Open AccessElectrochemical acetylcholinesterase biosensor for detection of cholinesterase inhibitors: study with eserine
Author(s) -
N. Lokar,
Vladimir S. Konko,
Damjana Drobne,
D. Vrtačnik
Publication year - 2019
Publication title -
informacije midem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.122
H-Index - 18
eISSN - 1855-4709
pISSN - 0352-9045
DOI - 10.33180/infmidem2018.406
Subject(s) - acetylcholinesterase , cholinesterase , chemistry , glutaraldehyde , physostigmine , acetylthiocholine , butyrylcholinesterase , biosensor , enzyme , chromatography , aché , immobilized enzyme , substrate (aquarium) , biochemistry , pharmacology , acetylcholine , medicine , biology , ecology
Cholinesterase inhibitors are widely used as pesticides, as chemical warfare agents and as drugs to treat symptoms ofAlzheimer’s disease. Therefore, it is a high need to develop methods for their detection which are fast, sensitive, and reliable. Thispaper reports a preliminary work in the development of an electrochemical biosensor based on acetylcholinesterase (AChE) which isconstructed by immobilization layers – cysteamine/glutaraldehyde/AChE on thin layer gold electrode for detection of cholinesteraseinhibitors. Eserine (physostigmine) was used as a test inhibitor. The enzyme immobilization efficacy was evaluated by measuringactivity of immobilized enzyme via Ellman’s method. The enzyme activity of the initial reduction of 33% in five days remained afterthat stable for at least one week. Chronoamperometric response to substrate acetylthiocholine chloride (ATCl) was assumed to followMichaelis-Menten kinetics. After exposure biosensor to 25 mM eserine for 10 min, 70% inhibition of enzyme was detected. Reactivationfactor of inhibited AChE was determined as 0.016 min-1.