Disclosed is an improved biofuel cell having a cathode comprising a dual function membrane, which contains an oxygen oxidoreductase enzyme immobilized within a buffered compartment of the membrane and an electron transport mediator which transfers electrons from an electron conducting electrode to t
Disclosed is an improved biofuel cell having a cathode comprising a dual function membrane, which contains an oxygen oxidoreductase enzyme immobilized within a buffered compartment of the membrane and an electron transport mediator which transfers electrons from an electron conducting electrode to the redox reaction catalyzed by the oxygen oxidoreductase enzyme. The improved biofuel cell also has an anode that contains an oxidoreductase enzyme that uses an organic fuel, such as alcohol, as a substrate. An electric current can flow between the anode and the cathode.
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1. A biocathode comprising: (a) an electron conductor;(b) an electron mediator;(c) at least one enzyme capable of reacting with a reduced form of the electron mediator and an oxidant to produce an oxidized form of the electron mediator and water; and(d) an electron conducting enzyme immobilization m
1. A biocathode comprising: (a) an electron conductor;(b) an electron mediator;(c) at least one enzyme capable of reacting with a reduced form of the electron mediator and an oxidant to produce an oxidized form of the electron mediator and water; and(d) an electron conducting enzyme immobilization material comprising an electrocatalyst, the enzyme immobilization material immobilizing and stabilizing the enzyme, the material being permeable to the oxidant, an oxidized form of the electrocatalyst being capable of gaining electrons from the electron conductor to produce a reduced form of the electrocatalyst that is capable of reacting with an oxidized form of the electron mediator to produce a reduced form of the electron mediator and an oxidized form of the electrocatalyst;wherein the enzyme is entrapped within pores of the enzyme immobilization material and the enzyme immobilization material comprises an ion exchange polymer comprising a hydrophobic cation. 2. A biocathode comprising: (a) an electron conductor;(b) an electron mediator;(c) at least one enzyme capable of reacting with a reduced form of the electron mediator and an oxidant to produce an oxidized form of the electron mediator and water; and(d) an electron conducting enzyme immobilization material comprising the electron mediator, the enzyme immobilization material immobilizing and stabilizing the enzyme, the material being permeable to the oxidant, an oxidized form of the electron mediator being capable of gaining electrons from the electron conductor to produce a reduced form of the electron mediator;wherein the enzyme is entrapped within pores of the enzyme immobilization material and the enzyme immobilization material comprises an ion exchange polymer comprising a hydrophobic cation. 3. A biocathode comprising: (a) an electron conductor;(b) an electron mediator;(c) at least one enzyme capable of reacting with a reduced form of the electron mediator and an oxidant to produce an oxidized form of the electron mediator and water; and(d) an electron conducting enzyme immobilization material comprising the electron mediator and an electrocatalyst, the enzyme immobilization material immobilizing and stabilizing the enzyme, the material being permeable to the oxidant, an oxidized form of the electrocatalyst being capable of gaining electrons from the electron conductor to produce a reduced form of the electrocatalyst that is capable of reacting with an oxidized form of the electron mediator to produce a reduced form of the electron mediator and an oxidized form of the electrocatalyst;wherein the enzyme is entrapped within pores of the enzyme immobilization material and the enzyme immobilization material comprises an ion exchange polymer comprising a hydrophobic cation. 4. A biocathode comprising: (a) an electron conductor;(b) an electron mediator;(c) at least one enzyme capable of reacting with a reduced form of the electron mediator and an oxidant to produce an oxidized form of the electron mediator and water; and(d) an electron conducting enzyme immobilization material comprising the electron mediator, the enzyme immobilization material immobilizing and stabilizing the enzyme, the material being permeable to the oxidant, an oxidized form of an electrocatalyst being capable of gaining electrons from the electron conductor to produce a reduced form of the electrocatalyst that is capable of reacting with an oxidized form of the electron mediator to produce a reduced form of the electron mediator and an oxidized form of the electrocatalyst;wherein the enzyme is entrapped within pores of the enzyme immobilization material and the enzyme immobilization material comprises an ion exchange polymer comprising a hydrophobic cation. 5. A biofuel cell for generating electricity comprising: a fuel fluid;an electron mediator;an anode capable of a reaction to oxidize the fuel fluid; anda biocathode of claim 1. 6. A biofuel cell for generating electricity comprising: a fuel fluid;an anode capable of a reaction to oxidize the fuel fluid; anda biocathode of claim 2. 7. A biofuel cell for generating electricity comprising: a fuel fluid;an anode capable of a reaction to oxidize the fuel fluid; anda biocathode of claim 3. 8. A biofuel cell for generating electricity comprising: a fuel fluid;an anode capable of a reaction to oxidize the fuel fluid; anda biocathode of claim 4. 9. The biocathode of claim 2 wherein the enzyme immobilization material comprises a perfluoro sulfonic acid-PTFE copolymer modified with a hydrophobic cation larger than NH4+ wherein the hydrophobic cation exchanges for protons as the counterion to the —SO3− groups of the perfluoro sulfonic acid-PTFE copolymer. 10. The biocathode of claim 2 wherein the electron conductor comprises a carbon-based material, a metallic conductor, a semiconductor, a metal oxide or a modified conductor. 11. The biocathode of claim 2 wherein the electron conductor comprises a carbon-based material. 12. The biocathode of claim 2 wherein the electron conductor comprises carbon cloth, carbon paper, carbon screen printed electrodes, carbon black, carbon powder, carbon fiber, single-walled carbon nanotubes, double-walled carbon nanotubes, multi-walled carbon nanotubes, carbon nanotube arrays, diamond-coated conductors, glass carbon, mesoporous carbon, graphite, uncompressed graphite worms, delaminated purified flake graphite, high performance graphite, highly ordered pyrolytic graphite, pyrolytic graphite or polycrystalline graphite. 13. The biocathode of claim 9 wherein the hydrophobic cation comprises an ammonium-based cation, quaternary ammonium cation, alkyltrimethylammonium cation, organic cation, phosphonium cation, triphenylphosphonium, pyridinium cation, imidazolium cation, hexdecylpyridinium, ethidium, viologen, methyl viologen, benzyl viologen, bis(triphenylphosphine)iminium metal complex, bipyridyl metal complex, phenanthroline-based metal complex, [Ru(bipyridine)3]2+ or [Fe(phenanthroline)3]3+. 14. The biocathode of claim 9 wherein the hydrophobic cation comprises a quaternary ammonium cation represented by formula 1 wherein R1, R2, R3 and R4 are independently hydrogen, hydrocarbyl, substituted hydrocarbyl or heterocyclo wherein at least one of R1, R2, R3 and R4 is other than hydrogen. 15. The biocathode of claim 14 wherein R1, R2, R3 and R4 are independently hydrogen, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl or tetradecyl wherein at least one of R1, R2, R3 and R4 is other than hydrogen. 16. The biocathode of claim 14 wherein R1, R2, R3 and R4 are the same and are methyl, ethyl, propyl, butyl, pentyl or hexyl. 17. The biocathode of claim 9 wherein the modified perfluoro sulfonic acid-PTFE copolymer is modified with tetrabutylammonium halide, triethylhexylammonium halide or trimethyldodecylammonium halide. 18. The biocathode of claim 9 wherein the modified perfluoro sulfonic acid-PTFE copolymer is modified with tetrabutylammonium halide or triethylhexylammonium halide. 19. The biocathode of claim 9 wherein the modified perfluoro sulfonic acid-PTFE copolymer is modified with triethylhexylammonium halide. 20. The biocathode of claim 2 wherein the enzyme comprises an oxidoreductase. 21. The biocathode of claim 2 wherein the enzyme comprises a glucose oxidase, alcohol-based oxidase or cholesterol-based oxidase. 22. The biocathode of claim 2 wherein the enzyme comprises laccase, cytochrome C oxidase, bilirubin oxidase or peroxidase. 23. The biocathode of claim 2 wherein the enzyme comprises an oxygen oxidoreductase having an optimum activity at a pH between about 6.5 and about 7.5. 24. The biocathode of claim 2 wherein the enzyme comprises bilirubin oxidase. 25. The biocathode of claim 2 wherein the electron mediator comprises a metalloprotein, a conjugated organic compound, a sugar, a sterol, a fatty acid or a coenzyme or substrate of an oxidase. 26. The biocathode of claim 2 wherein the oxidized form of the electron mediator comprises stellacyanin, bilirubin, glucose or cholesterol. 27. The biocathode of claim 2 wherein the oxidized form of the electron mediator comprises bilirubin. 28. The biocathode of claim 1 wherein the electrocatalyst for the electron mediator comprises organometallic cations with standard reduction potentials greater than +0.4 volts. 29. The biocathode of claim 1 wherein the electrocatalyst for the electron mediator comprises osmium, ruthenium, iron, nickel, rhodium, rhenium, or cobalt complexes. 30. The biocathode of claim 1 wherein the reduced form of the electrocatalyst for the electron mediator comprises Ru(phen)3+2, Fe(phen)3+2, Ru(bpy)3+2, Os(bpy)3+2 or Os(terpy)3+2. 31. The biocathode of claim 1 wherein the reduced form of the electrocatalyst for the electron mediator comprises Ru(bpy)3+2. 32. The biocathode of claim 1 wherein the concentration of the electrocatalyst is from about 100 mM to about 3M. 33. The biocathode of claim 1 wherein the concentration of the electrocatalyst is from about 250 mM to about 2.25M. 34. The biocathode of claim 1 wherein the concentration of the electrocatalyst is from about 500 mM to about 2M. 35. The biocathode of claim 1 wherein the concentration of the electrocatalyst is from about 1M to about 1.5M. 36. The biofuel cell of claim 6 wherein the oxidant comprises oxygen or peroxide. 37. The biofuel cell of claim 6 wherein the oxidant comprises oxygen. 38. The biofuel cell of claim 6 wherein the fuel fluid comprises ammonia, methanol, ethanol, propanol, isobutanol, butanol and isopropanol, allyl alcohols, aryl alcohols, glycerol, propanediol, mannitol, glucuronate, aldehyde, carbohydrates, glucose, glucose-1, D-glucose, L-glucose, glucose-6-phosphate, lactate, lactate-6-phosphate, D-lactate, L-lactate, fructose, galactose-1, galactose, aldose, sorbose, mannose, glycerate, coenzyme A, acetyl Co-A, malate, isocitrate, formaldehyde, acetaldehyde, acetate, citrate, L-gluconate, beta-hydroxysteroid, alpha-hydroxysteroid, lactaldehyde, testosterone, gluconate, fatty acids, lipids, phosphoglycerate, retinal, estradiol, cyclopentanol, hexadecanol, long-chain alcohols, coniferyl-alcohol, cinnamyl-alcohol, formate, long-chain aldehydes, pyruvate, butanal, acyl-CoA, steroids, amino acids, flavin, NADH, NADH2, NADPH, NADPH2 or hydrogen. 39. The biofuel cell of claim 6 wherein the fuel fluid comprises methanol, ethanol or propanol. 40. The biofuel cell of claim 6 wherein the fuel fluid comprises ethanol. 41. The biofuel cell of claim 6 wherein the anode is a bioanode. 42. The biofuel cell of claim 41 wherein the bioanode comprises a modified perfluoro sulfonic acid-PTFE copolymer. 43. The biofuel cell of claim 42 wherein the modified perfluoro sulfonic acid-PTFE copolymer is modified with tetrabutylammonium halide, triethylhexylammonium halide or trimethyldodecylammonium halide. 44. The biofuel cell of claim 42 wherein the modified perfluoro sulfonic acid-PTFE copolymer is modified with triethylhexylammonium halide. 45. The biofuel cell of claim 41 wherein the bioanode comprises a PQQ-dependent alcohol dehydrogenase which has a PQQ molecule electrostatically associated with it. 46. The biofuel cell of claim 45 wherein the bioanode and biocathode are not separated by a salt bridge or a polymer electrolyte membrane. 47. A method of generating electricity using the biofuel cell of claim 5 comprising (a) oxidizing the fuel fluid at the anode and reducing the oxidant at the biocathode;(b) oxidizing the reduced form of the electron mediator during the reduction of the oxidant at the biocathode;(c) oxidizing the electrocatalyst; and(d) reducing the electrocatalyst at the electron conductor. 48. A method of generating electricity using the biofuel cell of claim 6 comprising (a) oxidizing the fuel fluid at the anode and reducing the oxidant at the biocathode;(b) oxidizing the reduced form of the electron mediator during the reduction of the oxidant at the biocathode; and(c) reducing the electron mediator at the electron conductor. 49. The method of claim 47 wherein the oxidant is oxygen or a peroxide. 50. The method of claim 47 wherein the oxidant is oxygen. 51. The method of claim 47 wherein the fuel fluid comprises ammonia, methanol, ethanol, propanol, isobutanol, butanol and isopropanol, allyl alcohols, aryl alcohols, glycerol, propanediol, mannitol, glucuronate, aldehyde, carbohydrates, glucose, glucose-1, D-glucose, L-glucose, glucose-6-phosphate, lactate, lactate-6-phosphate, D-lactate, L-lactate, fructose, galactose-1, galactose, aldose, sorbose, mannose, glycerate, coenzyme A, acetyl Co-A, malate, isocitrate, formaldehyde, acetaldehyde, acetate, citrate, L-gluconate, beta-hydroxysteroid, alpha-hydroxysteroid, lactaldehyde, testosterone, gluconate, fatty acids, lipids, phosphoglycerate, retinal, estradiol, cyclopentanol, hexadecanol, long-chain alcohols, coniferyl-alcohol, cinnamyl-alcohol, formate, long-chain aldehydes, pyruvate, butanal, acyl-CoA, steroids, amino acids, flavin, NADH, NADH2, NADPH, NADPH2 or hydrogen. 52. The method of claim 47 wherein the fuel fluid comprises methanol, ethanol or propanol. 53. The method of claim 47 wherein the fuel fluid comprises ethanol. 54. The method of claim 47 wherein the enzyme comprises an oxidoreductase. 55. The method of claim 47 wherein the enzyme comprises a glucose oxidase, alcohol-based oxidase or cholesterol-based oxidase. 56. The method of claim 47 wherein the enzyme comprises laccase, cytochrome c oxidase, bilirubin oxidase or peroxidase. 57. The method of claim 47 wherein the enzyme comprises bilirubin oxidase. 58. The method of claim 47 wherein the electron mediator comprises a metalloprotein, a conjugated organic compound, a sugar, a sterol, a fatty acid or a coenzyme or substrate of an oxidase. 59. The method of claim 47 wherein the electron mediator comprises stellacyanin, bilirubin, glucose or cholesterol. 60. The method of claim 47 wherein the oxidized form of the electron mediator comprises bilirubin. 61. The method of claim 47 wherein the electrocatalyst for the electron mediator comprises organometallic cations with standard reduction potentials greater than +0.4 volts. 62. The method of claim 47 wherein the electrocatalyst for the electron mediator comprises osmium, ruthenium, iron, nickel, rhodium, rhenium, or cobalt complexes. 63. The method of claim 47 wherein the reduced form of the electrocatalyst for the electron mediator comprises Ru(phen)3+2, Fe(phen)3+2, Ru(bpy)3+2, Os(bpy)3+2 or Os(terpy)3+2. 64. The method of claim 47 wherein the reduced form of the electrocatalyst for the electron mediator comprises Ru(bpy)3+2.
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