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结构引导的代变异驱动α-1,3-基转移酶的双功能演变

Mengli Li¹, Ruoyu Jia¹, Jiawei Yao¹

¹State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi, Jiangsu 214122, China.

Journal of agricultural and food chemistry

|September 3, 2025

概括

研究人员使用计算设计和组合突变增强了α-1,3-Fucyltransferase (α-1,3-FucT) 酶活性和稳定性. 这提高了它在合成人乳寡糖3-糖 (3-FL) 的工业应用.

科学领域:

酶工程
生物催化
葡萄糖生物学

背景情况:

阿尔法-1,3-Fucyl转移酶 (α-1,3-FucT) 对于合成3 - fucosyllactose (3-FL),一种人乳寡糖是至关重要的.
α-1,3-FucT的工业应用受低活性和低热稳定性限制.

研究的目的:

为了提高α-1,3-FucT的催化性能和热稳定性.
开发一种有效的酶合理设计和工业酶工程策略.

主要方法:

用计算机辅助设计和贪的组合策略来优化酶.
四个计算工具确定了潜在的突变部位,随后进行了图书馆选.
一步一步的组合方法产生了有益的单个和四重突变.

主要成果:

六种单一突变显示出28%至84%更高的特异活性和更好的热稳定性.
一种四重突变 (K132I-E134M-N199F-K301P) 的酶活性增加了116%并延长了半衰期.
突变优化了基质结合和结构稳定性,通过重塑疏水性和结网络.

结论:

这项研究提出了使用组合突变进行合理酶设计的有效策略.
优化的α-1,3-FucT显示了显著的改进,为工业应用铺平了道路.
通过组合突变进行酶工程对于工业生物催化非常重要.

关键词:

催化作用分子修饰合理的设计热稳定性 α-1,3-基转移酶

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相关概念视频

In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.