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General Information
Manuscript title Nonlinear dependency of intracellular fluxes on growth rate in miniaturized continuous cultures of Escherichia coli.
PubMed ID 16461663
Journal Applied and Environmental Microbiology
Year 2006
Authors Annik Nanchen, Alexander Schicker and Uwe Sauer
Affiliations Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland
Keywords Escherichia coli, continuous culture, intracellular fluxes
Full text article Nanchen_2006.pdf
Project name not specified


Experiment Description
Organism Escherichia coli
Strain MG1655
Data type flux measurements
Data units (mmol/gDW.h)
Execution date not specified


Experimental Details
Temperature (°C) 37.0
pH 7.0
Carbon source glucose
Culture mode chemostat
Process condition aerobic
Dilution rate (h⁻¹) several dilution rates
Working volume (L) 0.010
Biomass concentration (g/L) 0.2-0.5
Medium composition

M9 medium contained (per liter of deionized water) 0.8 g NH4Cl, 0.5 g NaCl, 7.5 g Na2HPO4 · 2H2O, and 3.0 g KH2PO4. The following components were sterilized separately and then added (per liter of final volume): 2 ml of 1 M MgSO4, 1 ml of 0.1 M CaCl2, 0.3 ml of 1 mM filter-sterilized thiamine HCl, and 10 ml of a trace element solution containing (per liter) 1 g of FeCl3 · 6H2O, 0.18 g ZnSO4 · 7H2O, 0.12 g CuCl2 · 2H2O, 0.12 g MnSO4 · H2O, and 0.18 g CoCl2 · 6H2O. Sterilized glucose was added to a final concentration of 1 g per liter.

General protocol information Flux analysis method: 13C constrained MFA

Platform: GC-MS

Methods description - Notes

Metabolic-flux ratio analysis by GC-MS - Samples for gas chromatographymass spectrometry (GC-MS) analysis were prepared as described previously [1]. Briefly, the 13C-labeled chemostat cultures were harvested after the OD600 was stable for at least two volume changes (the min ...

------------References---------------
[1] Fischer, E., and U. Sauer. 2003. Metabolic flux profiling of Escherichia coli mutants in central carbon metabolism using GC-MS. Eur. J. Biochem. 270: 880–891. http://doi.org/dg3q64
[2] Fischer, E., N. Zamboni, and U. Sauer. 2004. High-throughput metabolic flux analysis based on gas chromatography-mass spectrometry derived 13C constraints. Anal. Biochem. 325:308–316. http://doi.org/c6kx35
[3] Zamboni, N., E. Fischer, and U. Sauer. 2005. FiatFlux—a software for metabolic flux analysis from 13C-glucose experiments. BMC Bioinformatics 6:209. http://doi.org/bj2zk2

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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: 2013-06-12 15:55:08 UTC

Updated: 2020-04-24 16:10:33 UTC

Version: 0

Status: (reviewed) 2013-12-06 17:17:38 UTC

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