罗塞塔驱动的meso-Diaminopimelate脱酶的快速进化,用于结构多样化的D-氨基酸生产
Hulin Qiu1, Wenzhong Gong1, Huilin Huang1
1School of Marine Sciences, Sun Yat-Sen University, Zhuhai, Guangdong 519080, China.
Journal of agricultural and food chemistry
|June 10, 2025
概括
罗塞塔驱动的进化快速增强D-氨基酸合成的中氨基酸脱酶 (DAPDH). 改造的酶表现出更好的活性和基质范围,使大容量D-氨基酸的有效生产成为可能.
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:
- 中氨基基酸脱酶 (DAPDH) 是对D-氨基酸合成有前途的.
- 目前的DAPDH表现出低的催化效率与非自然基质.
研究的目的:
- 开发一种快速有效的调整DAPDH基质特异性的方法.
- 设计具有对非自然基质增强活性的DAPDH变体.
主要方法:
- 使用罗塞塔驱动的进化框架.
- 构建了一个智能小突变图书馆,用于指导进化.
- 采用最小的选 (100-1,000变换剂) 在一到两轮.
主要成果:
- 增强DAPDH活性向甲酸 (1b) 增加至29.5单位/毫克和3-甲酸 (10b) 增加至5.1单位/毫克.
- 与之前报告的最佳突变相比,对1b的活性达到近10倍.
- 证明了在制剂尺度上大量D-氨基酸的高效合成,达到0.5M度,时空产量为128.46g/L/day.
结论:
- 基于罗塞塔的进化是加速酶工程的强大策略.
- 开发的框架为优化DAPDH和其他酶提供了一个多功能平台.
- 工程 DAPDH 能够有效地生产各种 D-氨基酸.
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