期刊
METABOLIC ENGINEERING
卷 21, 期 -, 页码 34-45出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2013.11.001
关键词
Schizosaccharomyces pombe; Protein secretion; C-13 metabolic flux analysis; Metabolic burden; Substrate mixtures; Macromolecular composition
资金
- BMBF (Federal Ministry of Education and Research, Germany, Project SWEEPRO) [FKZ 0315800A, 0315800B]
Protein secretion in yeast is generally associated with a burden to cellular metabolism. To investigate this metabolic burden in Schizosaccharonlyces 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 C-13-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 mitochonclrial 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. (C) 2013 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
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