Impact of protein uptake and degradation on recombinant protein secretion in yeast
Artikel i vetenskaplig tidskrift, 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

Protein production

metabolomics

Secretion

Endocytosis

extraction

Protein degradation

software

Saccharomyces cerevisiae

rates

topology

Författare

Keith Tyo

Chalmers, Kemi- och bioteknik, Livsvetenskaper, Systembiologi

Zihe Liu

Chalmers, Kemi- och bioteknik, Livsvetenskaper, Systembiologi

Ylva Magnusson

Chalmers

Dina Petranovic Nielsen

Chalmers, Kemi- och bioteknik, Livsvetenskaper, Systembiologi

Jens B Nielsen

Chalmers, Kemi- och bioteknik, Livsvetenskaper, Systembiologi

Applied Microbiology and Biotechnology

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

Vol. 98 16 7149-7159

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

Europeiska kommissionen (FP7), 2010-01-01 -- 2014-12-31.

Ämneskategorier

Industriell bioteknik

Mikrobiologi

DOI

10.1007/s00253-014-5783-7

Mer information

Senast uppdaterat

2018-09-10