结构分析和可变性景观引导的 ω-转氨酶工程,以提高催化性能
Puhong Yi1,2,3, Yue Xu1,2,3, Yuhua Hao1,2,3
1The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou 310014, P. R. China.
Journal of agricultural and food chemistry
|April 17, 2025
概括
ω-转氨酶 (ω-TAs) 是性氨基合成的关键生物催化剂. 通过远端突变I22D对Salmonella enterica ω-TA (SeTA) 进行工程,显著提高了其生产除草剂l-phosphinothricin (l-PPT) 的活性和稳定性.
科学领域:
- 生物催化剂是一种生物催化剂.
- 酵素工程是什么? 酶工程是什么
- 有机合成 有机合成
背景情况:
- ω-转氨酶 (ω-TAs) 对于性氨基的不对称合成至关重要的生物催化剂.
- 沙门氏体 ω-TA (SeTA) 是一种有前途的酶,用于生产除草剂l-phosphinothricin (l-PPT).
- 提高SeTA的催化活性和稳定性对于其工业应用至关重要.
研究的目的:
- 增强SeTA的催化活性和稳定性.
- 研究改善酶性能的结构和机制基础.
- 为其他 ω-TAs 的工程提供洞察力.
主要方法:
- 结构分析和SeTA的可变性景观建设.
- 局部定向突变发生,以产生远程突变.
- 酶活性测定和热稳定性测量 (半衰期,Tm).
- 分子动力学 (MD) 模拟以阐明全效应.
主要成果:
- 一个灵活的循环1区域被确定为催化活动的重要区域.
- 远端突变I22D (M1) 的特定活性增加了3.24倍.
- M1显著提高了热稳定性,在45°C和55°C的半衰期更长,化温度 (Tm) 更高.
- 医学模拟显示,I22D突变诱导了长距离的全效应,改变了活性部位口袋和基质道.
结论:
- 远端突变可以通过全osteric机制有效地提高 ω-TA 性能.
- 改造的SeTA突变体显示了提高的催化效率和稳定性.
- 这项研究为工业应用的 ω-TAs 的合理设计提供了宝贵的见解.
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