Modification of glycosyltransferase SrUGT76G1 based on computer-aided screening of mutation sites for efficient synthesis of rebaudioside M

文献类型： 外文期刊

第一作者： Zhao, Liang

作者： Zhao, Liang;Liu, Yaqi;Yang, Ting;Yang, Zichen;Zhao, Lei;Wang, Chengtao;Zhao, Liang;Liu, Yaqi;Yang, Ting;Yang, Zichen;Zhao, Lei;Wang, Chengtao;Zhao, Liang;Liu, Yaqi;Zhao, Lei;Li, Yingnan;Pan, Fei

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关键词： Stevia glycosides; Rebaudioside M; Glycosyltransferase; Molecular dynamics; Modification

期刊名称：FOOD BIOSCIENCE （ 影响因子：4.8； 五年影响因子：5.1 ）

ISSN： 2212-4292

年卷期： 2024 年 61 卷

页码：

收录情况： SCI

摘要： Steviol glycosides (SGs) from Stevia rebaudiana Bertoni is a widely used sweetener with the advantage of high sweet and low in calories. Stevioside (ST) and Rebaudioside A (Reb A) with a bitter aftertaste are the main sweetening components in SGs. Reb M is a highly potency sweetener with almost no bitter aftertaste but is found in low content in S . rebaudiana Bertoni. In this study, the pre- and post-catalytic differences of stevioside glycosyltransferase SrUGT76G1 were investigated through long-time molecular dynamics simulations. SrUGT76G1 was mutated to enhance its efficiency in catalyzing the synthesis of Reb D to Reb M by identifying the enzyme mutation site through an integrated three-in-one approach that combines hinge region analysis, coevolutionary conservation, and energy calculations. The k cat and k cat /K m values of the M88V mutant with Reb D as substrate were 2.18-fold and 4.85-fold higher than those of wild-type SrUGT76G1, respectively. In addition, the M88V mutant coupled with sucrose synthase StSUS1 increased the yield of Reb M synthesis from Reb D to 92.40%. The present study provides an effective method for rapid search for mutation sites in the enzyme and for increasing the of Reb M.

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