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General Information
| Manuscript title | Quantification of intracellular metabolites in Escherichia coli K12 using liquid chromatographic-electrospray ionization tandem mass spectrometric techniques. |
| PubMed ID | 11488613 |
| Journal | Analytical Biochemistry |
| Year | 2001 |
| Authors | Arne Buchholz, Ralf Takors, and Christian Wandrey |
| Affiliations | Institute of Biotechnology, Research Center Juelich, 52425 Juelich, Germany |
| Keywords | Escherichia coli, intracellular metabolites, ESI, extraction, LC-MS, quantification |
| Full text article | no file uploaded |
| Project name | not specified |
Experiment Description
| Organism | Escherichia coli |
| Strain | K-12 DSM |
| Data type | metabolites at steady-state |
| Data units | mM |
| Execution date | not specified |
Experimental Details
| Temperature (°C) | not specified |
| pH | not specified |
| Carbon source | glucose |
| Culture mode | chemostat |
| Process condition | aerobic |
| Dilution rate (h⁻¹) | 0.125 |
| Working volume (L) | 0.4 |
| Biomass concentration (g/L) | not specified |
| Medium composition | Minimal medium: 30 g/L glucose, 0.2 g/L NH4Cl, 2.0 g/L (NH4)2SO4, 3.25 g/L KH2PO4, 2.5 g/L K2HPO4, 1.5 g/L NaH2PO4, 0.5 g/L MgSO4; trace substances: 1.0E-2 g/L CaCl2, 5.0E-4 g/L ZnSO4, 2.5E-4 CuCl2, 2.5E-3 g/L MnSO4, 1.75E-3 g/L CoCl2, 1.25E-4 g/L H3BO3, 2.5E-3 g/L AlCl3, 5.0E-4 g/L Na2MoO4, 1.0E-2 g/L FeSO4. |
| General protocol information |
Sampling method:
Samples are taken after the cultures have remained constant at a cell density of 10.0 g/L for at least three residence times, thus ensuring steady-state conditions. Quenching procedure: Sampling was performed using precooled syringes containing 15 ml of the quenching fluid, 60%, v/v, aqueous methanol containing 70 mM Hepes (2-[4-(2-hydroxyethyl)- 1-piperazinyl]ethanesulfonic acid). Extraction technique: methanol-water Sample analyzing method: LC-ESI-MS |
| Methods description - Notes | Identification and Quantification of Metabolites - compounds were identified in cell extracts using retention time, m/z, specific fragmentation patterns, and spiking with known compounds. Quantification was accomplished via the [M-H]2 ion by applying the standard addition me ... ------------References------------
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Administrator KiMoSysFirst name: Administrator
Affiliation: INESC-ID/IST
Homepage: http://kdbio.inesc-id.pt/kimosys
Interests: mathematical modeling, accessible data, use of data
Created: 2013-05-29 15:18:00 UTC
Updated: 2020-04-24 16:10:32 UTC
Version: 0
Status: (reviewed) 2013-12-06 17:17:38 UTC
Views: 294
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