Chemiluminescence-based inhibition kinetics of alkaline phosphatase in the development of a pesticide biosensor

The use and application of the enzyme alkaline phosphatase in a chemiluminescence assay are discussed. The enzyme catalyzes the hydrolysis of a macrocyclic phosphate compound generating a chemiluminescence signal. On the basis of inhibition of this signal, a methodology for the detection and quantit...

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Bibliographic Details
Published in:Biotechnology progress 1995-11, Vol.11 (6), p.699-703
Main Authors: Ayyagari, M.S. (University of Massachusetts, Lowell.), Kamtekar, S, Pande, R, Marx, K.A, Kumar, J, Tripathy, S.K, Akkara, J, Kaplan, D.L
Format: Article
Language:English
Subjects:
1,2-DIOXETANE PHENYL PHOSPHATE
ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
Adamantane - analogs & derivatives
Adamantane - metabolism
ALKALINE PHOSPHATASE
Alkaline Phosphatase - antagonists & inhibitors
BIODETECTEUR
Biological and medical sciences
Biomass
BIOSENSORES
BIOSENSORS
Biotechnology
COMPOSE ORGANOPHOSPHORE
COMPUESTO ORGANICO DEL FOSFORO
ENZYMIC ACTIVITY
FOSFATASA ALCALINA
Fundamental and applied biological sciences. Psychology
Hydrolysis
Indicators and Reagents - metabolism
INHIBICION
INHIBITION
INSECTICIDAS
INSECTICIDE
INSECTICIDES
Insecticides - analysis
Kinetics
Luminescent Measurements
Methods. Procedures. Technologies
METIL PARATION
OPTICAL PROPERTIES
ORGANOPHOSPHORUS COMPOUNDS
PARATHION METHYL
PHOSPHATASE ALCALINE
PROPIEDADES OPTICAS
PROPRIETE OPTIQUE
Various methods and equipments
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Summary:The use and application of the enzyme alkaline phosphatase in a chemiluminescence assay are discussed. The enzyme catalyzes the hydrolysis of a macrocyclic phosphate compound generating a chemiluminescence signal. On the basis of inhibition of this signal, a methodology for the detection and quantitation of organophosphorus‐based pesticides has been developed. The methodology is studied with alkaline phosphatase in the bulk aqueous phase, and detection of the signal is accomplished by a simple optical setup. Parts per billion level detection of paraoxon and methyl parathion in bulk solutions is achieved. The technique is rapid and sensitive and is applicable to the detection of most organophosphorus‐based pesticides. The results from kinetic studies indicate a mixed type of inhibition of the enzyme by paraoxon and methyl parathion. The detection methodology forms an integral part of a biosensor under development and is adaptable to incorporating optical fibers for remote detection of pesticides.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp00036a015