Regiospecific flavonoid 7-O-methylation with Streptomyces avermitilis O-methyltransferase expressed in Escherichia coli.
O-Methylation, commonly found in synthesis of secondary metabolites of plants and micro-organisms, appears to transfer a methyl group to the hydroxyl group of the recipient which increases the hydrophobicity of the recipient. O-Methyltransferase (OMT), , was isolated and characterized from Streptomyces avermitilis MA-4680. Its amino acid sequence showed 68% similarity with antibiotic C-1027 OMT and 53% similarity with the carminomycin 4-OMT. was expressed in E. coli as a His-tag fusion protein and showed that the methyl was transferred onto the 7-hydroxyl group of the isoflavones, daidzein and genistein, and the flavones, kaempferol and quercetin, as well as the flavanone naringenin. NMR and liquid chromatography-mass spectrometry were used to confirm the location of the methyl group on the recipient compound of naringenin, which was biotransformed into sakuranetin by E. coli transformant expressing (E. coli Sa-2). Therefore, E. coli Sa-2 would be used for the synthesis of the antifungal flavonoid, sakuranetin, through biotransformation.
Kim
Bong-Gyu
BG
Bio/Molecular Informatics Center, Department of Molecular Biotechnology, Konkuk University, Seoul 143-701, Korea.
Jung
Bo-Ra
BR
Lee
Youngshim
Y
Hur
Hor-Gil
HG
Lim
Yoongho
Y
Ahn
Joong-Hoon
JH
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000235:genetics*; D044950:Flavanones / Q000378:metabolism; D005419:Flavonoids / Q000378:metabolism*; D015870:Gene Expression; D019833:Genistein / Q000378:metabolism; D007529:Isoflavones / Q000378:metabolism; D044949:Kaempferols / Q000378:metabolism; D008745:Methylation; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011794:Quercetin / Q000378:metabolism; D016415:Sequence Alignment; D013302:Streptomyces / Q000201:enzymology*
D044950:Flavanones; D005419:Flavonoids; D007529:Isoflavones; D044949:Kaempferols; C004742:daidzein; C006552:kaempferol; D011794:Quercetin; D019833:Genistein; D008780:Methyltransferases; C005273:naringenin