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A Simple Colorimetric Enzymatic-Assay for Okadaic Acid Detection Based on the Immobilization of Protein Phosphatase 2A in Sol-Gel

Author:
Hayat, Akhtar, Barthelmebs, Lise, Marty, Jean-Louis
Source:
Applied biochemistry and biotechnology 2012 v.166 no.1 pp. 47-56
ISSN:
0273-2289
Subject:
Dinophysis, Prorocentrum, colorimetry, detection limit, diarrhetic shellfish poisoning, enzyme activity, guanosine triphosphate, humans, okadaic acid, phosphoprotein phosphatase, polymers, silica, storage quality, toxins
Abstract:
Okadaic acid (OA), a lipophilic toxin, is produced by Dinophysis and Prorocentrum, and causes diarrheic shellfish poisoning to humans. The mechanism of OA action is based on the reversible inhibition of protein phosphatase type 2A (PP2A) by the toxin. Therefore, this inhibition could be used to develop assay for OA detection. In this work, a colorimetric test based on the PP2A inhibition was developed for OA detection. PP2A from GTP and Millipore was immobilized on silica sol-gel, and the detection was performed. A limit of detection of 0.29 and 1.14 μg/L was respectively observed for enzyme from GTP and Millipore. The immobilization technique provided a tool to preserve the enzymatic activity, which is very unstable in solution. The PP2A immobilized sol-gel exhibited a storage stability of near 5 months, when microtiter plate with enzyme-immobilized polymer was kept at −18C°. The combination of the simplicity of the colorimetric method, along with long storage stability achieved by sol-gel immobilization, demonstrated the potentiality of this technique to be used for commercial purpose.
Agid:
438946