在大肠杆菌中设计一种非氧化糖解途径,用于高水平的酸盐生产
Tingting Wang1, Lijuan Ding1,2, Huiying Luo1
1State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No.2 Yuanmingyuan West Road, Haidian district, Beijing, 100193, China.
Microbial cell factories
|August 22, 2024
概括
工程化大肠杆菌产生了110.2g/L的甲酸盐,甲基甲酸盐 (MMA) 的前体,使用一种抗菌株. 这种代谢工程的进步为MMA生产提供了一个可持续的替代方案.
科学领域:
- 生物技术是生物技术.
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
背景情况:
- 甲基甲酸盐 (MMA) 是一个关键的工业化学品,但传统的生产方法会给环境和健康带来风险.
- 甲酸是MMA生产的有前途的生物基前体,Escherichia coli被确定为高位的生产者.
- 大肠杆菌发酵的挑战包括有限的代谢工程研究和持续的菌体污染.
研究的目的:
- 从葡萄糖中改造一种耐菌体的大肠杆菌菌株,以提高从葡萄糖中产生的酸盐的产量.
- 为了优化酸盐生物合成途径并改善葡萄糖利用.
- 为可持续的MMA前体合成建立一个强大的微生物平台.
主要方法:
- 在BBa_J23100促进体下过度表达一个突变的Methanococcus jannaschii cimA基因 (MjcimA3.7),以建立甲酸盐路径.
- Ssp防御系统的基因组集成和菌体感染周期组件的突变,以赋予菌体耐药性.
- 工程非氧化糖解路径和去除乙酸合成以提高乙-CoA供应.
主要成果:
- 在大肠杆菌中构建了一个有效的酸生物合成途径.
- 成功开发了一种对菌体有耐药性的大肠杆菌菌株.
- 经过80个小时的食批发发酵后,该工程菌株获得了最高报告的甲酸标位110.2g/L.
- 从葡萄糖和生产力中获得的胺酸产量分别达到0.4 g/g和1.4 g/L·h.
结论:
- 开发的大肠杆菌菌株显示了迄今为止在这个宿主中报告的最高甲酸标位和生产率.
- 这种菌株为工业酸盐和MMA衍生品生产提供了具有成本效益和高效的平台.
- 采用的代谢工程策略突出了生物基化学制造业进一步进步的潜力.
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