Azine/hydrogel/nanotube composite-modified electrodes for NADH catalysis and enzyme immobilization

The development of new, efficient bioelectrodes is important to the improvement of biosensor and biofuel cell technology. NAD-dependent dehydrogenase enzymes represent a diverse field of oxidoreductase enzymes that can be used to create unique biosensors and biofuel cells, but require electrocatalysts to oxidize NADH in order to harvest the electrons efficiently from fuel oxidation. This study presents a new methodology for the co-immobilization of dehydrogenase enzymes, azine-based NADH electrocatalysts, carbon nanotubes, and polymer hydrogels. The easy “one-pot” mixing and casting procedure is shown to produce electrodes that can electro-oxidize NADH at low potentials. In situ electropolymerization of the azine dyes within the composites is shown to improve NADH sensitivity, but harms enzyme activity. Biosensors and biofuel cells are constructed with a model enzyme, glucose dehydrogenase, to show the application of this system in a glucose biosensor and biofuel cell. Glucose biosensors produced limiting current densities of 400 μA/cm2 and glucose/air-breathing biofuel cells produced power densities slightly greater than 100 μW/cm2.

[1]  Marguerite N. Germain,et al.  Structure and Electrochemical Properties of Electrocatalysts for NADH Oxidation , 2010 .

[2]  C. Mann,et al.  Electrochemical oxidation of primary aliphatic amines , 1967 .

[3]  Matsuhiko Nishizawa,et al.  Enzyme-based glucose fuel cell using Vitamin K3-immobilized polymer as an electron mediator , 2005 .

[4]  L. Miller,et al.  Oxidation of NADH by ferrocenium salts. Rate-limiting one-electron transfer , 1983 .

[5]  W. Blaedel,et al.  Study of the electrochemical oxidation of reduced nicotinamide adenine dinucleotide. , 1975, Analytical chemistry.

[6]  Kateryna Artyushkova,et al.  Anthracene-Modified Multi-Walled Carbon Nanotubes as Direct Electron Transfer Scaffolds for Enzymatic Oxygen Reduction , 2011 .

[7]  Theodore Kuwana,et al.  Electrocatalysis of dihydronicotinamide adenosine diphosphate with quinones and modified quinone electrodes , 1978 .

[8]  Shelley D. Minteer,et al.  Utilization of enzyme cascades for complete oxidation of lactate in an enzymatic biofuel cell , 2011 .

[9]  T. Ohsaka,et al.  A Miniature Glucose/O2 Biofuel Cell With a High Tolerance Against Ascorbic Acid , 2009 .

[10]  L. Gorton,et al.  A carbon paste electrode chemically modified with a phenothiazine polymer derivative for electrocatalytic oxidation of NADH: Preliminary study , 1993 .

[11]  P. Bartlett,et al.  Chapter 2 - The Application of Approximate Analytical Models in the Development of Modified Electrodes for NADH Oxidation , 1999 .

[12]  Shelley D. Minteer,et al.  Development of alcohol/O2 biofuel cells using salt-extracted tetrabutylammonium bromide/Nafion membranes to immobilize dehydrogenase enzymes , 2005 .

[13]  P. Elving,et al.  Nicotinamide-Nad Sequence: Redox Processes and Related Behavior: Behavior and Properties of Intermediate and Final Products , 1976 .

[14]  Robert L. Arechederra,et al.  Development of glycerol/O2 biofuel cell , 2007 .

[15]  Shelley D. Minteer,et al.  Bioelectrocatalytic oxidation of glucose in CNT impregnated hydrogels: Advantages of synthetic enzymatic metabolon formation , 2012 .

[16]  Robert L. Arechederra,et al.  Oxidation of Biofuels: Fuel Diversity and Effectiveness of Fuel Oxidation through Multiple Enzyme Cascades , 2010 .

[17]  L. Gorton,et al.  Electrochemical and SERS studies of chemically modified electrodes: Nile Blue A, a mediator for NADH oxidation , 1990 .

[18]  Joseph Wang,et al.  Carbon nanotube/teflon composite electrochemical sensors and biosensors. , 2003, Analytical chemistry.

[19]  S. Minteer,et al.  Comparison of electropolymerized thiazine dyes as an electrocatalyst in enzymatic biofuel cells and self powered sensors. , 2009, Journal of nanoscience and nanotechnology.

[20]  Shelley D. Minteer,et al.  Pyruvate/Air Enzymatic Biofuel Cell Capable of Complete Oxidation , 2009 .

[21]  W. Schuhmann,et al.  New amperometric dehydrogenase electrodes based on electrocatalytic NADH‐oxidation at poly (methylene blue)‐modified electrodes , 1994 .

[22]  Richard A. Durst,et al.  Mediator compounds for the electrochemical study of biological redox systems: a compilation , 1982 .

[23]  A. Karyakin,et al.  Electropolymerized Azines: A New Group of Electroactive Polymers , 1999 .

[24]  Yuehe Lin,et al.  Low-potential stable NADH detection at carbon-nanotube-modified glassy carbon electrodes , 2002 .

[25]  Lun Wang,et al.  Electrocatalytic activity of carbon spheres towards NADH oxidation at low overpotential and its applications in biosensors and biofuel cells , 2011 .

[26]  C. Mann Cyclic Stationary Electrode Voltammetry of Some Aliphatic Amines. , 1964 .

[27]  S. Minteer,et al.  Improving the microenvironment for enzyme immobilization at electrodes by hydrophobically modifying chitosan and Nafion® polymers , 2008 .

[28]  Evgeny Katz,et al.  Biofuel cell controlled by enzyme logic systems. , 2009, Journal of the American Chemical Society.

[29]  Mao-gen Zhang,et al.  Coimmobilization of dehydrogenases and their cofactors in electrochemical biosensors. , 2007, Analytical chemistry.

[30]  Shen-ming Chen,et al.  Reversible cyclic voltammetry of the NADH/NAD+ redox system on hybrid poly(luminol)/FAD film modified electrodes , 2006 .

[31]  S. Minteer,et al.  Citric acid cycle biomimic on a carbon electrode. , 2008, Biosensors & bioelectronics.

[32]  P. Elving,et al.  Mechanistic aspects of the electrochemical oxidation of dihydronicotinamide adenine dinucleotide (NADH) , 1980 .

[33]  Robert L. Arechederra,et al.  Evaluating Enzyme Cascades for Methanol/Air Biofuel Cells Based on NAD+‐Dependent Enzymes , 2010 .

[34]  M. Porter,et al.  Electrochemical oxidation of amine-containing compounds. A route to the surface modification of glassy carbon electrodes , 1994 .

[35]  Wolfgang Schuhmann,et al.  Electropolymerized Azines: Part II. In a Search of the Best Electrocatalyst of NADH Oxidation , 1999 .

[36]  Shelley D. Minteer,et al.  Enzymatic Biofuel Cell for Oxidation of Glucose to CO2 , 2012 .

[37]  S. Minteer,et al.  Bifunctional polyamines for the aqueous dispersion of carbon nanotubes and the formation of carbon nanotube-impregnated hydrogel composites , 2011 .

[38]  L. Gorton,et al.  Poly-phenothiazine derivative-modified glassy carbon electrode for NADH electrocatalytic oxidation , 2009 .

[39]  P. Elving,et al.  Nicotinamide-NAD sequence: redox process and related behavior, behavior and properties of intermediate and final products. [Use of pulse radiolysis in determining dimerization rates of free radicals] , 1976 .

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

[41]  Shelley D. Minteer,et al.  Complete Oxidation of Glycerol in an Enzymatic Biofuel Cell , 2009 .

[42]  Waldemar Gorski,et al.  Facilitation of NADH electro-oxidation at treated carbon nanotubes. , 2010, Analytical chemistry.

[43]  Malinauskas,et al.  Electropolymerization of Preadsorbed Layers of Some Azine Redox Dyes on Graphite. , 2000, Journal of colloid and interface science.

[44]  Hitoshi Muguruma,et al.  Amperometric biosensor based on multilayer containing carbon nanotube, plasma-polymerized film, electron transfer mediator phenothiazine, and glucose dehydrogenase. , 2012, Bioelectrochemistry.