期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 66, 期 50, 页码 13228-13235出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.8b05662
关键词
beta-glucosidase; thermostability; glucose tolerance; random mutagenesis; high-throughput screening; cellulase
资金
- National Natural Science Foundation of China [31770075, 31170117, 31870782]
- Science & Technology Projects of Guangzhou [201804010285]
- China Postdoctoral Science Foundation [2017M622859]
A high-performance beta-glucosidase for efficient cellulose hydrolysis needs to excel in thermostability, catalytic efficiency, and resistance to glucose inhibition. However, it is challenging to achieve superb properties in all three aspects in a single enzyme. In this study, a hyperactive and glucose-tolerant beta-glucosidase Ks5A7 was employed as the starting point. Four rounds of random mutagenesis were then performed, giving rise to a thermostable mutant 4R1 with five amino acid substitutions. The half-life of 4R1 at 50 degrees C is 8640-fold that of Ks5A7 (144 h vs 1 min). Meanwhile, 4R1 had a higher specific activity (374.26 vs 243.18 units.mg(-1)) than the wild type with a similar glucose tolerance. When supplemented to Celluclast 1.5L, the mutant significantly enhanced the hydrolysis of pretreated sugar cane bagasse, improving the released glucose concentration by 44%. With excellent performance in thermostability, activity, and glucose tolerance, 4R1 will serve as an exceptional catalyst for industrial applications.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据