High-level production of recombinant proteins in industrial microorganisms is often limited by the formation of misfolded proteins or protein aggregates, which consequently induce cellular stress responses. We hypothesized that in a yeast Alzheimer's disease (AD) model overexpression of amyloid-β peptides (Aβ42), one of the main peptides relevant for AD pathologies, induces similar phenotypes of cellular stress. Using this humanized AD model, we previously identified suppressors of Aβ42 cytotoxicity. Here we hypothesize that these suppressors could be used as metabolic engineering targets to alleviate cellular stress and improve recombinant protein production in the yeast Saccharomyces cerevisiae. Forty-six candidate genes were individually deleted and twenty were individually overexpressed. The positive targets that increased recombinant α-amylase production were further combined leading to an 18.7-fold increased recombinant protein production. These target genes are involved in multiple cellular networks including RNA processing, transcription, ER-mitochondrial complex, and protein unfolding. By using transcriptomics and proteomics analyses, combined with reverse metabolic engineering, we showed that reduced oxidative stress, increased membrane lipid biosynthesis and repressed arginine and sulfur amino acid biosynthesis are significant pathways for increased recombinant protein production. Our findings provide new insights towards developing synthetic yeast cell factories for biosynthesis of valuable proteins.

工业微生物中重组蛋白的高水平生产通常受到错误折叠蛋白质或蛋白质聚集体的形成的限制，从而诱导细胞应激反应。我们假设在酵母阿尔茨海默病 (AD) 模型中，淀粉样蛋白-β 肽 (Aβ42)（与 AD 病理相关的主要肽之一）的过表达会诱导相似的细胞应激表型。使用这种人源化的 AD 模型，我们之前确定了 Aβ42 细胞毒性的抑制因子。在这里，我们假设这些抑制因子可用作代谢工程靶点，以减轻细胞压力并改善酵母酿酒酵母中的重组蛋白生产. 46 个候选基因被单独删除，20 个单独过表达。增加重组 α-淀粉酶产量的阳性靶点进一步结合，导致重组蛋白产量增加 18.7 倍。这些靶基因参与多个细胞网络，包括 RNA 加工、转录、ER-线粒体复合物和蛋白质展开。通过使用转录组学和蛋白质组学分析，结合逆向代谢工程，我们发现减少氧化应激、增加膜脂生物合成和抑制精氨酸和硫氨基酸生物合成是增加重组蛋白产量的重要途径。我们的研究结果为开发用于生物合成有价值蛋白质的合成酵母细胞工厂提供了新的见解。