Synthetic organophosphorus (OP) compounds are highly poisonous substances, extensively applied in the agriculture field as pesticides and insecticides and used in the military as chemical warfare agents. The aim of the present study is to develop an eggshell membrane (natural polymer) based electrochemical biosensor for the detection of OP compounds. We have selected the eggshell membrane as a natural support for immobilization of microbial cells. The enzyme organophosphorus hydrolase (OPH) enzyme was produced in one of the soil borne bacteria Pseudomonas diminuta species, which is capable of hydrolyzing methyl parathion into dimethyl thiophosphoric acid and yellow colour detectable product p-nitrophenol. Whole Pseudomonas diminuta bacterial cells were immobilized on the eggshell membrane using glutaraldehyde as cross linking agent and placed on the working area of screen printed carbon electrode (SPCE). ESM with immobilized bacteria was placed on SPCE and it was connected to electrochemical work station and cyclic voltammograms were recorded. The potential +0.1 V was optimized to measure the changes in the current as per redox role of p-nitrophenol. There was increased oxidation current as the concentration of methyl parathion was increased. A single ESM with immobilized bacteria can be reused for 22 reactions and it was stabilized for 32 days at 4oC with 80% of initial enzyme activity.
Impact Factor
2022: 6.012
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