Steady state extracellular fluxes in anaerobic yeast Saccharomyces cerevisiae

experimentally measured extracellular fluxes in yeast Saccharomyces cerevisiae in anaerobic glucose limited chemostat (D=0.1 h-1) on minimal medium

Submitter: Maksim Zakhartsev

Assay type: Metabolite Profiling

Technology type: HPLC

Investigation: Steady state metabolic fluxes and metabolite co...

Study: Steady state fluxes in yeast Saccharomyces cere...

Measured steady state concentrations of intracellular metabolites in yeast Saccharomyces cerevisiae anaerobic chemostat

Steady state concentrations of intracellular metabolites in yeast Saccharomyces cerevisiae anaerobic chemostat at D = 0.1 h-1 on minimal medium. All metabolite concentrations are in mmol/L(CV).

Creator: Maksim Zakhartsev

Submitter: Maksim Zakhartsev

Simplified absolute metabolite quantification by gas chromatography–isotope dilution mass spectrometry on the basis of commercially available source material

Abstract (Expand)

In the field of metabolomics, GC–MS has rather established itself as a tool for semi-quantitative strategies like metabolic fingerprinting or metabolic profiling. Absolute quantification of intra- or … extracellular metabolites is nowadays mostly accomplished by application of diverse LC–MS techniques. Only few groups have so far adopted GC–MS technology for this exceptionally challenging task. Besides numerous and deeply investigated problems related to sample generation, the pronounced matrix effects in biological samples have led to the almost mandatory application of isotope dilution mass spectrometry (IDMS) for the accurate determination of absolute metabolite concentrations. Nevertheless, access to stable isotope labeled internal standards (ILIS), which are in many cases commercially unavailable, is quite laborious and very expensive. Here we present an improved and simplified gas chromatography–isotope dilution mass spectrometry (GC–IDMS) protocol for the absolute determination of intra- and extracellular metabolite levels. Commercially available 13C-labeled algal cells were used as a convenient source for the preparation of internal standards. Advantages as well as limitations of the described method are discussed.

Authors: Oliver Vielhauer, , Thomas Horn, Ralf Takors,

Date Published: 1st Dec 2011

Publication Type: Not specified

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