A mathematical model of metabolism and regulation provides a systems-level view of how Escherichia coli responds to oxygen

"Systems Understanding of Microbial Oxygen responses" (SUMO) investigates how Escherichia coli senses oxygen, or the associated changes in oxidation/reduction balance, via the Fnr and ArcA proteins, how these systems interact with other regulatory systems, and how the redox response of an E. coli population is generated from the responses of single cells. There are five sub-projects to determine system properties and behaviour and three sub-projects to employ different and complementary modelling
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Glucose transporter mutants were analyzed under aerobic and aerobic conditions in batch cultures with glucose as substrate. Acetate formation rates and glucose consumption rates were measured, as well as extracellular cAMP concentrations.

MG1655 and mutant strains with defects in glucose transport systems were analyzed in aerobic and anaerobic batch cultures.

ArcA phosphorylation in chemostat cultures grown at different aerobiosis levels was quantitated by Phos-tag SDS-PAGE gel analysis and subsequent immunodetection of ArcA.

This experiment uses a low-copy plasmid based system (MG1655 Δlac FF(-41.5)/RW50) for measuring FNR activity. Initial acetate calibration of the chemostat with the MG1655 Δlac strain was carried out, with β-galactosidase activity from the FF(-41.5)/RW50 reporter plasmid measured at 100%, 80%, 50%, 20% and 0% aerobiosis levels. Finally, the aerobiosis levels were re-determined by calculating the actual acetate flux in the sampled chemostat runs.

Note: the strain used (MG1655 Δlac) is not the same
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The task of this assay is to determine the impact of oxygen availability on the concentrations of metabolites from different central metabolic pathways. The focus lies on metabolites connected to glycolysis, tri-carbon-acid-cycle and energy metabolism. All strains have been cultured and analysed according to the SOPs listed below

This assay describes the determination of concentrations and ratio of metabolites of adenine nucleotides (NAD and NADH).
These metabolites have been extracted from Escherichia coli MG1655 and isgenic mutant strains.

This assay describes the determination of concentrations and ratio of metabolites of ubiquinones (oxidised and reduced form).
These metabolites have been extracted from Escherichia coli MG1655 and isgenic mutant strains.

This assay describes how to analyze gene expression rates via RT-PCR.

This document describes by-product formation rates measured in MG1655 at steady-state conditions in Infors-Multifors-Bioreactors.

This data file shows results from the different chemostat experiments.Gene expression rates are presented.

Measurement of by-product formation rates, substrate consumption rates and extracellular concentrations of cAMP under aerobic batch conditions with glucose as substrate.

Transcription of glucose uptake transport systems in a mutant background under anaerobic batch conditions.

Transcription of glucose uptake transport systems in a mutant background under aerobic batch conditions.

Measurement of by-product formation rates, substrate consumption rates and extracellular concentrations of cAMP under anaerobic batch conditions with glucose as substrate.

A .pdf files showing graphs of FNR activity at varying aerobiosis levels

Top graph shows no acetate re-calibration
Bottom graph shows data with acetate re-calibration

This .csv file shows FNR activity at different aerobiosis levels

Columns
a_old Desired aerobiosis level (Aerobiosis units or %AAU)
Bgal_activity Average FNR reporter activity (Miller units)
Bgal_STDEV STDEV of FNR reporter activities (Miller units)
rDOT(μM) Dissolved oxygen tension (μM)
DCW DCW (g/L)
[acetate] Extracellular acetate concentration (mM)
Qacetate Acetate flux (mmoles/h/gDCW)
a_new Actual aerobiosis level in sampled chemostat

his document contains data for quinone concentrations and ratios of ubichinon and menachinon.These data have been generated for analysis of MG1655 at steady-state conditions in Infors-Multifors-Bioreactors. All measurements have been done in Amsterdam at the group of Prof. Joost Teixeira de Mattos.

Duplicates of samples for quinones were taken after 20 hours of steady state conditions. Chemostat experiments were carried out in Infors-Bioreactors at 0, 20, 50, 80, 100 and 120% aerobiosis. All data
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This is the csv version of the file: 091016 quinones complete data magdeburg

This document contains data for quinone concentrations and ratios of ubichinon and menachinon.These data have been generated for analysis of MG1655 at steady-state conditions in Infors-Multifors-Bioreactors. All measurements have been done in Amsterdam at the group of Prof. Joost Teixeira de Mattos.

Duplicates of samples for quinones were taken after 20 hours of steady state conditions. Chemostat experiments were carried
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The model describes the catabolism of Escherichia coli and its regulation. The metabolic reactions are modeled by the thermokinetic model formalism. The model is simplified by assuming rapid equilibrium of many reactions. Regulation is modeled by phenomenological laws describing the activation or repression of enzymes and genes in dependence of metabolic signals. The model is intended to describe the behavior of E. coli in a chemostat culture in depedence on the oxygen supply.

The model is described
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