Overcoming the metabolic burden of protein secretion in Schizosaccharomyces pombe – A quantitative approach using 13C-based metabolic flux analysis

Tobias Klein, Sabrina Lange, Nadine Wilhelm, Matthias Bureik, Tae-Hoon Yang , Elmar Heinzle, Konstantin Schneider

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Protein secretion in yeast is generally associated with a burden to cellular metabolism. To investigate this metabolic burden in Schizosaccharomyces pombe, we constructed a set of strains secreting the model protein maltase in different amounts. We quantified the influence of protein secretion on the metabolism applying 13C-based metabolic flux analysis in chemostat cultures. Analysis of the macromolecular biomass composition revealed an increase in cellular lipid content at elevated levels of protein secretion and we observed altered metabolic fluxes in the pentose phosphate pathway, the TCA cycle, and around the pyruvate node including mitochondrial NADPH supply. Supplementing acetate to glucose or glycerol minimal media was found to improve protein secretion, accompanied by an increased cellular lipid content and carbon flux through the TCA cycle as well as increased mitochondrial NADPH production. Thus, systematic metabolic analyses can assist in identifying factors limiting protein secretion and in deriving strategies to overcome these limitations.
Original languageEnglish
JournalMetabolic Engineering
Volume21
Pages (from-to)34-45
Number of pages12
ISSN1096-7176
DOIs
Publication statusPublished - 2014
Externally publishedYes

Keywords

  • Schizosaccharomyces pombe
  • Protein secretion
  • 13C metabolic flux analysis
  • Metabolic burden
  • Substrate mixtures
  • Macromolecular composition

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