Evaluation of Supports and Methods for Immobilization of Enzyme Cyclodextringlycosyltransferase

  • Keli A. Sobral
  • Regina O. Rodrigues
  • Rogério D. Oliveira
  • José E. Olivo
  • Flávio F. de Moraes
  • Gisella M. Zanin
Part of the Applied Biochemistry and Biotechnology book series (ABAB)

Abstract

An experimental design with factorial planning was used for the immobilization of the enzyme cyclodextringlycosyltransferase (CGTase) from Bacillus firmus (strain no. 37) to select the best combination of support, method of immobilization, and conditions that gives primarily higher average values for the specific immobilized enzyme activity, and secondarily, higher average values for the percentage of protein fixation. The experimental design factors were as follows: supports—controlled-pore silica, chitosan, and alumina; immobilization methods—adsorption, and two covalent bonding methods, either with γ-aminopropyltriethoxysilane or hexa-methylenediamine (HEMDA); conditions—7°C without agitation and 26°C with stirring. The best combination of factors that lead to higher average values of the response variables was obtained with immobilization of CGTase in silica with HEMDA at 7°C. However, immobilization in chitosan at 7°C gave the highest immobilized CGTase specific activity, 0.25 μmole of ß-CD/(min·mg protein). Physical adsorption gave low specific enzyme activities, and, in general, a high load of enzyme leads to lower specific enzyme activity.

Index Entries

Cyclodextringlycosyltransferase immobilized enzyme controlled-pore silica alumina chitosan 

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References

  1. 1.
    Bulmus, V., Kesenci, K., and Piskin, E. (1998), Reactive Funct. Polym. 38, 1–9.CrossRefGoogle Scholar
  2. 2.
    Wojcik, A., Lobarzewski, J., and Blaszczynska, T. (1990), J. Chem. Technol. Biotechnol. 90, 287.Google Scholar
  3. 3.
    Hayashi, T., Hirayama, C., and Iwatsuki, M. (1992), J. Appl. Polym. Sci. 44, 143.CrossRefGoogle Scholar
  4. 4.
    Hayashi, T. and Ikada, Y. (1994), J. Appl. Polym. Sci. 42, 85.CrossRefGoogle Scholar
  5. 5.
    Tardioli, P. W., Zanin, G. M., and Moraes, F. F. (2000), Appl. Biochem. Biotechnol. 8486, 1003–1019.PubMedCrossRefGoogle Scholar
  6. 6.
    Bekers, O., Uijtendaal, E. V., Beijnen, J. H., Bult, A., and Underberg, W. J. M. (1991), Drug Dev. Ind. Pharm. 17, 1503–1549.CrossRefGoogle Scholar
  7. 7.
    Korokolvas, A. (1991), ENLACE Parmalab 2, 6–15.Google Scholar
  8. 8.
    Yang, C. P. and Su, C. S. (1989), J. Chem. Technol. Biotechnol. 46, 283–294.CrossRefGoogle Scholar
  9. 9.
    Abdel-Naby, M. A. (1999), Process Biochem. 34, 399–405.CrossRefGoogle Scholar
  10. 10.
    Szejtli, J. (1988), in Cyclodextrin Technology, Szejtli, J., ed., Kluwer, Dordrecht, The Netherlands, pp. 79–185.CrossRefGoogle Scholar
  11. 11.
    Matioli, G., Zanin G. M., Guimarães, F., and Moraes, F. F. (1998), Appl. Biochem. Biotechnol. 7072, 267–275.PubMedCrossRefGoogle Scholar
  12. 12.
    Pereira, E. B. (1999), MS thesis, UEM, Maringá, PR, Brazil.Google Scholar
  13. 13.
    Dixon, M. and Webb, E. C., (1979), Enzymes, 3rd ed., Chap. 7, Longman Group, London, UK, pp. 7–22.Google Scholar
  14. 14.
    Vikmon, M. (1982), in The First International Symposium on Cyclodextrin, Szejtli, J., ed., D. Reidel Publishing Company, Dordrecht, The Netherlands, pp. 60–74.Google Scholar
  15. 15.
    Hamon, V. and Moraes, F. F. (1990), Internal Report, Laboratorie de Technologie Enzymatique, Université de Technologie de Compiègne, Compiègne, France.Google Scholar
  16. 16.
    Bradford, M. (1976), Anal Biochem. 72, 248.PubMedCrossRefGoogle Scholar
  17. 17.
    Matioli, G., Zanin G. M., Guimarães, F., and Moraes, F. F. (2001), Appl Biochem. Biotechnol. 9193, 643–654.PubMedCrossRefGoogle Scholar
  18. 18.
    Bon, E., Freire, D. G., Mendes, M. F., and Soares, V. F. (1984), Biotechnol Bioeng. Symp. 14, 447–455.Google Scholar

Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Keli A. Sobral
    • 1
  • Regina O. Rodrigues
    • 1
  • Rogério D. Oliveira
    • 1
  • José E. Olivo
    • 1
  • Flávio F. de Moraes
    • 1
  • Gisella M. Zanin
    • 1
  1. 1.Chemical Engineering DepartmentState University of MaringáMaringá-PRBrazil

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