热稳定性
合理设计
突变体
地衣芽孢杆菌
淀粉酶
工业与生产工程
蛋白质工程
化学
比活度
酶
生物化学
生物
遗传学
基因
细菌
电气工程
工程类
枯草芽孢杆菌
作者
Xin Cui,Xin Yuan,Shunyi Li,Xinlin Hu,Jing Zhao,Guimin Zhang
标识
DOI:10.1007/s00449-022-02790-0
摘要
Higher activity and alkaline α-amylases are desired for textile desizing and detergent additive. Here, rational design was used to improve the specific activity and thermostability of the α-amylase BLA from Bacillus licheniformis. Seventeen mutants of BLA were designed based on sequence consensus analysis and folding free energy calculation, and characterized by measuring their respective activity and thermostability at pH 8.5. Among them, mutant Q360C exhibited nearly threefold improved activity than that of wild-type and retained a higher residual activity (75% vs 59% for wild-type) after preincubation at 70 ℃ for 30 min. The modeled structures and molecular dynamics simulations analysis demonstrated that the enhanced hydrophobic interaction near residue 360 and reduced disturbance to the conformation of catalytic residues are the possible reasons for the improved thermostability and activity of Q360C. The results suggest that 360th of BLA may act as a hotspot for engineering other enzymes in the GH13 superfamily.
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