Bioreaction Network Topology and Metabolic Flux Ratio Analysis by Biosynthetic Fractional 13C Labeling and Two-Dimensional NMR Spectroscopy

• Published in: Metabolic engineering Vol. 1; no. 3; pp. 189 - 197
• Main Authors: Szyperski, Thomas, Glaser, Ralf W, Hochuli, Michel, Fiaux, Jocelyne, Sauer, Uwe, Bailey, James E, Wüthrich, Kurt
• Format: Journal Article
• Language: English
• Published: Belgium Elsevier Inc 01.07.1999
• Subjects: 2D NMR; amino acid biosynthesis; Amino Acids - biosynthesis; Amino Acids - isolation & purification; Biomass; Biomedical Engineering; Bioreactors; biosynthetic fractional 13C labeling; Biotechnology; Carbon Isotopes; central carbon metabolism; Haloarcula - metabolism; in vivo flux ratios; Magnetic Resonance Spectroscopy - methods; metabolic flux balancing; METAFoR analysis; Software; central carbon metabolism; metabolic flux balancing; 2D NMR; in vivo flux ratios; METAFoR analysis; biosynthetic fractional 13C labeling; amino acid biosynthesis
• ISSN: 1096-7176; 1096-7184
• DOI: 10.1006/mben.1999.0116

Biosynthetically directed fractional 13C labeling of the proteinogenic amino acids is achieved by feeding a mixture of uniformly 13C-labeled and unlabeled carbon source compounds into a bioreaction network. Analysis of the resulting labeling pattern enables both a comprehensive characterization of the network topology and the determination of metabolic flux ratios. Attractive features with regard to routine applications are (i) an inherently small demand for 13C-labeled source compounds and (ii) the high sensitivity of two-dimensional [13C,1H]-correlation nuclear magnetic resonance spectroscopy for analysis of 13C-labeling patterns. A user-friendly program, FCAL, is available to allow rapid data analysis. This novel approach, which recently also has been employed in conjunction with metabolic flux balancing to obtain reliable estimates of in vivo fluxes, enables efficient support of metabolic engineering and biotechnology process design.