Biosensing nitrite using the system nitrite redutase/Nafion/methyl viologen--a voltammetric study.

This work describes the construction and voltammetric characterization of a nitrite biosensor based on a cytochrome c-type nitrite reductase (ccNiR) and the Nafion ionomeric matrix loaded with methyl viologen as redox mediator. Despite the potential electrostatic repulsions between the anionic substrate and the Nafion sulfonate groups, the resulting bioelectrode exhibited electrocatalytic activity toward nitrite. This phenomenon must be due to the nonuniformity of the enzyme/Nafion membrane, which allows the direct interaction between the substrate and numerous enzyme molecules. Nevertheless, the anionic nature of Nafion exerted a certain diffusion barrier to nitrite, as revealed by the unusually elevated limits of the linear dynamic range and k(m)(app). The irregularity of the composite membrane also contributed to slow down the rate of charge transfer throughout the Nafion polymer. The level of viologens incorporated within the Nafion membrane had a strong influence in the analytical parameters: as much mediator was present, lower was the sensitivity and wider was the linear range. For an optimized ratio enzyme/mediator the sensitivity was 445+/-8 mA M(-1)cm(-2), within the linear range 75-800 microM; the lowest detected nitrite concentration was 60 microM. The operational stability of the biosensor and the influence of some possible interferences were evaluated.

[1]  N V Thakor,et al.  In vivo nitric oxide sensor using non-conducting polymer-modified carbon fiber. , 1998, Biosensors & bioelectronics.

[2]  B. Strehlitz,et al.  A nitrite sensor based on a highly sensitive nitrite reductase mediator-coupled amperometric detection. , 1996, Analytical chemistry.

[3]  Robert Huber,et al.  The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties. , 2003, European journal of biochemistry.

[4]  D. Bélanger,et al.  Electrochemical and Enzymatic Studies of Electron Transfer Mediation by Ferrocene Derivatives with Nafion‐Glucose Oxidase Electrodes , 1999 .

[5]  H. Abruña,et al.  A nitrite biosensor based on a maltose binding protein nitrite reductase fusion immobilized on an electropolymerized film of a pyrrole-derived bipyridinium. , 1997, Analytical chemistry.

[6]  John H. T. Luong,et al.  A regenerable pseudo-reagentless glucose biosensor based on Nafion polymer and l,1'-dimethylferricinium mediator , 1995 .

[7]  I. A. Abreu,et al.  Nitrite reductase from Desulfovibrio desulfuricans (ATCC 27774)--a heterooligomer heme protein with sulfite reductase activity. , 1996, Biochemical and biophysical research communications.

[8]  T Matsumoto,et al.  A long-term lifetime amperometric glucose sensor with a perfluorocarbon polymer coating. , 2001, Biosensors & bioelectronics.

[9]  Jing-Juan Xu,et al.  Glucose biosensors prepared by electropolymerization of p-chlorophenylamine with and without Nafion , 2002 .

[10]  Robert B. Moore,et al.  State of understanding of nafion. , 2004, Chemical reviews.

[11]  W. C. Maskell,et al.  The electrochemical reduction of manganese dioxide in acidic solutions: Part III. voltammetric peak 3 , 1977 .

[12]  D. A. Russell,et al.  Optical biosensing of nitrite ions using cytochrome cd1 nitrite reductase encapsulated in a sol-gel matrix. , 2000, The Analyst.

[13]  C. R. Martin,et al.  Effect of hydrophobic interactions on the rates of ionic diffusion in nafion films at electrode surfaces , 1983 .

[14]  O. J. Murphy,et al.  Enzyme electrodes based on ionomer films coated on electrodes , 1993 .

[15]  G. Ellis,et al.  Nitrite and nitrate analyses: a clinical biochemistry perspective. , 1998, Clinical biochemistry.

[16]  Jiaqi Deng,et al.  An amperometric lactate sensor employing tetrathiafulvalene in Nafion film as electron shuttle , 1995 .

[17]  A. Hodges,et al.  DIFFUSION OF VIOLOGENS IN NAFION FILM STUDIED BY A COMBINED ELECTROCHEMICAL-SPECTROPHOTOMETRIC METHOD , 1991 .

[18]  C. Costa,et al.  Electrochemical studies of the hexaheme nitrite reductase from Desulfovibrio desulfuricans ATCC 27774. , 1993, European journal of biochemistry.

[19]  M. Arnold,et al.  Selectivity enhancement for glutamate with a Nafion/glutamate oxidase biosensor. , 1996, Talanta.

[20]  P. Ugo,et al.  Ion-exchange voltammetry and electrocatalytic sensing capabilities of cytochrome c at polyestersulfonated ionomer coated glassy carbon electrodes. , 2002, Biosensors & bioelectronics.

[21]  R. Compton,et al.  Detection and determination of nitrate and nitrite: a review. , 2001, Talanta.

[22]  M. Vidal,et al.  Optical biosensor based on nitrite reductase immobilised in controlled pore glass. , 2002, Biosensors & bioelectronics.

[23]  Takahiro Yamaguchi,et al.  Incorporation of 1,1'-dibenzyl-4,4'-bipyridinium cation into Nafion films and charge transport in loaded film electrodes , 2001 .

[24]  M. Abo,et al.  Development of dimethyl sulfoxide biosensor using a mediator immobilized enzyme electrode. , 2003, The Analyst.

[25]  M Scharf,et al.  Electrochemical studies on nitrite reductase towards a biosensor. , 1995, Biochemical and biophysical research communications.

[26]  A. M. Yacynych,et al.  Galactose biosensors using composite polymers to prevent interferences. , 1995, Biosensors & bioelectronics.