UDP-D-Glucose: Flavonol 3 -0 -and 7-O-Glucosyl Transferases from Young Leaves of Paederia scandens var. mairei

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Abstract
Two different O-glucosyl transferases (F3GT and F7GT ) catalyzing the transfer of ᴅ-glucose from UDP-ᴅ-glucose to the 3 and 7 positions of flavonol, respectively, were isolated from the young leaves of Paederia scandens var. mairei. F3GT and F7GT, which were recovered in about a 1:1 ratio in the activity, were purified by about 140- and 136-fold, respectively, by precipitation with ammonium sulfate followed by ion exchange chromatography and chromatofocusing. F3GT and F7GT both had a pH optimum of 7.5 in Tris-HCl buffer, and an Mr of 43 kDa. Neither F3GT nor F7GT had a Mg2+ requirement. Both were inhibited by each 1 mᴍ of Zn2+, Cu2+, N-ethylmaleimide and p-chloromercuribenzoate, and both were stimulated by 14 mᴍ 2-M E . F3GT and F7GT were different from each other in having an isoelectric point (pI) at pH 5.12 and 4.50, respectively. F3GT mediated the transfer of D-glucose exclusively to the 3-hydroxyl group of kaempferol and some flavonols, but neither the 7-O-glucosides nor the 3-O-glucosides of their flavonols were able to accept D-glucose. On the other hand, F7GT mediated the transfer of D-glucose exclusively to the 7-hydroxyl group of kaempferol and some flavonols, and in addition, the 3-O -glucosides of kaempferol and quercetin were able to accept D-glucose thoughn less efficiently. Consequently, the possibility of sequential steps of 3-O- and then 7-O-glucosylations of flavonols to give the 3,7-di-O-glucoside was discussed.

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