酵母的人性化生产复杂的终端化糖蛋白质
Stephen R Hamilton1, Robert C Davidson, Natarajan Sethuraman
1GlycoFi Inc., 21 Lafayette Street, Suite 200, Lebanon, NH 03766, USA.
概括
工程酵母Pichia pastoris生产人类糖蛋白与复杂的,化N-糖. 酵母糖化中的这一进步使得能够产生功能性重组蛋白质,如红蛋白.
科学领域:
- 生物技术是生物技术.
- 葡萄糖生物学 葡萄糖生物学
- 分子生物学分子生物学
背景情况:
- 皮奇亚牧草是一种用于重组蛋白质生产的常见系统.
- 原生酵母糖化与人类糖化不同,限制了治疗蛋白质的应用.
研究的目的:
- 为了设计Pichia pastoris用于生产类似人类的N-glycans.
- 在分泌的葡萄糖蛋白中达到高水平的终端化.
主要方法:
- 四个内源性酵母糖化基因的淘汰.
- 引入14个异质基因来模仿人类的N-糖基化途径.
- 培养工程酵母菌株用于糖蛋白生产.
主要成果:
- 成功消除了酵母特异性的糖化.
- 工程细胞系在复杂的葡萄糖蛋白上实现了>90%的终端化.
- 通过产生功能性重组性红色素来证明其实用性.
结论:
- 工程设计的Pichia pastoris系统有效地复制了人类的N-糖化.
- 该平台适用于生产复杂的,化糖蛋白,用于治疗应用.
相关概念视频
Oligosaccharide Assembly
Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...
Multiple sugar molecules that may or may...
Microbes in Food Production
Microbial fermentation is central to food biotechnology, enhancing flavor, texture, preservation, and stability. Fermentative microorganisms metabolize carbohydrates into organic acids, alcohols, and other metabolites that inhibit spoilage organisms and improve digestibility while contributing distinctive sensory qualities.In baking, amylases naturally present in flour hydrolyze starch into monosaccharides such as glucose, which Saccharomyces cerevisiae ferments anaerobically. Through...
Bioreactor Controls-III
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...
Production of Alcohol
Continuous fermentation is a key strategy in industrial ethanol production, particularly when efficiency, scalability, and high yields are essential. This approach allows for uninterrupted operation and optimized resource utilization. The primary feedstock, corn starch, undergoes enzymatic hydrolysis facilitated by α-amylase and glucoamylase. These enzymes break down the starch into fermentable sugars such as glucose, which are readily assimilated by fermentative microorganisms.Fermentation...
Production of Organic Acids
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...
Production of Pharmaceuticals
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 sterile, tightly...


