Mining and Engineering the Di- O -glycosylation Pattern of UGT72B1 for the Highly Efficient O -Glycosylation of Endogenous Quercetin

作者：Zhi Geng, Ting Zhao, Ke Li, Li-Ling Liang, Ming-Xuan Chen, Zhijing Zhou, Jun Dai, Zongjie Dai, Kai‐Zhi Jia · 发表于：Journal of Agricultural and Food Chemistry · 年份：2024 · DOI：10.1021/acs.jafc.4c08469 · 被引用次数：8 · 研究领域：Plant biochemistry and biosynthesis、Microbial Metabolic Engineering and Bioproduction、Microbial Metabolites in Food Biotechnology

Compared with mono- O -glycosylation, di- O -glycosylation endows the precursor with better performance. However, the mining and engineering of di- O -glycosylation patterns of glycosyltransferases are limited, hindering their synthetic applications. Here, an Arabidopsis xenobiotic-transforming glycosyltransferase, UGT72B1, was found to catalyze the glycosylation of endogenous quercetin and its monoglycosides, generating di- O- glucosides. Mutating M17/G18/Y315 into L/T/Q in UGT72B1 altered its regioselectivity toward quercetin 7- O -glucoside, enzymatically generating another 3,7-di- O -glycoside with up to a 100% conversion rate, and increased the sugar donor preference. Altering the regiospecificity of glycosyltransferases likely required coordination between the entrance and the active site, where the orientations of the sugar acceptors and donors shift to adopt a lower binding energy state. Moreover, quercetin 3,4′-di- O -β- d -glucoside and quercetin 3,7-di- O -β- d -glucoside synthesized were found to have the highest anti-inflammatory activities. Overall, this work presents an efficient strategy to engineer glycosylation patterns for the synthesis of quercetin di- O -β- d -glucosides to be used as food additives, therapeutics, and nutraceuticals.