The c-Met inhibitor influence of the input rup on the concentrations c(rup) in steady-state might be calculated from (27) where denotes the

Jacobian matrix (or elasticities) of the rates ri with respect to the concentrations cj: (28) In case the Jacobian D(c(rup)) is invertible, the dependency of the steady-state concentration c(rup) on the input rup Inhibitors,research,lifescience,medical is uniquely determined by (29) Equation (29) allows to calculate the slope for each component in dependence of the elasticities given in D. This will be important for the characterization of the steady-state solutions for given uptake rates. 3.2.3. Dynamic Network Analysis Based on the reaction scheme above, differential equations (o.d.e.) are set up and kinetic parameters

are either taken from network component analysis or estimated based on the experimental data. The dynamic system comprises differential equations for substrates (glucose, acetate), biomass, metabolites glucose Inhibitors,research,lifescience,medical 6-phosphate, fructose 1,6-bisphosphate, PEP and pyruvate. Furthermore, gene expression and control via FruR of the following enzymes is considered: phosphofruktokinase (PfkA), pyruvate kinase (pykF), and a lumped enzyme for glycolytic reactions. Simulation studies and parameter estimation are performed with MATLAB. Some of the enzymes of Inhibitors,research,lifescience,medical glycolysis Inhibitors,research,lifescience,medical are subject to transcriptional control by FruR. These enzymes are taken into account in the model with additional equations. Taking the simplified form

for the enzymes according to [3], the steady state value of an enzyme is proportional to the transcription factor activity that in turn is determined by the concentration of the metabolite fructose-1,6-bisphosphate (F16BP). Therefore, the steady state of the enzyme corresponds directly to ligand concentration and the respective parameter κi is determined with NCA (see Material and Methods): (30) with κi being the entry in the coupling matrix K. The complete dynamical system reads Inhibitors,research,lifescience,medical as follows: (31) (32) (33) (34) (35) (36) (37) (38) (39) tuclazepam (40) (41) In addition to the kinetic expressions given in the text, the following rate law is used to calculate the growth rate μ based on the yield coefficient Y: (42) Yield coefficient Y was determined as follows: for the seven experiments, the individual yield coefficients were determined by linear regression, finally a mean value for all experiments was calculated. The following kinetic parameters in Table 6 are used for dynamical simulation studies: Table 6 Summary of the kinetic parameters. gglc molecular weight for glucose. Basic units are OD (for biomass), μmol (for substrate), and hours (for time). a Taking a value of 0.32 g/OD (determined experimentally for a different study) this corresponds …