在Pseudomonas putida的工程菌株中通过增加马洛尼尔-CoA可用性来增强聚3-基酸盐的生物合成
Giusi Favoino1, Nicolas Krink1, Tobias Schwanemann2
1The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Lyngby, Denmark.
Microbial biotechnology
|November 6, 2024
概括
微生物细胞工厂可以使用malonyl-coenzyme A (CoA) 产生有价值的化合物. 研究人员对Pseudomonas putida进行了改造,以增加马洛尼尔-CoA水平,从而增强聚3-基酸盐的产生.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 微生物生物技术 微生物生物技术
背景情况:
- 马洛尼尔辅酶A (CoA) 是一种重要的代谢中间体,用于在微生物系统中合成各种化合物.
- 由于代谢竞争,马洛尼尔-CoA的有限可用性阻碍了微生物细胞工厂的效率.
- 伪虫 (Pseudomonas putida) 是一种生产附加值化学品的多功能宿主.
研究的目的:
- 为了设计一种Pseudomonas putida菌株,以增加细胞内马洛尼尔-CoA水平.
- 通过优化其生物合成和限制其消费来克服马洛尼尔-CoA稀缺性.
- 验证增强的马洛尼尔-CoA池,用于生产依赖马洛尼尔-CoA的生物制品.
主要方法:
- 减少基因组的Pseudomonas putida菌株的遗传修饰,涉及糖代谢,TCA循环和脂肪酸生物合成途径中的淘汰.
- 开发和应用一种酶合生物传感器 (基于RppA) 用于通过flaviolin生产在体内监测马洛尼尔-CoA度.
- 颜色测量选具有高马洛尼尔-CoA的菌株,并随后量化flaviolin.
主要成果:
- 工程菌株显示了显著增加的马洛尼尔-CoA度,这通过flaviolin量化证实了这一点.
- 通过向基因淘汰,四种工程菌株表现出增强的马洛尼尔-CoA可用性.
- 引入一种非正规的聚3-基酸生物合成途径导致了聚合物积累的增加.
结论:
- 该研究通过代谢工程成功提高了Pseudomonas putida中的马洛尼尔-CoA可用性.
- 基于RppA的生物传感器提供了一种有效的选方法,用于选过度生产马洛尼尔-CoA的菌株.
- 该策略验证了在工程微生物中提高各种马洛尼尔-CoA衍生化合物的产生潜力.
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