在大肠杆菌中工程酸盐生产的理性和半理性方法
Chenghu Chen1, Cong Gao1, Guipeng Hu2
1School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China.
ACS synthetic biology
|October 25, 2024
概括
我们通过优化酸盐合成酶 (Irp9) 和通过排泄蛋白修饰增强细胞耐受性来设计大肠杆菌以生产酸盐. 这导致酸盐的最大产量为3.72g/L,从而提升了微生物细胞工厂的能力.
科学领域:
- 生物技术是生物技术.
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
背景情况:
- 酸盐是一种关键的药物中间体,但微生物的生产受到低酶效率和酸盐毒性的阻碍.
- 现有的方法在大规模,经济高效的酸盐合成中面临挑战.
研究的目的:
- 在大肠杆菌中构建最短的酸盐生物合成途径.
- 提高酸盐合成酶的催化效率和改善微生物酸盐耐受性,以增加产量.
主要方法:
- 引入了酸盐合成酶Irp9进入一种产生l-氨酸的大肠杆菌菌株.
- 采用蛋白质工程来提高Irp9的催化效率.
- 综合适应进化和转录组分析,以了解酸盐耐受机制.
- 基于阐明的机制来增强酸盐生产的修改后的排泄蛋白.
主要成果:
- 在大肠杆菌中实现了最短的酸盐生物合成途径.
- 通过蛋白质工程将Irp9的催化效率提高了33.5%.
- 阐明了排泄蛋白在酸盐耐受性中的作用.
- 在震动瓶培养中达到3.72g/L的最大盐酸盐度.
结论:
- 流蛋白对于提高微生物细胞工厂的酸盐耐受性和生产率至关重要.
- 改造的大肠杆菌菌株和已识别的机制为绿色酸盐合成提供了一个有前途的平台.
- 这种方法对生产其他酸有潜在的应用.
相关概念视频
Bioreactor Controls-III
54
Strain improvement is a foundational strategy in industrial microbiology aimed at maximizing microbial productivity, particularly because natural isolates typically yield commercially valuable products in very low concentrations. Although optimizing the culture medium and environmental conditions can improve yields, these adjustments are inherently limited by the organism’s genetic potential. As a result, the focus shifts toward genetic modifications to enhance biosynthetic capacity. The...
54
Upstream Processing
85
Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
85
Production of Antibiotics
149
Penicillin, one of the earliest and most widely used antibiotics, is produced industrially by the filamentous fungus Penicillium chrysogenum. Large stirred-tank bioreactors ranging from tens to hundreds of thousands of liters maintain tightly controlled temperature, pH, and dissolved oxygen conditions to support fungal metabolism and maximize antibiotic yield. Penicillin is a secondary metabolite, synthesized primarily during the stationary growth phase, which requires a carefully managed...
149
Production of Pharmaceuticals
67
Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under...
67


