Interprotein Coupling Enhances the Electrocatalytic Efficiency of Tobacco Peroxidase Immobilized at a Graphite Electrode.
(2015) In Analytical Chemistry 87(21). p.10807-10814- Abstract
- Covalent immobilization of enzymes at electrodes via amide bond formation is usually carried out by a two-step protocol, in which surface carboxylic groups are first activated with the corresponding cross-coupling reagents and then reacted with protein amine groups. Herein, it is shown that a modification of the above protocol, involving the simultaneous incubation of tobacco peroxidase and the pyrolytic graphite electrode with the cross-coupling reagents produces higher and more stable electrocatalytic currents than those obtained with either physically adsorbed enzymes or covalently immobilized enzymes according to the usual immobilization protocol. The remarkably improved electrocatalytic properties of the present peroxidase biosensor... (More)
- Covalent immobilization of enzymes at electrodes via amide bond formation is usually carried out by a two-step protocol, in which surface carboxylic groups are first activated with the corresponding cross-coupling reagents and then reacted with protein amine groups. Herein, it is shown that a modification of the above protocol, involving the simultaneous incubation of tobacco peroxidase and the pyrolytic graphite electrode with the cross-coupling reagents produces higher and more stable electrocatalytic currents than those obtained with either physically adsorbed enzymes or covalently immobilized enzymes according to the usual immobilization protocol. The remarkably improved electrocatalytic properties of the present peroxidase biosensor that operates in the 0.3 V ≤ E ≤ 0.8 V (vs SHE) potential range can be attributed to both an efficient electronic coupling between tobacco peroxidase and graphite and to the formation of intra- and intermolecular amide bonds that stabilize the protein structure and improve the percentage of anchoring groups that provide an adequate orientation for electron exchange with the electrode. The optimized tobacco peroxidase sensor exhibits a working concentration range of 10-900 μM, a sensitivity of 0.08 A M(-1) cm(-2) (RSD 0.05), a detection limit of 2 μM (RSD 0.09), and a good long-term stability, as long as it operates at low temperature. These parameter values are among the best reported so far for a peroxidase biosensor operating under simple direct electron transfer conditions. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/8158922
- author
- Olloqui-Sariego, José Luis ; Zakharova, Galina S ; Poloznikov, Andrey A ; Calvente, Juan José ; Hushpulian, Dmitry M ; Gorton, Lo LU and Andreu, Rafael
- organization
- publishing date
- 2015
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical Chemistry
- volume
- 87
- issue
- 21
- pages
- 10807 - 10814
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- pmid:26437673
- wos:000364354900025
- scopus:84946430936
- pmid:26437673
- ISSN
- 1520-6882
- DOI
- 10.1021/acs.analchem.5b01710
- language
- English
- LU publication?
- yes
- id
- e87f7e5c-2d1d-41a8-aef7-05bfdcd7d4f6 (old id 8158922)
- date added to LUP
- 2016-04-01 10:00:38
- date last changed
- 2022-01-25 18:54:54
@article{e87f7e5c-2d1d-41a8-aef7-05bfdcd7d4f6,
abstract = {{Covalent immobilization of enzymes at electrodes via amide bond formation is usually carried out by a two-step protocol, in which surface carboxylic groups are first activated with the corresponding cross-coupling reagents and then reacted with protein amine groups. Herein, it is shown that a modification of the above protocol, involving the simultaneous incubation of tobacco peroxidase and the pyrolytic graphite electrode with the cross-coupling reagents produces higher and more stable electrocatalytic currents than those obtained with either physically adsorbed enzymes or covalently immobilized enzymes according to the usual immobilization protocol. The remarkably improved electrocatalytic properties of the present peroxidase biosensor that operates in the 0.3 V ≤ E ≤ 0.8 V (vs SHE) potential range can be attributed to both an efficient electronic coupling between tobacco peroxidase and graphite and to the formation of intra- and intermolecular amide bonds that stabilize the protein structure and improve the percentage of anchoring groups that provide an adequate orientation for electron exchange with the electrode. The optimized tobacco peroxidase sensor exhibits a working concentration range of 10-900 μM, a sensitivity of 0.08 A M(-1) cm(-2) (RSD 0.05), a detection limit of 2 μM (RSD 0.09), and a good long-term stability, as long as it operates at low temperature. These parameter values are among the best reported so far for a peroxidase biosensor operating under simple direct electron transfer conditions.}},
author = {{Olloqui-Sariego, José Luis and Zakharova, Galina S and Poloznikov, Andrey A and Calvente, Juan José and Hushpulian, Dmitry M and Gorton, Lo and Andreu, Rafael}},
issn = {{1520-6882}},
language = {{eng}},
number = {{21}},
pages = {{10807--10814}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Analytical Chemistry}},
title = {{Interprotein Coupling Enhances the Electrocatalytic Efficiency of Tobacco Peroxidase Immobilized at a Graphite Electrode.}},
url = {{http://dx.doi.org/10.1021/acs.analchem.5b01710}},
doi = {{10.1021/acs.analchem.5b01710}},
volume = {{87}},
year = {{2015}},
}