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General Information info

Manuscript title Metabolic flux analysis for a ppc mutant Escherichia coli based on 13C-labelling experiments together with enzyme activity assays and intracellular metabolite measurements.
PubMed ID 15158257
Journal FEMS Microbiology Letters
Year 2004
Authors Lifeng Peng, Marcos J. Arauzo-Bravo, Kazuyuki Shimizu
Affiliations Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Iizuka, Fukuoka 820-8502, Japan.
Keywords ppc mutant Escherichia coli, 2-Dimensional nuclear magnetic resonance, Gas chromatography-mass spectrometry, Metabolic flux analysis
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Experiment Description info

Organism Escherichia coli
Strain BW25113 and ppc mutant
Data type metabolites at steady-state
Data units mM
Execution date not specified

Experimental Details info

Temperature (0C) 37
pH 7.0
Carbon source glucose,
Culture mode chemostat
Process condition aerobic
Dilution rate (h-1) 0.2
Working volume (L) 1.0
Biomass concentration (g/L) Yield of biomass on glucose (g/g): 0,48 (WT) and 0,55 (ppc)
Medium composition

M9 minimal medium as described previously [3].

General protocol information Sampling method: Cells were harvested by centrifugation at 10,000 g for 10 min.

Quenching procedure: Washed twice with 100 mM Tris-HCl (pH 7.0) containing 20 mM KCl, 5 mM MnSO4, 2 mM DTT and 0.1 mM EDTA, and then resuspended in the same buffer (ca. 15 g wet cells in 50 ml buffer solution).

Extraction technique: sonication

Sample analyzing method: enzymatic

Methods description - Notes

Intracellular metabolite concentrations in quenched cell broth were measured by enzymatic procedures using a fluorescence spectrophotometer (F-4010, Hitachi Co.,
Japan) as described by Schaefer et al. [1] and Bergmeyer [2]. ----------------------------------References--------------------------------------
[1] Schaefer, U., Boos, W., Takors, R. and Weuster-Botz, D. (1999). Anal. Biochem. 270, 88–96. http://doi.org/dt38q9
[2] Bergmeyer, H.U. (1984) Methods of Enzymatic Analysis, 3rd edn,Vol. VI. Metabolites 1: Carbohydrates, pp. 220–227. VCH, Weinheim, Germany.
[3] Zhao, J. and Shimizu, K. (2002). J. Biotechnol. 101, 101–117. http://doi.org/cdjhrn

Data file
Downloadmetabolites KIMODATAID110_v1.xlsx
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Submission and curation info

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 2018-07-29 10:54:27 UTC

Updated 2018-07-29 10:55:30 UTC

Version 1

Status (reviewed) 2018-07-29 10:55:38 UTC




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