苯乙醇苷类化合物因具有抗氧化、抗肿瘤等生理活性在食品、医药等领域备受关注。糖基转移酶作为生物催化剂可用于各种糖苷产物的制备。为丰富苯乙醇苷类化合物的糖苷多样性,在红花钓钟柳转录组中筛选糖基转移酶候选基因并进行功能鉴定。该研究基于不同植物组织中基因差异表达分析筛选到7条候选基因,将其克隆到pET-MBP表达载体上,并导入大肠杆菌Escherichia.coli Transetta (DE3)异源表达后利用体外酶促反应进行其生物催化活性研究。获得一个新型糖基转移酶UGT712A2,可以催化桂叶苷B C4-OH的葡萄糖基化反应生成新的苯乙醇苷类化合物桂叶苷B-4-葡萄糖苷。同时,UGT712A2也可以催化毛蕊花糖苷、芦丁、胡黄连苷Ⅱ、7-去甲基软木花椒素等化合物的糖基化,具有一定的底物宽泛性。因此,UGT712A2可能是一种潜在的酶工具,用于苯乙醇苷类化合物和其他多种新型糖苷产物的制备,从而发现新的糖苷化合物应用于食品、医药领域。
Phenylethanoid glycosides are particularly intriguing due to their physiological properties as an antioxidant and an anticancer.Many glycoside compounds can be prepared using glycosyltransferases as biocatalysts.In this study, UDP glycosyltransferase candidate genes were screened from the plant transcriptome of Penstemon barbatus, and their functions were determined to enrich the glycoside variety of phenylethanoid glycosides.Seven candidate genes were chosen for this study after being analyzed for differential gene expression in various plant tissues.They were then cloned into a pET-MBP expression vector, put into Escherichia coli Transetta (DE3) for heterologous expression, and their biological catalytic activity was tested using in vitro enzyme catalysis.It was discovered that a novel glycosyltransferase, UGT712A2, could catalyze the glucosylation of osmanthuside B C4-OH to form the new phenylethanoid glycoside, osmanthuside B-4-glucoside.UGT712A2 has a substrate breadth and can also catalyze the glycosylation of substances including verbascoside, rutin, picroside II, and 7-demethylsuberosin.In light of this, UGT712A2 may be a useful enzyme for producing phenylethanoid glycosides as well as a number of other novel glycoside products, opening the door to the identification of new glycoside molecules with potential for use in both food and medicine.
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