RESEARCH ARTICLE


An Easy Method for Screening and Detection of Laccase Activity



Albino A. Dias*, António J.S. Matos, Irene Fraga, Ana Sampaio, Rui M.F. Bezerra
Centre for the Research and Technology of Agro-Environment and Biological Sciences - CITAB, Universidade de Trás-os-Montes e Alto Douro – UTAD, 5000-801 Vila Real, Portugal


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© 2017 Dias et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: (https://creativecommons.org/licenses/by/4.0/legalcode). This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Centre for the Research and Technology of Agro-Environment and Biological Sciences – CITAB, Universidade de Trás-os-Montes e Alto Douro – UTAD, 5000-801 Vila Real, Portugal, Tel: +351 259350250; E-mail: jdias@utad.pt


Abstract

Objective:

An instrument-free assay was developed for simultaneous detection of laccase activity in a large number of samples as diverse as screening of laccase-producing microbial cultures or chromatographic fractions.

Method:

Dried paper discs previously impregnated with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) were placed on a flat-bottom microplate (a simple way to avoid misidentification) and loaded with an aliquot from each sample.

Results:

Discs corresponding to samples containing laccase activity become green-bluish colored within first ten minutes of reaction, allowing direct detection through simple naked-eye inspection.

Conclusion:

As an example, this easy process was applied to the laccase purification in order to eliminate chromatographic fractions that did not contain laccase activity, thus reducing the number of spectrophotometric assays.

Keywords: ABTS, Laccase, White-rot fungi, Zymogram, Laccase purification, Laccase activity.