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Determination of intermediary metabolites in yeast. Critical examination of the effect of sampling conditions and recommendations for obtaining true levels

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Summary

The effect of sampling conditions on the levels of adenine nucleotides, pyridine nucleotides, glycolytic intermediates and related metabolites in yeast has been studied. A systematic examination of the conditions for harvesting has shown that it can be best accomplished by rapid filtration. Delays in the handling for removal of the medium, as is usual in the process of obtaining a number of data reported in the literature, lead to important changes in some of the metabolites examined. It is also shown that when a washing is imperative it can be carried out with a methanol-water mixture (50/50, v/v) cooled at −40° without loss of intracellular concentrations of non-readily diffusible metabolites.

On the basis of this experience the outline of a generally applicable procedure is presented.

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References

  1. Hanson, C. W. and Dworkin, M., 1974. J. Bacteriol. 118, 486–496.

    PubMed  Google Scholar 

  2. Lowry, O. H., Carter, J., Waed, J. B. and Glaser, L., 1971. J. Biol. Chem. 246, 6511–6521.

    PubMed  Google Scholar 

  3. Rothman, L. B. and Cahib, E., 1969. Biochemistry 8, 3332–3341.

    PubMed  Google Scholar 

  4. Polakis, E. S. and Bartley, W., 1966. Biochem. J. 99, 521–533.

    PubMed  Google Scholar 

  5. van Wijk, R., Slavenburg, J. H. and van de Poll, K. W., 1973. Proc. Koninkl. Nederl. Akademie van Wetenschappen, Series C 76, 512–527.

    Google Scholar 

  6. Gancedo, J. M. and Gancedo, C., 1973. Biochimie 55, 205–211.

    PubMed  Google Scholar 

  7. Olson, B. H. and Johnson, M. J., 1949. J. Bacteriol 57, 235–246.

    Google Scholar 

  8. Weibel, K. E., Mor, J. R. and Fiechter, A., 1974. Anal. Biochem. 58, 208–216.

    PubMed  Google Scholar 

  9. Bergmeyer, H. U., 1974. Methods in Enzymic Analysis, vol. III and IV, 2nd ed. Academic Press, New York.

    Google Scholar 

  10. Greenbaum, A. L., Clark, J. B. and McLean, P., 1965. Biochem. J. 95, 161–166.

    PubMed  Google Scholar 

  11. Conway, E. J. and Downey, M., 1950. Biochem. J. 47, 347–355.

    Google Scholar 

  12. Atkinson, D. E., 1969. Ann. Rev. Microbiol. 23, 47–68.

    Google Scholar 

  13. Heredia, C. F., Sols, A. and DelaFuente, G., 1968. Eur. J. Biochem. 5, 321–329.

    PubMed  Google Scholar 

  14. Holzer, H., Schultz, G. and Lynen, F., 1956. Biochem. Z. 328, 252–263.

    PubMed  Google Scholar 

Download references

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Authors and Affiliations

  1. Instituto de Enzimología del Consejo Superior de Investigaciones Científicas, Facultad de Medicina de la Universidad Autónoma, Madrid-34, Spain

    María J. Sáez & Rosario Lagunas

Authors
  1. María J. Sáez
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  2. Rosario Lagunas
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Sáez, M.J., Lagunas, R. Determination of intermediary metabolites in yeast. Critical examination of the effect of sampling conditions and recommendations for obtaining true levels. Mol Cell Biochem 13, 73–78 (1976). https://doi.org/10.1007/BF01837056

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  • DOI: https://doi.org/10.1007/BF01837056

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