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Carbon loss at high T

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SEEK ID: https://demo.fairdomhub.org/studies/9

Central Carbon Metabolism of Sulfolobus solfataricus

Projects: SulfoSys

Jacky Snoep

Experimentalists: Theresa Kouril

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Created: 29th Jan 2013 at 15:11

Last updated: 28th May 2015 at 10:50

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SulfoSys

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

Programme: SysMO

Public web page: http://sulfosys.com/

Central Carbon Metabolism of Sulfolobus solfataricus
SulfoSys

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.

Snapshots: Snapshot 1

Model reconstituted system
SulfoSys

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

Contributor: Jacky Snoep

Biological problem addressed: Model Analysis Type

Snapshots: No snapshots

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.

Contributor: Jacky Snoep

Assay type: Experimental Assay Type

Technology type: Progressive Curve Experiment

Snapshots: No snapshots

Model simulation and Exp data for reconstituted system
SulfoSys

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

Creator: Jacky Snoep

Contributor: Jacky Snoep

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Metabolite concentrations during reconstituted enzyme incubation
SulfoSys

The purified enzymes, PGK, GAPDH, TPI and FBPAase were incubated at 70 C en conversion of 3PG to F6P was followed

Creator: Jacky Snoep

Contributor: Jacky Snoep

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Mathematical model for the combined four enzyme system
SulfoSys

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

Creator: Jacky Snoep

Contributor: Jacky Snoep

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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, Jacky Snoep

Date Published: 22nd Aug 2013

Journal: FEBS J

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