Organophosphorus Hydrolase-Based Biosensor for Direct Detection of Paraoxon Using Silica-Coated Magnetic Nanoparticles

Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organoph...

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Published in:	Applied biochemistry and biotechnology Vol. 176; no. 2; pp. 359 - 371
Main Authors:	Khaksarinejad, Reza, Mohsenifar, Afshin, Rahmani-Cherati, Tavoos, Karami, Rezvan, Tabatabaei, Meisam
Format:	Journal Article
Language:	English
Published:	New York Springer-Verlag 01.05.2015 Springer US Springer Nature B.V
Subjects:	active sites Aryldialkylphosphatase - chemistry Bacterial Proteins - chemistry Biochemistry Biosensing Techniques - methods Biosensors Biotechnology Chemistry Chemistry and Materials Science Coefficient of variation Coumarins - chemistry Decontamination detection limit Kinetics Magnetite Nanoparticles - chemistry Nanoparticles paraoxon Paraoxon - analysis Pseudomonas - enzymology Silica Silicon Dioxide - chemistry Organophosphorous hydrolase Kinetics Nanobiosensor Nanomagnet-silica core-shell Paraoxon
ISSN:	0273-2289, 1559-0291, 1559-0291
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Abstract	Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme’s active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6–10 %), and the detection limit was as low as 5 × 10⁻⁶ μM.
AbstractList	Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme's active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6-10 %), and the detection limit was as low as 5×10^sup -6^ [mu]M. Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme's active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6-10%), and the detection limit was as low as 5 × 10(-6) μM. Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme’s active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6–10 %), and the detection limit was as low as 5 × 10⁻⁶ μM. Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme's active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6-10%), and the detection limit was as low as 5 × 10(-6) μM.Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme's active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6-10%), and the detection limit was as low as 5 × 10(-6) μM. Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme’s active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6–10 %), and the detection limit was as low as 5 × 10⁻⁶ μM. Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme’s active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6–10 %), and the detection limit was as low as 5 × 10 −6 μM.
Author	Rahmani-Cherati, Tavoos Tabatabaei, Meisam Karami, Rezvan Mohsenifar, Afshin Khaksarinejad, Reza
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Keywords	Organophosphorous hydrolase Kinetics Nanobiosensor Nanomagnet-silica core-shell Paraoxon
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10.1016/j.bios.2004.08.019
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Snippet	Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could...
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SubjectTerms	active sites Aryldialkylphosphatase - chemistry Bacterial Proteins - chemistry Biochemistry Biosensing Techniques - methods Biosensors Biotechnology Chemistry Chemistry and Materials Science Coefficient of variation Coumarins - chemistry Decontamination detection limit Kinetics Magnetite Nanoparticles - chemistry Nanoparticles paraoxon Paraoxon - analysis Pseudomonas - enzymology Silica Silicon Dioxide - chemistry
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Title	Organophosphorus Hydrolase-Based Biosensor for Direct Detection of Paraoxon Using Silica-Coated Magnetic Nanoparticles
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