GC‐MS Analysis of Amino Acids Rapidly Provides Rich Information for Isotopomer Balancing

@article{Dauner2000GCMSAO,
  title={GC‐MS Analysis of Amino Acids Rapidly Provides Rich Information for Isotopomer Balancing},
  author={Michael Dauner and Uwe H. Sauer},
  journal={Biotechnology Progress},
  year={2000},
  volume={16}
}
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… 
View on Wiley
lib3.dss.go.th
245 Citations
Highly Influential Citations
14
Background Citations
56
Methods Citations
39
Results Citations
1

245 Citations

Citation Type

Citation Type

New tools for mass isotopomer data evaluation in 13C flux analysis: Mass isotope correction, data consistency checking, and precursor relationships
TLDR
A new elegant method to correct MS measurement data for natural stable mass isotopes by infinite dimensional matrix calculus is presented and a new method for consistency checking of MS spectra that can be applied for automatic error recognition in high‐throughput flux analysis procedures is established.
Software LS-MIDA for efficient mass isotopomer distribution analysis in metabolic modelling
TLDR
The new software provides an open-source platform that easily and rapidly converts experimental MS patterns of labelled metabolites into isotopomer enrichments that are the basis for subsequent observation-driven analysis of pathways and fluxes, as well as for model-driven metabolic flux calculations.
Increasing the accuracy of mass isotopomer analysis through calibration curves constructed using biologically synthesized compounds.
TLDR
It is shown that the mass isotopomer distributions of the various biosynthesized amino acids are well determined and can be reasonably estimated based on proposed binomial approximation if the labeling state of the biomass reached an isotopic steady state.
Collisional fragmentation of central carbon metabolites in LC‐MS/MS increases precision of 13C metabolic flux analysis
TLDR
This catalogue will enable comprehensive in silico calculability analyses to identify the most sensitive measurements and direct experimental design and present an efficient analytical strategy to rapidly acquire large sets of 13C‐patterns by tandem MS, and an in‐depth analysis of the collisional fragmentation of primary intermediates.
Isotopomer distribution computation from tandem mass spectrometric data with overlapping fragment spectra
We present a method for determination of the isotopomer distributions of metabolites from the data generated by a tandem mass spectrometer. The method is an improvement over existing method as it is…
Determination of carbon labeling distribution of intracellular metabolites from single fragment ions by ion chromatography tandem mass spectrometry.
Isotope labeling pattern study of central carbon metabolites using GC/MS.
  • J. Jung, M. Oh
  • Chemistry
    Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
  • 2015
Microscale analysis of amino acids using gas chromatography-mass spectrometry after methyl chloroformate derivatization.
Low Labeling 13C Metabolic Flux Analysis of Saccharomyces cerevisiae Using Gas Chromatography–Combustion–Isotope Ratio Mass Spectrometry
TLDR
The GC–C–IRMS-based method involves low labeling (0.5 %) degree of expensive tracer substrate, and suits well for larger laboratory and industrial pilot-scale fermentations.
Isotopomer-based metabolomic analysis by NMR and mass spectrometry.
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 43 REFERENCES
Isotopomer analysis using GC-MS.
TLDR
The GC-MS method was used for analyzing the labeling patterns of amino acids from a batch cultivation of Penicillium chrysogenum grown on fully labeled glucose and showed no signs of any significant carbon isotope effects, and the measurements can be used without corrections for metabolic flux analysis.
Mass spectrometry for metabolic flux analysis.
TLDR
It is shown by simulation results for the central carbon metabolism that neglecting natural isotope distributions causes significant errors in intracellular flux distributions, and a matrix based general methodology is formulated to effectively handle mass distributions in simulation experiments.
Mass isotopomer distribution analysis at eight years: theoretical, analytic, and experimental considerations.
TLDR
Attention to potentially confounding factors and analytic performance is required for optimal application of MIDA, and guidelines for optimizing accuracy and reducing impact on biosynthetic parameters are suggested.
Correction of 13C mass isotopomer distributions for natural stable isotope abundance.
TLDR
Differences between predicted and measured unlabeled and 13C-labeled isotopomer distributions for hydroxamate di-t-butyl-dimethylsilyl (di-TBDMS) derivatized pyruvate were measured and implications including those on the calculation of glucose production due to gluconeogenesis in isolated perfused rat liver are discussed.
Bidirectional reaction steps in metabolic networks: IV. Optimal design of isotopomer labeling experiments.
TLDR
The positional enrichment, multiplet, or mass isotopomer measurements with the greatest information content for flux estimation are computed and various differently labeled input substrates are compared with respect to flux estimation (input design).
Modeling isotopomer distributions in biochemical networks using isotopomer mapping matrices.
TLDR
An isotopomer model has been implemented that describes label distribution in primary carbon metabolism, i.e., in a metabolic network including the Embden-Meyerhof-Parnas and pentose phosphate pathway, the tricarboxylic acid cycle, and selected anaplerotic reaction sequences.
Bidirectional reaction steps in metabolic networks: III. Explicit solution and analysis of isotopomer labeling systems.
TLDR
It turns out that isotopomers systems are not as complex as they appear at first glance, which enables some far-reaching conclusions to be drawn on the information potential of isotopomer experiments with respect to flux identification.
Determination of the fluxes in the central metabolism of Corynebacterium glutamicum by nuclear magnetic resonance spectroscopy combined with metabolite balancing
TLDR
In vivo one component within these anaplerotic reactions is a back flux from the carbon 4 units of the tricarboxylic acid cycle to the carbon 3 units of glycolysis of 30.6% and the total net flux of the anaplerosis reactions was quantitated as 38.0%.
Comparison of mass isotopomer dilution methods used to compute VLDL production in vivo.
TLDR
This paper takes a specific application, calculation of the fractional rate of incorporation of acetyl-CoA into very low density lipoprotein-bound palmitate, and compares the results obtained from all of the mass isotopomer methods using the same data set obtained in vivo in human subjects.
Biosynthetically directed fractional 13C-labeling of proteinogenic amino acids. An efficient analytical tool to investigate intermediary metabolism.
  • T. Szyperski
  • Biology, Chemistry
    European journal of biochemistry
  • 1995
TLDR
Biosynthetically directed fractional 13C labeling of amino acids provides an efficient analytical tool to quantitatively investigate glycolysis, pyruvate metabolism, pentose phosphate pathway, tricarboxylic acid cycle and C1 metabolism.
...
1
2
3
4
5
...