| Range |
Table - link
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| Organism |
Microbes |
| Reference |
Karen van Eunen & Barbara M. Bakker, The importance and challenges of in vivo-like enzyme kinetics, Perspectives in Science, Volume 1, Issues 1–6, May 2014, Pages 126–130, link pdf link p.127 table 1 |
| Primary Source |
García-Contreras R, Vos P, Westerhoff HV, Boogerd FC. Why in vivo may not equal in vitro - new effectors revealed by measurement of enzymatic activities under the same in vivo-like assay conditions. FEBS J. 2012 Nov279(22):4145-59. doi: 10.1111/febs.12007. AND Leroux AE, Haanstra JR, Bakker BM, Krauth-Siegel RL. Dissecting the catalytic mechanism of Trypanosoma brucei trypanothione synthetase by kinetic analysis and computational modeling. J Biol Chem. 2013 Aug 16 288(33):23751-64. doi: 10.1074/jbc.M113.483289. AND van Eunen K et al., Measuring enzyme activities under standardized in vivo-like conditions for systems biology. FEBS J. 2010 Feb277(3):749-60. doi: 10.1111/j.1742-4658.2009.07524.x. AND Goel A, Santos F, Vos WM, Teusink B, Molenaar D. Standardized assay medium to measure Lactococcus lactis enzyme activities while mimicking intracellular conditions. Appl Environ Microbiol. 2012 Jan78(1):134-43. doi: 10.1128/AEM.05276-11.PubMed ID22978366, 23814051, 20067525, 22020503
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| Comments |
P.127 right column top paragraph: "In vivo-like assay media have been developed for S. cerevisiae, L. lactis, E. coli and T. brucei. For E. coli and T. brucei, the assay medium was completely based on ion concentrations reported in the literature (primary sources García-Contreras et al., 2012 Leroux et al., 2013), while in S. cerevisiae and L. lactis the ion concentrations were determined by an elemental-composition analysis supplemented with published data (primary sources van Eunen et al., 2010 Goel et al., 2012). Table 1 shows the composition of the resulting assay buffers. The main differences were in the phosphate concentration and in the choice of the anion that compensates for the high cation concentration. The phosphate concentration in the cytosol depends strongly on the concentration of phosphate in the growth medium. The two assay media with the highest concentrations of phosphate, i.e. those for S. cerevisiae and E. coli, were based on cells that had been cultivated at a high concentration of phosphate (35–50 mM) in the growth medium. Thus, these high intracellular phosphate concentrations are not inherent properties of the organisms, but rather refer to the conditions under which they have been cultivated. This illustrates that physiological assay media should not only be tailored to the organism of interest, but also to the condition of interest." |
| Entered by |
Uri M |
| ID |
113251 |