Structural and biochemical studies of the glycosyltransferase Bs-YjiC from Bacillus subtilis

Bing Liu; Chang Zhao; Qianyin Xiang; Ninglin Zhao; Yunzi Luo; Rui Bao

doi:10.1016/j.ijbiomac.2020.10.238

Structural and biochemical studies of the glycosyltransferase Bs-YjiC from Bacillus subtilis

Int J Biol Macromol. 2021 Jan 1:166:806-817. doi: 10.1016/j.ijbiomac.2020.10.238. Epub 2020 Nov 2.

Authors

Bing Liu¹, Chang Zhao², Qianyin Xiang¹, Ninglin Zhao², Yunzi Luo³, Rui Bao⁴

Affiliations

¹ Department of Gastroenterology, West China Hospital, Sichuan University and Collaborative Innovation Center of Biotherapy, Chengdu 610041, China.
² Center of Infectious Diseases, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Center of Biotherapy, Chengdu 610041, China.
³ Department of Gastroenterology, West China Hospital, Sichuan University and Collaborative Innovation Center of Biotherapy, Chengdu 610041, China; Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China. Electronic address: yunzi.luo@tju.edu.cn.
⁴ Center of Infectious Diseases, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Center of Biotherapy, Chengdu 610041, China. Electronic address: baorui@scu.edu.cn.

PMID: 33152360
DOI: 10.1016/j.ijbiomac.2020.10.238

Abstract

Glycosylation possess prominent biological and pharmacological significance in natural product and drug candidate synthesis. The glycosyltransferase YjiC, discovered from Bacillus subtilis (Bs-YjiC), shows potential applications in drug development due to its wide substrate spectrums. In order to elucidate its catalytic mechanism, we solved the crystal structure of Bs-YjiC, demonstrating that Bs-YjiC adopts a typical GT-B fold consisting of a flexible N-domain and a relatively rigid C-domain. Structural analysis coupled with site-directed mutagenesis studies revealed that site Ser277 was critical for Nucleoside Diphosphate (NDP) recognition, while Glu317, Gln318, Ser128 and Ser129 were crucial for glycosyl moiety recognition. Our results illustrate the structural basis for acceptor promiscuity in Bs-YjiC and provide a starting point for further protein engineering of Bs-YjiC in industrial and pharmaceutical applications.

Keywords: Catalytic mechanism; Crystal structure; Glycosyltransferase; Nucleoside Diphosphate.

MeSH terms

Bacillus subtilis / enzymology*
Bacterial Proteins / chemistry*
Bacterial Proteins / metabolism
Catalytic Domain
Glycosyltransferases / chemistry*
Glycosyltransferases / metabolism
Protein Binding
Substrate Specificity

Substances

Bacterial Proteins
Glycosyltransferases