In vivo 13C NMR determines metabolic fluxes and steady state in linseed embryos

The analysis of time-course data from 13C-labelling experiments detected by in vivo NMR of developing linseed embryos is shown to be complementary to steady state metabolic flux analysis as it gives information on the isotopic and metabolic dynamics while reaching steady state and allows determinati...

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Veröffentlicht in:Phytochemistry (Oxford) 2007-08, Vol.68 (16), p.2341-2350
Hauptverfasser: Troufflard, Stéphanie, Roscher, Albrecht, Thomasset, Brigitte, Barbotin, Jean-Noël, Rawsthorne, Stephen, Portais, Jean-Charles
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Sprache:eng
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Zusammenfassung:The analysis of time-course data from 13C-labelling experiments detected by in vivo NMR of developing linseed embryos is shown to be complementary to steady state metabolic flux analysis as it gives information on the isotopic and metabolic dynamics while reaching steady state and allows determination of complementary fluxes. The dynamics of developing linseed embryo metabolism was investigated using 13C-labelling experiments where the real-time kinetics of label incorporation into metabolites was monitored in situ using in vivo NMR. The approach took advantage of the occurrence in this plant tissue of large metabolite pools – such as sucrose or lipids – to provide direct and quantitative measurement of the evolution of the labelling state within central metabolism. As a pre-requisite for the use of steady state flux measurements it was shown that isotopic steady state was reached within 3 h at the level of central intermediates whereas it took a further 6 h for the sucrose pool. Complete isotopic and metabolic steady state took 18 h to be reached. The data collected during the transient state where label was equilibrated but the metabolic steady state was incomplete, enabled the rates of lipid and sucrose synthesis to be measured in situ on the same sample. This approach is suitable to get a direct assessment of metabolic time-scales within living plant tissues and provides a valuable complement to steady state flux determinations.
ISSN:0031-9422
1873-3700
DOI:10.1016/j.phytochem.2007.04.018