A novel biosensor for the detection of organophosphorus (OP)-based pesticides using organophosphorus acid anhydrolase (O
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ENVIRONMENTAL BIOTECHNOLOGY
A novel biosensor for the detection of organophosphorus (OP)-based pesticides using organophosphorus acid anhydrolase (OPAA)-FL variant Monika Jain 1 & Priyanka Yadav 1 & Bhavana Joshi 1 & Abhijeet Joshi 1 & Prashant Kodgire 1 Received: 11 March 2020 / Revised: 28 October 2020 / Accepted: 8 November 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Indiscriminate use of organophosphorus (OP)-based insecticides is a great concern to human health because of bioaccumulationinduced health hazards. Potentially fatal consequences and limited treatment methods of OP poisoning necessitate the need for the development of reliable, selective, cost-effective, and sensitive methods of OP detection. To tackle this issue, the development of effective devices and methods is required to sensitively detect as well as degrade OPs. Enzymatic sensor systems have gained popularity due to high catalytic activity, enhanced detection limits, and high sensitivity with the environmentally benign operation. Organophosphorus acid anhydrolase (OPAA) from Alteromonas sp. JD6.5 is capable of hydrolyzing the P-F, P-O, P-S, and P-CN bonds, in OPs, including nerve agents of the G/V-series. Several mutants of OPAA are reported which have greater activity against various OPs. In this study, recombinant expression of the OPAA-FL variant in Escherichia coli was performed, purified, and subsequently tested for activity against ethyl paraoxon. OPAA-FL variant showed its optimum activity at pH 8.5 and 50 °C. Colorimetric and fluorometric assays were used for estimation of ethyl paraoxon based on p-nitrophenol and fluorescein isothiocyanate (FITC) fluorescence intensity, respectively. Colorimetric and fluorometric assay estimation indicates that ethyl paraoxon can be estimated in the linear range of 0.01 to 1 mM and 0.1 to 0.5 mM, with LOD values 0.04 mM and 0.056 mM, respectively. Furthermore, the OPAA-FL variant was immobilized into alginate microspheres for colorimetric detection of ethyl paraoxon and displayed a linear range of 0.025 to 1 mM with a LOD value of 0.06 mM. Key points • Biosensing of paraoxon with purified and encapsulated OPAA-FL variant. • Colorimetric and fluorometric biosensing assay developed using OPAA-FL variant for paraoxon. • First report on alginate encapsulation of OPAA-FL variant for biosensing of paraoxon.
Keywords Organophosphorus acid anhydrolase (OPAA)-FL variant . Acetylcholinesterase (AChE) . Paraoxon . Organophosphorus compounds (OPs)
Introduction An increase in the global population has led to greater demand for food supplies, which eventually demands increased * Abhijeet Joshi [email protected] * Prashant Kodgire [email protected] 1
Department of Biosciences and Biomedical Engineering, Indian Institute of Technology Indore, Khandwa Road, Indore, India
production from existing cultivable land. In recent times, massive agricultural expansion has necessitated an increase in pesticide and insecticide use in order to improve the productivity of the crops.
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