Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1990 Nov;56(11):3473–3477. doi: 10.1128/aem.56.11.3473-3477.1990

Immobilization of Chloroperoxidase on Aminopropyl-Glass

Tenshuk A Kadima 1, Michael A Pickard 1,*
PMCID: PMC184989  PMID: 16348352

Abstract

Chloroperoxidase (CPO) purified from Caldariomyces fumago CMI 89362 was covalently bound to aminopropyl-glass by using a modification of an established method. Acid-washed glass was derivatized by using aminopropyltriethoxysilane, and the enzyme was ionically bound at low ionic strength. Further treatment with glutaraldehyde covalently linked the enzyme to the glass beads in an active form. No elution of bound activity from glass beads could be detected with a variety of washings. The loading of enzyme protein to the glass beads was highest, 100 mg of CPO per g of glass, at high reaction ratios of CPO to glass, but the specific activity of the immobilized enzyme was highest, 36% of theoretical, at low enzyme-to-carrier ratios. No differences in the properties of the soluble and immobilized enzymes could be detected by a number of criteria: their pH-activity and pH-stability profiles were similar, as were their thermal stabilities. After five uses, the immobilized enzyme retained full activity between pH 6.0 and 6.7.

Full text

PDF
3473

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brotherton J. E., Emery A., Rodwell V. W. Characterization of sand as a support for immobilized enzymes. Biotechnol Bioeng. 1976 Apr;18(4):527–543. doi: 10.1002/bit.260180407. [DOI] [PubMed] [Google Scholar]
  2. Carmichael R. D., Jones A., Pickard M. A. Semicontinuous and Continuous Production of Chloroperoxidase by Caldariomyces fumago Immobilized in k-Carrageenan. Appl Environ Microbiol. 1986 Feb;51(2):276–280. doi: 10.1128/aem.51.2.276-280.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carmichael R. D., Pickard M. A. Continuous and Batch Production of Chloroperoxidase by Mycelial Pellets of Caldariomyces fumago in an Airlift Fermentor. Appl Environ Microbiol. 1989 Jan;55(1):17–20. doi: 10.1128/aem.55.1.17-20.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fang G. H., Kenigsberg P., Axley M. J., Nuell M., Hager L. P. Cloning and sequencing of chloroperoxidase cDNA. Nucleic Acids Res. 1986 Oct 24;14(20):8061–8071. doi: 10.1093/nar/14.20.8061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kadima T. A., Pickard M. A. A colorimetric assay for immobilized chloroperoxidase. Can J Microbiol. 1990 Apr;36(4):302–304. doi: 10.1139/m90-053. [DOI] [PubMed] [Google Scholar]
  6. Klibanov A. M., Tu T. M., Scott K. P. Peroxidase-catalyzed removal of phenols from coal-conversion waste waters. Science. 1983 Jul 15;221(4607):259–261. doi: 10.1126/science.221.4607.259-a. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Lambeir A. M., Dunford H. B. A kinetic and spectral study of the alkaline transitions of chloroperoxidase. Arch Biochem Biophys. 1983 Feb 1;220(2):549–556. doi: 10.1016/0003-9861(83)90446-0. [DOI] [PubMed] [Google Scholar]
  9. Manthey J. A., Hager L. P., McElvany K. D. Protein bromination by bromoperoxidase from Penicillus capitatus. Methods Enzymol. 1984;107:439–445. doi: 10.1016/0076-6879(84)07030-0. [DOI] [PubMed] [Google Scholar]
  10. Morris D. R., Hager L. P. Chloroperoxidase. I. Isolation and properties of the crystalline glycoprotein. J Biol Chem. 1966 Apr 25;241(8):1763–1768. [PubMed] [Google Scholar]
  11. Nuell M. J., Fang G. H., Axley M. J., Kenigsberg P., Hager L. P. Isolation and nucleotide sequence of the chloroperoxidase gene from Caldariomyces fumago. J Bacteriol. 1988 Feb;170(2):1007–1011. doi: 10.1128/jb.170.2.1007-1011.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Richards F. M., Knowles J. R. Glutaraldehyde as a protein cross-linkage reagent. J Mol Biol. 1968 Oct 14;37(1):231–233. doi: 10.1016/0022-2836(68)90086-7. [DOI] [PubMed] [Google Scholar]
  13. Thomas J. A., Morris D. R., Hager L. P. Chloroperoxidase. VII. Classical peroxidatic, catalatic, and halogenating forms of the enzyme. J Biol Chem. 1970 Jun;245(12):3129–3134. [PubMed] [Google Scholar]
  14. Weetall H. H. Covalent coupling methods for inorganic support materials. Methods Enzymol. 1976;44:134–148. doi: 10.1016/s0076-6879(76)44012-0. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES