13C-Labeled metabolic flux analysis of a fed-batch culture of elutriated Saccharomyces cerevisiae
| Autor: | Roeland Costenoble, Timo Barl, Dirk Müller, Walter M. van Gulik, Wouter A. van Winden, Joseph J. Heijnen, Matthias Reuss |
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| Rok vydání: | 2007 |
| Předmět: |
education.field_of_study
Carbon Isotopes Population Saccharomyces cerevisiae General Medicine Biology Pentose phosphate pathway Elutriation biology.organism_classification Applied Microbiology and Biotechnology Microbiology Fed-batch culture Cytosol Biochemistry Tandem Mass Spectrometry Metabolic flux analysis Culture Techniques Biophysics education Intracellular Metabolic Networks and Pathways Chromatography Liquid |
| Zdroj: | FEMS yeast research. 7(4) |
| ISSN: | 1567-1356 |
| Popis: | This study addresses the question of whether observable changes in fluxes in the primary carbon metabolism of Saccharomyces cerevisiae occur between the different phases of the cell division cycle. To detect such changes by metabolic flux analysis, a 13C-labeling experiment was performed with a fed-batch culture inoculated with a partially synchronized cell population obtained through centrifugal elutriation. Such a culture exhibits dynamic changes in the fractions of cells in different cell cycle phases over time. The mass isotopomer distributions of free intracellular metabolites in central carbon metabolism were measured by liquid chromatography-mass spectrometry. For four time points during the culture, these distributions were used to obtain the best estimates for the metabolic fluxes. The obtained flux fits suggested that the optimally fitted split ratio for the pentose phosphate pathway changed by almost a factor of 2 up and down around a value of 0.27 during the experiment. Statistical analysis revealed that some of the fitted flux distributions for different time points were significantly different from each other, indicating that cell cycle-dependent variations in cytosolic metabolic fluxes indeed occurred. |
| Databáze: | OpenAIRE |
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