Document Detail

(13)C-based metabolic flux analysis.
MedLine Citation:
PMID:  19478804     Owner:  NLM     Status:  MEDLINE    
Stable isotope, and in particular (13)C-based flux analysis, is the exclusive approach to experimentally quantify the integrated responses of metabolic networks. Here we describe a protocol that is based on growing microbes on (13)C-labeled glucose and subsequent gas chromatography mass spectrometric detection of (13)C-patterns in protein-bound amino acids. Relying on publicly available software packages, we then describe two complementary mathematical approaches to estimate either local ratios of converging fluxes or absolute fluxes through different pathways. As amino acids in cell protein are abundant and stable, this protocol requires a minimum of equipment and analytical expertise. Most other flux methods are variants of the principles presented here. A true alternative is the analytically more demanding dynamic flux analysis that relies on (13)C-pattern in free intracellular metabolites. The presented protocols take 5-10 d, have been used extensively in the past decade and are exemplified here for the central metabolism of Escherichia coli.
Nicola Zamboni; Sarah-Maria Fendt; Martin Rühl; Uwe Sauer
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Publication Detail:
Type:  Journal Article     Date:  2009-05-21
Journal Detail:
Title:  Nature protocols     Volume:  4     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2009  
Date Detail:
Created Date:  2009-05-29     Completed Date:  2009-08-10     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  878-92     Citation Subset:  IM    
Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland.
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MeSH Terms
Amino Acids / metabolism
Carbon Isotopes*
Escherichia coli / metabolism
Escherichia coli Proteins / metabolism
Gas Chromatography-Mass Spectrometry
Glucose / metabolism
Metabolic Networks and Pathways*
Metabolomics / methods*
Models, Biological
Reg. No./Substance:
0/Amino Acids; 0/Carbon Isotopes; 0/Escherichia coli Proteins; 50-99-7/Glucose

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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