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结构导向生物工程冷适应的Pseudomonas sp.的lid1的结构导向生物工程增强TB11酶的催化能力,以提高催化能力

Linlin Sha¹, Wen-Sen He², Tian Zheng³

¹College of Food and Health, Zhejiang Agriculture and Forest University, Hangzhou 311300, China; Zhejiang Provincial Key Laboratory of Characteristic Traditional Chinese Medicine Resource Protection and Innovative Utilization, Zhejiang Agriculture and Forest University, Hangzhou 311300, China.

International journal of biological macromolecules

|November 22, 2023

概括

使用结构导向方法对冷适应性酶 (CPE) 进行生物工程,提高了其水解活性和基质亲和力. 这种优化可以提高生物技术应用中的酶性能.

科学领域:

酶学是一种酶学.
蛋白质工程是指蛋白质工程.
生物催化剂是一种生物催化剂.

背景情况:

酶优化对于生物催化剂至关重要.
结构引导生物工程提供了一种合理的方法来增强酶特性.
适应寒冷的酶为工业应用带来了独特的挑战和机会.

研究的目的:

来自Pseudomonas sp.的冷适应性雌激酶 (CPE) 的生物工程. 为了提高催化性能.
调查lid1地区在CPE活动和稳定中的作用.
为了确定增强水解和基质亲和力的特定氨基酸替代物.

主要方法:

生物信息分析以指导lid1区域的蛋白质工程.
位点定向的突变发生,重点关注未保存的残留物Q55.
对短链和长链基质的酶活性测定.
热稳定性和运动分析 (基质亲和性).
在食品风味成分丰富中进行应用测试.

主要成果:

用变体 (Q55V,Q55N) 替换Q55显著增加了水解活性 (短链基质的2.0倍,长链基质的6.5倍),而不会损害热稳定性.
变种显示了Ca2+和有机溶剂的增强激活.
与Thermomyces lanuginosus lipase交换盖子降低了CPE的活性和稳定性.
动力学研究表明,对Q55变体 (Y,V,N,R) 的基质亲和力得到改善.
Q55V变种在单一奶油水解中有效丰富了风味成分.

结论:

Lid1区域的结构引导生物工程,特别是Q55替代,是提高冷适应酶性能的有效策略.
优化的CPE变种表现出卓越的水解活性,基质亲和力和稳定性,使其适合生物技术应用.
这项工作为为特定的工业需求量身定制冷适应酶提供了基础,例如在食品风味开发中.

关键词:

生物工程是生物工程.催化能力的催化能力.适应寒冷的雌激酶可以适应寒冷.

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