Partial Purification and Some Properties of Flavonol 3-O-Glycosyltransferases from Seedings of Vigna mungo, with Special Reference to the Formation of Kaempferol 3-O-Galactoside and 3-O-Glucoside
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概要
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A novel enzyme, UDP-D-galactose:flavonol 3-O-galactosyltransferase (F3GaT), catalyzing the transfer of D-galactose from UDP-D-galactose to the 3 position of 5,7,4'-trihydroxyfiavonol (kaempferol), was detected in and purified about 404-fold from seedlings of Vigna mungo by precipitation with ammonium sulfate, chromatography on Sephadex G-100 and chromatofocusing. The enzyme was separated by this procedure from a coexisting UDP-D-glucose:flavonol 3-0-glucosyltransferase (F3GT), which was simultaneously purified about 189-fold. F3GaT was isolated as a soluble enzyme with pH optima of 8.0 in imidazole-HCI buffer and 7.5 in histidine-HCl buffer. F3GT had the same pH optima. The M_r of both F3GaT and F3GT, which had isoelectric points of 5.1 and 6.1, respectively, was estimated by elution from a column of Sephadex G-100 to be about 43,000. The activities of F3GaT and F3GT were stimulated by 14 mM 2-mercapto-ethanol and strongly inhibited by 1 mM Cu^<2+>, 1 mM Zn^<2+>, and various reagents that react with sulfhydryl groups. Among various possible substrates for F3GaT that were tested, kaempferol, isorhamnetin and quercetin were the best. The K_m values for kaempferol and UDP-D-galactose were determined to be 0.40 μM and 125 μM, respectively. Similarly, F3GT had low K_m values of 0.69 μM for kaempferol and 1.67 mM for UDP-D-glucose. F3GaT and F3GT mediated the transfer of galactose and glucose, respectively, to the 3-hydroxyl groups exclusively of kaempferol, isorhamnetin and quercetin. Rhamnetin also functioned as a galactosyl acceptor though less efficiently.
- 日本植物生理学会の論文
著者
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Mato M
Department Of Biological Science Faculty Of Science Kumamoto University
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Mato Masami
Department Of Biological Science Faculty Of Science Kumamoto University
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Ishikura Nariyuki
Department Of Biological Science Faculty Of Science Kumamoto University
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