavone) and apigenin (4',5,7-dihydroxyisoflavone) to produce 3',4',5,7,-tetrahydroxyisoflavone, B-ring hydroxylated 5,7-dihydroxyflavone and 3',4',5,7,-tetrahydroxyflavone, respectively. Analyses of the reaction products were performed using HPLC, ESI-MS-MS and GC-MS and 1H NMR.
authors: School of Chemical and Biological Engineering, Seoul National University, Seoul, South Korea.
authors: Bishnu Prasad
authors: BP
authors: Pandey
authors: Nahum
authors: N
authors: Lee
authors: Kwon-Young
authors: KY
authors: Choi
authors: Eunok
authors: E
authors: Jung
authors: Da-Hye
authors: DH
authors: Jeong
authors: Byung-Gee
authors: BG
authors: Kim
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004579:Electron Transport; D005419:Flavonoids / Q000378:metabolism*; D006900:Hydroxylation; D007529:Isoflavones / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
chemical_list: D001426:Bacterial Proteins; D005419:Flavonoids; D007529:Isoflavones; D011994:Recombinant Proteins; C040641:genistin; C043561:chrysin; D003577:Cytochrome P-450 Enzyme System