Highly Sensitive Hydrogen Peroxide Biosensor Based on Tobacco Peroxidase Immobilized on p-Phenylenediamine Diazonium Cation Grafted Carbon Nanotubes : Preventing Fenton-like Inactivation at Negative Potential
(2021) In ChemElectroChem 8(13). p.2495-2504- Abstract
Herein, we present a novel electrode platform for H2O2 detection based on the immobilization of recombinant Tobacco Peroxidase (r-TOP) onto graphite electrodes (G) modified with p-phenylenediamine (p-PD) diazonium cation grafted multi-walled carbon nanotubes (MWCNTs). The employment of both p-phenylenediamine moieties and covalent cross-linking by using glutaraldehyde allowed us to enhance the sensitivity, stability, and selectivity toward H2O2 detection, as well as preventing enzyme inactivation due to the electro-Fenton reaction. This reaction continuously produces hydroxyl radicals, whose high and unselective reactivity is likely to reduce drastically the operating life of the biosensor.... (More)
Herein, we present a novel electrode platform for H2O2 detection based on the immobilization of recombinant Tobacco Peroxidase (r-TOP) onto graphite electrodes (G) modified with p-phenylenediamine (p-PD) diazonium cation grafted multi-walled carbon nanotubes (MWCNTs). The employment of both p-phenylenediamine moieties and covalent cross-linking by using glutaraldehyde allowed us to enhance the sensitivity, stability, and selectivity toward H2O2 detection, as well as preventing enzyme inactivation due to the electro-Fenton reaction. This reaction continuously produces hydroxyl radicals, whose high and unselective reactivity is likely to reduce drastically the operating life of the biosensor. The protection against the electro-Fenton reaction is mainly ascribed to a beneficial enzyme immobilization leading to a correct orientation achieved through cross-linking the enzyme in combination with interaction between the uncoupled -NH2 groups (mainly uncharged at pH 7, considering a pKa of 4.6) available on the electrode surface and the enzyme. In particular, the electrode based on the r-TOP/p-PD/MWCNTs/G platform showed a lower limit of detection of 1.8 μM H2O2, an extended linear range between 6 and 900 μM H2O2, as well as a significant increase in sensitivity (63.1±0.1 μA mM−1 cm−2) compared with previous work based on TOP. Finally, the r-TOP/p-PD/MWCNTs/G electrode was tested in several H2O2 spiked food samples as a screening analytical method for the detection of H2O2.
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- author
- Ciogli, Leonardo ; Zumpano, Rosaceleste ; Poloznikov, Andrey A. ; Hushpulian, Dmitry M. ; Tishkov, Vladimir I. ; Andreu, Rafael ; Gorton, Lo LU ; Mazzei, Franco ; Favero, Gabriele and Bollella, Paolo
- organization
- publishing date
- 2021
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- electro-Fenton reaction, hydrogen peroxide biosensor, modified electrodes, p-phenylenediamine (p-PD) diazonium cations, recombinant Tobacco Peroxidase (r-TOP)
- in
- ChemElectroChem
- volume
- 8
- issue
- 13
- pages
- 10 pages
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:85110546094
- ISSN
- 2196-0216
- DOI
- 10.1002/celc.202100341
- language
- English
- LU publication?
- yes
- id
- 69eb1a16-7be3-4060-abaa-68ba073cf0ff
- date added to LUP
- 2021-09-09 13:42:06
- date last changed
- 2022-04-27 03:47:54
@article{69eb1a16-7be3-4060-abaa-68ba073cf0ff, abstract = {{<p>Herein, we present a novel electrode platform for H<sub>2</sub>O<sub>2</sub> detection based on the immobilization of recombinant Tobacco Peroxidase (r-TOP) onto graphite electrodes (G) modified with p-phenylenediamine (p-PD) diazonium cation grafted multi-walled carbon nanotubes (MWCNTs). The employment of both p-phenylenediamine moieties and covalent cross-linking by using glutaraldehyde allowed us to enhance the sensitivity, stability, and selectivity toward H<sub>2</sub>O<sub>2</sub> detection, as well as preventing enzyme inactivation due to the electro-Fenton reaction. This reaction continuously produces hydroxyl radicals, whose high and unselective reactivity is likely to reduce drastically the operating life of the biosensor. The protection against the electro-Fenton reaction is mainly ascribed to a beneficial enzyme immobilization leading to a correct orientation achieved through cross-linking the enzyme in combination with interaction between the uncoupled -NH<sub>2</sub> groups (mainly uncharged at pH 7, considering a pK<sub>a</sub> of 4.6) available on the electrode surface and the enzyme. In particular, the electrode based on the r-TOP/p-PD/MWCNTs/G platform showed a lower limit of detection of 1.8 μM H<sub>2</sub>O<sub>2</sub>, an extended linear range between 6 and 900 μM H<sub>2</sub>O<sub>2</sub>, as well as a significant increase in sensitivity (63.1±0.1 μA mM<sup>−1</sup> cm<sup>−2</sup>) compared with previous work based on TOP. Finally, the r-TOP/p-PD/MWCNTs/G electrode was tested in several H<sub>2</sub>O<sub>2</sub> spiked food samples as a screening analytical method for the detection of H<sub>2</sub>O<sub>2</sub>.</p>}}, author = {{Ciogli, Leonardo and Zumpano, Rosaceleste and Poloznikov, Andrey A. and Hushpulian, Dmitry M. and Tishkov, Vladimir I. and Andreu, Rafael and Gorton, Lo and Mazzei, Franco and Favero, Gabriele and Bollella, Paolo}}, issn = {{2196-0216}}, keywords = {{electro-Fenton reaction; hydrogen peroxide biosensor; modified electrodes; p-phenylenediamine (p-PD) diazonium cations; recombinant Tobacco Peroxidase (r-TOP)}}, language = {{eng}}, number = {{13}}, pages = {{2495--2504}}, publisher = {{Wiley-Blackwell}}, series = {{ChemElectroChem}}, title = {{Highly Sensitive Hydrogen Peroxide Biosensor Based on Tobacco Peroxidase Immobilized on p-Phenylenediamine Diazonium Cation Grafted Carbon Nanotubes : Preventing Fenton-like Inactivation at Negative Potential}}, url = {{http://dx.doi.org/10.1002/celc.202100341}}, doi = {{10.1002/celc.202100341}}, volume = {{8}}, year = {{2021}}, }