食品与发酵工业2026，Vol.52Issue(6)：262-270,9.DOI:10.13995/j.cnki.11-1802/ts.043413
短管兔耳草葡萄糖基转移酶UGT74DU1的克隆与功能鉴定
Cloning and functional characterization of glucosyltransferase UGT74DU1 from Lagotis brevituba Maxim
摘要
Abstract
Glycosylation plays an important role in the structural diversification of plant natural products.Identifying efficient glycosyl-transferases is also a key step in synthesizing valuable glycosides.This research successfully isolated a glycosyltransferase gene from Lagotis brevituba Maxim,designated as UGT74DU1.UGT74DU1 comprises 1 413 base pairs,encoding a protein of 470 amino acid residues,with a calculated molecular mass of approximately 52.6 kDa.Bioinformatics analysis predicted an isoelectric point of 5.53.Phylogenetic and se-quence alignment analyses revealed that UGT74DU1 shares the closest evolutionary relationship with CrUGT74AN3,despite exhibiting on-ly 46.86％sequence similarity.UGT74DU1 was successfully expressed in Escherichia coli as soluble protein with the help of the chaperone plasmid pGro7.Subsequent functional characterization was performed through in vitro enzymatic assays.Through LC-MS analysis and NMR structural characterization,it was discovered that UGT74DU1 preferentially recognizes flavonoids such as quercetin,kaempferol,rutin,and isoquercitrin,catalyzing glucosylation at the C-7-OH position to produce flavonoid-7-O-glucosides.Furthermore,it could subsequently recognize the C-4'-OH position to generate flavonoid-4',7-O-diglucosides.Moreover,it could catalyze the glycosylation of phenylethanoid glycosides such as verbascoside,demonstrating a certain degree of substrate promiscuity.关键词
短管兔耳草/葡萄糖基转移酶/生物信息学分析/原核表达/功能验证Key words
Lagotis brevituba Maxim/glycosyltransferase/bioinformatics analysis/prokaryotic expression/functional verification引用本文复制引用
李凯薇,唐俊,李佳丽,范晓光,毕慧萍,刘涛..短管兔耳草葡萄糖基转移酶UGT74DU1的克隆与功能鉴定[J].食品与发酵工业,2026,52(6):262-270,9.基金项目
天津市合成生物技术创新能力提升行动项目(TSBICIP-KJGG-034) （TSBICIP-KJGG-034）
