Abstract
Pectinesterase was immobilized on a porous glass support, which surface was covered with glyceryl residues. The parameters of the immobilization were characterized with respect to the coupling method used as well as the support pore size. Chemical modification level resulted an important parameter in determining the activity of the immobilized derivative. Attachment of the enzyme through thiol groups gave the best results, whereas a nominal pore size of 20 nm seemed to be the most suitable for the demethoxylating activity of the enzyme on citrus pectin. Optimum conditions for activity as well as the inhibition constant for polygalacturonic acid did not change on immobilization, as the Michaelis constant did. Fluorescence spectra revealed a partial unfolding of the enzyme tertiary structure when immobilized.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Rouse, A. H. (1977),Citrus Science and Technology, Nagi, S., Shaw, Ph. E., and Veldhuis, N. K., eds., Avi, Westport, CT, pp. 110–120
Zaborsky, O. R. (1972),Immobilized Enzymes, Weast, R. C., ed., CRC Press, OH, p. 2.
Van Hodenhoven, F., de Witt, P., and Wisser, J. (1974),Carbohydr. Res. 34, 233–237.
Rexová-Benková, L., Mracková, M., and Babor, K. (1980),Collect. Czech. Chem. Commun. 45, 163–168.
Rexová-Benková, L. and Mracková-Dobrotová, M. (1981),Carbohydr. Res. 98, 115–118.
Rexová-Benková, L., Omelková J., and Kubánek, V. (1982),Collect. Czech. Chem. Commun. 47, 2716–2722.
Rexová-Benková, L., Omelková, J., Veruovic, B., and Kubánek, V. (1983),Biotechnol. Lett. 5, 737–740.
Hanish, W. H., Rickard, P. A. D., and Nyo, S. (1978),Biotechnol. Bioeng. 20, 95–100.
Weibel, M. K., Barrios, R., Delotto, R., and Humphrey, A. E. (1975),Biotechnol. Bioeng. 17, 85–98.
Vijayalakshmi, M. A., Picque, D., Jaumouille, R., and Sagard, E., (1980),Food Proc. Eng. 2, 152–158.
Vijayalakshmi, M. A., Baron, A., and Drilleau, J. F. (1982),Use of Enzymes in Food Technology, Dupuy, P., ed., Technique et Documentation Lavoisier, Paris, pp. 537–543.
Markovic, O. and Machova, E. (1985),Collect. Czech. Chem. Commun. 50, 2021–2027.
Anaya, M. C., López, M. C. A., and Arjona, J. L. (1982),Use of Enzymes in Food Technology, Dupuy, P., ed., Technique et Documentatiion Lavoisier, Paris, pp. 503–511.
Manjón, A., Llorca, F. I., Bonete, M. J., Bastida, J., and Iborra, J. L. (1985),Proc. Biochem. 20, 17–22.
Hartree, E. F. (1972),Anal. Biochem. 48, 422–427.
Kertesz, Z. I. (1956),Meth. Enzymol. 1, 158–166.
Glycophase G™. Pierce General Catalog (1976), Rockford, IL.
Enzacryl Carriers. Koch-Light Laboratories Technical Bulletin. Colnbrook, UK.
Srere, P. A., and Uyeda, K. (1976),Meth. Enzymol. 44, 11–18.
Markovic, I. (1983),Collect. Czech. Chem. Commun. 48, 668–674.
Romero, C., Manjon, A., and Iborra, J. L. (1988),Biotechnol. Lett. 10, 97–100.
BeMiller, J. N. (1986),Chemistry and Function of Pectins, Fishman, M. L. and Jen, J. J., eds., American Chemical Society, Washington, DC, pp. 2–12.
Lakowicz, J. R. (1983),Principles of Fluorescence Spectroscopy, Lakowicz, J. R., ed., Plenum, New York, pp. 341–381.
Engasser, J. M. and Horvath, C. (1973),J. Theor. Biol. 42, 137–155.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Borrego, F., Tari, M., Manjon, A. et al. Properties of pectinesterase immobilized on glycophase-coated controlled-pore glass. Appl Biochem Biotechnol 22, 129–140 (1989). https://doi.org/10.1007/BF02921740
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02921740