Direct electron transfer of horseradish peroxidase on Nafion-cysteine modified gold electrode

Abstract Direct electron transfer of horseradish peroxidase, immobilized on a functional membrane-modified gold electrode, was studied. The electrode showed a quasi-reversible electrochemical redox behavior with a formal potential of 60 mV (versus Ag/AgCl) in 20 mM potassium phosphate buffer solution at pH 7.0 and temperature 25 °C. The cathodic transfer coefficient was 0.42 and electron transfer rate constant was evaluated to be 1.6 s−1. Furthermore, the modified electrode was used as a biosensor and exhibited a satisfactory stability and sensitivity to H2O2. The linear range of this biosensor for H2O2 determination was from 5.0 × 10−6 to 1.5 × 10−4 M while its detection limit, based on a signal-to-noise ratio of 3, was 1.3 × 10−6 M. The apparent Michaelis–Menten constant ( K m app ) for immobilized HRP was calculated to be 1.6 × 10−4 M.

[1]  Hedayatollah Ghourchian,et al.  Direct electron transfer of redox proteins on a Nafion-cysteine modified gold electrode , 2006 .

[2]  A. Mauk,et al.  Cytochrome C: A Multidisciplinary Approach , 1996 .

[3]  Yuehe Lin,et al.  Enzyme nanoparticles-based electronic biosensor. , 2005, Chemical communications.

[4]  E. Laviron,et al.  Adsorption, autoinhibition and autocatalysis in polarography and in linear potential sweep voltammetry , 1974 .

[5]  Genxi Li,et al.  Direct Electrochemistry and Enhanced Catalytic Activity for Hemoglobin in a Sodium Montmorillonite Film , 2000 .

[6]  N. C. Veitch,et al.  Horseradish peroxidase: a modern view of a classic enzyme. , 2004, Phytochemistry.

[7]  Hongyuan Chen,et al.  Electrochemical study of a new methylene blue/silicon oxide nanocomposition mediator and its application for stable biosensor of hydrogen peroxide. , 2005, Biosensors & bioelectronics.

[8]  T. Ohsaka,et al.  Self-assembled monolayers suitable for electron-transfer promotion of copper, zinc-superoxide dismutase , 2004 .

[9]  Qiang Chen,et al.  An Unmediated Hydrogen Peroxide Sensor Based on a Hemoglobin-sds Film Modified Electrode , 2000 .

[10]  E. Laviron.  General expression of the linear potential sweep voltammogram in the case of diffusionless electrochemical systems , 1979 .

[11]  H. A. Harbury.  Oxidation-reduction potentials of horseradish peroxidase. , 1953, The Journal of biological chemistry.

[12]  James F. Rusling,et al.  Direct Electrochemistry of Myoglobin and Cytochrome P450cam in Alternate Layer-by-Layer Films with DNA and Other Polyions , 1998 .

[13]  H. Wang,et al.  Incorporation of horseradish peroxidase in a Kieselguhr membrane and the application to a mediator-free hydrogen peroxide sensor. , 2001, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[14]  S. Dong,et al.  A facile approach to immobilize protein for biosensor: self-assembled supported bilayer lipid membranes on glassy carbon electrode. , 2001, Biosensors & bioelectronics.

[15]  G Li,et al.  Electron-transfer reactivity and enzymatic activity of hemoglobin in a SP Sephadex membrane. , 2001, Analytical chemistry.

[16]  T. Ohsaka,et al.  Superoxide dismutase-based third-generation biosensor for superoxide anion. , 2002, Analytical chemistry.

[17]  T. Yonetani,et al.  Heme-modification studies on horseradish peroxidase. , 1972, Biochimica et biophysica acta.

[18]  S. Mazumdar,et al.  Direct electrochemistry of heme proteins: effect of electrode surface modification by neutral surfactants. , 2001, Bioelectrochemistry.

[19]  Genxi Li,et al.  Electron transfer reactivity and the catalytic activity of horseradish peroxidase incorporated in dipalmitoylphosphatidic acid films. , 2006, Bioelectrochemistry.

[20]  Roberto Santucci,et al.  Direct electrochemistry of membrane-entrapped horseradish peroxidase.: Part II: Amperometric detection of hydrogen peroxide , 1998 .

[21]  Genxi Li,et al.  Fabrication of Ultrathin, Protein-containing Films by Layer-by-Layer Assembly and Electrochemical Characterization of Hemoglobin Entrapped in the Film , 2003 .

[22]  James F. Rusling,et al.  Enhanced electron transfer for myoglobin in surfactant films on electrodes , 1993 .

[23]  G. S. Wilson,et al.  Biosensors : fundamentals and applications , 1987 .

[24]  T. Ohsaka,et al.  A carbon fiber microelectrode-based third-generation biosensor for superoxide anion. , 2005, Biosensors & bioelectronics.

[25]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[26]  Xinjian Liu,et al.  Enhanced electron-transfer reactivity of horseradish peroxidase in phosphatidylcholine films and its catalysis to nitric oxide. , 2004, Journal of biotechnology.

[27]  F. Teale,et al.  Cleavage of the haem-protein link by acid methylethylketone. , 1959, Biochimica et biophysica acta.

[28]  J. Rusling,et al.  Electron transfer between myoglobin and electrodes in thin films of phosphatidylcholines and dihexadecylphosphate. , 1997, Biophysical chemistry.

[29]  Q. Gao,et al.  An unmediated hydrogen peroxide biosensor based on hemoglobin incorporated in a montmorillonite membrane. , 2001, The Analyst.

[30]  T. Ohsaka,et al.  A facilitated electron transfer of copper--zinc superoxide dismutase (SOD) based on a cysteine-bridged SOD electrode. , 2002, Biochimica et biophysica acta.

[31]  H. Ju,et al.  Direct electrochemistry of horseradish peroxidase immobilized on a colloid/cysteamine-modified gold electrode. , 2000, Analytical biochemistry.

[32]  F. Armstrong,et al.  Electrocatalytic Voltammetry of Succinate Dehydrogenase: Direct Quantification of the Catalytic Properties of a Complex Electron-Transport Enzyme , 1996 .

[33]  H. Heering,et al.  Interpreting the Catalytic Voltammetry of Electroactive Enzymes Adsorbed on Electrodes , 1998 .

[34]  N. Hu,et al.  Direct electron transfer for hemoglobin in biomembrane-like dimyristoyl phosphatidylcholine films on pyrolytic graphite electrodes. , 1999, Bioelectrochemistry and bioenergetics.

[35]  Huangxian Ju,et al.  Hydrogen peroxide sensor based on horseradish peroxidase-labeled Au colloids immobilized on gold electrode surface by cysteamine monolayer , 1999 .

[36]  Qin Xu,et al.  Porous Gold-Nanoparticle−CaCO3 Hybrid Material: Preparation, Characterization, and Application for Horseradish Peroxidase Assembly and Direct Electrochemistry , 2006 .

[37]  Genxi Li,et al.  Enhanced peroxidase activity of hemoglobin in a DNA membrane and its application to an unmediated hydrogen peroxide biosensor. , 2003, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[38]  N. Hu,et al.  Direct electrochemistry and electrocatalysis with horseradish peroxidase in Eastman AQ films. , 2001, Bioelectrochemistry.

[39]  Yoon-Bo Shim,et al.  Direct electrochemistry of horseradish peroxidase bonded on a conducting polymer modified glassy carbon electrode. , 2003, Biosensors & bioelectronics.

[40]  E. Ferapontova.  Direct peroxidase bioelectrocatalysis on a variety of electrode materials , 2004 .

[41]  L. Gorton,et al.  Direct electron transfer between heme-containing enzymes and electrodes as basis for third generation biosensors , 1999 .

[42]  J. Rusling,et al.  PROTON-COUPLED ELECTRON TRANSFER FROM ELECTRODES TO MYOGLOBIN IN ORDERED BIOMEMBRANE-LIKE FILMS , 1997 .

[43]  Genxi Li,et al.  Nitric Oxide Biosensors Based on Hb/Phosphatidylcholine Films , 2002, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[44]  Yan Liu,et al.  Direct electrochemistry of horseradish peroxidase immobilized on gold colloid/cysteine/nafion-modified platinum disk electrode , 2006 .

[45]  James F. Rusling,et al.  Electroactive Myoglobin Films Grown Layer-by-Layer with Poly(styrenesulfonate) on Pyrolytic Graphite Electrodes , 2000 .

[46]  Ali Akbar Saboury,et al.  Nafion‐Methylene Blue Functional Membrane and Its Application in Chemical and Biosensing , 2007 .

[47]  H. Ghourchian,et al.  Anthraquinone 2-carboxylic acid as an electron shuttling mediator and attached electron relay for horseradish peroxidase , 2005 .

[48]  J. Kong,et al.  Characterization of the direct electron transfer and bioelectrocatalysis of horseradish peroxidase in DNA film at pyrolytic graphite electrode , 2000 .

[49]  Fraser A. Armstrong,et al.  Reaction of complex metalloproteins studied by protein-film voltammetry , 1997 .

[50]  G. S. Wilson,et al.  Rotating ring-disk enzyme electrode for biocatalysis kinetic studies and characterization of the immobilized enzyme layer , 1980 .