Efficient Synthesis of Glycodiversified Nucleoside Analogues by a Thermophilic Promiscuous Glycosyltransferase

作     者：Wang, Zhuqing Li, Jiahui Wang, Xueyun Jin, Boyu Zhou, Liwei Zhao, Zili Gu, Meijia Song, Xuemin Wang, Jiahong Deng, Zixin Wu, Shuwen Zhang, Zhengyu Chen, Wenqing 

作者机构：Wuhan Univ Zhongnan Hosp TaiKang Ctr Life & Med Sci Sch Pharmaceut SciDept Anesthesiol Wuhan 430071 Peoples R China Wuhan Univ Key Lab Combinatorial Biosynth & Drug Discovery Minist Educ Wuhan 430071 Peoples R China Wuhan Univ Coll Life Sci State Key Lab Virol Wuhan 430072 Peoples R China Nanjing Forestry Univ Int Innovat Ctr Forest Chem & Mat Nanjing 210037 Peoples R China 

出 版 物：《ACS CATALYSIS》 (ACS Catal.)

年 卷 期：2025年第15卷第2期

页      面：1217-1229页

核心收录：

学科分类：07[理学] 0703[理学-化学] 

基　　金：Hubei Provincial Department of Education [2021YFC2100600] National Key R&D Program of China [32170026, 32470047, 32371303] National Natural Science Foundation of China Hubei Province's Outstanding Medical Academic Leader Program (Chutian Talent Plan) 

主　　题：glycosyltransferase promiscuity purine nucleosideanalogues 3 '-O-beta-glucosylribavirin biocatalysis 

摘      要：3 -O-beta-Glucosyl purine-related nucleosides are actinobacterial natural products with intricate structures, in which the glucosyl attachment to a nucleoside scaffold is governed by a glycosyltransferase. However, the molecular logic and engineered application of the glycosyltransferase have nearly remained unexplored. Here, we report the discovery, characterization, and exploitation of the thermophilic glycosyltransferase ScaGT. We uncover that ScaGT and its homologue AvpGT indicate prominent promiscuity against both sugar donors and a variety of nucleosides. Remarkably, we have solved the ternary complex structure of AvpGT, unveiling that it employs an unpreceded twin-tyrosine gate mechanism for substrate recognition and promiscuity, and we have also realized directed biosynthesis of diversified purine nucleoside analogues with unexpectedly enhanced titer via introduction of the external scaGT or avpGT. Moreover, we reveal that 3 -O-beta-glucosyl ribavirin exhibits significantly enhanced antiviral activities, thereof showing its promising application potentials. Finally, we have further achieved the gram-scale production of 3 -O-beta-glucosyl ribavirin by both biocatalytic and fermentation-feeding strategies. These findings expand the biochemical repertoire regarding glycosyltransferase reactions and provide the basis for rapid mining and rational engineering of more related glycosyltransferases toward synthetic biology applications.