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General Information
Manuscript title Intracellular metabolite pool changes in response to nutrient depletion induced metabolic switching in Streptomyces coelicolor.
PubMed ID not specified
Journal Metabolites
Year 2012
Authors Alexander Wentzel, Harvard Sletta, Stream Consortium, Trond E. Ellingsen and Per Bruheim
Affiliations Department of Biotechnology, SINTEF Materials and Chemistry, Sem Saelandsvei 2a, N-7465 Trondheim, Norway and Department of Biotechnology, Norwegian University of Science and Technology, Sem Saelandsvei 6/8, N-7491 Trondheim, Norway
Keywords Streptomyces coelicolor, transition phase, metabolic switching metabolite profiling
Full text article Wentzel_2012.pdf
Project name not specified

Experiment Description
Organism Streptomyces coelicolor
Strain M145 (W.T.) and phoP mutant (INB201)
Data type time-series data of metabolites
Data units µM
Execution date not specified

Experimental Details
Temperature (°C) 30.0
pH not specified
Carbon source glucose
Culture mode batch
Process condition aerobic
Dilution rate (h⁻¹)
Working volume (L) 1.8
Biomass concentration (g/L) see worksheets
Medium composition

(SSBM-P) medium for studying the effect of phosphate depletion: Na-glutamate, 55.2 g/L; D-glucose, 40 g/L; MgSO4, 2.0 mM; phosphate, 4.6 mM; supplemented minimal medium trace element solution SMM-TE [39], 8 mL/L and TMS1, 5.6 mL/L. TMS1 consisted of FeSO4 × 7 H2O, 5 g/L; CuSO4 × 5 H2O, 390 mg/L; ZnSO4 × 7 H2O, 440 mg/L; MnSO4 × H2O, 150 mg/L; Na2MoO4 × 2 H2O, 10 mg/L; CoCl2 × 6 H2O, 20 mg/L, and HCl, 50 ml/L. (SSBM-E) medium for studying the effect of L-glutamate depletion: identical to SSBM-P except for the concentrations of Na-glutamate and phosphate adjusted to be 15 g/L and 9.2 mM, respectively.

General protocol information Sampling method: Samples for metabolite profiling were withdrawn from the cultivations in 1–2 h time intervals.

Quenching procedure: Quenching procedure: 5 mL culture sample was withdrawn from the fermentation vessel and immediately applied to a 0.8 μm Supor 800 filter (Pall). On the filter disc, the cells were subsequently washed twice with one volume 2.63% (w/v) NaCl solution each.

Extraction technique: methanol

Sample analyzing method: GC-MS, LC-MS

Methods description - Notes

Metabolite Extraction - samples stored at −80 °C were completely thawed on an ethanol bath at −23 °C. An internal standard mix was added to each 25 mL sample (biomass from 5 mL sample on filter in 25 mL 60% methanol solution) yielding final concentrations of 3.34 mM D3-alani ...

[1] Villas-Boas, S.G.; Delicado, D.G.; Akesson, M.; Nielsen, J. Simultaneous analysis of amino and nonamino organic acids as methyl chloroformate derivatives using gas chromatography-mass spectrometry. Anal. Biochem. 2003, 322, 134–138.
[2] Luo, B.; Groenke, K.; Takors, R.; Wandrey, C.; Oldiges, M. Simultaneous determination of multiple intracellular metabolites in glycolysis, pentose phosphate pathway and tricarboxylic acid cycle by liquid chromatography-mass spectrometry. J. Chromatogr. A 2007, 1147, 153–164.

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Data file
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KIMODATAID76_M145_SSBM-P_timeseries.csv Preview
KIMODATAID76_M145_SSBM-E_timeseries.csv Preview
KIMODATAID76_INB201_SSBM-P_timeseries.csv Preview
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Submission and curation

Entered by: Administrator KiMoSysFirst name: Administrator
Affiliation: INESC-ID/IST
Interests: mathematical modeling, accessible data, use of data

Created: 2013-06-28 14:43:07 UTC

Updated: 2020-04-24 16:10:34 UTC

Version: 0

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

Views: 153

Downloads: 30

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