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通过虚拟和突变发生和合理设计,在工程 d-氨基酸氧化酶中进行优质基质转化

Heng Tang^1,2, Hong-Li Zhu^1,2, Jin-Qiao Zhao^1,2

¹Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, P. R. China.

Biotechnology journal

|July 17, 2024

概括

研究人员增强了d-氨基酸氧化酶 (DAAO) 酶活性,以产生纯的l-glufosinate (l-PPT). 一种新的合理设计程序产生了一种突变物,其酶活性增加了5000%以上,优化了d-glufosinate (d-PPT) 的氧化.

科学领域:

生物催化剂是一种生物催化剂.
酶工程是什么? 酶工程是什么?
蛋白质设计蛋白质设计

背景情况:

D-氨基酸氧化酶 (DAAO) 对于通过d-glufosinate (d-PPT) 氧化合成光学纯的l-glufosinate (l-PPT) 是至关重要的.
提高DAAO的效率和d-PPT的特异性对于工业应用至关重要.

研究的目的:

设计一种Rasamsonia emersonii DAAO (ReDAAO) 变种,其对d-PPT的活性增强.
阐明ReDAAO活动的结构性决定因素,并制定合理的设计策略.

主要方法:

使用Caver 3.0识别基质结合口袋和氨酸扫描以确定关键残留物.
采用虚拟和突变发生法 (VSM) 来预测有益突变和用于酶查的实验验证.
在四轮中代组合了有益的突变,以实现显著的活动增强.

主要成果:

确定了影响ReDAAO活性和基质结合的关键残留物.
VSM和代突变组合导致了一个ReDAAO突变 (N53V/F57Q/V94R/V242R).
与野生类型相比,最终突变的酶活性增加了5097%.

结论:

合理的酶设计,以VSM等计算方法为指导,可以显著提高生物催化剂性能.
突变的代组合是优化酶活性的可行策略.
开发的ReDAAO突变为高效的l-PPT生产提供了一个有前途的工具.

关键词:

艾拉宁扫描可以扫描.d-氨基酸氧化酶的使用.分子动力学模拟模拟合理的设计理性的设计.虚拟和突变的突变发生.

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