Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor
Organophosphates are organic substances that contain a phosphoryl or a thiophosphoryl bond. They are mainly used around the world as pesticides, but can also be used as chemical warfare agents. Their detection is normally entrusted to techniques like GC- and LC-MS that, although sensitive, do not al...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2015-02-01
|
| Series: | Sensors |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1424-8220/15/2/3932 |
| _version_ | 1818013905810620416 |
|---|---|
| author | Paola Carullo Giovanni Paolo Cetrangolo Luigi Mandrich Giuseppe Manco Ferdinando Febbraio |
| author_facet | Paola Carullo Giovanni Paolo Cetrangolo Luigi Mandrich Giuseppe Manco Ferdinando Febbraio |
| author_sort | Paola Carullo |
| collection | DOAJ |
| description | Organophosphates are organic substances that contain a phosphoryl or a thiophosphoryl bond. They are mainly used around the world as pesticides, but can also be used as chemical warfare agents. Their detection is normally entrusted to techniques like GC- and LC-MS that, although sensitive, do not allow their identification on site and in real time. We have approached their identification by exploiting the high-affinity binding of these compounds with the esterase 2 from Alicyclobacillus acidocaldarius. Using an in silico analysis to evaluate the binding affinities of the enzyme with organophosphate inhibitors, like paraoxon, and other organophosphate compounds, like parathion, chlorpyriphos, and other organophosphate thio-derivatives, we have designed fluorescence spectroscopy experiments to study the quenching of the tryptophan residues after esterase 2 binding with the organophosphate pesticides. The changes in the fluorescence signals permitted an immediate and quantitative identification of these compounds from nano- to picomolar concentrations. A fluorescence based polarity-sensitive probe (ANS) was also employed as a means to understand the extent of the interactions involved, as well as to explore other ways to detect organophosphate pesticides. Finally, we designed a framework for the development of a biosensor that exploits fluorescence technology in combination with a sensitive and very stable bio-receptor. |
| first_indexed | 2024-04-14T06:39:07Z |
| format | Article |
| id | doaj.art-449efd4bcf8c43bb9afc736159721563 |
| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-04-14T06:39:07Z |
| publishDate | 2015-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Sensors |
| spelling | doaj.art-449efd4bcf8c43bb9afc7361597215632022-12-22T02:07:23ZengMDPI AGSensors1424-82202015-02-011523932395110.3390/s150203932s150203932Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic SensorPaola Carullo0Giovanni Paolo Cetrangolo1Luigi Mandrich2Giuseppe Manco3Ferdinando Febbraio4The Institute of Protein Biochemistry (IBP) of the Italian National Research Council (CNR), Via P. Castellino 111, 80131 Naples, ItalyThe Institute of Protein Biochemistry (IBP) of the Italian National Research Council (CNR), Via P. Castellino 111, 80131 Naples, ItalyThe Institute of Protein Biochemistry (IBP) of the Italian National Research Council (CNR), Via P. Castellino 111, 80131 Naples, ItalyThe Institute of Protein Biochemistry (IBP) of the Italian National Research Council (CNR), Via P. Castellino 111, 80131 Naples, ItalyThe Institute of Protein Biochemistry (IBP) of the Italian National Research Council (CNR), Via P. Castellino 111, 80131 Naples, ItalyOrganophosphates are organic substances that contain a phosphoryl or a thiophosphoryl bond. They are mainly used around the world as pesticides, but can also be used as chemical warfare agents. Their detection is normally entrusted to techniques like GC- and LC-MS that, although sensitive, do not allow their identification on site and in real time. We have approached their identification by exploiting the high-affinity binding of these compounds with the esterase 2 from Alicyclobacillus acidocaldarius. Using an in silico analysis to evaluate the binding affinities of the enzyme with organophosphate inhibitors, like paraoxon, and other organophosphate compounds, like parathion, chlorpyriphos, and other organophosphate thio-derivatives, we have designed fluorescence spectroscopy experiments to study the quenching of the tryptophan residues after esterase 2 binding with the organophosphate pesticides. The changes in the fluorescence signals permitted an immediate and quantitative identification of these compounds from nano- to picomolar concentrations. A fluorescence based polarity-sensitive probe (ANS) was also employed as a means to understand the extent of the interactions involved, as well as to explore other ways to detect organophosphate pesticides. Finally, we designed a framework for the development of a biosensor that exploits fluorescence technology in combination with a sensitive and very stable bio-receptor.http://www.mdpi.com/1424-8220/15/2/3932biosensororganophosphatesfluorescence spectroscopythermostable esterase 2nerve agentspesticides |
| spellingShingle | Paola Carullo Giovanni Paolo Cetrangolo Luigi Mandrich Giuseppe Manco Ferdinando Febbraio Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor Sensors biosensor organophosphates fluorescence spectroscopy thermostable esterase 2 nerve agents pesticides |
| title | Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor |
| title_full | Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor |
| title_fullStr | Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor |
| title_full_unstemmed | Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor |
| title_short | Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor |
| title_sort | fluorescence spectroscopy approaches for the development of a real time organophosphate detection system using an enzymatic sensor |
| topic | biosensor organophosphates fluorescence spectroscopy thermostable esterase 2 nerve agents pesticides |
| url | http://www.mdpi.com/1424-8220/15/2/3932 |
| work_keys_str_mv | AT paolacarullo fluorescencespectroscopyapproachesforthedevelopmentofarealtimeorganophosphatedetectionsystemusinganenzymaticsensor AT giovannipaolocetrangolo fluorescencespectroscopyapproachesforthedevelopmentofarealtimeorganophosphatedetectionsystemusinganenzymaticsensor AT luigimandrich fluorescencespectroscopyapproachesforthedevelopmentofarealtimeorganophosphatedetectionsystemusinganenzymaticsensor AT giuseppemanco fluorescencespectroscopyapproachesforthedevelopmentofarealtimeorganophosphatedetectionsystemusinganenzymaticsensor AT ferdinandofebbraio fluorescencespectroscopyapproachesforthedevelopmentofarealtimeorganophosphatedetectionsystemusinganenzymaticsensor |