Structural and biochemical studies of the glycosyltransferase Bs-YjiC from Bacillus subtilis
Structural and biochemical studies of the glycosyltransferase Bs-YjiC from Bacillus subtilis
复制标题
枯草芽孢杆菌糖基转移酶 Bs-YjiC 的结构和生化研究。
DOI：
10.1016/j.ijbiomac.2020.10.238
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发表时间：
2021-01-01
影响因子：
8.2
通讯作者：
Bao, Rui
中科院分区：
文献类型：
--
作者：
Liu, Bing;Zhao, Chang;Bao, Rui
糖基化在天然产物和候选药物的合成中具有重要的生物学和药理学意义。从枯草芽孢杆菌（Bacillus subtilis, Bs-YjiC）中发现的糖基转移酶由于其广泛的底物光谱，在药物开发中具有潜在的应用前景。为了阐明其催化机理，我们对Bs-YjiC的晶体结构进行了求解，结果表明Bs-YjiC采用了典型的GT-B折叠结构，由一个柔性的n域和一个相对刚性的c域组成。结构分析和定点突变研究表明，Ser277位点是二磷酸核苷（NDP）识别的关键位点，而Glu317、Gln318、Ser128和Ser129位点是糖基片段识别的关键位点。我们的研究结果阐明了Bs-YjiC中受体混杂的结构基础，并为Bs-YjiC在工业和制药应用中的进一步蛋白质工程提供了起点。(C) 2020 Elsevier B.V.版权所有
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. (C) 2020 Elsevier B.V. All rights reserved.