title: Design of mutants for enhanced thermostability of β-glycosidase BglY from Thermus thermophilus.
abstract: 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 β-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 °C, and complete lack of substrate inhibition toward the substrate p-nitrophenyl-β-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.
authors: Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China.
authors: Zhuo-Lin
authors: ZL
authors: Yi
authors: Shuai-Bing
authors: SB
authors: Zhang
authors: Xiao-Qiong
authors: XQ
authors: Pei
authors: Zhong-Liu
authors: ZL
authors: Wu
mesh_terms: D004789:Enzyme Activation; D004795:Enzyme Stability; D024861:Genetic Enhancement / Q000379:methods*; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D013696:Temperature; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics*; D001617:beta-Glucosidase / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics
chemical_list: D001617:beta-Glucosidase