A Novel Glucose-Tolerant GH1 beta-Glucosidase and Improvement of Its Glucose updates Tolerance Using Site-Directed Mutation

文献类型： 外文期刊

第一作者： Sun, Jingjing

作者： Sun, Jingjing;Wang, Wei;Hao, Jianhua;Sun, Jingjing;Wang, Wei;Hao, Jianhua;Ying, Yu;Hao, Jianhua

作者机构：

关键词： Marine bacterium; beta-Glucosidase; Glucose tolerance; Site-directed mutation; Ethanol tolerance

期刊名称：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY （ 影响因子：2.926； 五年影响因子：2.685 ）

ISSN： 0273-2289

年卷期： 2020 年 192 卷 3 期

页码：

收录情况： SCI

摘要： A novel GH1 beta-glucosidase gene (bgla) from marine bacterium was sequenced and expressed in Escherichia coli. After purification by Ni2+ affinity chromatography, the recombinant protein was characterized. The purified recombinant enzyme showed maximum activity at 40 degrees C, pH 7.5 and was stable between temperatures that range from 4 to 30 degrees C and over the pH range of 6-10. The enzyme displayed a high tolerance to glucose and maximum stimulation at the presence of 100 mM glucose. To improve glucose tolerance of the enzyme, a site-directed mutation (f171w) was introduced into beta-glucosidase. The recombinant F171W showed a higher glucose tolerance than the wild type and maintained more than 40% residual activity at the presence of 4 M glucose. Additionally, the recombinant enzymes showed notable tolerance to ethanol. These properties suggest the enzymes may have potential applications for the fermentation of lignocellulosic sugars and the production of biofuels.

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