Bao Jichen, Huang Mingtao, Petranovic Dina, Nielsen Jens
Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.
Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, Gothenburg, Sweden.
Appl Environ Microbiol. 2017 Jun 30;83(14). doi: 10.1128/AEM.03400-16. Print 2017 Jul 15.
The yeast is widely used to produce biopharmaceutical proteins. However, the limited capacity of the secretory pathway may reduce its productivity. Here, we increased the secretion of a heterologous α-amylase, a model protein used for studying the protein secretory pathway in yeast, by moderately overexpressing , which is involved in protein translocation from the endoplasmic reticulum to the Golgi apparatus. The moderate overexpression of increased α-amylase secretion by generating more endoplasmic reticulum exit sites. The production of reactive oxygen species resulting from the heterologous α-amylase production was reduced. A genome-wide expression analysis indicated decreased endoplasmic reticulum stress in the strain that moderately overexpressed , which was consistent with a decreased volume of the endoplasmic reticulum. Additionally, fewer mitochondria were observed. Finally, the moderate overexpression of was shown to improve the secretion of two other recombinant proteins, endoglucanase I and glucan-1,4-α-glucosidase, indicating that this mechanism is of general relevance. There is an increasing demand for recombinant proteins to be used as enzymes and pharmaceuticals. The yeast is a cell factory that is widely used to produce recombinant proteins. Our study revealed that moderate overexpression of increased recombinant protein secretion in This new strategy can be combined with other targets to engineer cell factories to efficiently produce protein in the future.
酵母被广泛用于生产生物制药蛋白。然而，分泌途径的有限能力可能会降低其生产力。在这里，我们通过适度过表达参与蛋白质从内质网转运到高尔基体的[具体基因名称未给出]，提高了一种用于研究酵母中蛋白质分泌途径的模型蛋白——异源α-淀粉酶的分泌量。[具体基因名称未给出]的适度过表达通过产生更多内质网出口位点增加了α-淀粉酶的分泌。异源α-淀粉酶产生所导致的活性氧物质的产生减少。全基因组表达分析表明，适度过表达[具体基因名称未给出]的菌株中内质网应激降低，这与内质网体积减小一致。此外，观察到的线粒体较少。最后，[具体基因名称未给出]的适度过表达被证明可改善另外两种重组蛋白——内切葡聚糖酶I和葡聚糖-1,4-α-葡萄糖苷酶的分泌，表明这种机制具有普遍相关性。对用作酶和药物的重组蛋白的需求日益增加。酵母是一种广泛用于生产重组蛋白的细胞工厂。我们的研究表明，[具体基因名称未给出]的适度过表达增加了酵母中重组蛋白的分泌。这种新策略可与其他靶点结合，在未来设计细胞工厂以高效生产蛋白质。
