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通过基于β转的氨基酸偏好的多个计算机辅助理性设计来提高碳氧酶A的热稳定性

Yue Ming¹, Haoxiang Zhang¹, Zitong Zhao¹

¹College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.

International journal of biological macromolecules

|June 17, 2023

概括

计算机辅助设计增强了碳氧酶A (CPA) 的热稳定性,这是一种对毒素A (OTA) 降解至关重要的酶. 这种蛋白质工程策略改善了CPA.

科学领域:

生物化学和分子生物学
酶工程是什么? 酶工程是什么?
蛋白质工程是指蛋白质工程.

背景情况:

碳氧酸酶A (CPA) 具有高效的水解能力和毒素A (OTA) 降解活性,使其在食品和生物应用中具有价值.
目前,CPA的低温稳定性限制了其工业用途,特别是在高温酶反应中.

研究的目的:

通过合理的蛋白质设计,增强碳氧酶A (CPA) 的热稳定性.
扩大CPA的工业适用性,特别是用于Ochratoxin A (OTA) 降解.

主要方法:

用分子动力学 (MD) 模拟来预测针对性修改的灵活循环.
三个计算程序 (Rosetta,FoldX,PoPMuSiC) 选了基于氨基酸偏好的变体在β轮.
MD模拟验证了两个变体 (R124K和S134P) 的热稳定性得到改善.

主要成果:

S134P变体在45°C时半衰期增加4.2分钟 (t_1/2),化温度上升1.9°C (T_m).
与野生类型的CPA相比,R124K变种在45°C时表现出7.4分钟长的t_1/2,4.1°C高的半失活温度 (T₅₀¹⁰),以及T_m的1.2°C增加.
结构分析阐明了工程变体中增强的热稳定性背后的机制.

结论:

计算机辅助的理性设计,专注于β转时的氨基酸偏好,有效地提高了CPA的热稳定性.
这一策略增强了CPA在OTA降解中的工业适用性,并为设计菌毒素降解酶提供了有价值的方法.

关键词:

氨基酸偏好氨基酸偏好碳氧胺酶A是一种碳氧酶.理性的设计理性的设计.热稳定性热稳定性

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