热稳定性辅酶-酶,通过蛋白质和发酵工程有效生物合成2-罗利
Bicheng Yu1, Wei Song1, Shenjie Wang1
1School of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, China.
ACS synthetic biology
|April 8, 2025
概括
这项研究设计了一种更具热稳定的酶,EcCaiC,用于2-pyrrolidone生物合成. 改进的酶和工程细菌在生物反应器发酵中实现了高产量,建立了一个可持续的生产平台.
科学领域:
- 生物技术是生物技术.
- 生物化学工程 生物化学工程
- 酶工程是什么? 酶工程是什么?
背景情况:
- 2-罗利是一种关键的工业中间产品,但传统的合成对环境造成了沉重的负担.
- 目前的生物合成途径受到协酶A酶 (CaiC) 温度稳定性较差的限制.
研究的目的:
- 通过蛋白质工程来提高大肠杆菌共酶A联酶 (EcCaiC) 的热稳定性和催化效率.
- 开发一种高效的微生物发酵工艺,用于生产2-pyrrolidone.
主要方法:
- 使用FoldX和Rosetta进行EcCaiC的蛋白质工程,用于虚拟突变发生.
- 在Corynebacterium glutamicum中通过向基因淘汰来表达工程EcCaiC.
- 在5升生物反应器中进行料批发发酵.
主要成果:
- 工程突变M3在55°C的半衰期增加了7.86倍,催化效率提高了52.8%.
- 改造品种的2-pyrrolidone产量为58.28g/L,葡萄糖转化率为0.32g/g.
- 在料批发发酵中达到0.97g/L/h的高生产率.
结论:
- 蛋白质工程成功地提高了EcCaiC的热稳定性和2 - 罗利胺生物合成的效率.
- 建立了一个有效和强大的生物合成平台来生产2-pyrrolidone.
- 这项工作为可持续的工业规模2-pyrrolidone制造提供了基础.
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