期刊
BIORESOURCE TECHNOLOGY
卷 129, 期 -, 页码 629-633出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2012.12.098
关键词
beta-Glycosidase; Rational design; Substrate inhibition; Thermostability
资金
- National Natural Science Foundation of China [21072183]
- Province Science Foundation of Sichuan, China [2010SZ0128]
- Open Fund of Key Laboratory of Environmental and Applied Microbiology [Y1C5101106]
- West Light Foundation of the Chinese Academy of Sciences
Three design strategies, based on rational and semi-rational approaches, were employed to investigate the functional impact of thermostability-related amino acid substitutions in the beta-glycosidase BglY from Thermus thermophilus. Five beneficial mutations were identified, of which 1 mutation was located in the active cavity of the enzyme and contributed to the released substrate inhibition. Combining all 5 beneficial substitutions resulted in the mutant HF5 with a 4.7-fold increase in half-life, with thermal inactivation at 93 degrees C, and complete lack of substrate inhibition toward the substrate p-nitrophenyl-beta-D-glucopyranoside at lower reaction temperatures. The results of this study provide valuable information on amino acid substitutions related to thermostability and substrate inhibition of BglY. (C) 2012 Elsevier Ltd. All rights reserved.
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