Impact of protein uptake and degradation on recombinant protein secretion in yeast
Journal article, 2014

Protein titers, a key bioprocessing metric, depend both on the synthesis of protein and the degradation of protein. Secreted recombinant protein production in Saccharomyces cerevisiae is an attractive platform as minimal media can be used for cultivation, thus reducing fermentation costs and simplifying downstream purification, compared to other systems that require complex media. As such, engineering S. cerevisiae to improve titers has been then the subject of significant attention, but the majority of previous efforts have been focused on improving protein synthesis. Here, we characterize the protein uptake and degradation pathways of S. cerevisiae to better understand its impact on protein secretion titers. We do find that S. cerevisiae can consume significant (in the range of 1 g/L/day) quantities of whole proteins. Characterizing the physiological state and combining metabolomics and transcriptomics, we identify metabolic and regulatory markers that are consistent with uptake of whole proteins by endocytosis, followed by intracellular degradation and catabolism of substituent amino acids. Uptake and degradation of recombinant protein products may be common in S. cerevisiae protein secretion systems, and the current data should help formulate strategies to mitigate product loss.

saccharomyces-cerevisiae

metabolomics

Protein production

Saccharomyces cerevisiae

extraction

Protein degradation

software

rates

topology

Secretion

Endocytosis

Author

Keith Tyo

Chalmers, Chemical and Biological Engineering, Life Sciences, System Biology

Zihe Liu

Chalmers, Chemical and Biological Engineering, Life Sciences, System Biology

Ylva Magnusson

Chalmers, Chemical and Biological Engineering, Life Sciences, System Biology

Dina Petranovic Nielsen

Chalmers, Chemical and Biological Engineering, Life Sciences, System Biology

Jens B Nielsen

Novo Nordisk

Chalmers, Chemical and Biological Engineering, Life Sciences, System Biology

Applied Microbiology and Biotechnology

0175-7598 (ISSN) 1432-0614 (eISSN)

Vol. 98 16 7149-7159

Industrial Systems Biology of Yeast and A. oryzae (INSYSBIO)

European Commission (FP7), 2010-01-01 -- 2014-12-31.

Subject Categories

Industrial Biotechnology

Microbiology

DOI

10.1007/s00253-014-5783-7

More information

Latest update

12/4/2018