JP5205818B2 - Fuel cells and electronics - Google Patents

Description

  The present invention relates to a fuel cell in which an enzyme, a microorganism or a cell and an electron mediator are used for at least one of a positive electrode and a negative electrode, and an electronic device using the fuel cell.

The fuel cell has a structure in which a positive electrode (oxidant electrode) and a negative electrode (fuel electrode) face each other with an electrolyte (proton conductor) interposed therebetween. In the conventional fuel cell, the fuel (hydrogen) supplied to the negative electrode is oxidized and separated into electrons and protons (H ⁺ ), the electrons are transferred to the negative electrode, and H ⁺ moves through the electrolyte to the positive electrode. In the positive electrode, this H ⁺ reacts with oxygen supplied from the outside and electrons sent from the negative electrode through the external circuit to generate H ₂ O.

  In this way, fuel cells are highly efficient power generators that directly convert the chemical energy of fuel into electrical energy, and the chemical energy of fossil energy such as natural gas, oil, and coal can be used regardless of where and when it is used. Moreover, it can be extracted as electric energy with high conversion efficiency. For this reason, research and development of fuel cells for large-scale power generation has been actively conducted. For example, a fuel cell is mounted on the space shuttle, and it has proven that it can supply water for crew members at the same time as electric power, and that it is a clean power generator.

Furthermore, in recent years, fuel cells having a relatively low operating temperature range from room temperature to about 90 ° C., such as solid polymer fuel cells, have been developed and attracting attention. For this reason, not only large-scale power generation applications but also applications to small systems such as automobile power supplies and portable power supplies such as personal computers and mobile devices are being sought.
Thus, the fuel cell can be used in a wide range of applications from large-scale power generation to small-scale power generation, and has attracted much attention as a highly efficient power generation device. However, in fuel cells, natural gas, petroleum, coal, etc. are usually used as fuel by converting them into hydrogen gas using a reformer, which consumes limited resources and needs to be heated to a high temperature. There are various problems such as the need for an expensive noble metal catalyst such as (Pt). Even when hydrogen gas or methanol is used directly as a fuel, care must be taken when handling it.

Accordingly, attention has been paid to the fact that biological metabolism performed in living organisms is a highly efficient energy conversion mechanism, and proposals have been made to apply this to fuel cells. The biological metabolism here includes respiration, photosynthesis and the like performed in microbial somatic cells. Biological metabolism has the characteristics that the power generation efficiency is extremely high and the reaction proceeds under mild conditions of about room temperature.
For example, respiration takes nutrients such as sugars, fats, and proteins into microorganisms or cells, and these chemical energies are converted to carbon dioxide (glycolytic and tricarboxylic acid (TCA) cycles through a number of enzymatic reaction steps. In the process of generating CO ₂ ), nicotinamide adenine dinucleotide (NAD ⁺ ) is reduced to reduced nicotinamide adenine dinucleotide (NADH) to convert it into redox energy, that is, electric energy, and further, an electron transfer system In this mechanism, the electric energy of NADH is directly converted into electric energy of proton gradient, and oxygen is reduced to generate water. The electrical energy obtained here produces ATP from adenosine diphosphate (ADP) via adenosine triphosphate (ATP) synthase, and this ATP is used for reactions necessary for the growth of microorganisms and cells. Used. Such energy conversion occurs in the cytosol and mitochondria.

In addition, photosynthesis captures light energy and reduces nicotinamide adenine dinucleotide phosphate (NADP ⁺ ) via an electron transfer system to reduce nicotinamide adenine dinucleotide phosphate (NADPH), thereby converting it into electrical energy. In the process of conversion, it is a mechanism that oxidizes water to produce oxygen. This electric energy takes in CO ₂ and is used for carbon fixation reaction, and is used for carbohydrate synthesis.
As a technique for utilizing the above-described biometabolism for a fuel cell, a microbial cell has been reported in which an electron generated in a microorganism is taken out of the microorganism and an electric current is obtained by passing the electron to an electrode (for example, a patent). Reference 1).

On the other hand, microorganisms and cells have many unnecessary reactions in addition to the intended reaction such as conversion from chemical energy to electrical energy. In the above method, electric energy is consumed for unwanted reactions and sufficient energy conversion efficiency is achieved. Therefore, in order to improve this, a fuel cell (biofuel cell) that performs only a desired reaction using an enzyme has been proposed (see, for example, Patent Documents 2 to 11). In this biofuel cell, a fuel is decomposed by an enzyme and separated into protons and electrons, and those using alcohols such as methanol and ethanol or monosaccharides such as glucose have been developed as fuel.
In the positive electrode and negative electrode of the above-described conventional biofuel cell and microbial cell, an electron mediator (electron transfer material) is used in order to smoothly exchange electrons between the enzyme or microorganism used as a catalyst and the electrode. (For example, refer patent documents 1-11.).

JP 2000-133297 A JP 2003-282124 A JP 2004-71559 A JP 2005-13210 A JP 2005-310613 A JP 2006-24555 A JP 2006-49215 A JP 2006-93090 A JP 2006-127957 A JP 2006-156354 A JP 2007-12281 A

  However, when an electron mediator is used for the positive electrode and the negative electrode of a biofuel cell and a microbial cell, if the electron mediator used for the positive electrode moves to the negative electrode side and conversely the electron mediator used for the negative electrode moves to the positive electrode side, the biofuel It is known to cause a decrease in output and electric capacity of batteries and microbial batteries. Conventionally, in order to avoid this problem, it has been necessary to immobilize an electron mediator on the positive electrode and the negative electrode. However, since the electron mediator is generally a low molecule, elution is completely suppressed, and electrons are connected to the positive electrode and the negative electrode. It is not easy to maintain the state in which the mediator is immobilized for a long time. For this reason, it has been difficult to sufficiently suppress the decrease in output and the decrease in electric capacity of the biofuel cell and the microbial cell.

Therefore, the problem to be solved by the present invention is that when an enzyme, a microorganism or a cell and an electron mediator are used for at least one of the positive electrode and the negative electrode, the effect that the electron mediator moves from one of the positive electrode and the negative electrode to the other is effective. It is an object of the present invention to provide a high-performance fuel cell that can be prevented in an effective manner and can sufficiently suppress a decrease in output and a decrease in electric capacity.
Another problem to be solved by the present invention is to provide an electronic device using the excellent fuel cell as described above.

In order to solve the above problem, the first invention is:
In a fuel cell having a structure in which a positive electrode and a negative electrode are opposed via a proton conductor, and an enzyme, a microorganism or a cell and an electron mediator are used for at least one of the positive electrode and the negative electrode,
The proton conductor has a charge having the same sign as the charge of the oxidized or reduced form of the electron mediator.

Typically, the proton conductor includes a polymer having a charge of the same sign as the charge of the oxidized or reduced form of the electron mediator, such as a polyanion or polycation, so that the proton conductor is an oxidized or reduced form of the electron mediator. The charge of the proton conductor has the same sign as the charge of the oxidized or reduced form of the electron mediator by other methods. Good. Specifically, when the charge of the oxidized or reduced form of the electron mediator used for at least one of the positive electrode and the negative electrode is a negative charge, a polymer having a negative charge, such as a polyanion, is included in the proton conductor. When the oxidant or reductant of the electron mediator has a positive charge, a polymer having a positive charge, such as a polycation, is included in the proton conductor. Examples of the polyanion include Nafion (trade name, DuPont, USA), which is an ion exchange resin having a fluorine-containing carbon sulfonic acid group, dichromate ion (Cr ₂ O ₇ ²⁻ ), paramolybdate ion ([ Mo ₇ O ₂₄ ] ⁶⁻ ), polyacrylic acid (for example, sodium polyacrylate (PAAcNa)), or the like can be used. For example, poly-L-lysine (PLL) can be used as the polycation.
Preferably, an enzyme, a microorganism, or a cell and an electron mediator are immobilized on at least one of the positive electrode and the negative electrode. Various conventionally known methods can be used for immobilization.

The enzyme, microorganism or cell used for at least one of the positive electrode and the negative electrode, and preferably immobilized, may be various and is selected as necessary.
Specifically, the enzyme used for the negative electrode includes, for example, an oxidase that promotes and decomposes monosaccharides in the case of using a monosaccharide such as glucose as a fuel. A coenzyme oxidase which converts the coenzyme reduced by the enzyme back into an oxidant. By the action of the coenzyme oxidase, electrons are generated when the coenzyme returns to the oxidized form, and the electrons are transferred from the coenzyme oxidase to the electrode via the electron mediator. For example, NAD ⁺ -dependent glucose dehydrogenase (GDH) is used as the oxidase, nicotinamide adenine dinucleotide (NAD ⁺ ) is used as the coenzyme, and diaphorase is used as the coenzyme oxidase.

  When using polysaccharides (a broadly defined polysaccharide, which refers to all carbohydrates that produce two or more monosaccharides by hydrolysis, including oligosaccharides such as disaccharides, trisaccharides, and tetrasaccharides) as fuel, Preferably, in addition to the above-mentioned oxidase, coenzyme oxidase, coenzyme and electron mediator, a degradation enzyme that promotes degradation such as hydrolysis of polysaccharides and produces monosaccharides such as glucose is also immobilized. . Specific examples of the polysaccharide include starch, amylose, amylopectin, glycogen, cellulose, maltose, sucrose, and lactose. These are a combination of two or more monosaccharides, and any polysaccharide contains glucose as a monosaccharide of the binding unit. Note that amylose and amylopectin are components contained in starch, and starch is a mixture of amylose and amylopectin. When glucoamylase is used as a polysaccharide-degrading enzyme and glucose dehydrogenase is used as an oxidase that degrades monosaccharides, polysaccharides that can be degraded to glucose by glucoamylase, such as starch, amylose, amylopectin, glycogen As long as it contains any one of maltose, it is possible to generate electricity using this as fuel. Glucoamylase is a degrading enzyme that hydrolyzes α-glucan such as starch to produce glucose, and glucose dehydrogenase is an oxidase that oxidizes β-D-glucose to D-glucono-δ-lactone. Preferably, the degradation enzyme that decomposes the polysaccharide is also immobilized on the negative electrode, and the polysaccharide that will eventually become the fuel is also immobilized on the negative electrode.

  Further, when starch is used as fuel, it is possible to use a gelatinized fuel obtained by gelatinizing starch. In this case, it is preferable to use a method in which gelatinized starch is brought into contact with a negative electrode on which an enzyme or the like is immobilized, or is immobilized on the negative electrode together with the enzyme or the like. When such an electrode is used, the starch concentration on the negative electrode surface can be maintained at a higher level than when starch dissolved in the solution is used, the enzymatic degradation reaction becomes faster, and the output is improved. The fuel handling is easier than in the case of a solution, the fuel supply system can be simplified, and the fuel cell does not need to be used upside down, which is very advantageous when used for mobile devices, for example. .

  Basically, any electron mediator may be used, but a compound having a quinone skeleton, particularly, a compound having a naphthoquinone skeleton is preferably used. As the compound having a naphthoquinone skeleton, various naphthoquinone derivatives can be used. Specifically, for example, 2-amino-1,4-naphthoquinone (ANQ), 2-amino-3-methyl-1 2, 4-naphthoquinone (AMNQ), 2-methyl-1,4-naphthoquinone (VK3), 2-amino-3-carboxy-1,4-naphthoquinone (ACNQ), and the like are used. As a compound having a quinone skeleton, for example, anthraquinone or a derivative thereof can be used in addition to a compound having a naphthoquinone skeleton. In addition to the compound having a quinone skeleton, the electron mediator may contain one or two or more other compounds that function as an electron mediator, if necessary. As a solvent used when a compound having a quinone skeleton, particularly a compound having a naphthoquinone skeleton, is immobilized on the negative electrode, acetone is preferably used. Thus, by using acetone as a solvent, the solubility of the compound having a quinone skeleton can be increased, and the compound having a quinone skeleton can be efficiently immobilized on the negative electrode. If necessary, the solvent may contain one or two or more other solvents other than acetone.

In one example, the negative electrode is 2-methyl-1,4-naphthoquinone (VK3) as an electron mediator, reduced nicotinamide adenine dinucleotide (NADH) as a coenzyme, glucose dehydrogenase and coenzyme oxidase as oxidases The diaphorase as is immobilized, and preferably 1.0 (mol): 0.33-1.0 (mol): (1.8-3.6) × 10 ⁶ (U): (0. It is fixed at a ratio of 85 to 1.7) × 10 ⁷ (U). However, U (unit) is an index indicating enzyme activity, and indicates the degree to which 1 μmol of substrate reacts per minute at a certain temperature and pH.

On the other hand, the enzyme used for the positive electrode includes oxygen reductase. As this oxygen reductase, for example, bilirubin oxidase (BOD), laccase, ascorbate oxidase and the like can be used. As the electron mediator, for example, hexacyanoferrate ion (Fe (CN) ₆ ^{3− / 4−} ), which is a polyvalent anion generated by ionizing potassium hexacyanoferrate in a solution, can be used.

  Various materials can be used as an immobilizing material for immobilizing an enzyme, a coenzyme, an electron mediator, etc. on the negative electrode or the positive electrode, preferably poly-L-lysine (PLL). A polyion complex formed using a polycation or a salt thereof and a polyanion or a salt thereof such as polyacrylic acid (for example, sodium polyacrylate (PAAcNa)) can be used, and an enzyme is contained inside the polyion complex. , Coenzymes, electron mediators, and the like can be included.

  When microorganisms or cells are used for the positive electrode or the negative electrode, conventionally known microorganisms or cells can be used as the microorganisms or cells (for example, see Patent Document 1). Microorganisms or cells used for the negative electrode may have any function as an electron donor that donates electrons to the electrode via an electron mediator. Microorganisms or cells used for the positive electrode may be electrons from the electrode. Any material may be used as long as it has a function as an electron acceptor that is received via an electron mediator. For example, microorganisms having a function as an electron donor include various conventionally known microorganisms such as bacteria belonging to each genus such as Saccharomyces, Hansenula, Candida, Micrococcus, Staphylococcus, filamentous fungi, yeast, and genetic manipulation. The produced microorganism can also be used and is not particularly limited. Examples of cells having a function as an electron donor or electron acceptor include various types of cells such as algae such as Synechococcus sp. And Anebaena variabilis, protozoa, erythrocytes, leukocytes, cultured cells, and animal and plant cells. There is no particular restriction.

The proton conductor transports protons (H ⁺ ) generated in the negative electrode to the positive electrode, and is made of a material that has no electron conductivity and can transport H ⁺ . As the proton conductor, an electrolyte is usually used. Various electrolytes can be used as the electrolyte, and are selected as necessary. Specifically, for example, a perfluorocarbon sulfonic acid (PFS) resin film, a copolymer film of a trifluorostyrene derivative, Polybenzimidazole membrane impregnated with phosphoric acid, aromatic polyetherketone sulfonic acid membrane, PSSA-PVA (polystyrene sulfonate polyvinyl alcohol copolymer), PSSA-EVOH (polystyrene sulfonate ethylene vinyl alcohol copolymer), Examples thereof include an ion exchange resin having a fluorocarbon sulfonic acid group (for example, Nafion (trade name, DuPont, USA)).

The above electrolyte generally contains a buffer substance (buffer solution). This is because the enzyme, microorganism or cell used as a catalyst is very sensitive to the pH of the solution, and therefore the buffer is used to control the enzyme, microorganism or cell near the pH at which it is likely to function. The concentration of the buffer substance is preferably 0.2M or more and 2M or less, more preferably 0.4M or more and 2M or less, and further preferably 0.8M or more and 1.2M or less from the viewpoint of obtaining a sufficiently high buffer capacity. It is. In this way, when the concentration of the buffer substance contained in the electrolyte surrounding the enzyme, microorganism or cell is 0.2 M or more and 2.5 M or less, the increase or decrease of the proton due to the proton-mediated enzyme reaction or the like during high output operation. Even if it occurs inside the electrode or in the immobilized membrane of the enzyme, a sufficient buffering action can be obtained, and the deviation from the optimum pH can be suppressed sufficiently small. You can fully demonstrate your abilities. Buffer substances, in general, as long as _a pK _a of 5 to 9, may it be used What Specific examples and, dihydrogen phosphate ion (H ₂ PO ₄ ^- ), 2-amino-2-hydroxymethyl-1,3-propanediol (abbreviated to Tris), 2- (N-morpholino) ethanesulfonic acid (MES), cacodylic acid, carbonic acid (H ₂ CO ₃ ), hydrogen citrate Ions, N- (2-acetamido) iminodiacetic acid (ADA), piperazine-N, N′-bis (2-ethanesulfonic acid) (PIPES), N- (2-acetamido) -2-aminoethanesulfonic acid ( ACES), 3- (N-morpholino) propanesulfonic acid (MOPS), N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES), N-2-hydroxyethylpiperazine- '-3-propanesulfonic acid (HEPPS), N- [tris (hydroxymethyl) methyl] glycine (abbreviation tricine), glycylglycine, N, N-bis (2-hydroxyethyl) glycine (abbreviation bicine), etc. . Examples of the substance that generates dihydrogen phosphate ions (H ₂ PO ₄ ⁻ ) include sodium dihydrogen phosphate (NaH ₂ PO ₄ ) and potassium dihydrogen phosphate (KH ₂ PO ₄ ). As the buffer substance, a compound containing an imidazole ring is also preferable. Specifically, the compound containing an imidazole ring includes imidazole, triazole, pyridine derivative, bipyridine derivative, imidazole derivative (histidine, 1-methylimidazole, 2-methylimidazole, 4-methylimidazole, 2-ethylimidazole, imidazole-2 -Ethyl carboxylate, imidazole-2-carboxaldehyde, imidazole-4-carboxylic acid, imidazole-4,5-dicarboxylic acid, imidazol-1-yl-acetic acid, 2-acetylbenzimidazole, 1-acetylimidazole, N-acetyl Imidazole, 2-aminobenzimidazole, N- (3-aminopropyl) imidazole, 5-amino-2- (trifluoromethyl) benzimidazole, 4-azabenzimidazole, 4-aza-2-mercapto Benzimidazole, benzimidazole, 1-benzylimidazole, 1-butylimidazole) and the like. The pH of the electrolyte containing the buffer substance is preferably around 7, but may generally be any of 1-14.

  As a material of the positive electrode or the negative electrode, a conventionally known material such as a carbon-based material can be used, and a skeleton composed of a porous material and a carbon-based material covering at least a part of the surface of the skeleton are mainly used. It is possible to use a porous conductive material including the material described above. This porous conductive material can be obtained by coating at least a part of the surface of the skeleton made of the porous material with a material mainly composed of a carbon-based material. The porous material constituting the skeleton of the porous conductive material may be basically any material as long as the skeleton can be stably maintained even if the porosity is high. It does not matter whether or not there is sex. As the porous material, a material having high porosity and high conductivity is preferably used. Specifically, as such a porous material having a high porosity and high conductivity, a metal material (metal or alloy), a carbon-based material with a strong skeleton (improved brittleness), or the like is used. Can do. When a metal material is used as the porous material, various options are conceivable because the metal material has different state stability depending on the usage environment such as pH and potential of the solution. For example, nickel, copper, silver, gold A foam metal such as nickel-chromium alloy and stainless steel or a foam alloy is one of the easily available materials. As the porous material, a resin material (for example, a sponge-like material) can be used in addition to the metal material and the carbon-based material. The porosity and pore diameter (minimum pore diameter) of this porous material are in balance with the thickness of the material mainly composed of a carbon-based material that is coated on the surface of the skeleton made of this porous material. Is determined according to the required porosity and pore diameter. The pore diameter of the porous material is generally 10 nm to 1 mm, typically 10 nm to 600 μm. On the other hand, it is necessary to use a material that covers the surface of the skeleton and has conductivity and is stable at an assumed operating potential. Here, a material mainly composed of a carbon-based material is used as such a material. Carbon-based materials generally have a wide potential window, and many are chemically stable. Specifically, the carbon-based material is mainly composed of a carbon-based material, and the carbon-based material is the main component, and the secondary material is selected according to the characteristics required for the porous conductive material. Some materials contain a small amount of material. Specific examples of the latter material include a material whose electrical conductivity has been improved by adding a metal or other highly conductive material to the carbon-based material, or a polytetrafluoroethylene-based material or the like added to the carbon-based material. Thus, it is a material imparted with a function other than conductivity, such as imparting surface water repellency. There are various types of carbon-based materials, but any carbon-based material may be used. In addition to carbon alone, carbon may be added with other elements. The carbon-based material is particularly preferably a fine powder carbon material having high conductivity and a high surface area. Specific examples of the carbon-based material include materials imparted with high conductivity such as KB (Ketjen Black) and functional carbon materials such as carbon nanotubes and fullerenes. As a coating method of the material mainly composed of the carbon-based material, any coating method can be used as long as the surface of the skeleton made of the porous material can be coated by using an appropriate binder as necessary. Also good. The pore diameter of the porous conductive material is selected to a size that allows a solution containing a substrate or the like to easily enter and exit through the pores, and is generally 9 nm to 1 mm, more typically 1 μm to 1 mm, and more generally Specifically, it is 1 to 600 μm. A state in which at least a part of the surface of the skeleton made of a porous material is coated with a material mainly composed of a carbon-based material, or a surface of at least a part of the skeleton made of a porous material is mainly composed of a carbon-based material In the state coated with the material, it is desirable to prevent all the holes from communicating with each other or clogging with a material mainly composed of a carbon-based material.

  This fuel cell can be used for almost anything that requires electric power, and can be of any size. For example, electronic devices, mobile objects (automobiles, motorcycles, aircraft, rockets, spacecrafts, etc.), power devices, construction machinery It can be used for machine tools, power generation systems, cogeneration systems, etc. The output, size, shape, type of fuel, etc. are determined depending on the application.

The second invention is
In an electronic device using one or more fuel cells,
At least one of the fuel cells is
In a fuel cell having a structure in which a positive electrode and a negative electrode are opposed via a proton conductor, and an enzyme, a microorganism or a cell and an electron mediator are used for at least one of the positive electrode and the negative electrode,
The proton conductor has a charge of the same sign as the charge of the oxidized or reduced form of the electron mediator.

This electronic device may basically be any type, and includes both portable and stationary types. Specific examples include cell phones, mobile devices, robots, personal computers, and the like. Computers, game devices, in-vehicle devices, home appliances, industrial products, etc.
In the second invention, what has been described in relation to the first invention is valid as long as it is not against the nature thereof.

  In the present invention configured as described above, the proton conductor has the same sign as the charge of the oxidized or reduced form of the electron mediator, so that the charge of the proton conductor and the oxidized or reduced form of the electron mediator Repulsive force works between the electric charge of For this reason, it becomes difficult for the electron mediator to move to the proton conductor side, and it is possible to effectively suppress the electron mediator from passing through the proton conductor and moving to the opposite side.

  According to this invention, when an enzyme, a microorganism, or a cell and an electron mediator are used for at least one of the positive electrode and the negative electrode, it is possible to effectively prevent the electron mediator from moving from one of the positive electrode and the negative electrode to the other. Therefore, a decrease in output and a decrease in electric capacity can be sufficiently suppressed, and a fuel cell having excellent performance can be obtained. By using such an excellent fuel cell, a high-performance electronic device can be realized.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
FIG. 1 schematically shows a biofuel cell according to a first embodiment of the present invention. In this biofuel cell, glucose is used as the fuel. FIG. 2 schematically shows details of the configuration of the negative electrode of the biofuel cell, an example of an enzyme group immobilized on the negative electrode, and an electron transfer reaction by the enzyme group.
As shown in FIG. 1, this biofuel cell has a structure in which a negative electrode 1 and a positive electrode 2 face each other with an electrolyte layer 3 conducting only protons. The negative electrode 1 decomposes glucose supplied as fuel with an enzyme to extract electrons and generate protons (H ⁺ ). The positive electrode 2 generates water by protons transported from the negative electrode 1 through the electrolyte layer 3, electrons transmitted from the negative electrode 1 through an external circuit, and oxygen in the air, for example.

The negative electrode 1 is composed of, for example, an electrode 11 (see FIG. 2) made of porous carbon or the like, an enzyme involved in glucose decomposition, and a coenzyme in which a reductant is generated in an oxidation reaction in the glucose decomposition process (for example, , NAD ⁺ , NADP ^{+ and the} like), a coenzyme oxidase (eg, diaphorase) that oxidizes a reduced form of the coenzyme (eg, NADH, NADPH, etc.) An electron mediator that receives electrons and passes them to the electrode 11 is configured to be fixed by a fixing material made of, for example, a polymer.

As an enzyme involved in the degradation of glucose, for example, glucose dehydrogenase (GDH) can be used. In the presence of this oxidase, for example, β-D-glucose can be oxidized to D-glucono-δ-lactone.
Furthermore, this D-glucono-δ-lactone can be decomposed into 2-keto-6-phospho-D-gluconate by the presence of two enzymes, gluconokinase and phosphogluconate dehydrogenase (PhGDH). Can do. That is, D-glucono-δ-lactone is converted to D-gluconate by hydrolysis, and D-gluconate is converted to adenosine triphosphate (ATP) and adenosine diphosphate (ADP) in the presence of gluconokinase. And then phosphorylated to 6-phospho-D-gluconate. This 6-phospho-D-gluconate is oxidized to 2-keto-6-phospho-D-gluconate by the action of the oxidase PhGDH.

In addition to the above decomposition process, glucose can also be decomposed to CO ₂ by utilizing sugar metabolism. The decomposition process utilizing sugar metabolism is roughly divided into glucose decomposition and pyruvic acid generation by a glycolysis system and a TCA cycle, which are widely known reaction systems.
The oxidation reaction in the monosaccharide decomposition process is accompanied by a coenzyme reduction reaction. This coenzyme is almost determined by the acting enzyme. In the case of GDH, NAD ⁺ is used as the coenzyme. That is, when β-D-glucose is oxidized to D-glucono-δ-lactone by the action of GDH, NAD ⁺ is reduced to NADH to generate H ⁺ .

The produced NADH is immediately oxidized to NAD ⁺ in the presence of diaphorase (DI), generating two electrons and H ⁺ . Therefore, two electrons and two H ⁺ are generated by one-step oxidation reaction per glucose molecule. In the two-stage oxidation reaction, a total of four electrons and four H ⁺ are generated.
The electrons generated in the above process are transferred from diaphorase to the electrode 11 through the electron mediator, and H ⁺ is transported to the positive electrode 2 through the electrolyte layer 3.

  The electron mediator transfers electrons to and from the electrode 11, and the output voltage of the fuel cell depends on the redox potential of the electron mediator. That is, in order to obtain a higher output voltage, it is preferable to select an electron mediator having a more negative potential on the negative electrode 1 side. However, the reaction affinity of the electron mediator to the enzyme, the rate of electron exchange with the electrode 11, an inhibitor (light, Structural stability against oxygen) must also be considered. From such a viewpoint, 2-amino-3-carboxy-1,4-naphthoquinone (ACNQ), vitamin K3, and the like are preferable as the electron mediator acting on the negative electrode 1. In addition, for example, compounds having a quinone skeleton, metal complexes such as osmium (Os), ruthenium (Ru), iron (Fe), cobalt (Co), viologen compounds such as benzyl viologen, compounds having a nicotinamide structure, riboflavin structure A compound having a nucleotide, a compound having a nucleotide-phosphate structure, or the like can also be used as an electron mediator.

The above enzyme, coenzyme, and electron mediator are optimal for the enzyme by using a buffer solution such as a phosphate buffer solution or a Tris buffer solution contained in the electrolyte layer 3 so that the electrode reaction can be performed efficiently and constantly. It is preferably maintained at a low pH, for example, around pH 7. For this reason, in this first embodiment, at least the electrolyte layer 3 around the enzyme immobilized on the negative electrode 1 and the positive electrode 2 has a buffer solution of 0.2 M or more and 2.5 M or less, preferably The concentration is 0.2 M or more and 2 M or less, more preferably 0.4 M or more and 2 M or less, and further preferably 0.8 M or more and 1.2 M or less. By doing so, a high buffering capacity can be obtained, and the original ability of the enzyme can be fully exhibited even during high power operation of the fuel cell. As the phosphate buffer, for example, NaH ₂ PO ₄ or KH ₂ PO ₄ is used. Further, the ionic strength (IS) is too large or too small to adversely affect the enzyme activity, but considering the electrochemical response, it should be an appropriate ionic strength, for example, about 0.3. Is preferred. However, pH and ionic strength have optimum values for each enzyme used, and are not limited to the values described above.

  The enzyme, coenzyme, and electron mediator are preferably immobilized on the electrode 11 using an immobilizing material in order to efficiently capture an enzyme reaction phenomenon occurring in the vicinity of the electrode as an electric signal. Furthermore, the enzyme reaction system of the negative electrode 1 can be stabilized by immobilizing the enzyme and the coenzyme for decomposing the fuel on the electrode 11. As such an immobilizing material, for example, a combination of glutaraldehyde (GA) and poly-L-lysine (PLL) or a combination of sodium polyacrylate (PAAcNa) and poly-L-lysine (PLL). These may be used, these may be used alone, or other polymers may be used. By using an immobilization material combining glutaraldehyde and poly-L-lysine, it becomes possible to greatly improve the enzyme immobilization ability of each, and to obtain an excellent enzyme immobilization ability as a whole immobilization material. Can do. In this case, the optimum composition ratio between glutaraldehyde and poly-L-lysine varies depending on the enzyme to be immobilized and the substrate of the enzyme, but generally an arbitrary composition ratio may be used. As a specific example, a glutaraldehyde aqueous solution (0.125%) and a poly-L-lysine aqueous solution (1%) are used, and the ratio thereof is 1: 1, 1: 2, 2: 1, or the like.

In FIG. 2, for example, glucose dehydrogenase (GDH) is an enzyme involved in the degradation of glucose, NAD ⁺ is a coenzyme that produces a reductant in the oxidation reaction in the glucose degradation process, and a reductant of coenzyme. The case where the coenzyme oxidase that oxidizes a certain NADH is diaphorase (DI), and the electron mediator that receives electrons from the coenzyme oxidase accompanying the oxidation of the coenzyme and passes it to the electrode 11 is ACNQ is shown.

The positive electrode 2 is obtained by immobilizing an oxygen reductase and an electron mediator that transfers electrons between the electrodes on an electrode made of, for example, porous carbon. As the oxygen reductase, for example, bilirubin oxidase (BOD), laccase, ascorbate oxidase and the like can be used. As the electron mediator, for example, hexacyanoferrate ions generated by ionization of potassium hexacyanoferrate can be used. This electron mediator is preferably immobilized at a sufficiently high concentration, for example, 0.64 × 10 ⁻⁶ mol / mm ² or more on average.
In the positive electrode 2, in the presence of oxygen reductase, oxygen in the air is reduced by H ⁺ from the electrolyte layer 3 and electrons from the negative electrode 1 to generate water.

The electrolyte layer 3 is a proton conductor that transports H ⁺ generated in the negative electrode 1 to the positive electrode 2 and is made of a material that does not have electronic conductivity and can transport H ⁺ . At least the surface of the electrolyte layer 3 on the positive electrode 2 side is negatively charged and has a negative charge. Specifically, for example, at least a part of the electrolyte layer 3 on the positive electrode 2 side includes a polyanion having a negative charge, or the entire electrolyte layer 3 includes a polyanion having a negative charge. As this polyanion, for example, Nafion (trade name, DuPont, USA) which is an ion exchange resin having a fluorine-containing carbon sulfonic acid group is used.

In the fuel cell configured as described above, when glucose is supplied to the negative electrode 1, the glucose is decomposed by a decomposing enzyme including an oxidase. Since oxidase is involved in the monosaccharide decomposition process, electrons and H ⁺ can be generated on the negative electrode 1 side, and a current can be generated between the negative electrode 1 and the positive electrode 2.

Here, in order to verify that when the electrolyte layer 3 has a charge having the same sign as the charge of the oxidized or reduced form of the electron mediator, the electron mediator can be prevented from passing through the electrolyte layer 3. The results of the comparative experiment will be described.
First, two commercially available glassy carbon (GC) electrodes (diameter 3 mm) were prepared, and both were polished and cleaned. Next, 5 μl of a commercially available Nafion emulsion (20%) as a polyanion was added to one glassy carbon electrode and dried. Next, the two glassy carbon electrodes are immersed in a 1 mM hexacyanoferrate ion (polyvalent anion) aqueous solution (50 mM NaH ₂ PO ₄ / NaOH buffer, pH 7), and cyclic voltammetry at a sweep rate of 20 mVs ⁻¹ . (CV) was performed. The result is shown in FIG. 3A. FIG. 3B shows an enlarged CV curve in the case of using the glassy carbon electrode to which Nafion is added in FIG. 3A. As can be seen from FIGS. 3A and B, in the glassy carbon electrode to which Nafion was added, the redox peak current caused by the hexacyanoferrate ion as an electron mediator was less than 1/20 of that of the glassy carbon electrode to which Nafion was not added. became. This indicates that the hexacyanoferrate ion, which is a polyvalent anion having a negative charge, is not diffusing and permeating to Nafion, which is a polyanion having a negative charge.

Next, commercially available carbon felt (BRAY made by TORAY) was used as porous carbon, and this carbon felt was cut into 1 cm square, soaked with 80 μl of hexacyanoferrate ion (1M), and dried. Two electrodes thus produced were used as test electrodes. As shown in FIG. 4, a film-like separator 16 (corresponding to the electrolyte layer 3) is placed on the test electrode 15, and a working electrode 17 is provided so as to face the test electrode 15 with the separator 16 interposed therebetween. As the working electrode 17, a commercially available carbon felt (B0050 made by TORAY) cut into 1 cm square is used. The separator 16 and the working electrode 17 are obtained by dissolving hexacyanoferrate ions as an electron mediator in a buffer solution 18 of 0.4 M NaH ₂ PO ₄ (pH 7) (illustration of a container containing the buffer solution 18 is omitted). ). As the separator 16, cellophane having no charge and Nafion (pH 7) which is a polyanion having a negative charge were used. Electrons that have passed through the separator 16 from the test electrode 15 by performing cyclic voltammetry 5 minutes, 1 hour, and 2 hours after contacting the separator 16 with the buffer solution 18 (electrolytic solution) in which hexacyanoferrate ions are dissolved. The redox peak values of mediators, that is, hexacyanoferrate ions were compared. The counter electrode 19 and the reference electrode 20 were immersed in the buffer solution 18, and an electrochemical measurement device (not shown) was connected to the working electrode 17, the counter electrode 19, and the reference electrode 20. Pt line was used as the counter electrode 19 and Ag | AgCl was used as the reference electrode 20. The measurement was performed at atmospheric pressure, and the measurement temperature was 25 ° C. The measurement results when Nafion is used as the separator 16 are shown in FIG. Moreover, the measurement result at the time of using a cellophane as the separator 16 is shown in FIG. As can be seen from FIGS. 5 and 6, when cellophane is used as the separator 16, a redox peak of hexacyanoferrate ions is observed as soon as 5 minutes after the start of measurement. While the reduction peak value increases, when Nafion is used as the separator 16, no redox peak of hexacyanoferrate ion is observed even after 2 hours have passed since the measurement was started. From this, it is confirmed that when cellophane is used as the separator 16, hexacyanoferrate ions permeate through the separator 16, but when Nafion is used as the separator 16, hexacyanoferrate ions do not permeate the separator 16. It was done.

Next, the result of electrochemical measurement at a single electrode of the enzyme / electron mediator immobilized electrode used as the negative electrode 1 will be described.
The enzyme / electron mediator immobilized electrode was prepared as follows.
First, various solutions were prepared as follows. As a buffer solution, a 100 mM sodium dihydrogen phosphate (NaH ₂ PO ₄ ) buffer solution (IS = 0.3, pH = 7.0) was used.
Diaphorase (DI) (EC1.6.99.-, manufactured by Unitika, B1D111) was weighed 5 to 10 mg and dissolved in 1.0 ml of a buffer solution to obtain a DI enzyme buffer solution ((1)).
Glucose dehydrogenase (GDH) (NAD-dependent type, EC1.1.1.17, manufactured by Toyobo Co., Ltd., GLD-311) was weighed in an amount of 10 to 15 mg, dissolved in 1.0 ml of buffer solution, and GDH enzyme buffer solution ((2)) It was.
The buffer solution for dissolving the enzyme is preferably refrigerated until just before, and the enzyme buffer solution is preferably stored as refrigerated as much as possible.
3DH to 60.0 mg of NADH (manufactured by Sigma Aldrich, N-8129) was weighed and dissolved in 0.1 ml of buffer solution to obtain NADH buffer solution ((3)).
An appropriate amount of poly-L-lysine hydrobromide (PLL) (manufactured by Wako, 164-16961) was weighed and dissolved in ion-exchanged water so as to be 1 to 2 wt% to obtain a PLL aqueous solution ((4)). .
10-50 mg of 2-amino-1,4-naphthoquinone (ANQ) (synthetic product) was weighed and dissolved in 1 ml of an acetone solution to obtain an ANQ acetone solution ((5)).
Sodium polyacrylate (PAAcNa) (manufactured by Aldrich, 041-00595) was weighed in an appropriate amount and dissolved in ion-exchanged water so as to be 0.01 to 0.1 wt% to obtain a PAAcNa aqueous solution ((6)).

The various solutions prepared as described above were porous using a microsyringe in the amounts shown below in the order of (5), (1), (3), (2), (4), (6). After coating on quality carbon (made by Tokai Carbon, 0.5 × 0.5 cm ² , thickness 2 mm), it was appropriately dried to prepare an enzyme / electron mediator fixed electrode.
DI enzyme buffer solution (1): 10 μl
GDH enzyme buffer solution (2): 10 μl
NADH buffer solution (3): 10 μl
PLL aqueous solution (4): 10 μl
ANQ acetone solution (5): 7 μl
PAAcNa aqueous solution (6): 4 μl

Using the measurement solution shown in Table 1, the enzyme / electron mediator-immobilized electrode thus prepared was set to 0.1 V with respect to the reference electrode Ag | AgCl, a potential sufficiently higher than the redox potential of the electron mediator, and the chronoampero Measurement was performed. The glucose concentration of the fuel was 0.4M. FIG. 7 shows the results of chronoamperometry measured by changing the concentration of NaH ₂ PO ₄ buffer. Moreover, the result of the chronoamperometry measured by changing the ionic strength of the NaH ₂ PO ₄ buffer is shown in FIG.

As can be seen from FIG. 7 and Table 1, in Comparative Example 1 where the NaH ₂ PO ₄ concentration was 0, that is, no buffer solution was added, the initial current was higher than that in Comparative Example 2 where the NaH ₂ PO ₄ concentration was 0.1M. The current is almost zero after 1800 seconds.
Further, when Comparative Example 2 is compared with Examples 1, 2, and 3, the NaH ₂ PO ₄ concentration is 0.2 to 2.5 M, and in particular 0.4 to 2.0 M, so that the current value is Comparative Example 2. It can be seen that it increases several times or more, especially at 1M.
On the other hand, when the NaH ₂ PO ₄ concentration is 3M as in Comparative Example 3, the obtained current value decreases to the same extent as in Comparative Example 1.

Further, as can be seen from FIG. 8 and Table 1, in Comparative Examples 2, 4, and 5, when the NaH ₂ PO ₄ concentration was kept constant at 0.1 M and only the ionic strength was increased, However, the current after 1800 seconds tended to decrease.
Furthermore, enzyme activity was measured by ultraviolet-visible spectroscopy (UV-vis) method. The results are shown in FIGS. Here, FIGS. 9 and 10 show changes in the reaction rate (v ₀ ) of the reaction by diaphorase (DI) with respect to the NaH ₂ PO ₄ concentration and ionic strength, respectively. However, the concentration of DI is 2.5 nM, the concentration of ANQ is 2.2 mM, and the concentration of NADH is 40 mM. FIG. 11 and FIG. 12 show changes in the reaction rate (v ₀ ) of the reaction by glucose dehydrogenase (GDH) with respect to the NaH ₂ PO ₄ concentration and ionic strength, respectively. However, the concentration of GDH is 1.59 nM, the concentration of glucose is 133 mM, and the concentration of NADH is 1.50 mM. According to the measurement results shown in FIGS. 9 to 12, there was no significant increase in enzyme activity accompanying the increase in NaH ₂ PO ₄ concentration, but there was a tendency to decrease slightly. From this, it can be considered that the increase in the current value accompanying the increase in the NaH ₂ PO ₄ concentration is due to the effect of suppressing the pH change inside the electrode or the immobilized film.

On the other hand, even when bilirubin oxidase (BOD) was immobilized on the positive electrode 2 as an oxygen reductase, the current value was maintained and improved with increasing NaH ₂ PO ₄ concentration. FIG. 13 and FIG. 14 show the results of measuring enzyme activity by UV-vis method in this case. Here, FIG. 13 and FIG. 14 show changes in the reaction rate (v ₀ ) of the reaction by BOD with respect to the NaH ₂ PO ₄ concentration and ionic strength, respectively. However, the concentration of BOD is 2.4 nM and the concentration of bilirubin is 1.8 mg / ml. According to the measurement results shown in FIG. 13 and FIG. 14, in the positive electrode 2 as well, a large increase in enzyme activity accompanying an increase in NaH ₂ PO ₄ concentration was not observed, but a tendency to decrease slightly was observed.
As a cause of obtaining the above result, it is conceivable that proton movement in the electrode or in the immobilized membrane is slow.

Next, when BOD is immobilized on the positive electrode 2 as an oxygen reductase and imidazole and hydrochloric acid are mixed and adjusted to pH 7 as a buffer solution, the current value is maintained and improved as the buffer solution concentration increases. Was noticeable. Table 2 and FIG. 15 show the results of chronoamperometry measured by changing the concentration of imidazole in this case. FIG. 16 shows the buffer concentration dependence of the current value (the value after 3600 seconds in Table 2 and FIG. 15). Table 2, FIG. 15 and FIG. 16 also show the results when NaH ₂ PO ₄ buffer and 10% acetate buffer are used as the buffer. As shown in FIG. 17, this measurement was performed with a film-like cellophane 21 placed on the positive electrode 2 and a buffer solution 22 in contact with the cellophane 21. As the positive electrode 2, an enzyme / electron mediator fixed electrode prepared as follows was used. First, commercially available carbon felt (BORAY made by TORAY) was used as porous carbon, and this carbon felt was cut into 1 cm square. Next, 80 μl of hexacyanoferrate ion (100 mM), 80 μl of poly-L-lysine (1 wt%), 80 μl of BOD solution (50 mg / ml) are sequentially infiltrated into the above carbon felt and dried to dry the enzyme / An electron mediator fixed electrode was obtained. Two sheets of the enzyme / electron mediator-immobilized electrode produced in this way were stacked to make a positive electrode 2.

As can be seen from Table 2 and FIG. 15, when the NaH ₂ PO ₄ concentration is 1.0 M, an initial current is generated, but after 3600 seconds, the current is greatly reduced. On the other hand, when the imidazole concentration is 0.4M, 1.0M and 2.0M, almost no decrease in current is observed even after 3600 seconds. In the case of 10% acetate buffer (pH = 2.8), the initial current is low, but the current at the same level as in the case where the imidazole concentration is 2.0 M is maintained even after 3600 seconds. As can be seen from FIG. 16, the current value increases linearly with respect to the concentration when the imidazole concentration is in the range of 0.2 to 2.5M. Further, the NaH ₂ PO ₄ buffer and imidazole buffer, near both pK _a of 7, even though almost be oxygen solubility same, if the imidazole is present in a buffer of the same concentration, a large oxygen reduction current obtained It was.

Chronoamperometry was performed for 3600 seconds as described above, and then cyclic voltammetry (CV) between −0.3 and +0.6 V was performed. The result is shown in FIG. However, in this measurement, as shown in FIG. 19, a positive electrode 2 composed of an enzyme / electron mediator-immobilized electrode similar to the above was used as a working electrode, and this was placed on a PTFE (polytetrafluoroethylene) membrane 23. The test was performed in a state where the buffer solution 22 was brought into contact with 2. The counter electrode 24 and the reference electrode 25 were immersed in the buffer solution 22, and an electrochemical measurement device (not shown) was connected to the positive electrode 2, the counter electrode 24, and the reference electrode 25 as working electrodes. Pt line was used as the counter electrode 24 and Ag | AgCl was used as the reference electrode 25. The measurement was performed at atmospheric pressure, and the measurement temperature was 25 ° C. As the buffer solution 22, two types of imidazole / hydrochloric acid buffer solution (pH 7, 1.0 M) and NaH ₂ PO ₄ / NaOH buffer solution (pH 7, 1.0 M) were used.
From FIG. 18, it can be seen that when an imidazole / hydrochloric acid buffer (pH 7, 1.0 M) is used as the buffer 22, extremely good CV characteristics are obtained.
From the above, it was confirmed that the imidazole buffer solution has an advantage even if the measurement system is changed.

A specific configuration example of this biofuel cell is shown in FIGS. 20A and 20B.
As shown in FIGS. 20A and 20B, this biofuel cell includes a negative electrode 1 composed of an enzyme / electron mediator-immobilized carbon electrode in which an enzyme or an electron mediator is immobilized on a carbon felt of 0.25 cm ² with an immobilizing material; A positive electrode 2 composed of an enzyme / electron mediator-immobilized carbon electrode in which an enzyme or an electron mediator is immobilized on a carbon felt of 25 cm ² with an immobilizing material is opposed to an electrolyte layer 3 containing a buffer substance. doing. In this case, Ti current collectors 41 and 42 are placed under the positive electrode 2 and the negative electrode 1, respectively, so that current can be easily collected. Reference numerals 43 and 44 denote fixed plates. These fixing plates 43 and 44 are fastened to each other by screws 45, and the positive electrode 2, the negative electrode 1, the electrolyte layer 3, and the Ti current collectors 41 and 42 are sandwiched between them. One surface (outer surface) of the fixing plate 43 is provided with a circular recess 43a for taking in air, and a plurality of holes 43b penetrating to the other surface are provided in the bottom surface of the recess 43a. These holes 43 b serve as air supply paths to the positive electrode 2. On the other hand, a circular recess 44a for fuel loading is provided on one surface (outer surface) of the fixing plate 44, and a number of holes 44b penetrating to the other surface are provided on the bottom surface of the recess 44a. These holes 44 b serve as fuel supply paths to the negative electrode 1. A spacer 46 is provided on the periphery of the other surface of the fixing plate 44, and when the fixing plates 43 and 44 are fastened to each other with screws 45, the interval between them becomes a predetermined interval. .
As shown in FIG. 16B, a load 47 is connected between the Ti current collectors 41, 42, and a glucose / buffer solution is put as fuel in the recess 44 a of the fixed plate 44 to generate power.

  As described above, according to the first embodiment, since the electrolyte layer 3 has the same sign as the charge of the oxidized or reduced form of the electron mediator used for the positive electrode 2 and the negative electrode 1, It is possible to effectively prevent one electron mediator of the negative electrode 1 from passing through the electrolyte layer 3 and moving to the other of the positive electrode 2 and the negative electrode 1. For this reason, the fall of the output of a biofuel cell and the fall of an electric capacity can fully be suppressed. In addition, when the concentration of the buffer substance (buffer solution) contained in the electrolyte layer 3 is not less than 0.2M and not more than 2.5M, a sufficient buffer capacity can be obtained. Even at this time, the pH of the electrolyte layer 3 around the enzyme immobilized on the positive electrode 2 and the negative electrode 1 can be maintained near the optimum pH, and the ability inherent to the enzyme can be fully exhibited. . As a result, a high-performance biofuel cell capable of high output operation can be realized. This biofuel cell is suitable for application to power sources of various electronic devices, mobile objects, power generation systems, and the like.

Next explained is a biofuel cell according to the second embodiment of the invention.
21A, B and C and FIG. 22 show the biofuel cell, FIGS. 21A, B and C show a top view, a cross-sectional view and a back view of the biofuel cell, and FIG. 22 shows the components of the biofuel cell. It is a disassembled perspective view shown disassembled.

  As shown in FIGS. 21A, B and C and FIG. 22, in this biofuel cell, the positive electrode 2, the electrolyte layer 3 are placed in the space formed between the positive electrode current collector 51 and the negative electrode current collector 52. The negative electrode 1 is accommodated between the positive electrode current collector 51 and the negative electrode current collector 52 above and below the negative electrode 1. The positive electrode current collector 51, the negative electrode current collector 52, the positive electrode 2, the electrolyte layer 3, and the adjacent one of the negative electrode 1 are in close contact with each other. In this case, the positive electrode current collector 51, the negative electrode current collector 52, the positive electrode 2, the electrolyte layer 3, and the negative electrode 1 have a circular planar shape, and the entire biofuel cell also has a circular planar shape.

  The positive electrode current collector 51 is for collecting current generated at the positive electrode 2, and current is taken out from the positive electrode current collector 51. The negative electrode current collector 52 is for collecting current generated in the negative electrode 1. The positive electrode current collector 51 and the negative electrode current collector 52 are generally formed of a metal, an alloy, or the like, but are not limited thereto. The positive electrode current collector 51 is flat and has a substantially cylindrical shape. The negative electrode current collector 52 is also flat and has a substantially cylindrical shape. Then, the edge of the outer peripheral portion 51a of the positive electrode current collector 51 includes a ring-shaped gasket 56a made of an insulating material such as silicone rubber and a ring-shaped hydrophobic resin 56b such as polytetrafluoroethylene (PTFE). The space for accommodating the positive electrode 2, the electrolyte layer 3, and the negative electrode 1 is formed by caulking the outer peripheral portion 52 a of the negative electrode current collector 52. The hydrophobic resin 56b is provided in a space surrounded by the positive electrode 2, the positive electrode current collector 51, and the gasket 56a, in close contact with the positive electrode 2, the positive electrode current collector 51, and the gasket 56a. The hydrophobic resin 56b can effectively suppress excessive penetration of fuel into the positive electrode 2 side. The end of the electrolyte layer 3 extends to the outside of the positive electrode 2 and the negative electrode 1 and is sandwiched between the gasket 56a and the hydrophobic resin 56b. The positive electrode current collector 51 has a plurality of oxidant supply ports 51b on the entire bottom surface, and the positive electrode 2 is exposed inside these oxidant supply ports 51b. FIG. 21C and FIG. 22 show 13 circular oxidant supply ports 51b, but this is only an example, and the number, shape, size, and arrangement of the oxidant supply ports 51b can be selected as appropriate. it can. The negative electrode current collector 52 also has a plurality of fuel supply ports 52b on the entire upper surface thereof, and the negative electrode 1 is exposed inside these fuel supply ports 52b. FIG. 22 shows seven circular fuel supply ports 52b, but this is only an example, and the number, shape, size, and arrangement of the fuel supply ports 52b can be appropriately selected.

The negative electrode current collector 52 has a cylindrical fuel tank 57 on the surface opposite to the negative electrode 1. The fuel tank 57 is formed integrally with the negative electrode current collector 52. In the fuel tank 57, a fuel (not shown) to be used, for example, a glucose solution or a solution obtained by adding an electrolyte to the glucose solution is placed. A cylindrical lid 58 is detachably attached to the fuel tank 57. For example, the lid 58 is fitted into the fuel tank 57 or screwed. A circular fuel supply port 58 a is formed at the center of the lid 58. The fuel supply port 58a is sealed, for example, by attaching a seal seal (not shown).
The configuration of the biofuel cell other than the above is the same as that of the first embodiment as long as it does not contradict its properties.

Next, an example of a method for manufacturing this biofuel cell will be described. This manufacturing method is shown in FIGS.
As shown in FIG. 23A, first, a cylindrical positive electrode current collector 51 having one end opened is prepared. A plurality of oxidant supply ports 51 b are formed on the entire bottom surface of the positive electrode current collector 51. A ring-shaped hydrophobic resin 56b is placed on the outer peripheral portion of the bottom surface inside the positive electrode current collector 51, and the positive electrode 2, the electrolyte layer 3, and the negative electrode 1 are sequentially stacked on the central portion of the bottom surface.

  On the other hand, as shown in FIG. 23B, a cylindrical fuel tank 57 is integrally formed on a cylindrical negative electrode current collector 52 with one end open. A plurality of fuel supply ports 52 b are formed on the entire surface of the negative electrode current collector 52. A gasket 56 a having a U-shaped cross section is attached to the edge of the outer peripheral surface of the negative electrode current collector 52. Then, the negative electrode current collector 52 is placed on the negative electrode 1 with its open side down, and the positive electrode 2, the electrolyte layer 3, and the negative electrode 1 are interposed between the positive electrode current collector 51 and the negative electrode current collector 52. Between.

  Next, as shown in FIG. 23C, the positive electrode 2, the electrolyte layer 3 and the negative electrode 1 sandwiched between the positive electrode current collector 51 and the negative electrode current collector 52 are placed on a caulking machine base 61. Then, the negative electrode current collector 52 is pressed by the pressing member 62 so that the positive electrode current collector 51, the positive electrode 2, the electrolyte layer 3, the negative electrode 1, and the negative electrode current collector 52 are adjacent to each other. The edge of the outer peripheral portion 51b of the positive electrode current collector 51 is caulked against the outer peripheral portion 52b of the negative electrode current collector 52 through the gasket 56a and the hydrophobic resin 56b. When this caulking is performed, the gasket 56a is gradually crushed so that there is no gap between the positive electrode current collector 51 and the gasket 56a and between the negative electrode current collector 52 and the gasket 56a. At this time, the hydrophobic resin 56b is also gradually compressed so as to be in close contact with the positive electrode 2, the positive electrode current collector 51, and the gasket 56a. By doing so, a space for accommodating the positive electrode 2, the electrolyte layer 3, and the negative electrode 1 is formed inside the positive electrode current collector 51 and the negative electrode current collector 52 while being electrically insulated from each other by the gasket 56 a. Is done. Thereafter, the caulking tool 63 is raised.

Thus, as shown in FIG. 23D, a biofuel cell in which the positive electrode 2, the electrolyte layer 3 and the negative electrode 1 are housed in the space formed between the positive electrode current collector 51 and the negative electrode current collector 52 is manufactured. The
Next, a lid 58 is attached to the fuel tank 57, fuel and electrolyte are injected from the fuel supply port 58a of the lid 58, and then the fuel supply port 58a is closed by attaching a hermetic seal. However, the fuel and electrolyte may be injected into the fuel tank 57 in the step shown in FIG. 23B.

In this biofuel cell, when, for example, a glucose solution is used as the fuel to be placed in the fuel tank 57, the negative electrode 1 decomposes the supplied glucose with an enzyme to extract electrons and generate H ⁺ . The positive electrode 2 generates water from H ⁺ transported from the negative electrode 1 through the electrolyte layer 3, electrons sent from the negative electrode 1 through an external circuit, and oxygen in the air, for example. An output voltage is obtained between the positive electrode current collector 51 and the negative electrode current collector 52.

As shown in FIG. 24, mesh electrodes 71 and 72 may be formed on the positive electrode current collector 51 and the negative electrode current collector 52 of this biofuel cell, respectively. In this case, external air enters the oxidant supply port 51 b of the positive electrode current collector 51 through the hole of the mesh electrode 71, and fuel enters the fuel tank 57 from the fuel supply port 58 a of the lid 58 through the hole of the mesh electrode 72. .
FIG. 25 shows a case where two biofuel cells are connected in series. In this case, the mesh electrode 73 is disposed between the positive electrode current collector 51 of one biofuel cell (the upper biofuel cell in the figure) and the lid 58 of the other biofuel cell (the lower biofuel cell in the figure). Between. In this case, external air enters the oxidant supply port 51 b of the positive electrode current collector 51 through the hole of the mesh electrode 73. The fuel can be supplied using a fuel supply system.
FIG. 26 shows a case where two biofuel cells are connected in parallel. In this case, the fuel tank 57 of one biofuel cell (the upper biofuel cell in the figure) and the fuel tank 57 of the other biofuel cell (the lower biofuel cell in the figure) are connected to the fuel of the lid 58. The supply ports 58 a are brought into contact with each other so as to coincide with each other, and the electrodes 74 are drawn out from the side surfaces of these fuel tanks 57. Further, mesh electrodes 75 and 76 are formed on the positive electrode current collector 51 of the one biofuel cell and the positive electrode current collector 51 of the other biofuel cell, respectively. These mesh electrodes 75 and 76 are connected to each other. External air enters the oxidizing agent supply port 51 b of the positive electrode current collector 51 through the holes of the mesh electrodes 75 and 76.

According to the second embodiment, when the fuel tank 57 is removed, the same advantages as those of the first embodiment can be obtained in the coin-type or button-type biofuel cell. In this biofuel cell, the positive electrode 2, the electrolyte layer 3 and the negative electrode 1 are sandwiched between the positive electrode current collector 51 and the negative electrode current collector 52, and the edge of the outer peripheral portion 51 a of the positive electrode current collector 51 is attached to the gasket 56. By caulking to the outer peripheral portion 52a of the negative electrode current collector 52 through this, in this biofuel cell, each constituent element can be brought into close contact with each other, so that variations in output can be prevented. The battery solution such as fuel and electrolyte can be prevented from leaking from the interface between the constituent elements. In addition, this biofuel cell has a simple manufacturing process. In addition, the biofuel cell can be easily downsized. Furthermore, this biofuel cell uses a glucose solution or starch as the fuel, and by selecting the pH of the electrolyte used to be around 7 (neutral), it is safe even if the fuel or electrolyte leaks to the outside.
In addition, it is necessary to add a fuel and an electrolyte at the time of production in an air cell that is currently in practical use, and it is difficult to add it after the production, whereas in this biofuel cell, a fuel and an electrolyte are added after the production. Therefore, biofuel cells are easier to manufacture than air cells currently in practical use.

Next explained is a biofuel cell according to the third embodiment of the invention.
As shown in FIG. 27, in the third embodiment, the fuel tank 57 provided integrally with the negative electrode current collector 52 is removed from the biofuel cell according to the second embodiment, and the positive electrode current collector 51 is further removed. And a negative electrode current collector 52 formed with mesh electrodes 71 and 72, respectively, and the biofuel cell is placed on the fuel 57a placed in an open fuel tank 57 with the negative electrode 1 side down and the positive electrode 2 side up. Use it in a floating state.
Except for the above, the third embodiment is the same as the first embodiment as long as it does not contradict its nature.
According to the third embodiment, the same advantages as those of the first embodiment can be obtained.

Next explained is a biofuel cell according to the fourth embodiment of the invention. The biofuel cell according to the second embodiment is a coin type or a button type, whereas this biofuel cell is a cylindrical type.
28A and B and FIG. 29 show this biofuel cell, FIG. 28A is a front view of this biofuel cell, FIG. 28B is a longitudinal sectional view of this biofuel cell, and FIG. 29 shows each component of this biofuel cell. It is a disassembled perspective view shown disassembled.

  As shown in FIGS. 28A and 28 and FIG. 29, in this biofuel cell, a cylindrical negative electrode current collector 52, a negative electrode 1, an electrolyte layer 3, a positive electrode 2 and a cylindrical negative electrode current collector A positive electrode current collector 51 is sequentially provided. In this case, the fuel holding portion 77 is a space surrounded by the cylindrical negative electrode current collector 52. One end of the fuel holding portion 77 protrudes to the outside, and a lid 78 is attached to this one end. Although not shown, the negative electrode current collector 52 on the outer periphery of the fuel holding portion 77 has a plurality of fuel supply ports 52b formed on the entire surface. The electrolyte layer 3 has a bag shape surrounding the negative electrode 1 and the negative electrode current collector 52. A portion between the electrolyte layer 3 and the negative electrode current collector 52 at one end of the fuel holding portion 77 is sealed by, for example, a seal member (not shown), and the fuel does not leak to the outside from this portion. Yes.

  In this biofuel cell, fuel and electrolyte are put in the fuel holding unit 77. These fuel and electrolyte reach the negative electrode 1 through the fuel supply port 52b of the negative electrode current collector 52 and penetrate into the gap of the negative electrode 1, thereby being stored inside the negative electrode 1. . In order to increase the amount of fuel that can be stored in the negative electrode 1, the porosity of the negative electrode 1 is desirably 60% or more, but is not limited thereto.

In this biofuel cell, a gas-liquid separation layer may be provided on the outer peripheral surface of the positive electrode current collector 51 in order to improve durability. As a material of this gas-liquid separation layer, for example, a waterproof moisture-permeable material (a material obtained by combining a film obtained by stretching polytetrafluoroethylene and a polyurethane polymer) (for example, Gore-Tex manufactured by WL Gore & Associates) Product name)). In order to make the components of the biofuel cell uniformly adhere to each other, it is preferable that the elastic rubber (even in the form of a band) has a network structure that allows air to pass from the outside to the outside or inside of the gas-liquid separation layer. A sheet form is also possible, and the whole components of the biofuel cell are tightened.
Except for the above, the fourth embodiment is the same as the first and second embodiments as long as it does not contradict its nature.
According to the fourth embodiment, the same advantages as those of the first and second embodiments can be obtained.

Next explained is a biofuel cell according to the fifth embodiment of the invention.
The biofuel cell according to the fifth embodiment is the same as the biofuel cell according to the first embodiment except that a porous conductive material as shown in FIG. 30 is used as the material of the electrode 11 of the negative electrode 1. It has a configuration.
FIG. 30A schematically shows the structure of the porous conductive material, and FIG. 30B is a cross-sectional view of the skeleton of the porous conductive material. As shown in FIGS. 30A and 30B, this porous conductive material includes a skeleton 81 made of a porous material having a three-dimensional network structure, and a carbon-based material 82 that covers the surface of the skeleton 81. This porous conductive material has a three-dimensional network structure in which a large number of holes 83 surrounded by a carbon-based material 82 correspond to a network. In this case, these holes 83 communicate with each other. The form of the carbon-based material 82 is not limited and may be any of a fibrous shape (needle shape) and a granular shape.

As the skeleton 81 made of a porous material, a foam metal or a foam alloy such as nickel foam is used. The porosity of the skeleton 81 is generally 85% or more, more typically 90% or more, and the pore diameter is typically, for example, 10 nm to 1 mm, more typically 10 nm to 600 μm, and more generally 1 to 600 μm, typically 50 to 300 μm, more typically 100 to 250 μm. As the carbon-based material 82, for example, a highly conductive material such as ketjen black is preferable, but a functional carbon material such as carbon nanotube or fullerene may be used.
The porosity of the porous conductive material is generally 80% or more, more typically 90% or more, and the diameter of the hole 83 is typically, for example, 9 nm to 1 mm, more generally 9 nm to It is 600 μm, more generally 1 to 600 μm, typically 30 to 400 μm, more typically 80 to 230 μm.

Next, a method for producing this porous conductive material will be described.
As shown in FIG. 31A, first, a skeleton 81 made of a foam metal or a foam alloy (for example, foam nickel) is prepared.
Next, as shown in FIG. 31B, a carbon-based material 82 is coated on the surface of the skeleton 81 made of the foam metal or foam alloy. As this coating method, a conventionally known method can be used. For example, the carbon-based material 82 is coated by spraying an emulsion containing carbon powder, a suitable binder, or the like onto the surface of the skeleton 81 by spraying. The coating thickness of the carbon-based material 82 is determined in accordance with the porosity and the pore diameter required for the porous conductive material in consideration of the porosity and the pore diameter of the skeleton 81 made of the foam metal or foam alloy. In this coating, a large number of holes 83 surrounded by the carbon-based material 82 are communicated with each other.
In this way, the target porous conductive material is manufactured.

  According to the fifth embodiment, the porous conductive material obtained by coating the surface of the skeleton 81 made of foam metal or foam alloy with the carbon-based material 82 has a sufficiently large diameter of the hole 83 and a rough three-dimensional network shape. While having a structure, it has high strength and high conductivity, and a necessary and sufficient surface area can be obtained. For this reason, the negative electrode 1 formed by forming an electrode 81 using this porous conductive material and immobilizing an enzyme, a coenzyme, an electron mediator, etc. on the electrode 81 has a high efficiency in enzyme metabolism reaction on the negative electrode 1. Or the enzyme reaction phenomenon occurring in the vicinity of the electrode 11 can be efficiently captured as an electric signal, and is stable regardless of the use environment, and is a high-performance biofuel cell Can be realized.

Next explained is a biofuel cell according to the sixth embodiment of the invention.
In this biofuel cell, starch, which is a polysaccharide, is used as a fuel. In addition, with the use of starch as fuel, glucoamylase, which is a degrading enzyme that decomposes starch into glucose, is also immobilized on the negative electrode 11.
In this biofuel cell, when starch is supplied as fuel to the negative electrode 1 side, this starch is hydrolyzed into glucose by glucoamylase, and this glucose is further decomposed by glucose dehydrogenase, accompanied by an oxidation reaction in this decomposition process. NAD ⁺ is reduced to produce NADH, which is oxidized by diaphorase and separated into two electrons, NAD ⁺ and H ⁺ . Therefore, two electrons and two H ⁺ are generated by one-step oxidation reaction per molecule of glucose. In the two-stage oxidation reaction, a total of 4 electrons and 4 H ⁺ are generated. The electrons thus generated are transferred to the electrode 11 of the negative electrode 1, and H ⁺ moves to the positive electrode 2 through the electrolyte layer 3. In the positive electrode 2, this H ⁺ reacts with oxygen supplied from the outside and electrons sent from the negative electrode 1 through an external circuit to generate H ₂ O.
Other than the above, the biofuel cell according to the first embodiment is the same.
According to the sixth embodiment, the same advantages as in the first embodiment can be obtained, and the amount of power generation can be increased by using starch as the fuel, compared with the case where glucose is used as the fuel. The advantage that it can be obtained.

As mentioned above, although embodiment of this invention was described concretely, this invention is not limited to the above-mentioned embodiment, The various deformation | transformation based on the technical idea of this invention is possible.
For example, the numerical values, structures, configurations, shapes, materials, and the like given in the above-described embodiments are merely examples, and different numerical values, structures, configurations, shapes, materials, and the like may be used as necessary.

1 is a schematic diagram showing a biofuel cell according to a first embodiment of the present invention. It is a basic diagram which shows typically the detail of a structure of the negative electrode of the biofuel cell by 1st Embodiment of this invention, an example of the enzyme group fix | immobilized by this negative electrode, and the electron transfer reaction by this enzyme group. It is a basic diagram which shows the result of the cyclic voltammetry performed in order to verify the permeation | transmission prevention effect of the electron mediator in the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measuring system used for the cyclic voltammetry performed in order to verify the permeation | transmission prevention effect of the electron mediator in the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the result of the cyclic voltammetry performed in order to verify the permeation | transmission prevention effect of the electron mediator in the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the result of the cyclic voltammetry performed in order to verify the permeation | transmission prevention effect of the electron mediator in the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the result of the chronoamperometry performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the result of the chronoamperometry performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measurement result of the enzyme activity performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measurement result of the enzyme activity performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measurement result of the enzyme activity performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measurement result of the enzyme activity performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measurement result of the enzyme activity performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measurement result of the enzyme activity performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the result of the chronoamperometry performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the relationship between the buffer solution density | concentration obtained from the result of the chronoamperometry performed for evaluation of the biofuel cell by 1st Embodiment of this invention, and the current density obtained. FIG. 16 is a schematic diagram showing a measurement system used for the measurement of chronoamperometry shown in FIG. 15. It is a basic diagram which shows the result of the cyclic voltammetry performed for evaluation of the biofuel cell by 1st Embodiment of this invention. It is a basic diagram which shows the measuring system used for the measurement of cyclic voltammetry shown in FIG. It is a basic diagram which shows the specific structural example of the biofuel cell by 1st Embodiment of this invention. It is the top view, sectional drawing, and back view which show the biofuel cell by the 2nd Embodiment of this invention. It is a disassembled perspective view which shows the biofuel cell by 2nd Embodiment of this invention. It is a basic diagram for demonstrating the manufacturing method of the biofuel cell by 2nd Embodiment of this invention. It is a basic diagram for demonstrating the 1st example of the usage method of the biofuel cell by the 2nd Embodiment of this invention. It is a basic diagram for demonstrating the 2nd example of the usage method of the biofuel cell by the 2nd Embodiment of this invention. It is a basic diagram for demonstrating the 3rd example of the usage method of the biofuel cell by 2nd Embodiment of this invention. It is a basic diagram which shows the biofuel cell by 3rd Embodiment of this invention, and its usage. It is the front view and longitudinal cross-sectional view which show the biofuel cell by 4th Embodiment of this invention. It is a disassembled perspective view which shows the biofuel cell by 4th Embodiment of this invention. It is the basic diagram and sectional drawing for demonstrating the structure of the porous conductive material used for the electrode material of a negative electrode in the biofuel cell by the 5th Embodiment of this invention. It is a basic diagram for demonstrating the manufacturing method of the porous electrically conductive material used for the electrode material of a negative electrode in the biofuel cell by the 5th Embodiment of this invention.

Explanation of symbols

  DESCRIPTION OF SYMBOLS 1 ... Negative electrode, 2 ... Positive electrode, 3 ... Electrolyte layer, 11 ... Electrode, 41, 42 ... Ti collector, 43, 44 ... Fixing plate, 47 ... Load, 51 ... Positive electrode collector, 51b ... Oxidant supply port 52 ... Negative electrode current collector, 52b ... Fuel supply port, 56a ... Gasket, 56b ... Hydrophobic resin, 57 ... Fuel tank, 58 ... Lid, 71, 72, 73 ... Mesh electrode, 77 ... Fuel holding part, 78 ... Lid, 81 ... skeleton, 82 ... carbon material, 83 ... hole

Claims (11)

The positive electrode and the negative electrode have a structure facing each other through a proton conductor, and at least one of the positive electrode and the negative electrode includes an enzyme, a microorganism or a cell, and an electron mediator ,
The proton conductor have a the oxidized form or the reduced form of the charge of the same sign of charge of the electron mediator,
The proton conductor includes a polyanion or polycation having a charge of the same sign as the charge of the oxidized or reduced form of the electron mediator,
A fuel cell , wherein the proton conductor comprises an electrolyte containing a buffer solution in which imidazole and hydrochloric acid are mixed . The fuel cell according to claim 1, wherein the concentration of the imidazole in the buffer solution is 0.1 to 4.0M. The fuel cell according to claim 1 or 2, wherein the enzyme includes an oxidase that is immobilized on the negative electrode and promotes and decomposes monosaccharides. The fuel cell according to claim 3, wherein the enzyme contains a coenzyme oxidase that returns a coenzyme reduced by oxidation of the monosaccharide to an oxidant and passes electrons to the negative electrode via an electron mediator. The oxidized form of the above coenzyme is NAD ⁺ The fuel cell according to claim 4, wherein the coenzyme oxidase is diaphorase. The oxidase is NAD ⁺ The fuel cell according to claim 3, which is a dependent glucose dehydrogenase. The fuel according to claim 1 or 2, wherein the enzyme includes a degrading enzyme immobilized on the negative electrode that promotes the degradation of the polysaccharide to produce a monosaccharide, and an oxidase that promotes the oxidation of the produced monosaccharide and decomposes the monosaccharide. battery. The degrading enzyme is glucoamylase, and the oxidase is NAD ⁺ The fuel cell according to claim 7, which is a dependent glucose dehydrogenase. The fuel cell according to any one of claims 1 to 8, wherein the enzyme includes an oxygen reductase immobilized on the positive electrode. The fuel cell according to claim 9, wherein the oxygen reductase is bilirubin oxidase. Using one or more fuel cells,
At least one of the fuel cells is
The positive electrode and the negative electrode have a structure facing each other through a proton conductor, and at least one of the positive electrode and the negative electrode includes an enzyme, a microorganism or a cell, and an electron mediator,
The proton conductor has a charge of the same sign as the charge of the oxidized or reduced form of the electron mediator;
The proton conductor includes a polyanion or polycation having a charge of the same sign as the charge of the oxidized or reduced form of the electron mediator,
An electronic device in which the proton conductor is made of an electrolyte containing a buffer solution in which imidazole and hydrochloric acid are mixed.

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