定制Pichia pastoris的氨基酸代谢,用于重组蛋白质生产
Hannes Rußmayer1,2, Markus Buchetics1,2, Matthias Mattanovich1,3,4
1Department of Biotechnology, Institute of Microbiology and Microbial Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
Biotechnology journal
|September 5, 2023
概括
工程酵母氨基酸代谢促进了重组蛋白质的生产. 过度表达GCN4调节剂显著增加了2.6倍的碳酸酶 (CES) 标位,突出显示了氨基酸可用性的重要性.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 代谢工程是代谢工程.
背景情况:
- 氨基酸是蛋白质必不可少的构建基.
- 再组合蛋白质的生产可以影响氨基酸生物合成.
- 氨基酸的可用性会影响蛋白质的产量.
研究的目的:
- 研究工程氨基酸代谢如何影响Pichia pastoris.中的重组蛋白质生产.
- 为了确定增强蛋白质产量的特定基因和调节者.
- 为了验证生物反应器培养中的发现.
主要方法:
- 代谢组和转录组的系统级分析.
- 氨基酸代谢基因和GCN4转录因子的构成性过度表达.
- 测量复合蛋白 (碳酸酶) 生产.
- 料批发发酵用于扩大规模的验证.
主要成果:
- 过度表达氨酸和芳香氨基酸通路基因增加了模型蛋白质的生产.
- 过度表达GCN4调节器产生了最显著的蛋白质生产增加.
- 细胞内氨基酸水平的增加与改善的重组蛋白质生产直接相关.
- 过度表达GCN4导致食批量培养中的氧化酶标位增加了2.6倍.
结论:
- 工程氨基酸生物合成途径,特别是通过GCN4调节,是一种可行的策略,可以增强Pichia pastoris的重组蛋白质生产.
- 增加细胞内氨基酸的可用性是提高蛋白质产量的关键因素.
- 对于代谢工程来说,GCN4是一个强大的目标,可以促进异质蛋白质的表达.
相关概念视频
Amino Acid Catabolism
50
Microorganisms rely on proteins as an essential carbon and energy source, particularly in environments with limited polysaccharides or lipids. However, proteins are too large to cross the plasma membrane unaided, necessitating enzymatic degradation. Microbes secrete extracellular proteases and peptidases that hydrolyze proteins into peptides, which can then be transported across the membrane. Once inside the cell, intracellular proteases degrade these peptides into free amino acids, which...
50
Amino Acid Biosynthetic Pathways
35
Amino acid biosynthesis is essential for cell growth, protein synthesis, and metabolic regulation. Cells generate essential and non-essential amino acids from metabolic intermediates to sustain vital biological functions. These intermediates originate from key metabolic pathways: glycolysis, the tricarboxylic acid (TCA) cycle, and the pentose phosphate pathway. Important precursors include α-ketoglutarate, pyruvate, oxaloacetate, phosphoenolpyruvate, and erythrose-4-phosphate, which...
35
The Central Dogma
21.8K
The central dogma explains the flow of genetic information from DNA nucleotides to the amino acid sequence of proteins.
RNA is the Missing Link Between DNA and Proteins
In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is...
RNA is the Missing Link Between DNA and Proteins
In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is...
21.8K


