Different routes of protein folding contribute to improved protein production in saccharomyces cerevisiae
Artikel i vetenskaplig tidskrift, 2020

Protein folding is often considered the flux controlling process in protein synthesis and secretion. Here, two previously isolated Saccharomyces cerevisiae strains with increased α-amylase productivity were analyzed in chemostat cultures at different dilution rates using multi-omics data. Based on the analysis, we identified different routes of the protein folding pathway to improve protein production. In the first strain, the increased abundance of proteins working on the folding process, coordinated with upregulated glycogen metabolism and trehalose metabolism, helped increase α-amylase productivity 1.95-fold compared to the level in the original strain in chemostat culture at a dilution rate of 0.2/h. The second strain further strengthened the folding precision to improve protein production. More precise folding helps the cell improve protein production efficiency and reduce the expenditure of energy on the handling of misfolded proteins. As calculated using an enzyme-constrained genome-scale metabolic model, the second strain had an increased productivity of 2.36-fold with lower energy expenditure than that of the original under the same condition. Further study revealed that the regulation of N-glycans played an important role in the folding precision control and that overexpression of the glucosidase Cwh41p can significantly improve protein production, especially for the strains with improved folding ca-pacity but lower folding precision. Our findings elucidated in detail the mechanisms in two strains having improved protein productivity and thereby provided novel insights for industrial recombinant protein production as well as demonstrating how multi-omics analysis can be used for identification of novel strain-engineering targets. IMPORTANCE Protein folding plays an important role in protein maturation and se-cretion. In recombinant protein production, many studies have focused on the folding pathway to improve productivity. Here, we identified two different routes for improving protein production by yeast. We found that improving folding precision is a better strategy. Dysfunction of this process is also associated with several aberrant protein-associated human diseases. Here, our findings about the role of glucosidase Cwh41p in the precision control system and the characterization of the strain with a more precise folding process could contribute to the development of novel therapeutic strategies.

Multi-omics analysis

Protein secretory pathway

Protein folding precision

Protein production

Constraint-based modeling

Författare

Qi Qi

Novo Nordisk Fonden

Chalmers, Biologi och bioteknik, Systembiologi

Feiran Li

Novo Nordisk Fonden

Chalmers, Biologi och bioteknik, Systembiologi

Rosemary Yu

Novo Nordisk Fonden

Martin Engqvist

Chalmers, Biologi och bioteknik, Systembiologi

Verena Siewers

Novo Nordisk Fonden

Chalmers, Biologi och bioteknik, Systembiologi

Johannes Fuchs

Göteborgs universitet

Jens B Nielsen

Danmarks Tekniske Universitet (DTU)

Novo Nordisk Fonden

BioInnovation Institute

Chalmers, Biologi och bioteknik, Systembiologi

mBio

2161-2129 (ISSN) 2150-7511 (eISSN)

Vol. 11 6 1-12 e02743-20

Ämneskategorier

Biokemi och molekylärbiologi

Mikrobiologi

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

Styrkeområden

Hälsa och teknik

DOI

10.1128/mBio.02743-20

PubMed

33173005

Mer information

Senast uppdaterat

2023-05-26