Enhancing the endo-activity of the thermophilic chitinase to yield chitooligosaccharides with high degrees of polymerization

文献类型： 外文期刊

第一作者： Zhang, Ruohan

作者： Zhang, Ruohan;Zhang, Honglian;Tu, Tao;Luo, Huiying;Yao, Bin;Tian, Jian;Huang, Huoqing;Guan, Feifei;Tian, Xiaoqian;Zhang, Yan;Wu, Ningfeng;Tian, Jian;Tian, Xiaoqian;Zhang, Yan;Sun, Jilu

作者机构：

关键词： Chitin; Chitinase; Endo-activity; Mutants; Functional oligosaccharides

期刊名称：BIORESOURCES AND BIOPROCESSING （ 影响因子：4.6； 五年影响因子：6.1 ）

ISSN：

年卷期： 2024 年 11 卷 1 期

页码：

收录情况： SCI

摘要： Thermophilic endo-chitinases are essential for production of highly polymerized chitooligosaccharides, which are advantageous for plant immunity, animal nutrition and health. However, thermophilic endo-chitinases are scarce and the transformation from exo- to endo-activity of chitinases is still a challenging problem. In this study, to enhance the endo-activity of the thermophilic chitinase Chi304, we proposed two approaches for rational design based on comprehensive structural and evolutionary analyses. Four effective single-point mutants were identified among 28 designed mutations. The ratio of (GlcNAc)3 to (GlcNAc)2 quantity (DP3/2) in the hydrolysates of the four single-point mutants undertaking colloidal chitin degradation were 1.89, 1.65, 1.24, and 1.38 times that of Chi304, respectively. When combining to double-point mutants, the DP3/2 proportions produced by F79A/W140R, F79A/M264L, F79A/W272R, and M264L/W272R were 2.06, 1.67, 1.82, and 1.86 times that of Chi304 and all four double-point mutants exhibited enhanced endo-activity. When applied to produce chitooligosaccharides (DP >= 3), F79A/W140R accumulated the most (GlcNAc)4, while M264L/W272R was the best to produce (GlcNAc)3, which was 2.28 times that of Chi304. The two mutants had exposed shallower substrate-binding pockets and stronger binding abilities to shape the substrate. Overall, this research offers a practical approach to altering the cutting pattern of a chitinase to generate functional chitooligosaccharides.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序