Assembly of alternating polymerized mediator, polymerized coenzyme, and enzyme modified electrode by layer-by-layer adsorption technique

Abstract The polymerized mediator (polyethyleneimine ferrocene), and polymerized NADP + (Alg–NADP + ) were prepared and their application to biosensing was investigated. An amperometric biosensor was fabricated utilizing two enzymes, glutathione reductase (GR), and glucose-6-phosphate dehydrogenase (G6PDH) with polymerized NADP + as part of a layer-by-layer adsorption (LBL) technique on a chemically modified carbon electrode. An electrocatalytic steady-state current as a result of the oxidation of glucose-6-phosphate (G6P) was observed using the bi-enzyme electrode in the presence of free NADP + . The current response of the modified electrode depended on the number of enzyme layers, the polymerized mediator and the sequence of adsorption. The optimum multilayer assembly sequence consisted of the polymerized mediator, GR, and G6PDH from the innermost to outermost layer. In the presence of free NADP + , the bi-enzyme biosensor with immobilized Alg–NADP + showed higher response for G6P than that without Alg–NADP + .

[1]  Haitao Zheng,et al.  Layer-by-layer assembly of enzymes and polymerized mediator on electrode surface by electrostatic adsorption , 2004 .

[2]  M. Senda,et al.  Electron-transfer function of NAD+-immobilized alginic acid. , 1996, Biochimica et biophysica acta.

[3]  Lo Gorton,et al.  Electrocatalytic oxidation of NAD(P) H at mediator-modified electrodes. , 2002, Journal of biotechnology.

[4]  Katsuhiko Ariga,et al.  A careful examination of the adsorption step in the alternate layer-by-layer assembly of linear polyanion and polycation , 1999 .

[5]  A. English,et al.  NAD(P)H Sensors Based on Enzyme Entrapment in Ferrocene-Containing Polyacrylamide-Based Redox Gels , 1998 .

[6]  Arne Torstensson,et al.  Catalytic oxidation of reduced nicotinamide adenine dinucleotide by graphite electrodes modified with adsorbed aromatics containing catechol functionalities , 1981 .

[7]  F. Scheller,et al.  A Bienzyme Carbon Paste Electrode for the Sensitive Detection of NADPH and the Measurement of Glucose‐6‐phosphate Dehydrogenase , 1999 .

[8]  N. Mano,et al.  Electrodes modified with nitrofluorenone derivatives as a basis for new biosensors. , 2001, Biosensors & bioelectronics.

[9]  Katsuhiko Ariga,et al.  ASSEMBLY OF MULTICOMPONENT PROTEIN FILMS BY MEANS OF ELECTROSTATIC LAYER-BY-LAYER ADSORPTION , 1995 .

[10]  L. Kubota,et al.  Ferrocenecarboxylic acid adsorbed on Nb2O5 film grafted on a SiO2 surface: NADH oxidation study , 2001 .

[11]  K. Kano,et al.  Electrochemical Oxidation of NADH Catalyzed by Diaphorase Conjugated with Poly-1-vinylimidazole Complexed with Os(2,2′-dipyridylamine)2Cl , 2002 .

[12]  Mikio Sakakibara,et al.  Electrochemical reduction of immobilized NADP+ on a polymer modified electrode with a co-polymerized mediator , 2002 .

[13]  K. Kano,et al.  Mediated bioelectrocatalysis based on nad-related enzymes with reversible characteristics , 1998 .

[14]  E. Calvo,et al.  Donnan permselectivity in layer-by-layer self-assembled redox polyelectrolye thin films. , 2002, Journal of the American Chemical Society.

[15]  H. Abruña,et al.  Analytical strategies for amperometric biosensors based on chemically modified electrodes. , 1998, Biosensors & bioelectronics.

[16]  J. Marty,et al.  Reagentless ethanol sensor based on a NAD-dependent dehydrogenase , 1997 .

[17]  A. Bassi,et al.  Mediated, amperometric biosensor for glucose-6-phosphate monitoring based on entrapped glucose-6-phosphate dehydrogenase, Mg2+ ions, tetracyanoquinodimethane, and nicotinamide adenine dinucleotide phosphate in carbon paste. , 1999, Analytical biochemistry.

[18]  A. Cass,et al.  Ferricinium ion as an electron acceptor for oxido-reductases , 1985 .

[19]  C. Danilowicz,et al.  Layer-by-layer self-assembly of glucose oxidase and Os(Bpy)2CIPyCH2NH-poly(allylamine) bioelectrode. , 2001, Analytical chemistry.

[20]  K. Ariga,et al.  Activity and stability of glucose oxidase in molecular films assembled alternately with polyions. , 1999, Journal of bioscience and bioengineering.

[21]  P. Tuñón,et al.  Amperometric biosensors based on NAD(P)‐dependent dehydrogenase enzymes , 1997 .

[22]  C Nicolini,et al.  Cholesterol biosensors prepared by layer-by-layer technique. , 2001, Biosensors & bioelectronics.

[23]  Jia-cong Shen,et al.  Chemically modified electrode via layer-by-layer deposition of glucose oxidase (GOD) and polycation-bearing Os complex , 1998 .