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General Information
Manuscript title Metabolic Fluxes during Strong Carbon Catabolite Repression by Malate in Bacillus subtilis.
PubMed ID 19917605
Journal The Journal of Biological Chemistry
Year 2009
Authors Roelco J. Kleijn, Joerg M. Buescher, Ludovic Le Chat, Matthieu Jules, Stephane Aymerich and Uwe Sauer
Affiliations Institute of Molecular System Biology, ETH Zurich, CH-8093 Zurich, Switzerland
Keywords Metabolism, Metabolism/Gluconeogenesis, Metabolism/Intermediary, Metabolism/Regulation, Metabolism/Tricarboxylic Acid Cycle, Methods/Mass Spectrometry, flux analysis, metabolomics
Full text article Kleijn_2010.pdf
Project name not specified

Experiment Description
Organism Bacillus subtilis
Strain Wild-type BSB168 trp+
Data type metabolites at steady-state
Data units mM
Execution date not specified

Experimental Details
Temperature (°C) 37
pH not specified
Carbon source glucose, glucose + malate, malate
Culture mode batch
Process condition aerobic
Dilution rate (h⁻¹) -
Working volume (L) 0.03
Biomass concentration (g/L) 0.5
Medium composition

M9 minimal medium (per liter): 8.5 g of Na2HPO4.2H20, 3 g of KH2PO4, 1 g of NH4Cl, 0.5 g of NaCl. The following components were sterilized separately and then added (per liter of final medium): 1 ml of 0.1 m CaCl2.2H2O, 1 ml of 1 m MgSO4.7H2O, 1 ml of 50 mm FeCl3.6H2O, and 10 ml of trace salt solution. The trace salts solution contained (per liter): 170 mg of ZnCl2, 100 mg of MnCl2.4H2O, 60.0 mg of CoCl2.6H2O, 60.0 mg of Na2MoO4.2H2O, and 43.0 mg CuCl2.2H2O. When preparing the medium, the base salts were added first followed by CalCl2, MgSO4, FeCl3, and finally the trace elements.

General protocol information Sampling method: Liquid culture was grown to OD650 of ~0.1, at which time it was transferred to filter culture as follows: for each filter culture, 5 mL of liquid culture was passed through an 82mm diameter round nylon filter and placed cell-side up onto a agarose plate

Quenching procedure: rapid centrifugation method where 1 ml of culture broth was transferred into a 1.5-ml tube and centrifuged for 15 s at 14,000×g in a tabletop centrifuge. The supernatant was decanted, and the pellet was frozen in liquid nitrogen.

Extraction technique: hot ethanol

Sample analyzing method: LC-MS, GC-TOF

Methods description - Notes

GC-TOF Workflow: Dried aliquots were derivatized with 15 μl of either TMS-reagent (N-methyl-N-(trimethylsilyl)trifluoroacetamide, Fluka, Buchs, Switzerland) or TBDMS reagent (N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide, Fluka), and aliquots of 5 μl were injected int ...

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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: 2018-07-09 16:17:27 UTC

Updated: 2020-04-24 16:10:35 UTC

Version: 2

Status: (reviewed) 2018-07-09 16:17:31 UTC

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Downloads: 26

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