Regiospecific flavonoid 7-0-methylation with Streptomyces avermitilis O-methyltransferase expressed in Escherichia coli

Abstract

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-Methyltransf erase (OMT), SaOMT-2, 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. SaOMT-2 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 SaOMT-2 (E. coli Sa-2). Therefore, E coli Sa-2 would be used for the synthesis of the antifungal flavonoid, sakuranetin, through biotransformation.