To develop a convenient and efficient means for organophosphate (OP) insecticide detection, a simple, cost-effective, and easy-to-use absorbance-based sensing device was generated using methyl parathion hydrolase fused with glutathione-S-transferase (MPH-GST) covalently immobilized onto a chitosan film-coated microplate.
With methyl parathion (MP) as a representative substrate, this MPH-GST sensing microplate had the detection limit of 0.1 µM and the linear range of 0.1–50 µM. Despite its highest stability at 4 °C, it was considerably stable at 25 °C with high activity for 30 days. It was also most stable at pH 8.0 and could be efficiently reused up to 100 rounds. The device revealed a high percentage of recovery for tap water spiked with a known concentration of MP, which was also comparable to the result obtained from gas chromatography-mass spectrometry. It also showed a high recovery of 82–100% with MP spiked agricultural products and satisfactory results with non-spiked samples. This immobilized enzyme sensing system was more sensitive and efficient than the whole cell system from our previous work.
All of the advantages of the MPH-GST sensing microplate developed have rendered it suitable for rapid and convenient OP screening, and for being a bio-element for fabricating a potential optical biosensor in the future.
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Methyl parathion hydroloase
Methyl parathion hydrolase fused with glutathione-S-transferase
Gas chromatography–mass spectrometry
Limit of detection
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We thank the Department of Biotechnology, Faculty of Engineering and Industrial Technology, Silpakorn University and the Division of Science and Technology, Faculty of Science and Technology, Phetchaburi Rajabhat University for providing facilities and equipment necessary for this research. We also thank Miss Tipaporn Subsomboon for her helpful technical support and Ms. Jongkonrat Wichitwechkarn for proofreading and editing the manuscript.
This research was funded by Research and Researcher for Industry (RRI) grant, Thailand Research Fund, Contract No. PHD59I0024. Mixberry Co., Ltd. joined in part in the RRI funding program. The research work was also financially supported by the Research Network for Higher Education in Lower Central Region of Thailand through Silpakorn University Research, Innovation and Creativity Administration Office, a part of which was jointly funded by A4 Fruit Trading Co., Ltd.
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Rapichai, W., Chaichalerm, S., Mearnchu, J. et al. MPH-GST sensing microplate for easy detection of organophosphate insecticides. Biotechnol Lett (2021). https://doi.org/10.1007/s10529-021-03078-1
- Methyl parathion hydrolase
- Covalent immobilization
- Sensing microplate