Bioreaction network topology and metabolic flux ratio analysis by biosynthetic fractional 13C labeling and two-dimensional NMR spectroscopy.
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.
Szyperski
T
T
Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Hönggerberg, Zürich, Switzerland. szypersk@acsu.buffalo.edu
Glaser
R W
RW
Hochuli
M
M
Fiaux
J
J
Sauer
U
U
Bailey
J E
JE
Wüthrich
K
K
D000596:Amino Acids / Q000096:biosynthesis* / Q000302:isolation & purification; D018533:Biomass; D001698:Biomedical Engineering; D019149:Bioreactors; D001709:Biotechnology; D002247:Carbon Isotopes; D019613:Haloarcula / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D012984:Software
D000596:Amino Acids; D002247:Carbon Isotopes