*Corresponding author:
Fabiane Caxico de Abreu, Institute of Chesmitry and Biotechnology, Federal University of Alagoas Maceió, AL, 57072-900, BrazilReceived: August 23, 2018; Published: August 28, 2018
DOI: 10.26717/BJSTR.2018.08.001655
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Background: AChE is a serine hydrolase responsible for terminating neurotransmission by hydrolyzing the acetylcholine released on synaptic cleft. Studies of AChE as a target of pesticide toxicity have yielded several practical outcomes and are the basis for constructing biosensors. These devices are primarily designed to determine and quantify the inhibition of AChE by toxic chemicals.
Objective: to construct a biosensor based on acetylcholinesterase from the brain of the Prochilodus brevis fish, and to use the same as biomarker of agrochemicals that inhibit the enzyme.
Methods: Acetylcholinesterase was isolated from curimatã fish brain (Prochilodus Brevis) and partially purified using amonium sulfate precipitation followed by size-exclusion chromatography (ChE1). AChE from curimatã fish brain was directly immobilizes on the surface of glassy carbon electrode modified with multi-walled carbon nanotubes.
Results: Acetylcholinesterase was characterized as having a specific activity of 0.194U/mg. The optimum activity was found at pH 8.5, phosphate buffer 0.7μM, 28ºC and exhibited a thermostability at 37°C. The glassy carbon modified electrode exhibits excellent electrocatalytic activity to the increase of thiocholine, with a linear response in the 50μM to 0.85mM concentration range, with a 73μM limit of detection and with a 240μM limit of quantification.
Conclusion: The AChE from curimatã fish brain allowed the modification of the glassy carbon electrode providing a potential sensor detection system that can be used for determination of thiocholine (pesticides) in real samples of environmental importance.
Abstract | Introduction | Materials and Methods | Results and Discussion | Conclusion | Acknowledgement | References|