Department of Biology and Biological Engineering, Chalmers University of Technology, SE41296, Gothenburg, Sweden.
Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE41296, Gothenburg, Sweden.
Nat Commun. 2017 Oct 25;8(1):1131. doi: 10.1038/s41467-017-00999-2.
The biotech industry relies on cell factories for production of pharmaceutical proteins, of which several are among the top-selling medicines. There is, therefore, considerable interest in improving the efficiency of protein production by cell factories. Protein secretion involves numerous intracellular processes with many underlying mechanisms still remaining unclear. Here, we use RNA-seq to study the genome-wide transcriptional response to protein secretion in mutant yeast strains. We find that many cellular processes have to be attuned to support efficient protein secretion. In particular, altered energy metabolism resulting in reduced respiration and increased fermentation, as well as balancing of amino-acid biosynthesis and reduced thiamine biosynthesis seem to be particularly important. We confirm our findings by inverse engineering and physiological characterization and show that by tuning metabolism cells are able to efficiently secrete recombinant proteins. Our findings provide increased understanding of which cellular regulations and pathways are associated with efficient protein secretion.
生物技术产业依赖细胞工厂来生产药物蛋白，其中有几种是最畅销的药物。因此，人们非常关注如何提高细胞工厂生产蛋白的效率。蛋白分泌涉及许多细胞内过程，其背后的许多机制仍不清楚。在这里，我们使用 RNA-seq 来研究突变酵母菌株中蛋白分泌的全基因组转录反应。我们发现，许多细胞过程必须进行调整以支持有效的蛋白分泌。特别是，改变的能量代谢导致呼吸作用减少和发酵作用增加，以及氨基酸生物合成的平衡和硫胺素生物合成的减少似乎尤为重要。我们通过反向工程和生理特征确认了我们的发现，并表明通过调节代谢，细胞能够有效地分泌重组蛋白。我们的研究结果增加了对与有效蛋白分泌相关的细胞调节和途径的理解。
