4.3 Article

ABTS assay of phenol oxidase activity in soil

Journal

JOURNAL OF MICROBIOLOGICAL METHODS
Volume 71, Issue 3, Pages 319-324

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2007.09.020

Keywords

ABTS; laccase; peroxidase; phenol oxidase activity; soil enzymology

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Phenol oxidases (PO) are involved in degradation of many recalcitrant aromatic compounds and may be sensitive to some pollutants. Hence, their activities may be a useful indicator for evaluating soil quality and health. To this end, the aim of this study was to develop a simple method to assay PO activity directly in bulk samples by spectrophotometric test using 2,2'-azinobis-(-3 ethylbenzothiazoline-6-sulfononic acid) diammonium salt (ABTS) as the substrate. Three Mediterranean soils were used as models. For each soil, we studied the kinetic parameters and the effects of certain factors (i.e. amount of soil, PH, temperature, incubation time and substrate concentration) in order to determine the optimum conditions for the ABTS assay. Results showed that PO attain their optimum activities when incubating 0.1 g of soil at 30 degrees C for 5 min with 10 ml of a Modified Universal Buffer (MUB) at pH 2 and 200 mu I of a 0. 1 M ABTS solution. (c) 2007 Elsevier B.V. All rights reserved.

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