通过局部定向突变发生在水友醇中的乙基兰雌激酶的增强活性和稳定性
Henry Madubuike1, Natalie Ferry1
1School of Science Engineering and Environment, University of Salford, Manchester M5 4WT, UK.
Molecules (Basel, Switzerland)
|November 14, 2023
概括
改造的乙基兰化酶 (BaAXE) 在有机溶剂中显示出增强的性能和稳定性. 引入带电残留的合理设计改善了酶活性和可用于可持续生物催化剂的工业应用.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 工业应用中的蛋白质工程.
- 可持续的化学可持续的化学
背景情况:
- 对环保合成的日益增长的需求需要具有高工业性能的生物催化剂.
- 目前的生物催化剂在工业条件下通常表现出有限的性能.
- 酶工程对于开发创新和可持续的生物催化工艺至关重要.
研究的目的:
- 为了提高乙西兰酶 (BaAXE) 在水友性有机溶剂中的性能.
- 研究合理设计,特别是引入带电残留物对酶稳定性和活性的影响.
- 为更广泛的工业生物催化剂应用增强BaAXE.
主要方法:
- 合理的设计方法包括将BaAXE结构分为基板结合和影响溶剂的区域.
- 在特定的溶剂影响区域 (A12D,Q143E) 进行局部定向的突变发生,以产生突变MUT12,MUT143和MUT12-143.3.
- 在不同酒精度下评估酶动力学 (Km) 和性能.
- 计算分析以了解结构-功能关系.
主要成果:
- 与野生类型BaAXE相比,突变MUT12和MUT143的Km值明显较低.
- 改造突变体在低度的乙醇和甲醇中表现出更好的活性和稳定性.
- 与MUT143.3不同的是,MUT12和MUT12-143在40%的甲醇中保持了相当大的活性 (>70%),而MUT143.
- 计算分析揭示了高性能突变物中的新型分子间相互作用.
结论:
- 引入充电残留物的合理设计有效地提高了有机溶剂中的酶性能.
- 工程 BaAXE 变种显示出对工业生物催化剂的承诺,提供更好的稳定性和活性.
- 这项研究强调了蛋白质表面修饰对于酶适应非原生环境的重要性.
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