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Functional characterization, structural basis, and protein engineering of a rare flavonoid 2'-O-glycosyltransferase from Scutellaria baicalensis

文献类型：期刊论文

作者Wang, Zilong8; Du, Xueqing6,7; Ye, Guo8; Wang, Haotian8; Liu, Yizhan8; Liu, Chenrui8; Li, Fudong4,5; Agren, Hans3; Zhou, Yang2; Li, Junhao3
刊名ACTA PHARMACEUTICA SINICA B
出版日期2024-08-01
卷号14期号:8页码:3746-3759
关键词Glycosyltransferase Regio-selectivity Crystal structure De-glycosylation Catalytic mechanisms
ISSN号2211-3835
DOI10.1016/j.apsb.2024.04.001
通讯作者Li, Junhao(junhao.li@physics.uu.se) ; He, Chao(chaohe@ahu.edu.cn) ; Guo, De-An(daguo@simm.ac.cn) ; Ye, Min(yemin@bjmu.edu.cn)
英文摘要Glycosylation is an important post-modification reaction in plant secondary metabolism, and contributes to structural diversity of bioactive natural products. In plants, glycosylation is usually catalyzed by UDP-glycosyltransferases. Flavonoid 2'-O-glycosides are rare glycosides. However, no UGTs have been reported, thus far, to specifically catalyze 2'-O-glycosylation of flavonoids. In this work, UGT71AP2 was identified from the medicinal plant Scutellaria baicalensis as the first flavonoid 2'-O-glycosyltransferase. It could preferentially transfer a glycosyl moiety to 2'-hydroxy of at least nine flavonoids to yield six new compounds. Some of the 2'-O-glycosides showed noticeable inhibitory activities against cyclooxygenase 2. The crystal structure of UGT71AP2 (2.15 A & ring; ) was solved, and mechanisms of its regio-selectivity was interpreted by pKa calculations, molecular docking, MD simulation, MM/GBSA binding free energy, QM/MM, and hydrogen-deuterium exchange mass spectrometry analysis. Through structure-guided rational design, we obtained the L138T/V179D/ M180T mutant with remarkably enhanced regio-selectivity (the ratio of 7-O-glycosylation O-glycosylation byproducts decreased from 48% to 4%) and catalytic efficiency of 2'-O-glycosylation '- O-glycosylation (kcat/Km, k cat / K m , 0.23 L/(s$mmol), 12-fold higher than the native). Moreover, UGT71AP2 also possesses moderate UDP-dependent de- glycosylation activity, and is a dual function glycosyltransferase. This work provides an efficient bio- catalyst and sets a good example for protein engineering to optimize enzyme catalytic features through rational design. 2024 The Authors. Published by Elsevier B.V. on behalf of Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
WOS关键词NATURAL-PRODUCT GLYCOSYLTRANSFERASE ; CRYSTAL-STRUCTURES ; GLYCOSYLATION ; BIOSYNTHESIS ; INSIGHTS ; MODEL ; CONSTITUENTS ; PROMISCUITY ; SPECIFICITY ; GLYCOSIDES
资助项目National Key Research and Development Program of China[2023YFA0914100] ; China National Postdoctoral Program for Innovation Talents[BX20220022] ; National Natural Science Foundation of China[82304326] ; Natural Science Foundation of Anhui Province, China[2008085MC92]
WOS研究方向Pharmacology & Pharmacy
语种英语
WOS记录号WOS:001293132000001
出版者INST MATERIA MEDICA, CHINESE ACAD MEDICAL SCIENCES
源URL[http://119.78.100.183/handle/2S10ELR8/312803]  
专题中国科学院上海药物研究所
通讯作者Li, Junhao; He, Chao; Guo, De-An; Ye, Min
作者单位1.Chinese Acad Sci, Shanghai Res Ctr Modernizat Tradit Chinese Med, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China
2.Jinan Univ, Sch Pharm, Guangzhou 510632, Peoples R China
3.Uppsala Univ, Dept Phys & Astron, SE-75120 Uppsala, Sweden
4.Univ Sci & Technol China, Sch Life Sci, Hefei 230026, Peoples R China
5.Univ Sci & Technol China, Natl Sci Ctr Phys Sci Microscale, Div Mol & Cell Biophys, Hefei 230026, Peoples R China
6.Anhui Univ, Sch Life Sci, Hefei 230601, Peoples R China
7.Anhui Univ, Anhui Key Lab Modern Biomfg, Hefei 230601, Peoples R China
8.Peking Univ, Sch Pharmaceut Sci, State Key Lab Nat & Biomimet Drugs, Beijing 100191, Peoples R China
推荐引用方式
GB/T 7714		 Wang, Zilong,Du, Xueqing,Ye, Guo,et al. Functional characterization, structural basis, and protein engineering of a rare flavonoid 2'-O-glycosyltransferase from Scutellaria baicalensis[J]. ACTA PHARMACEUTICA SINICA B,2024,14(8):3746-3759.
APA Wang, Zilong.,Du, Xueqing.,Ye, Guo.,Wang, Haotian.,Liu, Yizhan.,...&Ye, Min.(2024).Functional characterization, structural basis, and protein engineering of a rare flavonoid 2'-O-glycosyltransferase from Scutellaria baicalensis.ACTA PHARMACEUTICA SINICA B,14(8),3746-3759.
MLA Wang, Zilong,et al."Functional characterization, structural basis, and protein engineering of a rare flavonoid 2'-O-glycosyltransferase from Scutellaria baicalensis".ACTA PHARMACEUTICA SINICA B 14.8(2024):3746-3759.

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来源：上海药物研究所

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