Liquid chromatography–tandem quadrupole mass spectrometry and comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry measurement of targeted metabolites of Methylobacterium extorquens AM1 grown on two different carbon sources

Complementary methods using liquid chromatography–tandem quadrupole mass spectrometry (LC–MS/MS) and comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry (GC × GC–TOF-MS) were developed and applied to determine targeted metabolites involved in central carbon metabolism [...

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Bibliographic Details
Published in:Journal of Chromatography A 2009-04, Vol.1216 (15), p.3280-3289
Main Authors: Yang, Song, Sadilek, Martin, Synovec, Robert E., Lidstrom, Mary E.
Format: Article
Language:English
Subjects:
bacteria
Bacteriology
Biological and medical sciences
carbon
Central carbon metabolism
Fundamental and applied biological sciences. Psychology
GC × GC–TOF-MS
glyoxylic acid
hydrophilic interaction chromatography
LC–MS/MS
mass spectrometry
metabolism
metabolites
Metabolomics
Methylobacterium extorquens
Methylobacterium extorquens AM1
Microbiology
Morphology, structure, chemical composition
nucleotides
reversed-phase liquid chromatography
serine
succinic acid
Targeted metabolite
tricarboxylic acid cycle
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Summary:Complementary methods using liquid chromatography–tandem quadrupole mass spectrometry (LC–MS/MS) and comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry (GC × GC–TOF-MS) were developed and applied to determine targeted metabolites involved in central carbon metabolism [including tricarboxylic acid cycle, serine cycle, ethylmalonyl-coenzyme A (ethylmalonyl-CoA) pathway and poly-β-hydroxybutyrate cycle] of the bacterium Methylobacterium extorquens AM1 grown on two carbon sources, ethylamine (C2) and succinate (C4). Nucleotides, acyl-CoAs and a few volatile metabolites in cell extracts of M. extorquens AM1 were readily separated using either hydrophilic interaction liquid chromatography or reversed-phase liquid chromatography, and detected with good sensitivity by MS/MS. However, volatile intermediates within a low mass range (
ISSN:0021-9673
1873-3778
DOI:10.1016/j.chroma.2009.02.030