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General Information
Manuscript title Absolute Metabolite Concentrations and Implied Enzyme Active Site Occupancy in Escherichia coli.
PubMed ID 19561621
Journal Nature Chemical Biology
Year 2009
Authors Bryson D Bennett, Elizabeth H Kimball, Melissa Gao, Robin Osterhout, Stephen J Van Dien, and Joshua D Rabinowitz
Affiliations Department of Chemistry and Lewis-Sigler Institute for Integrative Genomics, Princeton
Keywords not specified
Full text article no file uploaded
Project name not specified


Experiment Description
Organism Escherichia coli
Strain WT K-12 strain NCM3722 of E. coli
Data type metabolites at steady-state
Data units mM
Execution date not specified


Experimental Details
Temperature (°C) 37
pH not specified
Carbon source glucose, glycerol, acetate
Culture mode batch
Process condition aerobic
Dilution rate (h⁻¹) -
Working volume (L) 0.05
Biomass concentration (g/L) Cultures were grown to OD650 of 0.35.
Medium composition

Gutnick minimal complete medium with 4g/L of glucose, glycerol, or acetate as the carbon source. Complete media formulation was 4.7g/L KH2PO4, 13.5g/L K2HPO4, 1 g/L K2SO4, 0.1 g/L MgSO4·7H2O, and final concentrations of 10 mM NH4Cl, and 4g/L of the carbon source (acetate was added as 6.65 g/L of potassium acetate)

General protocol information Sampling method: This liquid culture was grown to OD650~0.1, at which time it was transferred to filter culture as follows:for each filter culture, 5 mL of liquid culture was passed through an 82 mm diameter round nylon filter and the placed cell-side up onto a plate.

Quenching procedure: Cells were filter (0.45-µm), washed with 10 mL of Milli-Q water and plunged into 1 mL of methanol containing 10µL of 0.56 mM PIPES.

Extraction technique: acidic acetonitrile-methanol

Sample analyzing method: NMR, LC-ESI-MS

Methods description - Notes

See more details in Supplementary Methods online.

Data file
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Submission and curation

Entered by: Administrator KiMoSysFirst name: Administrator
Affiliation: INESC-ID/IST
Homepage: http://kdbio.inesc-id.pt/kimosys
Interests: mathematical modeling, accessible data, use of data

Created: 2018-07-06 10:59:59 UTC

Updated: 2020-04-24 16:10:35 UTC

Version: 2

Status: (reviewed) 2018-07-06 11:01:53 UTC

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