Suppressors of amyloid-β toxicity improve recombinant protein production in yeast by reducing oxidative stress and tuning cellular metabolism
Journal article, 2022

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.

Protein misfolding and aggregation

Cell engineering

Yeast cell factories

Cell stress

Amyloid-β

Author

Xin Chen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Novo Nordisk Foundation

Xiaowei Li

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Boyang Ji

BioInnovation Institute

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Yanyan Wang

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Olena Ishchuk

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Egor Vorontsov

University of Gothenburg

Dina Petranovic Nielsen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Novo Nordisk Foundation

Verena Siewers

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Novo Nordisk Foundation

Martin Engqvist

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Metabolic Engineering

1096-7176 (ISSN) 1096-7184 (eISSN)

Vol. 72 311-324

Subject Categories

Cell Biology

Biochemistry and Molecular Biology

Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

DOI

10.1016/j.ymben.2022.04.005

More information

Latest update

5/26/2023