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GC-MS analysis of amino acids rapidly provides rich information for isotopomer balancing.
Journal article

GC-MS analysis of amino acids rapidly provides rich information for isotopomer balancing.

  • Dauner M Institute of Biotechnology, ETH Zürich, CH 8093 Zürich, Switzerland.
  • Sauer U
  • 2000-08-10
Published in:
  • Biotechnology progress. - 2000
Amino Acids
Biomass
Escherichia coli
Gas Chromatography-Mass Spectrometry
Models, Chemical
Serum Albumin, Bovine
English Gas chromatography-mass spectrometry (GC-MS) is a rapid method that provides rich information on isotopomer distributions for metabolic flux analysis. First, we established a fast and reliable experimental protocol for GC-MS analysis of amino acids from total biomass hydrolyzates, and common experimental pitfalls are discussed. Second, a suitable interface for the use of mass isotopomer data is presented. For this purpose, a general, matrix-based correction procedure that accounts for naturally occurring isotopes is introduced. Simulated and experimentally determined mass distributions of unlabeled amino acids showed a deviation of less than 3% for 89% of the analyzed fragments. Third, to investigate general properties of GC-MS-based isotopomer balancing, altered flux ratios through glycolysis and pentose phosphate pathway and/or exchange fluxes were simulated. Different fluxes were found to exert specific and significant influence on the mass isotopomer distributions, thus indicating that GC-MS data contain valuable information for metabolic flux analysis. Fourth, comparison of different methods revealed that GC-MS analysis provides the largest number of independent constraints on amino acid isotopomer abundance, followed by correlation spectroscopy and fractional enrichment analysis by nuclear magnetic resonance.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://folia.unifr.ch/global/documents/153984
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