Application of 2D-TOCSY NMR to the measurement of specific(13C-enrichments in complex mixtures of 13C-labeled metabolites.
A 2D-NMR method based on zero-quantum filtered (ZQF-) TOtal Correlation SpectroscopY (TOCSY) was applied to measure 13C-enrichments in complex mixtures of 13C-labeled metabolites generated in carbon-labeling experiments. Using ZQF-TOCSY, more than 30 13C-enrichments could be potentially measured from the analysis of a biomass hydrolyzate prepared from Escherichia coli cells grown on a mixture of 20% [U-13C]-glucose and 80% [1-13C]-glucose, without need for separation of metabolites. The method is applicable to biomass hydrolyzates, cell extracts, and other complex biological samples. It is also applicable to any combination of labeled substrates and provides a basis for examining non-steady-state conditions.
Massou
Stéphane
S
MetaSys, Laboratoire d'Ingénierie des Systèmes Biologiques et Procédés, UMR INSA/CNRS 5504 INRA 792, LBB/INSA, 135 Avenue de Rangueil, 31077 Toulouse Cedex 4, France.
Nicolas
Cécile
C
Letisse
Fabien
F
Portais
Jean-Charles
JC
D018533:Biomass; D002247:Carbon Isotopes / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry*; D005947:Glucose / Q000032:analysis* / Q000737:chemistry; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D012680:Sensitivity and Specificity
D002247:Carbon Isotopes; D005947:Glucose