Advertisement

Biotechnology Letters

, Volume 22, Issue 2, pp 105–108 | Cite as

A simple method for monitoring chromatography column eluates for laccase activity during enzyme purification

  • Robert F.H. Dekker
  • Khu Yee Ling
  • Aneli M. Barbosa
Article

Abstract

A simple and economical method is described that allows rapid detection of laccase activity in chromatography column fractions during enzyme purification. Aliquots of column eluants are applied to filter paper coated with 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) containing a numbered grid, and incubated at ambient temperature for 20 min. Indications of enzyme activity are simply observed by a colour change. This method avoids having to manually assay each fraction of a chromatographic run for enzyme activity.

ABTS Botryosphaeria chromogenic substrates laccases zymogram method 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Barbosa AM, Dekker RFH, Hardy GE (1996) Veratryl alcohol as an inducer of laccase by an ascomycete, Botryosphaeria sp., when screened on the polymeric dye Poly R-478. Lett. Appl.Microbiol. 23: 93–96.Google Scholar
  2. Coll PM, Fernández-Abalos JM, Villanueva JR, Santamaría R, Pérez P (1993) Purification and characterization of a phenol oxidase (laccase) from the lignin-degrading basidiomycete PM1 (CECT 2971). Appl. Environ. Microbiol. 59: 2607–2613.Google Scholar
  3. Dean JFD, Eriksson KEL (1994) Laccase and the deposition of lignin in vascular plants. Holzforschung. (Suppl.) 48: 21–33.Google Scholar
  4. Eggert C, Temp U, Eriksson KEL (1996) The ligninolytic system of the white rot fungus Pycnoporus cinnabarinus: purification and characterization of the laccase. Appl. Environ. Microbiol. 62: 1151–1158.Google Scholar
  5. Harkin JM, Larsen MJ, Obst JR (1974) Use of syringaldazine for detection of laccase in sporophores of wood rotting fungi. Mycologia 66: 469–476.Google Scholar
  6. Smith M, Thurston CF, Wood DA (1997) Fungal laccases: role in delignification and possible industrial applications. In: Messerschmidt A, ed. Multi-Copper Oxidases. Singapore: World Scientific, pp. 201–224.Google Scholar
  7. Sundman V, Nase L (1971) A simple plate test for direct visualization of biological lignin degradation. Pap. Timber 53: 65–71.Google Scholar
  8. Thurston CF (1994) The structure and function of fungal laccases. Microbiology 140: 19–26.Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Robert F.H. Dekker
    • 1
    • 2
  • Khu Yee Ling
    • 1
    • 2
  • Aneli M. Barbosa
    • 1
    • 2
  1. 1.Division of Science, BiotechnologyMurdoch UniversityMurdochAustralia
  2. 2.Departamento de Alimentos e MedicamentosUniversidade Estadual de LondrinaLondrina–PRBrazil (Fax

Personalised recommendations