DataEntryID 37
    General information
      Manuscript title: Is the glycolytic flux in Lactococcus lactis primarily controlled by the redox charge? Kinetics of NAD(+) and NADH pools determined in vivo by 13C NMR.
      PubMed ID: http://www.ncbi.nlm.nih.gov/pubmed/12011086
      Journal: The Journal of Biological Chemistry
      Year: 2002
      Authors: Ana Rute Neves, Rita Ventura, Nahla Mansour, Claire Shearman, Michael J. Gasson, Christopher Maycock, Ana Ramos and Helena Santos
      Affiliations: Instituto de Teconcologia Química e Biológica, Universidade Nova de Lisboa and Instituto de Biologia Experimental e Tecnológico, Oeiras, Portugal
      Keywords: Lactococcus lactis, metabolism of glucose, NMR, in vivo
      Full text article: https://kimosys.org/rails/active_storage/blobs/eyJfcmFpbHMiOnsibWVzc2FnZSI6IkJBaHBBbWNFIiwiZXhwIjpudWxsLCJwdXIiOiJibG9iX2lkIn19--e19913b0748db4c39d648f28bb62d90fa8a03c9c/Neves_2002.pdf
      Project name: PneumoSyS

    Experiment description
      Organism: Lactococcus lactis
      Strain: MG1363
      Data type: time-series data of metabolites
      Data units: not specified
      Execution date: not specified

    Experimental details
      Temperature (°C): 30.0
      pH: 6.5
      Carbon source: glucose
      Culture mode: batch
      Process condition: anaerobic
      Dilution rate (h⁻¹): not specified
      Working volume: 0.050 L
      Biomass concentration (g/L): not specified
      Medium composition: Chemical-defined medium
      General protocol information: Sampling Method:  Cells were harvested in logarithmic growth phase (A 600 = 2.2); Quenching: Centrifuges, washed, and suspended to a protein concentration of 16,5 mg/ml in 50mM Kpi or Mes/KOA buffer, pH 6,5, for 13C or 31P NMR respectively.; Extraction list: boiling ethanol; Analysis list: NMR;
      Methods description: NMR Experiments - cells were harvested in mid-logarithmic growth phase (A600 = 2.2), centrifuged, washed twice, and suspended to a protein concentration of 16.5 mg/ml in 50 mM KPi or Mes/KOH buffer, pH 6.5, for 13C or 31P NMR experiments, respectively.
Quantification of Products - Lactate, acetoin, acetate, 2,3-butanediol, ethanol, and formate were quantified in NMR sample extracts by 1H NMR [1]. The concentration of minor products (e.g. pyruvate, ethanol, diacetyl) and metabolic intermediates that remained inside the cells (PEP, 3-PGA) was determined from the analysis of 13C spectra of NMR sample extracts as previously described [2]. Intracellular metabolite concentrations were calculated using a value of 2.9 
l/mg protein for the intracellular volume of L. lactis [3]. All NMR spectra of living cells were run at 30 °C with a quadruple-nucleus probe head on a Bruker DRX500 spectrometer. Acquisition of 31P NMR and 13C NMR spectra was performed as described by Neves et al. [2]; however, acquisition parameters for 13C NMR were modified as follows: data size, 16,000; recycle delay, 0.3 s; number of transients, 480. Carbon and phosphorus chemical shifts are referenced to the resonances of external methanol or H3PO4 (85%) designated at 49.3 ppm and 0.0 ppm, respectively.

---------References---------
[1] de Vos, W.M (1999). Curr. Opin, Microbiol. 2, 289-295. http://doi.org/dztzb8
[2] Neves, A. R., Ramos, A., Nunes, M. C., Kleerebezem, M., Hugenholtz, J., de Vos, W. M., Almeida, J., and Santos, H. (1999). Biotechnol. Bioeng. 64, 200–212. http://doi.org/fqrv8j
[3] Poolman, B., Smid, E. J., Veldkamp, H., and Konings, W. N. (1987). J. Bacteriol. 169, 1460–1468.
      Data file: http://kimosys.org/repository/37/download?parameter=1126; 
      Alternative formats: http://kimosys.org/repository/37/attached_files/19/download; http://kimosys.org/repository/37/attached_files/18/download; http://kimosys.org/repository/37/attached_files/17/download; http://kimosys.org/repository/37/attached_files/16/download; http://kimosys.org/repository/37/attached_files/15/download; 

    Submission and curation
      Entered by: Administrator KiMoSys
      Created: 2013-04-24 13:08:01 UTC
      Updated: 2020-04-24 16:10:29 UTC
      Version: 1
      Status: (reviewed) 2014-05-01 22:49:12 UTC
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