General Information info
|Manuscript title||Differential glucose repression in common yeast strains in response to HXK2 deletion.|
|Journal||FEMS Yeast Research|
|Authors||Anne Kümmel, Jennifer Christina Ewald, Sarah‐Maria Fendt, Stefan Jasper Jol, Paola Picotti, Ruedi Aebersold, Uwe Sauer, Nicola Zamboni, Matthias Heinemann|
|Affiliations||Institute of Molecular Systems Biology, ETH Zurich|
|Keywords||glucose repression; hexokinase 2; FY4; CEN.PK; PKA; metabolomics|
|Full text article||Kummel_2010.pdf|
|Project name||not specified|
Experiment Description info
|Strain||CEN.PK 113-7D, CEN.PK/JT4, FY4|
|Data type||metabolites at steady-state|
|Execution date||not specified|
Experimental Details info
|Dilution rate (h-1)||-|
|Working volume (L)||not specified|
|Biomass concentration (g/L)||Calculated using an earlier determined OD-to-biomass DW correlation coefficient of 0.486 gDWL/OD for FY4 and 0.52 gDWL/OD for CEN.PK strains.|
Minimal defined medium with glucose as sole carbon source. Prepared from autoclaved salt and glucose solutions and sterile filtered solutions of vitamins and trace metals to reach concentrations as described in Verduyn et al. .
|General protocol information||
Intracellular metabolite concentrations of two biological and two technical replicates were determined from samples withdrawn from culture at an OD of approximately 1.5. Samples of 1–4mL were taken at each sampling time point.
Quenching procedure: quenched in methanol at -40 ºC. For the determination of the cAMP concentrations, a sample volume of 10mL was taken. After centrifuging for 3 min at 15 550 g in a rotor precooled to -9 ºC, the samples were frozen at -40 ºC.
Extraction technique: hot ethanol
Sample analyzing method: LC-MS, GC-TOF
|Methods description - Notes||
For quantification by GC-TOF, two sample aliquots were derivatized with either TMS-agent [N-methyl-N-(trimethylsilyl) trifluoroacetamide, Fluka] or TBDMS agent (N tertbutyldimethylsilyl-N-methyltrifluoroacetamide, Fluka). The samples were separated via GC on an HP5-MS column (Hewlett-Packard, length 30m ID 0.25 film 0.25 mm) and injected for MS analysis into a TOF spectrometer (Pegasus III, Leco). Detailed information on process parameters are described in Ewald et al. .
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