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General Information
Manuscript title Analysis of Escherichia coli Anaplerotic Metabolism and Its Regulation Mechanisms From the Metabolic Responses to Altered Dilution Rates and Phosphoenolpyruvate Carboxykinase Knockout.
PubMed ID 12966569
Journal Biotechnology and Bioengineering
Year 2003
Authors Yang C, Hua Q, Baba T, Mori H, Shimizu K
Affiliations Metabolome Unit, Institute for Advanced Biosciences, Keio University, Tsuruoka 997-0017, Japan.
Keywords Escherichia coli; metabolic flux; 13C labeling; anaplerotic reaction; phosphoenolpyruvate carboxykinase; in vivo regulation
Full text article Yang2003.pdf
Project name not specified


Experiment Description
Organism Escherichia coli
Strain W3110 and pck mutant
Data type metabolites at steady-state
Data units mM
Execution date not specified


Experimental Details
Temperature (°C) 37
pH 7.0
Carbon source glucose,
Culture mode chemostat
Process condition aerobic
Dilution rate (h⁻¹) 0.1, 0.32, 0.55 (WT ) and 0.1 (pck)
Working volume (L) 1.0
Biomass concentration (g/L) Yield glu,X (g/g) = 0.40 ± 0.02 (D = 0.1h-1), 0.44 ± 0.02 (D = 0.32h-1) and 0.48 ± 0.03 (D = 0.55h-1) for WT; 0.46 ± 0.02 (D = 0.1h-1) for pck
Medium composition

Medium containing (per liter): 5.0 g of glucose, 1.0 g of NH4Cl, 2.7 g of (NH4)2SO4,6.8gofNa2HPO4,3.0gofKH2PO4, 0.6 g of NaCl, 0.2 g ofMgSO4 7H2O, 1.0 µg of thiamine HCl, 2.0 µL of polypropylene glycol 2000 as an antiform agent, and 10 mL of trace element solution [6].

General protocol information Sampling method: not described

Quenching procedure: To rapidly quench the cell metabolism, 5 mL of culture suspension was cooled to 0°C in a −50°C methanol bath within 15–20 s. Cells were separated from the culture medium by centrifugation, and resuspended in cold 100% methanol immediately.

Extraction technique: chloroform

Sample analyzing method: enzymatic

Methods description - Notes

To rapidly quench the cell metabolism, 5 mL of culture suspension was cooled to 0°C in a −50°C methanol bath within 15–20 s. Cells were separated from the culture medium by centrifugation, and resuspended in cold 100% methanol immediately. Intracellular metabolites were extr ...

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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: 2018-07-31 15:50:28 UTC

Updated: 2020-04-24 16:10:36 UTC

Version: 0

Status: (reviewed) 2018-07-31 15:51:08 UTC

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