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
作者： Sun, Jingjing 1 ; Wang, Wei 1 ; Ying, Yu 3 ; Hao, Jianhua 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Sustainable Dev Polar Fishery, Minist Agr & Rural Affairs, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Drugs & Bioprod, Qingdao 266071, Peoples R China
3.Qingdao Inst Food & Drug Control, Qingdao 266071, Peoples R China
4.Jiangsu Collaborat Innovat Ctr Exploitat & Utiliz, Lianyungang 222005, Peoples R China
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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