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

• 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.

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

NATURVETENSKAP  -- Biologi -- Cellbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Cell Biology (hsv//eng)

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)

Nyckelord

Amyloid-β

Cell engineering

Cell stress

Protein misfolding and aggregation

Yeast cell factories

Amino acids

Biochemistry

Biosynthesis

Cytology

Metabolic engineering

Metabolism

Neurodegenerative diseases

Peptides

Recombinant proteins

Transcription

Transcription factors

Yeast

Alzheimers disease

Cellular stress

Disease models

Protein aggregation

Protein misfolding

Recombinant protein productions

Yeast cell

Yeast cell factory

Glycoproteins

amylase

amyloid beta protein

amyloid beta protein[1-42]

arginine

membrane lipid

mitochondrial protein

recombinant protein

sulfur amino acid

Article

cell metabolism

gene deletion

gene expression

gene overexpression

genetic transcription

lipogenesis

mitochondrion

nonhuman

oxidative stress

phenotype

protein expression

protein unfolding

proteomics

RNA processing

Saccharomyces cerevisiae

transcriptomics

Protein misfolding and aggregation

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)