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. 2020 Jun 23:27:e00495.
doi: 10.1016/j.btre.2020.e00495. eCollection 2020 Sep.

Easy-to-use and reliable absorbance-based MPH-GST biosensor for the detection of methyl parathion pesticide

Witsanu Senbua  1 , Jhirat Mearnchu  2 , Jesdawan Wichitwechkarn  1
Affiliations

Easy-to-use and reliable absorbance-based MPH-GST biosensor for the detection of methyl parathion pesticide

Witsanu Senbua et al. Biotechnol Rep (Amst). .

Abstract

Due to high contamination of organophosphate (OP) insecticides in agricultural products and the environment, efficient and convenient devices for their monitoring are necessary. Here, a simple, inexpensive, efficient, and easy-to-use absorbance-based biosensor was fabricated utilizing recombinant methyl parathion hydrolase fused with glutathione-S-transferase (MPH-GST), covalently immobilized onto a chitosan film-coated polystyrene microplate, for the detection of methyl parathion (MP) as a representative of OPs. Having been connected to the transducer system designed to work through an Arduino microcontroller, the biosensor could detect MP as efficiently as the conventional methods, with the detection limit of 0.1 μM, the lowest value ever reported for this method. It was stable at 25 °C for 30 days, could function 100 rounds repetitively, and yielded high recovery with real samples. Hence, this simply designed MPH-GST biosensor could be an easy and inexpensive alternative for efficient OP screening at site to help control its contamination.

Keywords: AChE, acetylcholinesterase; Absorbance-based biosensor; CP, chlopyrifos; MP, methyl parathion; MPH, methyl parathion hydroloase; MPH-GST; MPH-GST, methyl parathion hydrolase fused with glutathione-S-transferase; Methyl parathion; Methyl parathion hydrolase; OPH, organophosphate hydrolase; Organophosphate; PX, paraoxon.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
MP degradation reaction by MPH.
Fig. 2
Fig. 2
Schematic operation of the optical transducer system.
Fig. 3
Fig. 3
Circuit diagram of optical transducer system.
Fig. 4
Fig. 4
MPH activity assay of the MPH-GST sensing device. (a) immobilized MPH-GST well; (b) blank well.
Fig. 5
Fig. 5
Effect of chitosan concentration on the activity of the MPH-GST biosensing component.
Fig. 6
Fig. 6
Effect of chitosan volume on the activity of the MPH-GST biosensing component.
Fig. 7
Fig. 7
Effect of temperature (a) and pH (b) on the activity of MPH-GST biosensor.
Fig. 8
Fig. 8
Calibration curve of the MPH-GST biosensor.
Fig. 9
Fig. 9
Temperature stability of the MPH-GST biosensor.
Fig. 10
Fig. 10
pH stability of the MPH-GST biosensor.
Fig. 11
Fig. 11
Reusability of the MPH-GST biosensor for MP detection.
See this image and copyright information in PMC

References

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