Direct electrochemistry of horseradish peroxidase bonded on a conducting polymer modified glassy carbon electrode.

Direct electron transfer process of immobilized horseradish peroxidase (HRP) on a conducting polymer film, and its application as a biosensor for H2O2, were investigated by using electrochemical methods. The HRP was immobilized by covalent bonding between amino group of the HRP and carboxylic acid group of 5,2':5',2"-terthiophene-3'-carboxylic acid polymer (TCAP) which is present on a glassy carbon (GC). A pair of redox peaks attributed to the direct redox process of HRP immobilized on the biosensor electrode were observed at the HRPmid R:TCAPmid R:GC electrode in a 10 mM phosphate buffer solution (pH 7.4). The surface coverage of the HRP immobilized on TCAPmid R:GC was about 1.2 x 10(-12) mol cm(-2) and the electron transfer rate (ks) was determined to be 1.03 s(-1). The HRPmid R:TCAPmid R:GC electrode acted as a sensor and displayed an excellent specific electrocatalytic response to the reduction of H2O2 without the aid of an electron transfer mediator. The calibration range of H2O2 was determined from 0.3-1.5 mM with a good linear relation.

[1]  Wolfgang Schuhmann,et al.  Amperometric enzyme biosensors based on optimised electron-transfer pathways and non-manual immobilisation procedures. , 2002, Journal of biotechnology.

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

[3]  S. Cosnier Biomolecule immobilization on electrode surfaces by entrapment or attachment to electrochemically polymerized films. A review. , 1999, Biosensors & bioelectronics.

[4]  W. Schuhmann,et al.  Minizymes. A new strategy for the development of reagentless amperometric biosensors based on direct electron-transfer processes , 1997 .

[5]  Gordon G. Wallace,et al.  Conducting electroactive polymer-based biosensors , 1999 .

[6]  Jenny Emnéus,et al.  Kinetic models of horseradish peroxidase action on a graphite electrode , 1995 .

[7]  K. E. Everse,et al.  Peroxidases in chemistry and biology , 1990 .

[8]  Y. Shim,et al.  Simple preparation of terthiophene-3′-carboxylic acid and characterization of its polymer , 2002 .

[9]  S. Dong,et al.  The electrochemical study of oxidation-reduction properties of horseradish peroxidase , 1997 .

[10]  Wolfgang Schuhmann,et al.  Electron transfer principles in amperometric biosensors: direct electron transfer between enzymes and electrode surface , 1996 .

[11]  E. Ferapontova,et al.  Effect of cysteine mutations on direct electron transfer of horseradish peroxidase on gold. , 2002, Biosensors & bioelectronics.

[12]  Takeshi Yamauchi,et al.  Glucose-sensing characteristics of conducting polymer bound with glucose oxidase , 1999 .

[13]  C. Danilowicz,et al.  Electrical Communication between Electrodes and Enzymes Mediated by Redox Hydrogels. , 1996, Analytical chemistry.

[14]  C. Lowe,et al.  Immobilization of glucose oxidase in ferrocene-modified pyrrole polymers. , 1988, Analytical chemistry.

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

[16]  M. Watanabe,et al.  Amperometric biosensor for polyphenol based on horseradish peroxidase immobilized on gold electrodes , 2001 .

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

[18]  W. Schuhmann,et al.  A reagentless glucose biosensor based on glucose oxidase entrapped into osmium-complex modified polypyrrole films , 2001 .

[19]  M. R. Tarasevich,et al.  192 - Electrochemical Properties of Peroxidase , 1978 .

[20]  N. Oyama,et al.  Kinetic analysis of electron transfer from a graphite coating to horseradish peroxidase , 1998 .

[21]  Adam Heller,et al.  Electrical communication between redox centers of glucose oxidase and electrodes via electrostatically and covalently bound redox polymers , 1989 .

[22]  Wolfgang Schuhmann,et al.  Electron-transfer pathways in amperometric biosensors. Ferrocene-modified enzymes entrapped in conducting-polymer layers , 1995 .

[23]  A. Malinauskas Electrocatalysis at conducting polymers , 1999 .

[24]  C. Visy,et al.  Detection of uric acid with a new type of conducting polymer-based enzymatic sensor by bipotentiostatic technique , 1999 .

[25]  A Heller,et al.  Cross-linked redox gels containing glucose oxidase for amperometric biosensor applications. , 1990, Analytical chemistry.