Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Determination of the Intracellular Concentrations of Metabolites in Escherichia coli Collected during the Exponential and Stationary Growth Phases using Liquid Chromatography-Mass Spectrometry

  • Park, Chang-Hun (Department of Chemical and Biomolecular Engineering, Sogang University) ;
  • Park, Chang-Hun (Department of Chemistry, Sogang University) ;
  • Lee, Youn-Jin (Department of Chemistry, Sogang University) ;
  • Lee, Sang-Yup (Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology) ;
  • Oh, Han-Bin (Department of Chemistry, Sogang University) ;
  • Lee, Jin-Won (Department of Chemical and Biomolecular Engineering, Sogang University)
  • Received : 2010.09.30
  • Accepted : 2010.12.03
  • Published : 2011.02.20
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Abstract

In the present study, we demonstrate that SRM LC-MS/MS method developed by Luo et al. (ref. 10) can be successfully applied to the quantitative analysis of intracellular metabolites in E. coli that are collected at the exponential and stationary growth phases. A focus is given on measuring the changes in the concentrations of intracellular metabolites in batch cultures, which were induced during both the dynamically changing exponential and stationary growth phases. The following intracellular metabolites are quantified in the exponential and stationary phases of E. coli growth, using the SRM mode of a triple quadrupole mass spectrometer: glucose-1-phosphate, fructose-1,6-bisphosphate, phosphoenolpyruvate, pyruvate, acetyl-coenzyme A, 6-phosphogluconate, ribulose-5-phosphate, xylulose-5-phosphate, erythrose-4-phosphate. The determined intracellular metabolite concentration profiles are shown to be in a good agreement with the growth profiles of E. coli, which clearly indicates that SRM LC-MS/MS can be successfully used for following the metabolite changes induced at different growth stages.

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References

  1. Fiehn, O. Comparative and Functional Genomics 2001, 2, 155. https://doi.org/10.1002/cfg.82
  2. Fiehn, O.; Kopka, J.; Dormann, P.; Altmann T.; Trethewey, R. N.; Willmitzer, L. Nature Biotechnol. 2000, 18, 1157.
  3. Gipson, G. T.; Tatsuoka, K. S.; Sokhansanj, B. A.; Ball, R. J.; Connor, S. C. Metabolomics 2008, 4, 94. https://doi.org/10.1007/s11306-007-0096-9
  4. Hollywood, K.; Brison, D. R.; Goodacre, R. Proteomics 2006, 6, 4716. https://doi.org/10.1002/pmic.200600106
  5. Dettmer, K.; Aronov, P. A.; Hammock, B. D. Mass Spectrometry Reviews 2007, 26, 51. https://doi.org/10.1002/mas.20108
  6. Shulaev, V. Briefings in Bioinformatics 2006, 7, 128. https://doi.org/10.1093/bib/bbl012
  7. Buchholz, A.; Hurlebaus, J.; Wandrey, C.; Takors, R. Biomolecular Engineering 2002, 19, 5. https://doi.org/10.1016/S1389-0344(02)00003-5
  8. Dunn, W. B.; Ellis, D. I. Trends in Analytical Chemistry 2005, 24, 285. https://doi.org/10.1016/j.trac.2004.11.021
  9. Moco, S.; Bino, R. J.; De Vos, R. C. H.; Vervoort, J. Trends in Analytical Chemistry 2007, 26, 855. https://doi.org/10.1016/j.trac.2007.08.003
  10. Luo, B.; Groenke, K.; Takors, R.; Wandrey, C.; Oldiges, M. Journal of Chromatography A 2007, 1147, 153. https://doi.org/10.1016/j.chroma.2007.02.034
  11. Coulier, L.; Bas, R.; Jespersen, S.; Verheij, E.; Van der werf, M. J.; Hankemeier, T. Analytical Chemistry 2006, 78, 6573. https://doi.org/10.1021/ac0607616
  12. Huck, J. H. J.; Struys, E. A.; Verhoeven, N. M.; Jakobs C.; Van der knap, M. S. Clinical Chemistry 2003, 49, 1375. https://doi.org/10.1373/49.8.1375
  13. Sekiguchi, Y.; Mitsuhashi, N.; Kokaji, T.; Miyakoda, Hi.; Mimura, T. Journal of Chromatography A 2005, 1085, 131. https://doi.org/10.1016/j.chroma.2005.01.098
  14. Van dam, J. C.; Eman, M. R.; Frank, J.; Lange, H. C.; Van dedem, G. W. K.; Heijnen S. J. Anal. Chim. Acta 2002, 460, 209.
  15. Chassgnolie, C.; Noisommit-Rizzi, N.; Schmid, J. W.; Mauch, K.; Reuss, M. Biotechnol. Bioeng. 2002, 79, 53. https://doi.org/10.1002/bit.10288
  16. Winder, C. L.; Dunn, W. B.; Schuler, S.; Broadhurst, D.; Jarvis, R.; Stephens, G. M.; Goodacre, R. Analytical Chemistry 2008, 80, 2939. https://doi.org/10.1021/ac7023409
  17. Maharjan, R. P.; Ferenci, T. Analytical Biochemistry 2003, 313, 145 https://doi.org/10.1016/S0003-2697(02)00536-5
  18. Miller, J. C.; Miller, J. N. Statistics for Analytical Chemistry; Ellis Horwood Ltd.: Chichester, UK, 1988.
  19. Rao, D. G. In Introduction to Biochemical Engineering; McGraw- Hill: New delhi, India, 2006; p169.
  20. Soga, T.; Ueno, Y.; Naraoka, H.; Ohashi, Y.; Tomita, M.; Nishioka, T. Analytical Chemistry 2002, 74, 2233. https://doi.org/10.1021/ac020064n
  21. Pramanik, J.; Keasling, J. D. Biotechnol Bioeng. 1997, 56, 399
  22. Kolter, R.; Siegele, D. A.; Torma, A. Annual Reviews Microbiol. 1993, 47, 855. https://doi.org/10.1146/annurev.mi.47.100193.004231

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