Cysteine Engineering of an Endo-polygalacturonase from Talaromyces leycettanus JCM 12802 to Improve Its Thermostability

文献类型： 外文期刊

第一作者： Wang, Sheng

作者： Wang, Sheng;Su, Xiaoyun;Wang, Yaru;Zhang, Jie;Luo, Huiying;Yao, Bin;Huang, Huoqing;Tu, Tao;Wang, Sheng;Meng, Kun;Hakulinen, Nina

作者机构：

关键词： endo-polygalacturonase; protein engineering; cysteine; thermostability

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.279； 五年影响因子：5.269 ）

ISSN： 0021-8561

年卷期： 2021 年 69 卷 22 期

页码：

收录情况： SCI

摘要： Thermostable enzymes have many advantages for industrial applications. Therefore, in this study, computer-aided design technology was used to improve the thermostability of a highly active endo-polygalacturonase from Talaromyces leycettanus JCM12802 at an optimal temperature of 70 degrees C. The melting temperature and specific activity of the obtained mutant T316C/G344C were increased by 10 degrees C and 36.5%, respectively, compared with the wild-type enzyme. The crystal structure of the T316C/G344C mutant showed no formation of a disulfide bond between the introduced cysteines, indicating a different mechanism than the conventional mechanism underlying improved enzyme thermostability. The cysteine substitutions directly formed a new alkyl hydrophobic interaction and caused conformational changes in the side chains of the adjacent residues Asn315 and Thr343, which in turn caused a local reconstruction of hydrogen bonds. This method greatly improved the thermostability of the enzyme without affecting its activity; thus, our findings are of great significance for both theoretical research and practical applications.

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