工程Pichia pastoris用于高效的De Novo合成2'-Fucosyllactose的工程
Hao Fang1,2,3, Jialun Gao4,5, Liang Yu6
1Center for Future Foods, Muyuan Laboratory, 110 Shangding Road, Zhengzhou 450016, Henan Province, China.
Journal of agricultural and food chemistry
|March 28, 2025
概括
皮奇亚牧草被改造为产生2'-Fucosyllactose (2'-FL),这是一个关键的人类牛奶寡糖化物 (HMO). 代谢工程实现了创纪录的22.35g/L标位,证明了工业HMO生产的潜力.
科学领域:
- 生物技术是生物技术.
- 合成生物学 合成生物学
- 微生物工程 微生物工程
背景情况:
- 2′-糖乳糖 (2′-FL) 是最丰富的母乳寡糖化物 (HMO),对婴儿营养至关重要.
- 皮奇亚牧草是工业生物技术应用的成熟,安全的宿主.
研究的目的:
- 为了设计Pichia pastoris用于2′-FL.的新生合成.
- 通过代谢工程和酶工程优化2'-FL的生产.
主要方法:
- 使用CRISPR-Cas9基因组编辑来构建2'-FL生物合成途径.
- 代谢工程策略增强了前体供应,GTP和NADPH再生.
- 进行了关键酶FucT2的半导体设计.
主要成果:
- 最初的2′-FL产量达到了1.01g/L.
- 工程工作,包括引入SUMO/Ub标签和添加传送器,改进标题.
- 在3升发酵器中进行的最终缩放产生了创纪录的22.35g/L的2'FL.
结论:
- 皮奇亚牧草是一种可行的底盘,用于高效的HMO生产.
- 应用的代谢和酶工程策略显著提高了2′-FL产量.
- 这项工作为2′-FL的工业生物合成建立了一个强大的平台.
相关概念视频
Bioreactor Controls-III
53
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...
53
Upstream Processing
77
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...
77
Production of Organic Acids
72
Lactic acid, an important organic acid extensively applied in food, pharmaceutical, and biodegradable polymer industries, is primarily produced via microbial fermentation. This method is favored over chemical synthesis due to its environmental sustainability and capacity for enantiomerically pure product formation. Among various microbial processes, the fermentation of starch-based substrates stands out due to the abundance and renewability of raw materials like corn and potatoes.Hydrolysis of...
72
Production of Antibiotics
82
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...
82
Production of Pharmaceuticals
64
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...
64


