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Applied Biochemistry and Biotechnology

, Volume 13, Issue 1, pp 15–27 | Cite as

Liquid—liquid extraction of lactate dehydrogenase from muscle using polymer-bound triazine dyes

  • G. Johansson
  • M. Joelsson
Original Articles

Abstract

An extract from porcine muscle containing soluble enzymes has been partitioned between the two liquid phases of an aqueous, biphasic system consisting of dextran, polyethylene glycol, and water. The influence of polymer-bound triazine dyes (Cibacron blue F3G-A and Procion yellow HE-3G) on the partition of lactate dehydrogenase and total protein was studied. The effects of pH and concentrations of polymers and buffer on this so-called affinity partitioning were also determined. The two-phase systems were used in extraction procedures for purification of lactate dehydrogenase to a specific activity of 456–494 U (7.6–8.4 μkat) per mg protein. The use of these systems for extraction of enzymes in technical scale is discussed.

Index entries

Lactate dehydrogenase liquid-liquid extraction of affinity partitioning liquid-liquid extraction two-phase systems triazine dyes, polymer bound, influence of on partitioning of lactate dehydrogenase and total protein 

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References

  1. 1.
    Johansson, G., Hartman, A., and Albertsson, P.-Å. (1973),Eur. J. Biochem. 33, 379.CrossRefGoogle Scholar
  2. 2.
    Kula, M.-R., Kroner, K. H., and Hustedt, H. (1982), inAdvances in Biochemical Engineering, vol. 24, Fiechter, A., ed., Springer-Verlag, Berlin, pp. 73–118.Google Scholar
  3. 3.
    Johansson, G., and Joelsson, M. (1985),Enzyme Microb. Technol,7, 629.CrossRefGoogle Scholar
  4. 4.
    Shanbhag, V. P., and Johansson, G. (1974),Biochem. Biophys. Res. Commun. 61, 1141.CrossRefGoogle Scholar
  5. 5.
    Axelsson, C.-G., and Shanbhag, V. P. (1976),Eur. J. Biochem. 71, 419.CrossRefGoogle Scholar
  6. 6.
    Kroner, H. K., Cordes, A., Schelper, A., Morr, M., Bückmann, A. F., and Kula, M.-R. (1982), inAffinity Chromatography and Related Techniques, Gribnau, T. C. J., Visser, J., and Nivard, R. J. F., eds., Elsevier, Amsterdam, pp. 491–501.Google Scholar
  7. 7.
    Johansson, G., Kopperschläger, G., and Albertsson, P.-Å. (1983),Eur. J. Biochem. 131, 589.CrossRefGoogle Scholar
  8. 8.
    Kopperschläger, G., Lorenz, G., and Usbeck, E. (1983),J. Chromatogr. 259, 97.CrossRefGoogle Scholar
  9. 9.
    Dean, P. D. G., and Watson, D. H. (1979),J. Chromatogr. 165, 301.CrossRefGoogle Scholar
  10. 10.
    Johansson, G., and Andersson, M. (1984),J. Chromatogr. 291, 175.CrossRefGoogle Scholar
  11. 11.
    Lowe, C. R., and Pearson, J. C. (1984),Methods Enzymol. 104, 97.Google Scholar
  12. 12.
    Johansson, G., and Andersson, M. (1984),J. Chromatogr. 303, 39.CrossRefGoogle Scholar
  13. 13.
    Cordes, A. (1985).Affinitätsverteilung—Theoretische Aspekte sowie die Auslegung eines darauf basierenden Verfahrens zur Reinigung der Formiat-Dehydrogenase von Candida boidinii. Thesis, Braunschweig.Google Scholar
  14. 14.
    Kopperschläger, G., and Johansson, G. (1982),Anal. Biochem. 124, 117.CrossRefGoogle Scholar
  15. 15.
    Johansson, G. (1984),Methods Enzymol. 104, 356.Google Scholar
  16. 16.
    Johansson, G., and Joelsson, M. (1985),Biotechnol. Bioeng. 27, 621.CrossRefGoogle Scholar
  17. 17.
    Bergmeyer, H. U. (ed.) (1970),Methoden der Enzymatischen Analyse, vol. 1, 2 Ed., Verlag Chemie, Weinheim/Bergstr.Google Scholar
  18. 18.
    Scopes, R. K. (1975),Methods Enzymol. 42, 127.CrossRefGoogle Scholar
  19. 19.
    Cohen, P. P. (1955),Methods Enzymol. 2, 178.CrossRefGoogle Scholar
  20. 20.
    Bradford, M. M. (1976),Anal. Biochem. 72, 248.CrossRefGoogle Scholar
  21. 21.
    Albertsson, P.-Å. (1986),Partition of Cell Particles and Macromolecules, 3 Ed., Wiley, New York.Google Scholar
  22. 22.
    Albertsson, P.-Å., Andersson, B., Larsson, C., and Åkerlund, H.-E. (1981),Methods Biochem. Anal 28, 115.CrossRefGoogle Scholar
  23. 23.
    Laemmli, U. V. (1970),Nature 227, 359.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1986

Authors and Affiliations

  • G. Johansson
    • 1
  • M. Joelsson
    • 1
  1. 1.Department of Biochemistry, Chemical CenterUniversity of LundLundSweden

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