Design of mutants for enhanced thermostability of ¦Â-glycosidase BglY from Thermus thermophilus

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• 作者：Zhuo-Lin Yi^a ; ^b ; Shuai-Bing Zhang^a ; ^b ; ^c ; ^{zhangshuaibing1986@yahoo.com.cn} ; Xiao-Qiong Pei^a ; ^b ; Zhong-Liu Wu^a ; ^b ; ^{wuzhl@cib.ac.cn}
• 关键词：¦Â-Glycosidase ; Rational design ; Substrate inhibition ; Thermostability
• 刊名：Bioresource Technology
• 出版年：2013
• 出版时间：February, 2013
• 年：2013
• 卷：129
• 期：Complete
• 页码：629-633
• 全文大小：343 K

文摘

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 ¦Â-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 ¡ãC, and complete lack of substrate inhibition toward the substrate p-nitrophenyl-¦Â-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.