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• W2063763893 abstract "A novel approach to 13C metabolic flux analysis (MFA) is presented using cytosolic metabolite pool sizes and their 13C labeling data from an isotopically non-stationary 13C labeling experiment (INST-CLE). The procedure is demonstrated with an E. coli wild type strain grown at fed batch conditions. The intra cellular labeling dynamics are excited by a sudden step increase of the 13C portion in the substrate feed. Due to unchanged saturation of the substrate uptake system, the metabolic fluxes remain constant during the following sampling time period of only 16 s, in which 20 samples are taken by an automated rapid sampling device immediately stopping metabolism by methanol quenching. Subsequent cell disruptive sample preparation and LC–MS/MS enabled simultaneous determination of pool sizes and mass isotopomers of intra cellular metabolites requiring detection limits in the nM range. Based on this data the new computational flux analysis tool 13CFLUX/INST is used to determine the intra cellular fluxes based on a complex carbon labeling network model. The measured data is in good agreement with the model predictions, thus proving the applicability of the new isotopically non-stationary 13C metabolic flux analysis (INST-13C-MFA) concept. Moreover, it is shown that significant new information with respect to flux identifiability, non-measurable pool sizes, data consistency, or large storage pools can be taken from the novel kind of experimental data. This offers new insight into the biological operation of the metabolic network in vivo." @default.
• W2063763893 created "2016-06-24" @default.
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• W2063763893 date "2007-04-01" @default.
• W2063763893 modified "2023-10-17" @default.
• W2063763893 title "Metabolic flux analysis at ultra short time scale: Isotopically non-stationary 13C labeling experiments" @default.
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• W2063763893 doi "https://doi.org/10.1016/j.jbiotec.2006.11.015" @default.
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