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Using multiple tracers for 13C metabolic flux analysis.

Maciek R Antoniewicz1

  • 1Metabolic Engineering and Systems Biology Laboratory, Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA. mranton@udel.edu

Methods in Molecular Biology (Clifton, N.J.)
|February 19, 2013
PubMed
Summary
This summary is machine-generated.

This study provides guidelines for (13)C-Metabolic flux analysis (MFA) using multiple isotopic tracers. This technique quantifies intracellular metabolic fluxes in living cells for systems biology and metabolic engineering.

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Area of Science:

  • Biotechnology
  • Systems Biology
  • Metabolic Engineering

Background:

  • Intracellular metabolic fluxes are crucial for understanding cell physiology.
  • (13)C-Metabolic flux analysis ((13)C-MFA) quantifies these fluxes in living cells.
  • Understanding metabolic fluxes aids metabolic engineering and systems biology modeling.

Purpose of the Study:

  • To provide guidelines for performing (13)C-MFA studies.
  • To detail methods for using multiple isotopic tracers.
  • To illustrate applications in microbial systems with mixed carbon sources.

Main Methods:

  • Selection of appropriate isotopic tracers.
  • Conducting isotopic labeling experiments.
  • Measuring isotopic labeling distributions in metabolic products.
  • Estimating metabolic fluxes via least-squares regression.
  • Assessing goodness of fit and confidence intervals.

Main Results:

  • Guidelines for designing (13)C-MFA studies with multiple tracers are presented.
  • The methodology is demonstrated with microbes utilizing glucose and xylose mixtures.
  • The approach is adaptable for other biological systems, including mammalian cells.

Conclusions:

  • Multiple isotopic tracers enhance (13)C-MFA for complex biological systems.
  • The described procedures are applicable to diverse cellular contexts.
  • (13)C-MFA is a valuable tool for quantitative systems biology and metabolic engineering.