Central Carbon Metabolism of Sulfolobus solfataricus

Silicon cell model for the central carbohydrate metabolism of the archaeon Sulfolobus solfataricus under temperature variation

Mathematical model of a subset of reactions comprising the three most temperature sensitive intermediates of the gluconeogenic pathway in S. solfataricus

By varying the input or output flux via PGK and FBPAase manipulation respectively the pathway efficiency can be optimised.

Kinetic characterisation of Phosphoglycerokinase

Kinetic characterization of the FBPA/ase with respect to GAP en DHAP saturation.

GAP and DHAP concentrations were measured over time at 70C.

Combined plot fo the model simulations and the experimental data for the PGK and FBPAase titrations.

Model simulation showing the effect of varying PGK or FBPAase concentration on the efficiency for 3PG to F6P conversion.

Experimental data for the reconstituted system are plotted together with the model prediction

Saturation of PGK with 3PG

The PGK, GAPDH, TPI and FBPAase were modelled together using the individual rate equations. Closed system.

An exponential decay model was fitted to the experimental data set of GAP degradation at 70C. The model is uploaded as an SBML file. A data file showing the model fit to the experimental data set is included. The rate constant for GAP degradation at 70C was estimated to be 0.056 1/min, equivalent to a half life time of 12.4 min.

An exponential decay model was fitted to the experimental data set of DHAP degradation at 70C. The model is uploaded as an SBML file. A data file showing the model fit to the experimental data set is included. The rate constant for DHAP degradation at 70C was estimated to be 0.0225 1/min, equivalent to a half life time of 30.8 min.

Mathematical model for FBPAase kinetics

Mathematical model for GAPDH kinetics, saturation for GAP, BPG, NADP, NADPH, Pi.

Cloning and heterologous expression of gluconeogenic enzymes in E. coli

Preparation of cell free extracts of the recombinant E. coli strains expressing the respective S. solfataricus enzymes.

The recombinant enzymes were purified from the cell free extracts of E. coli expressing the respective enzymes.