Biosensors based on novel peroxidases with improved properties in direct and mediated electron transfer

Lindgren, Annika; Ruzgas, Tautgirdas; Gorton, Lo; Csöregi, Elisabeth; Ardila, Gerardo Bautista; Sakharov, Ivan Yu.; Gazaryan, I. G.

doi:10.1016/s0956-5663(00)00110-x

Cited by 134 publications

(71 citation statements)

References 26 publications

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“…HRP entrapped in modified organic polymer films, composite materials, immobilized or covalently attached onto graphite electrodes, exhibited high stability and activity in organic solvents such as acetonitrile, chloroform, acetone and a range of alcohols. Whereas HRP exhibits quite high activity and stability, other peroxidases with improved activity and stability properties are now available [26]. A novel peroxidase isolated from the royal palm tree (RPTP, pI 3.5) possesses higher thermal and pH stability than all the peroxidases reported to date and an additional tolerance to high concentrations of H 2 O 2 (which acts as a killer substrate for peroxidases) [27].…”

Section: Introductionmentioning

confidence: 99%

Bioelectrocatalysis of Plant Peroxidases Immobilized on Graphite in Aqueous and Mixed Solvent Media

et al. 2005

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The bioelectrocatalytic function of two plant peroxidases, horseradish peroxidase (HRP) and a newly purified royal palm tree-leaf peroxidase (RPTP), was studied on graphite electrodes in aqueous buffer solutions, over the pH range 7.0 -3.0, and in aqueous buffer solutions, at pH 6.0, containing different amounts of a polar organic cosolvent, i.e., ethanol and acetonitrile. The kinetics of direct and of mediated (using catechol as mediator) reduction of H 2 O 2 at the HRP-and RPTP-modified graphite electrodes were analyzed amperometrically at À 50 mV(vs. Ag j AgCl j 0.1 M KCl) in a flow-through wall-jet electrochemical system. Values of the apparent rate constant of the heterogeneous electron transfer between the enzyme and graphite, k s , the rate constant for enzymatic reduction of H 2 O 2 , k 1 , and the rate constant of mediated reduction of H 2 O 2 , k 3, were determined using the modified Koutecky-Levich approach. Analysis of the variation of the rate constants and the response of the peroxidase-modified electrodes to H 2 O 2 with pH and content of the organic cosolvent demonstrated that maximal peroxidase bioelectrocatalytic activity occurred at pH 5.0 -6.0 and at concentrations of ethanol of 10 -20% v/v. At a lower pH, higher concentrations of ethanol, and in aqueous solutions of acetonitrile, the bioelectrocatalytic activity of the peroxidase-modified electrodes drastically decreased. However, contrary to the data for homogeneous catalysis, both peroxidases were bioelectrocatalytically active even in 95% organic co-solvents, thus demonstrating a stabilizing effect of the enzyme immobilization on the bioelectrocatalytic performance of the peroxidases. RPTP immobilized on graphite demonstrated lower overall activity but a higher pH-and organic cosolvent-stability than HRP.

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Section: Introductionmentioning

confidence: 99%

Bioelectrocatalysis of Plant Peroxidases Immobilized on Graphite in Aqueous and Mixed Solvent Media

et al. 2005

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“…Hydrogen peroxide also has acted as an active chemical reagent for preparation of many chemicals like hydrazine and as detoxicating organic pollutants such as formaldehyde and phenol. 11,71 The integrations of new materials and fabrication strategies with conventional voltammetric techniques have given birth to the new amperometric biosensors. 49,61,65 The use of amperometric biosensors based on horseradish peroxidase provided an alternative process for the measurement of hydrogen peroxide.…”

Section: ·5 Biocomposite Electrodesmentioning

confidence: 99%

“…49,61,65 The use of amperometric biosensors based on horseradish peroxidase provided an alternative process for the measurement of hydrogen peroxide. 10,57,61,71 The design and construction of rigid matrix materials utilized enzyme-graphite-polymer composites. The rigidity of the biocomposite did ensure a simple and lasting immobilization of the enzyme.…”

Section: ·5 Biocomposite Electrodesmentioning

confidence: 99%

Recent Developments, Characteristics, and Potential Applications of Electrochemical Biosensors

2004

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The objective of this study is to analyze the technical importance, performance, techniques, advantages, and disadvantages of the biosensors in general and of the electrochemical biosensors in particular. A product of reaction diffuses to the transducer in the first generation biosensors (based on Clark biosensors). The mediated biosensors or second generation biosensors use specific mediators between the reaction and the transducer to improve sensitivity. The second generation biosensors involve two steps: first, there is a redox reaction between enzyme and substrate that is reoxidized by the mediator, and eventually the mediator is oxidized by the electrode. No normal product or mediator diffusion is directly involved in the third generation biosensors, direct biosensors. Based on the type of transducer, current biosensors are divided into optical, mass, thermal, and electrochemical sensors. They are used in medical diagnostics, food quality controls, environmental monitoring, and other applications. These biosensors are also grouped under two broad categories of sensors: direct and indirect detection systems. Moreover, these systems could be further grouped into continuous or batch operation. Therefore, amperometric biosensors and their current applications are focused on more in detail since they are the most commonly used biosensors in monitoring and diagnosing tests in clinical analysis. Problems related to the commercialization of medical, environmental, and industrial biosensors as well as their performance characteristics, their competitiveness in comparison to the conventional analytical tools, and their costs determine the future development of these biosensors.

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“…8,9 The latter approach is particularly attractive due to simplicity and reliability of measurements, and the possibility for miniaturization. Electrochemical biosensors commonly rely on the enzyme, horseradish peroxidase (HRP)-an oxidoreductase enzyme that catalyzes reduction of H 2 O 2 .…”

Section: Introductionmentioning

confidence: 99%

Electrochemical biosensors for on-chip detection of oxidative stress from immune cells

et al. 2011

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Seamless integration of biological components with electrochemical sensors is critical in the development of microdevices for cell analysis. The present paper describes the integration miniature Au electrodes next to immune cells (macrophages) in order to detect cell-secreted hydrogen peroxide (H2O2). Photopatterning of poly(ethylene glycol) (PEG) hydrogels was used to both immobilize horseradish peroxidase molecules onto electrodes and to define regions for cell attachment in the vicinity of sensing electrodes. Electrodes micropatterned in such a manner were enclosed inside poly(dimethylsiloxane) fluid conduits and incubated with macrophages. The cells attached onto the exposed glass regions in the vicinity of the electrodes and nowhere else on the non-fouling PEG hydrogel surface. A microfluidic device was converted into an electrochemical cell by placing flow-through Ag/AgCl reference and Pt wire counter electrodes at the outlet and inlet, respectively. This microdevice with integrated H2O2-sensing electrodes had sensitivity of 27 μA/cm2 mM with a limit of detection of 2 μM. Importantly, this microdevice allowed controllable seeding of macrophages next to electrodes, activation of these cells and on-chip monitoring of H2O2 release in real time. In the future, this biosensor platform may be utilized for monitoring of macrophage responses to pathogens or for the study of inflammatory signaling in micropatterned cell cultures.

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Biosensors based on novel peroxidases with improved properties in direct and mediated electron transfer

Cited by 134 publications

References 26 publications

Bioelectrocatalysis of Plant Peroxidases Immobilized on Graphite in Aqueous and Mixed Solvent Media

Bioelectrocatalysis of Plant Peroxidases Immobilized on Graphite in Aqueous and Mixed Solvent Media

Recent Developments, Characteristics, and Potential Applications of Electrochemical Biosensors

Electrochemical biosensors for on-chip detection of oxidative stress from immune cells

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