A model of yeast glycolysis based on a consistent kinetic characterisation of all its enzymes.
http://www.ncbi.nlm.nih.gov/pubmed/23831062
FEBS Letters
2013
Kieran Smallbone, Hanan L. Messiha, Kathleen M. Carroll, Catherine L. Winder, Naglis Malys, Warwick B. Dunn, Ettore Murabito, Neil Swainston, Joseph O. Dada, Farid Khan, Pınar Pir, Evangelos Simeonidis, Irena Spasić, Jill Wishart, Dieter Weichart, Neil W. Hayes, Daniel Jameson, David S. Broomhead, Stephen G. Oliver, Simon J. Gaskell, John E.G. McCarthy, Norman W. Paton, Hans V. Westerhoff, Douglas B. Kell, Pedro Mendes
Manchester Centre for Integrative Systems Biology, Manchester Institute of Biotechnology, The University of Manchester, UK
Glycolysis, Systems biology, Enzyme kinetic, Isoenzyme, Modelling
https://kimosys.org/rails/active_storage/blobs/eyJfcmFpbHMiOnsibWVzc2FnZSI6IkJBaHBBcjRFIiwiZXhwIjpudWxsLCJwdXIiOiJibG9iX2lkIn19--83aa95be6c5b8986eda3d15002240aa59b0cfd4b/Smallbone_2013.pdf
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Saccharomyces cerevisiae
Y23925
metabolites at steady-state
molecules/cell
not specified
not specified °C
not specified
glucose
chemostat
aerobic
µmax h⁻¹
not specified L
monitored by measuring the electrical capacitance of the culture. g/L
not specified
Sampling Method: 10 ml of culture.; Quenching: 10 ml of culture solution into 40 ml of a 60:40 methanol/water solution [1] stored at a temperature of −47 °C [2]. Immediate separation of cells was performed by applying centrifugation (4000 g for 5 min) followed by removal of the quenching solution. ; Extraction list: freezing-thawing in methanol, methanol-water; Analysis list: GC-MS, LC-MS;
Samples were analysed using two analytical platforms. Fructose‐1,6‐bisphosphate was quantified applying ultra-performance liquid chromatography mass spectrometry (UPLC‐MS) (Waters Acquity UPLC coupled to a ThermoFisher hybrid electrospray LTQ‐Orbitrap mass spectrometer (ThermoFisher Scientific, http://www.thermofisher.com/)) and the standard addition method for accurate quantification. Other metabolites were quantified applying GC–MS (Agilent 6890 Gas Chromatograph coupled to a Leco Pegasus III electron impact‐time‐of‐flight mass spectrometer (Leco, http://www.leco.com/)) and the external calibration method for accurate quantification. The concentrations determined in the extracted samples were reported as the number of molecules per cell. The gas chromatographic separation of 2‐phosphoglycerate and 3‐phosphoglycerate is technically demanding and was not achieved during this study. Therefore, the combined concentration of both metabolites was reported rather than separate concentrations for each metabolite. Concentrations were calculated relative to an effective cytoplasmic volume of 5 fl.
-----------------References--------------------
[1] W.B. Dunn , C.L. Winder. Methods Enzymol., 500, (2011), 277– 297. http://doi.org/dfwnv3 [2] M. Martins , W. Sha , C. Evans , S. Martino-Catt , P. Mendes , V. Shulaev. Yeast, 24, (2007), 181– 188. http://doi.org/c2qv8f
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