An introduction to 13C metabolic flux analysis.

In recent years, metabolic flux analysis has become one of the major tools in metabolic engineering (1, 2, 3). The aim of MFA is the detailed quantification of all metabolic fluxes in the central metabolic pathways of a microorganism. The result is a flux map that shows the distribution of anabolic and catabolic fluxes over the metabolic network. Figure 1 shows such a flux distribution for the simple example discussed throughout the text. More complex examples for quite different microorganisms can be found in (4, 5, 6, 7, 8, 9). Based on such a flux analysis, the result of a genetic manipulation can be judged or possible targets for future genetic modifications might be identified. Thus MFA is also an important tool for genetic engineering.

[1]  H. Förstel,et al.  Stable isotopes : proceedings of the 4th international conference, Jülich, March 23-26, 1981 , 1982 .

[2]  Anne Lohrli.  Chapman and Hall , 1985 .

[3]  W. Holms,et al.  The central metabolic pathways of Escherichia coli: relationship between flux and control at a branch point, efficiency of conversion to biomass, and excretion of acetate. , 1986, Current topics in cellular regulation.

[4]  Karl Schügerl,et al.  Bioreaction Engineering: Modeling and Control , 1987 .

[5]  J. Bailey,et al.  Toward a science of metabolic engineering , 1991, Science.

[6]  William H. Press,et al.  The Art of Scientific Computing Second Edition , 1998 .

[7]  A. D. de Graaf,et al.  Flux partitioning in the split pathway of lysine synthesis in Corynebacterium glutamicum. Quantification by 13C- and 1H-NMR spectroscopy. , 1993, European journal of biochemistry.

[8]  W. Eisenreich,et al.  Retrobiosynthetic analysis of carbon fixation in the phototrophic eubacterium Chloroflexus aurantiacus. , 1993, European journal of biochemistry.

[9]  G. Stephanopoulos,et al.  Metabolic flux distributions in Corynebacterium glutamicum during growth and lysine overproduction , 2000, Biotechnology and bioengineering.

[10]  B. Palsson,et al.  Metabolic Flux Balancing: Basic Concepts, Scientific and Practical Use , 1994, Bio/Technology.

[11]  T. Szyperski.  Biosynthetically Directed Fractional 13C‐labeling of Proteinogenic Amino Acids , 1995 .

[12]  G. Stephanopoulos,et al.  Intracellular flux analysis in hybridomas using mass balances and in vitro 13C nmr , 1995, Biotechnology and bioengineering.

[13]  T. Szyperski.  Biosynthetically directed fractional 13C-labeling of proteinogenic amino acids. An efficient analytical tool to investigate intermediary metabolism. , 1995, European journal of biochemistry.

[14]  H Sahm,et al.  Determination of the fluxes in the central metabolism of Corynebacterium glutamicum by nuclear magnetic resonance spectroscopy combined with metabolite balancing , 1996, Biotechnology and bioengineering.

[15]  Arne Elofsson.  Bioinformatics: From nucleic acids and proteins to cell metabolism: Edited by D. Schomburg and U. Lessel, VCH; Weinheim-New York, 1995. viii + 195 pp. DM 148.00 (hb). ISBN 3-527-30072-4 , 1996 .

[16]  W. Wiechert,et al.  Bidirectional reaction steps in metabolic networks: I. Modeling and simulation of carbon isotope labeling experiments. , 1997, Biotechnology and bioengineering.

[17]  U. Sauer,et al.  Metabolic fluxes in riboflavin-producing Bacillus subtilis , 1997, Nature Biotechnology.

[18]  W. Wiechert,et al.  Bidirectional reaction steps in metabolic networks: II. Flux estimation and statistical analysis. , 1997, Biotechnology and bioengineering.

[19]  G. Stephanopoulos,et al.  Metabolic Engineering: Principles And Methodologies , 1998 .

[20]  T Szyperski,et al.  13C-NMR, MS and metabolic flux balancing in biotechnology research , 1998, Quarterly Reviews of Biophysics.

[21]  B. Christensen,et al.  Isotopomer analysis using GC-MS. , 1999, Metabolic engineering.

[22]  E. Heinzle,et al.  Mass spectrometry for metabolic flux analysis. , 1999, Biotechnology and bioengineering.

[23]  W Wiechert,et al.  Bidirectional reaction steps in metabolic networks: IV. Optimal design of isotopomer labeling experiments. , 1999, Biotechnology and bioengineering.

[24]  W. Wiechert,et al.  Metabolic state of Zymomonas mobilis in glucose-, fructose-, and xylose-fed continuous cultures as analysed by 13C- and 31P-NMR spectroscopy , 1999, Archives of Microbiology.

[25]  A J Sinskey,et al.  Metabolite and isotopomer balancing in the analysis of metabolic cycles: II. Applications. , 1999, Biotechnology and bioengineering.

[26]  W. Wiechert,et al.  Bidirectional reaction steps in metabolic networks: III. Explicit solution and analysis of isotopomer labeling systems. , 1999, Biotechnology and bioengineering.

[27]  J. Nielsen,et al.  Quantitative analysis of metabolic fluxes in Escherichia coli, using two-dimensional NMR spectroscopy and complete isotopomer models. , 1999, Journal of biotechnology.

[28]  G. Stephanopoulos.  Metabolic fluxes and metabolic engineering. , 1999, Metabolic engineering.

[29]  W. Wiechert,et al.  In Vivo Quantification of Parallel and Bidirectional Fluxes in the Anaplerosis of Corynebacterium glutamicum * , 2000, The Journal of Biological Chemistry.

[30]  H Sahm,et al.  Determination of full 13C isotopomer distributions for metabolic flux analysis using heteronuclear spin echo difference NMR spectroscopy. , 2000, Journal of biotechnology.

[31]  W. Wiechert.  13C metabolic flux analysis. , 2001, Metabolic engineering.

[32]  J E Bailey,et al.  Metabolic flux analysis with a comprehensive isotopomer model in Bacillus subtilis. , 2001, Biotechnology and bioengineering.

[33]  W Wiechert,et al.  A universal framework for 13C metabolic flux analysis. , 2001, Metabolic engineering.

[34]  W Wiechert,et al.  Metabolic isotopomer labeling systems. Part I: global dynamic behavior. , 2001, Mathematical biosciences.