Stationary versus non-stationary (13)C-MFA: a comparison using a consistent dataset

Journal of Biotechnology
Stephan NoackWolfgang Wiechert

Abstract

Besides the well-established (13)C-metabolic flux analysis ((13)C-MFA) which characterizes a cell's fluxome in a metabolic and isotopic stationary state a current area of research is isotopically non-stationary MFA. Non-stationary (13)C-MFA uses short-time isotopic transient data instead of long-time isotopic equilibrium data and thus is capable to resolve fluxes within much shorter labeling experiments. However, a comparison of both methods with data from one single experiment has not been made so far. In order to create a consistent database for directly comparing both methods a (13)C-labeling experiment in a fed-batch cultivation with a Corynebacterium glutamicum lysine producer was carried out. During the experiment the substrate glucose was switched from unlabeled to a specifically labeled glucose mixture which was immediately traced by fast sampling and metabolite quenching. The time course of labeling enrichments in intracellular metabolites until isotopic stationarity was monitored by LC-MS/MS. The resulting dataset was evaluated using the classical as well as the isotopic non-stationary MFA approach. The results show that not only the obtained relative data, i.e. intracellular flux distributions, but also the more info...Continue Reading

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Citations

Jul 7, 2011·Applied Microbiology and Biotechnology·Katharina Nöh, Wolfgang Wiechert
May 7, 2013·Metabolic Engineering·Xuewen ChenYair Shachar-Hill
Mar 1, 2012·Journal of Experimental Botany·John O'GradyJohn A Morgan
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