Skip to main content
SpringerLink
Log in

Matrix-bounded enzyme: Potato esterase entrapped in polyacrylamide gel after gel electrophoresis

Matrix-gebundenes Enzym: Kartoffelesterase, inkludiert in Polyacrylamidgel nach Gelelektrophorese

  • Original Papers
  • Published:
Fresenius' Zeitschrift für analytische Chemie Aims and scope Submit manuscript

Zusammenfassung

Von den Kartoffelesterasen wurde ein Isoenzym durch Gel-Elektrophorese in Polyacrylamid (PAA) getrennt. Das entsprechende Gelstück mit dem durch das Netzwerk stabilisierten Enzym tränkte man mit AA und Vernetzer (BIS) unterschiedlicher Konzentration und polymerisierte mit Licht das Enzym ein. Die Mischungen AA/BIS sind in einem bestimmten Verhältnis (6∶0,36) optimal., eine Ausbeute von 77% der Anfangsaktivität kann erreicht werden. Einige Eigenschaften (pHi, K m, MG u.ä.) werden beschrieben, insbesondere ist die 100fach bessere Lagerstabilität hervorzuheben.

Summary

An isoenzyme of potato esterase has been entrapped in situ after polyacrylamide (PAA)-electrophoresis, using PAA of different concentrations and crosslinking. Best results in respect to diffusion, flow rates and yield (77%) have been found for a gel containing 6% acrylamide with 6% methylen-bis-acrylamide, penetrating a gel of cyanogum with esterase. The latter is stabilized during polymerization by the gel. Properties of the entrapped enzyme (pHi, K m, MW etc.) have been determined, 100 fold better storage stability is achieved.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

References

  1. Bernfeld, P., Bieber, R. E.: Arch. Biochem. Biophys. 131, 587 (1969).

    Article  CAS  Google Scholar 

  2. Bernfeld, P., Bieber, R. E., MacDonnell, P. C.: Arch. Biochem. Biophys. 127, 779 (1968).

    Article  CAS  Google Scholar 

  3. Bernfeld, P., Wan, J.: Science 142, 678 (1963).

    Article  CAS  Google Scholar 

  4. Brown, H. D., Patel, A. B., Chattopadhyay, S. K.: J. Chromatog. 35, 103 (1968).

    Article  CAS  Google Scholar 

  5. Degani, Y., Miron, T.: Biochim. Biophys. Acta 212, 362 (1970).

    Article  CAS  Google Scholar 

  6. Dinish, K. N.: Entrapping of potato esterase in polyacrylamide gel. Vortrag, Kolloquium 7th December 1971, Biologische Bundesanstalt, Braunschweig.

  7. Green, M. N., Tsou, K.-C., Bressler, R., Seligmann, A. M.: Arch. Biochem. Biophys. 57, 458 (1955).

    Article  CAS  Google Scholar 

  8. Hicks, G. P., Updike, S. J.: Anal. Chem. 38, 726 (1966).

    Article  CAS  Google Scholar 

  9. Katchalski, E.: Chem. Eng. News, 15th Febr. 1971, p. 87, 3rd Jan. 1972, p. 24.

  10. Katchalski, E., Silman, I., Goldman, R.: Advances in Enzymology 34, 445 (1971).

    CAS  Google Scholar 

  11. Koenig, R., Stegemann, H., Francksen, H., Paul., H. L.: Biochim. Biophys. Acta 207, 184 (1970).

    Article  CAS  Google Scholar 

  12. Loeschcke, V., Stegemann, H.: Z. Naturforsch. 21b, 879 (1966).

    Google Scholar 

  13. Macko, V., Stegemann, H.: Z. Physiol. Chem. 350, 917 (1969).

    CAS  Google Scholar 

  14. Main, A. R.: Biochem. J. 74, 10 (1960); ctd. in: Th. E. Barman, Enzymes, Vol. II, p. 502. Berlin-Heidelberg-New York: Springer 1969.

    Article  CAS  Google Scholar 

  15. Mosbach, K., Larsson, P.-O.: Biotechnol. Bioeng. 12, 19 (1970); Mosbach, K., Mosbach, R.: Acta Chem. Scand. 20, 2807 (1966).

    Article  CAS  Google Scholar 

  16. Orth, H. D., Brümmer, W.: Angew. Chem. 84, 319 (1972).

    Article  Google Scholar 

  17. Shapiro, A. L., Viñuela, E., Maizel, J. V.: Biochem. Biophys. Res. Commun. 28, 815 (1967).

    Article  CAS  Google Scholar 

  18. Shivaram, K. N.: Eigenschaften von inkludierter Kartoffelesterase. Vortrag, Kolloquium 7th December 1971, Biologische Bundesanstalt, Braunschweig.

  19. Shivaram, K. N., Dinish, K. N., Stegemann, H.: Entrapping of Potato Esterase in Polyacrylamide(PAA)-Gel after PAA-Gel Eiektrophoresis. XXth Colloquium Protides of the Biological Fluids, Brugge (Belgium), 3rd to 7th May 1972. Abstract No. 115.

  20. Stegemann, H.: Z. Physiol. Chem. 319, 64 (1960).

    Article  CAS  Google Scholar 

  21. Stegemann, H.: Z. Anal. Chem. 252, 165 (1970).

    Article  CAS  Google Scholar 

  22. Stegemann, H.: Z. Physiol. Chem. 348, 951 (1967); Liefländer, M., Stegemann, H.: Z. Physiol. Chem. 349, 157 (1968).

    CAS  Google Scholar 

  23. Wieland, T., Determann, H., Bünnig, K.: Z. Naturforsch. 21b, 1003 (1966).

    Google Scholar 

  24. Wieland, T.: Nature New Biology, News section 236, 98 (1972).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biologische Bundesanstalt, Institut für Biochemie, Braunschweig

    K. N. Dinish, K. N. Shivaram & H. Stegemann

Authors
  1. K. N. Dinish
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K. N. Shivaram
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H. Stegemann
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

We are indebted to Professor Dr. G. Schuhmann, President of the BBA, for the encouragement to this work.

For SDS-gel eleetrophoresis and the photographic work we thank Mrs. Knackstedt and Miss Krögerrecklenfort.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Dinish, K.N., Shivaram, K.N. & Stegemann, H. Matrix-bounded enzyme: Potato esterase entrapped in polyacrylamide gel after gel electrophoresis. Z. Anal. Chem. 263, 27–32 (1973). https://doi.org/10.1007/BF00424326

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00424326

Search

Navigation