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• W2115026369 abstract "Abstract The novel concept of isotopic dynamic 13 C metabolic flux analysis (ID‐ 13 C MFA) enables integrated analysis of isotopomer data from isotopic transient and/or isotopic stationary phase of a 13 C labeling experiment, short‐time experiments, and an extended range of applications of 13 C MFA. In the presented work, an experimental and computational framework consisting of short‐time 13 C labeling, an integrated rapid sampling procedure, a LC‐MS analytical method, numerical integration of the system of isotopomer differential equations, and estimation of metabolic fluxes was developed and applied to determine intracellular fluxes in glycolysis, pentose phosphate pathway (PPP), and citric acid cycle (TCA) in Escherichia coli grown in aerobic, glucose‐limited chemostat culture at a dilution rate of D = 0.10 h −1 . Intracellular steady state concentrations were quantified for 12 metabolic intermediates. A total of 90 LC‐MS mass isotopomers were quantified at sampling times t = 0, 91, 226, 346, 589 s and at isotopic stationary conditions. Isotopic stationarity was reached within 10 min in glycolytic and PPP metabolites. Consistent flux solutions were obtained by ID‐ 13 C MFA using isotopic dynamic and isotopic stationary 13 C labeling data and by isotopic stationary 13 C MFA (IS‐ 13 C MFA) using solely isotopic stationary data. It is demonstrated that integration of dynamic 13 C labeling data increases the sensitivity of flux estimation, particularly at the glucose‐6‐phosphate branch point. The identified split ratio between glycolysis and PPP was 55%:44%. These results were confirmed by IS‐ 13 C MFA additionally using labeling data in proteinogenic amino acids (GC‐MS) obtained after 5 h from sampled biomass. Biotechnol. Bioeng. 2008;99: 1170–1185. © 2007 Wiley Periodicals, Inc." @default.
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• W2115026369 date "2007-10-30" @default.
• W2115026369 modified "2023-10-15" @default.
• W2115026369 title "Metabolic flux analysis inEscherichia coli by integrating isotopic dynamic and isotopic stationary13C labeling data" @default.
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• W2115026369 doi "https://doi.org/10.1002/bit.21675" @default.
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