Metabolic profiling of the central metabolic pathways in Escherichia coli
Julia Hiller, doctoral thesis Technische Universität München, 2006
The identification of the controlling metabolic reactions in micro-organisms under process conditions is necessary for a target-oriented bioprocess development. Objective of this study was thus the identification of in-vivo enzyme kinetics of the central metabolic pathways in Escherichia coli. A new sampling device with in-situ inactivation of cell metabolism inside a bioreactor was developed and applied for the measurement of intracellular metabolite concentrations under defined process conditions. The corresponding intracellular metabolic fluxes were estimated based on a stoichiometric model of the central metabolic pathways. In-vivo kinetics of specific enzymes of the central metabolic pathways were identified by making use of the estimated metabolite concentrations and fluxes at different steady states. Thus, for the first time, metabolic control analysis based on in-vivo data was enabled.
- Hiller J, Franco-Lara E, Papaioannou V, Weuster-Botz D (2007): Fast sampling and quenching procedures for microbial metabolic profiling. Biotechnol Let 29: 1161-1167.
- Hiller J, Franco-Lara E, Weuster-Botz D (2007): Metabolic profiling of Escherichia coli cultivations: evaluation of extraction and metabolite analysis procedures. Biotechnol Let 29: 1169-1178.