在接近零的生长速度下,重组科马加泰拉 phaffii的蛋白质生产动态和生理适应
Corinna Rebnegger1,2,3, Benjamin L Coltman1,2, Viktoria Kowarz1,2
1CD-Laboratory for Growth-Decoupled Protein Production in Yeast at Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria.
Microbial cell factories
|February 8, 2024
概括
酵母可以生产蛋白质,即使在极其缓慢的生长速度. 这项研究使用了保留剂培养,以了解重组科马加塔 phaffii如何适应接近零增长,揭示了将增长与蛋白质生产脱的战略.
科学领域:
- 生物技术是生物技术.
- 微生物生理学 微生物生理学
- 合成生物学 合成生物学
背景情况:
- 酵母的特定生产率 (qP) 通常与生长率 (μ) 相对应.
- 了解低μ的局限性对于解策略至关重要.
- 对于接近零的μ的生产动态和生理学的知识很少.
研究的目的:
- 在接近零增长的Komagataella phaffii中研究重组蛋白质生产动态.
- 了解K. phaffii在长期缓慢生长的情况下的生理适应.
- 探讨作为一种将生长与生产脱的工具的收缩机种植.
主要方法:
- 重组K. phaffii的有氧,葡萄糖限制的保持剂培养,分泌一个VHH抗体.
- 监测细胞生长,活力和VHH生产.
- 转录基因和脂质基因分析以评估细胞适应.
主要成果:
- 实现了大约两个月的翻倍时间 (μ ≈ 0.00047 h−1).
- 保持高细胞活力,并观察到新的VHH合成和分泌 (qP ≈ 0.019 mg g-1 h-1).
- 观察到细胞组成的显著变化,包括储存脂质和碳水化合物的增加,表明为长期生存做好了准备.
结论:
- 保持剂种植对于研究接近零的生长条件是有效的.
- 确定了脱增长和蛋白质生产的潜在工程目标.
- 获得了关于K. phaffii生理适应极度缓慢生长的见解.
相关概念视频
Stringent Response in E. coli
Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...
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...
Upstream Processing
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...
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...


