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Manuscript title Differential glucose repression in common yeast strains in response to HXK2 deletion.
PubMed ID 20199578
Journal FEMS Yeast Research
Year 2010
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 Downloadarticle Kummel_2010.pdf
Project name not specified

Experiment Description info

Organism Saccharomyces cerevisiae
Strain CEN.PK 113-7D, CEN.PK/JT4, FY4
Data type metabolites at steady-state
Data units mM
Execution date not specified

Experimental Details info

Temperature (0C) 30
pH 5.0
Carbon source glucose,
Culture mode batch
Process condition aerobic
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.
Medium composition

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. [1].

General protocol information Sampling method: Intracellular metabolite concentrations of two biological and two technical replicates were determined from samples withdrawn from culture at an OD of approximately 1.5.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. [2].

-------------References------------
[1] Verduyn C, Postma E, ScheffersWA &Van Dijken JP (1992) Effect of benzoic acid on metabolic fluxes in yeasts: a continuousculture study on the regulation of respiration and alcoholic fermentation. Yeast 8: 501–517. http://doi.org/fgpqk3 [2] Ewald JC, Heux S & Zamboni N (2009) High-throughput quantitative metabolomics: workflow for cultivation, quenching, and analysis of yeast in a multiwell format. Anal Chem 81: 3623–3629. http://doi.org/chgtcq

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Entered by Administrator KiMoSysFirst name: Administrator
Affiliation: INESC-ID/IST
Homepage: http://kdbio.inesc-id.pt/kimosys
Interests: mathematical modeling, accessible data, use of data

Created 2018-07-07 19:01:50 UTC

Updated 2018-07-07 21:16:38 UTC

Version 1

Status (reviewed) 2018-07-07 21:17:37 UTC




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