Improved Isotopic Profiling by Pure Shift Heteronuclear 2D J -Resolved NMR Spectroscopy

Archive ouverte

Sinnaeve, Davy | Dinclaux, Mickael | Cahoreau, Edern | Millard, Pierre | Portais, Jean-Charles | Letisse, Fabien | Lippens, Guy

Edité par CCSD ; American Chemical Society -

International audience. Quantitative information on the carbon isotope content of metabolites is essential for flux analysis. Whereas this information is in principle present in proton NMR spectra through both direct and long-range heteronuclear coupling constants, spectral overlap and homonuclear coupling constants both hinder its extraction. We demonstrate here how pure shift 2D J-resolved NMR spectroscopy can simultaneously remove the homonuclear couplings and separate the chemical shift information from the heteronuclear coupling patterns. We demonstrate the power of this method on cell lysates from different bacterial cultures and investigate in detail the branched chain amino acid biosynthesis.

Consulter en ligne

Suggestions

Du même auteur

Increasing field strength versus advanced isotope labeling for NMR based fluxomics

Archive ouverte | Dinclaux, Mickael | CCSD

International audience. Nuclear magnetic resonance (NMR)-based fluxomics seeks to measure the incorporation of isotope labels in selected metabolites to follow kinetically the synthesis of the latter. It can however...

Exploring the Glucose Fluxotype of the E. coli y-ome Using High-Resolution Fluxomics

Archive ouverte | Bergès, Cécilia | CCSD

International audience. We have developed a robust workflow to measure high-resolution fluxotypes (metabolic flux phenotypes) for large strain libraries under fully controlled growth conditions. This was achieved by...

Acetate fluxes in Escherichia coli are determined by the thermodynamic control of the Pta-AckA pathway.

Archive ouverte | Enjalbert, Brice | CCSD

Escherichia coli excretes acetate upon growth on fermentable sugars, but the regulation of this production remains elusive. Acetate excretion on excess glucose is thought to be an irreversible process. However, dynamic (13)C-metab...

Chargement des enrichissements...