More efficient barley malting under catalyst: Thermostability improvement of a beta-1,3-1,4-glucanase through surface charge engineering with higher activity

文献类型： 外文期刊

第一作者： Zhang, Wenxin

作者： Zhang, Wenxin;Zhang, Yixin;Lu, Yu;Herman, Richard Ansah;Zhang, Sheng;Hu, Yanghao;Zhao, Weiguo;Wang, Jun;You, Shuai;Zhao, Weiguo;Wang, Jun;You, Shuai

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关键词： ?-1; 3-1; 4-Glucanase; Surface charge engineering; Thermostability; Catalytic activity; Molecular dynamics simulation; Malting and brewing

期刊名称：ENZYME AND MICROBIAL TECHNOLOGY （ 影响因子：3.705； 五年影响因子：3.989 ）

ISSN： 0141-0229

年卷期： 2023 年 162 卷

页码：

收录情况： SCI

摘要： beta-1,3-1,4-Glucanase is an indispensable biocatalyst in barley brewing industry for its crucial effect in reducing the viscosity of mash. However, the unsatisfactory thermostability greatly limited its application performance. In this study, structure-based surface charge engineering was conducted aiming at thermostability improvement of BisGlu16B, a highly active beta-1,3-1,4-glucanase from Bispora sp. MEY-1. By applying the enzyme thermal stability system (ETSS), residues D47, D213, and D253 were inferred to be critical sites for thermal properties. Single (D47A, D213A, and D253A) and combination (D47A/D213A/D253A) mutants were generated and compared with BisGlu16B. Among all improved mutants, D47A/D213A/D253A outstanded in thermostability. In com-parison with BisGlu16B, its T50 and Tm were respectively increased by 7.0 degrees C and 4.3 degrees C, while the t1/2 at 70 degrees C was 8.1 times that of the wild type. Furthermore, the catalytic activity of D47A/D213A/D253A also increased by 42.5%, compared with BisGlu16B (42,900 +/- 300 U/mg vs. 30,100 +/- 800 U/mg). Comparing with BisGlu16B and commercial enzyme treatment groups, under simulated malting conditions, efficiency improvement was observed in decreasement of viscosity (35.5% and 90.7%) and filtration time (30.9% and 34.6%) for D47A/ D213A/D253A treatment group. Molecular dynamics simulation showed that mutation sites A47, A213, and A253 increased the protein rigidity by lowering the overall root mean square deviation (RMSD). This study may bring optimization of technology and improvement of producing efficiency to the present brewing industry.

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