Advances in analysis of microbial metabolic fluxes via (13)C isotopic labeling

Mass Spectrom Rev. 2009 Mar-Apr;28(2):362-75. doi: 10.1002/mas.20191.

Abstract

Metabolic flux analysis via (13)C labeling ((13)C MFA) quantitatively tracks metabolic pathway activity and determines overall enzymatic function in cells. Three core techniques are necessary for (13)C MFA: (1) a steady state cell culture in a defined medium with labeled-carbon substrates; (2) precise measurements of the labeling pattern of targeted metabolites; and (3) evaluation of the data sets obtained from mass spectrometry measurements with a computer model to calculate the metabolic fluxes. In this review, we summarize recent advances in the (13)C-flux analysis technologies, including mini-bioreactor usage for tracer experiments, isotopomer analysis of metabolites via high resolution mass spectrometry (such as GC-MS, LC-MS, or FT-ICR), high performance and large-scale isotopomer modeling programs for flux analysis, and the integration of fluxomics with other functional genomics studies. It will be shown that there is a significant value for (13)C-based metabolic flux analysis in many biological research fields.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Review

MeSH terms

  • Bacterial Physiological Phenomena*
  • Bacterial Proteins / metabolism*
  • Bioreactors / microbiology*
  • Carbon / metabolism*
  • Carbon Isotopes / analysis
  • Carbon Isotopes / chemistry
  • Isotope Labeling / methods
  • Isotope Labeling / trends
  • Mass Spectrometry / methods*
  • Mass Spectrometry / trends*
  • Multienzyme Complexes / metabolism*

Substances

  • Bacterial Proteins
  • Carbon Isotopes
  • Multienzyme Complexes
  • Carbon