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Model simulation and Exp data for reconstituted system
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Experimental data for the reconstituted system are plotted together with the model prediction

SEEK ID: https://demo.fairdomhub.org/data_files/52?version=1

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Created: 2nd Feb 2013 at 20:55

Last updated: 28th May 2015 at 08:50

Last used: 26th Sep 2021 at 19:26

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

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An investigation in the CCM of S. solfataricus with a focus on the unique temperature adaptations and regulation; using a combined modelling and experimental approach.

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Carbon loss at high T
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No description specified

Person responsible: Jacky Snoep

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Reconstituted system reference state
SulfoSys

The four purified enzymes were incubated in assay buffer and consumption of 3PG and production of F6P were measured in time, together with GAP and DHAP concentrations.

Submitter: Jacky Snoep

Assay type: Experimental Assay Type

Technology type: Progressive Curve Experiment

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Model reconstituted system
SulfoSys

Mathematical model for the reconstituted system with PGK, GAPDH, TPI and FBPAase.

Submitter: Jacky Snoep

Biological problem addressed: Model Analysis Type

Snapshots: No snapshots

Intermediate instability at high temperature leads to low pathway efficiency for an in vitro reconstituted system of gluconeogenesis in Sulfolobus solfataricus
SulfoSys

Abstract (Expand)

Four enzymes of the gluconeogenic pathway in Sulfolobus solfataricus were purified and kinetically characterized. The enzymes were reconstituted in vitro to quantify the contribution of temperature … instability of the pathway intermediates to carbon loss from the system. The reconstituted system, consisting of phosphoglycerate kinase, glyceraldehyde 3-phosphate dehydrogenase, triose phosphate isomerase and the fructose 1,6-bisphosphate aldolase/phosphatase, maintained a constant consumption rate of 3-phosphoglycerate and production of fructose 6-phosphate over a 1-h period. Cofactors ATP and NADPH were regenerated via pyruvate kinase and glucose dehydrogenase. A mathematical model was constructed on the basis of the kinetics of the purified enzymes and the measured half-life times of the pathway intermediates. The model quantitatively predicted the system fluxes and metabolite concentrations. Relative enzyme concentrations were chosen such that half the carbon in the system was lost due to degradation of the thermolabile intermediates dihydroxyacetone phosphate, glyceraldehyde 3-phosphate and 1,3-bisphosphoglycerate, indicating that intermediate instability at high temperature can significantly affect pathway efficiency.

Authors: T. Kouril, D. Esser, J. Kort, H. V. Westerhoff, B. Siebers, J. L. Snoep

Date Published: 22nd Aug 2013

Publication Type: Not specified

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