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
Identifiers and Pagination:Year: 2017
First Page: 89
Last Page: 93
Publisher Id: TOBIOTJ-11-89
Article History:Received Date: 14/03/2017
Revision Received Date: 21/07/2017
Acceptance Date: 13/08/2017
Electronic publication date: 14/09/2017
Collection year: 2017
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.
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.
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.
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.
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.