JP2019213470A - Gas discharging device and energy generating device - Google Patents

Abstract

To provide a gas discharging device capable of discharging gas by using enzyme reaction, or an energy generating device equipped with the gas discharging device.SOLUTION: A gas discharging device comprises a first enzyme immobilizing member, a second enzyme immobilizing member, and solution containing at least either one of substrate selected from the group consisting of a substrate of enzyme immobilized to the first enzyme immobilizing member and a substrate of enzyme immobilized to the second enzyme immobilizing member, in which the first enzyme immobilizing member and the second enzyme immobilizing member are in contact with the solution, and the device discharges gas to the opposite side of a side contacting the solution in the second enzyme immobilizing member.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a gas release device and an energy generation device.

In recent years, using a membrane, an electrode, or the like on which an enzyme is immobilized, and a substrate of the enzyme, energy generation using a compound generated by an enzyme reaction, detection of a compound, and the like have been performed.
Patent Document 1 discloses a method in which an electrode for hydrogen peroxide detection and a porous membrane formed by immobilizing an enzyme are fixed, and a redox enzyme having oxygen as a hydrogen acceptor is fixed to the electrode for hydrogen peroxide detection of the porous membrane. An enzyme electrode is described in which catalase is immobilized on the other side.

JP-A-56-142449

Conventionally, in various industries, gas (gas) is supplied.
Further, it is considered that, for example, if gas can be supplied to conventional materials, equipment, devices, and the like used in the atmosphere, more useful effects may be obtained.
For example, as one example, an “air battery” that generates energy using oxygen in the atmosphere has been developed. As the air battery, for example, an air zinc battery using oxygen and zinc in the atmosphere, an air bio battery using oxygen in the air and enzymes of a living body, and generating energy by a redox reaction of a biological component such as sugar, and the like. No.
It is considered that by supplying oxygen to the air battery using oxygen in the atmosphere, an effect of improving power generation output can be obtained.

An object of one embodiment of the present disclosure is to provide a gas release device capable of releasing a gas (eg, oxygen) using an enzymatic reaction, or an energy generation device including the gas release device. It is to be.
Another object of another embodiment of the present disclosure is to provide an energy generation device capable of generating energy using an oxygen and enzyme reaction.

Means for solving the above problems include the following aspects.
<1> A first enzyme-immobilized member, a second enzyme-immobilized member, an enzyme substrate immobilized on the first enzyme-immobilized member, and an enzyme immobilized on the second enzyme-immobilized member. A solution containing at least one substrate selected from the group consisting of enzyme substrates,
The first enzyme-immobilized member and the second enzyme-immobilized member are in contact with the solution,
Releasing gas to the opposite side of the second enzyme immobilization member from the side in contact with the solution,
Outgassing device.
<2> The enzymes immobilized on the first enzyme-immobilized member are glucose oxidase and pyranose oxidase, and the enzyme immobilized on the second enzyme-immobilized member is catalase. The gas releasing device according to <1>, wherein the contained substrate is D-glucose, and the gas is oxygen.
<3> An energy generation device comprising: the gas release device according to <1> or <2>; and an energy generation unit that generates energy by using gas released by the gas release device.
<4> The energy generating section includes a positive electrode serving as a first enzyme-immobilized electrode, a negative electrode serving as a second enzyme-immobilized electrode, and an electrolyte containing an enzyme substrate immobilized on the negative electrode. The energy generator according to <3>, wherein the enzyme immobilized on the positive electrode is in contact with the electrolyte, and the enzyme immobilized on the negative electrode is in contact with the electrolyte to generate power.
<5> a positive electrode that is a first enzyme-immobilized electrode;
A negative electrode that is a second enzyme-immobilized electrode;
An electrolyte containing an enzyme substrate immobilized on the negative electrode,
A gas-liquid separator capable of transmitting gas without allowing the electrolyte to leak out,
An energy generating device, wherein an enzyme immobilized on the positive electrode contacts the electrolyte and the gas-liquid separating member, and the negative electrode contacts the electrolyte to generate electric power.

According to one embodiment of the present disclosure, there is provided a gas release device capable of releasing a gas using an enzyme reaction, or an energy generation device including the gas release device.
According to another aspect of the present disclosure, there is provided an energy generation device capable of generating energy using an oxygen and enzyme reaction.

FIG. 1 is a schematic cross-sectional view illustrating an example of a gas supply device 10 that is an embodiment of a gas release device according to the present disclosure. FIG. 1 is a schematic cross-sectional view illustrating an example of an air bio battery 100 that is an embodiment of an energy generation device according to the present disclosure. FIG. 2 is a schematic cross-sectional view illustrating an example of a battery including a gas supply device and a battery unit that generates energy by using gas supplied by the gas supply device. It is a photograph of the gas supply device produced in the example. It is a schematic sectional drawing which shows the attachment direction of the 1st enzyme immobilization film | membrane in the gas supply apparatus described in FIG. 4, and the 2nd enzyme immobilization film | membrane. It is a graph which shows the evaluation result of the gas supply capability of the gas supply device produced in the example. 6 is a graph showing evaluation results of power generation characteristics of batteries 2 and batteries 4 manufactured in Examples.

Hereinafter, the contents of the present disclosure will be described in detail. The description of the constituent elements described below may be made based on typical embodiments of the present disclosure, but the present disclosure is not limited to such embodiments.
In addition, in this specification, "-" which shows a numerical range is used by the meaning which includes the numerical value described before and behind that as a lower limit and an upper limit.
In the present disclosure, “mass%” and “weight%” are synonymous, and “part by mass” and “part by weight” are synonymous.
In the drawings, the same components will be described with the same reference numerals. Further, reference numerals in the drawings may be omitted.
In the present disclosure, when referring to the amount of each component in the composition, when there are a plurality of substances corresponding to each component in the composition, unless otherwise specified, a plurality of substances present in the composition Means the total amount.
Furthermore, in the present disclosure, a combination of two or more preferred embodiments is a more preferred embodiment.
Hereinafter, the present disclosure will be described in detail.

(Gas release device)
The gas release device according to the present disclosure includes a first enzyme-immobilized member, a second enzyme-immobilized member, an enzyme substrate immobilized on the first enzyme-immobilized member, and the second enzyme-immobilized member. A solution containing at least one substrate selected from the group consisting of enzyme substrates immobilized on the immobilization member (hereinafter, also referred to as a “specific solution”). A second enzyme-immobilized member is in contact with the solution, and releases gas to a side of the second enzyme-immobilized member opposite to a side in contact with the solution. A gas release device according to the present disclosure releases a gas to a side of the second enzyme immobilization member opposite to a side in contact with the solution using an enzyme reaction.
The gas release device according to the present disclosure absorbs gas from the side of the first enzyme immobilization member opposite to the side in contact with the solution, and is in contact with the solution in the second enzyme immobilization member. The gas supply device is preferably a gas supply device that discharges gas to the opposite side and supplies the gas to a required target.
The gas to be absorbed and the gas to be supplied may be the same or different, but are preferably the same.
Here, since the enzymatic reaction is often an irreversible reaction, even if the absorbed gas and the supplied gas are the same gas, the gas is reacted against the concentration gradient of the gas. It will be possible to supply.
Further, the gas release device according to the present disclosure depends on the design. For example, by using an easily available specific solution (for example, a glucose solution or the like), gas supply can be easily put to practical use. Is considered to be very useful. As the glucose solution or the like, for example, a body fluid such as blood, sweat, tissue fluid, or the like may be used. For example, it is considered that the affinity is excellent when used in combination with a bioimplantable battery or a battery described later. Further, application to animals, insects, plants, and the like can be considered, and further, use of beverages such as juices and food can be considered.
Hereinafter, typical aspects of the gas release device according to the present disclosure will be described with reference to the drawings, but the gas release device according to the present disclosure is not limited to the drawings.
In the description in the text, reference numerals in the drawings may be omitted.

FIG. 1 is a schematic cross-sectional view illustrating an example of a gas supply device 10 that is an embodiment of a gas release device according to the present disclosure.
The gas supply device 10 shown in FIG. 1 includes a first enzyme-immobilized film 12 (an example of a first enzyme-immobilized member) and a second enzyme-immobilized film 22 (an example of a second enzyme-immobilized member). And a specific solution 30.
In the gas supply device 10 shown in FIG. 1, the first enzyme-immobilized membrane 12 (the second enzyme-immobilized membrane 22) is made of a first support 14 (permeating the specific solution 30) in consideration of the strength of the membrane. It is fixed by a second support 24). However, when there is no problem in the membrane strength, or when a plate-like enzyme immobilizing member having a predetermined strength is used, the first support 14 (the second support 24) may be omitted.
In the gas supply device 10 shown in FIG. 1, the first enzyme-immobilized membrane 12, the first support 14, the specific solution 30, the second support 24, and the second enzyme-immobilized membrane 22 are It is supported by 40. The shape of the case 40 is not particularly limited.
In the gas supply device 10 shown in FIG. 1, for example, the gas is absorbed from the side (opening 42) of the first enzyme-immobilized membrane 12 opposite to the side in contact with the specific solution 30, and the second enzyme-immobilized membrane 12 Gas can be released to the opposite side (opening 46) of the oxide film 22 from the side in contact with the specific solution 30. The gas released to the opening 46 is supplied to the outside through the opening 44.

<Gas type>
The gas supply device 10 shown in FIG. 1 appropriately supplies the enzyme immobilized on the first enzyme immobilized film 12, the enzyme immobilized on the second enzyme immobilized film 22, the substrate contained in the specific solution 30, and the like. By changing, it is possible to select the gas to be supplied.
The gas supplied by the gas supply device 10 includes oxygen (O ₂ ), hydrogen (H ₂ ), carbon dioxide (CO ₂ ), and the like. From the viewpoint of industrial availability, for example, oxygen is used. Preferably, there is.
For example, in the gas supply device 10, the enzymes immobilized on the first enzyme immobilization membrane 12 are glucose oxidase (Glucose oxidase, GOD) and pyranose oxidase (POX). An embodiment in which D-glucose is used and the enzyme immobilized on the second enzyme-immobilized membrane 22 is catalase (CAT) is preferable. Hereinafter, the gas supply device of the above embodiment is also referred to as “gas supply device A”. According to the gas supply device A, it is possible to provide a gas supply device that supplies oxygen.
Details of the enzymatic reaction caused in the gas supply device A are shown below.

In the gas supply device A, the above-mentioned enzyme reactions (1) to (3) occur in the first enzyme-immobilized membrane 12, and convert oxygen to hydrogen peroxide.
Thereafter, the hydrogen peroxide generated in the enzyme reactions (1) to (3) moves to the second enzyme-immobilized membrane 22 through the specific solvent 30, and the ( The oxygen is finally supplied from the opening 44 by being converted into oxygen by the enzymatic reaction of 4).
Since the reactions (1) to (4) are enzymatic reactions and are irreversibly close reactions, the supply of oxygen is performed more efficiently than the oxygen concentration of the opening 42 in the gas supply device A. It is considered that the reaction proceeds even when the oxygen concentration is high.
That is, it is possible to make the oxygen concentration of the opening 46 higher than the oxygen concentration of the opening 42 in the gas supply device A. In other words, according to the gas supply device 10 according to the present disclosure, it can be said that the gas can be transported between the opening 42 and the opening 46 against the gas concentration gradient.
Further, pyranose oxidase is immobilized on the first enzyme-immobilized membrane 12 in addition to glucose oxidase. Therefore, the enzyme reactions (2) and (3) occur in addition to the enzyme reaction (1), and more hydrogen peroxide is generated. As a result, a higher concentration of oxygen can be generated by the enzymatic reaction of (4).

  The combination of the enzyme immobilized on the first enzyme immobilization member, the substrate contained in the specific solution, the enzyme immobilized on the second enzyme immobilization member, and the released gas are shown in Table 1 below. The combinations described are exemplified, but are not limited thereto.

The enzyme immobilized on the first enzyme-immobilized member (for example, the first enzyme-immobilized membrane 12) and the enzyme immobilized on the second enzyme-immobilized member (for example, the second enzyme-immobilized membrane 22). The method for selecting the enzyme species of the enzyme is not particularly limited, and may be appropriately selected according to the target supplied gas.
For example, as the enzyme immobilized on the first enzyme-immobilized membrane 12, the substrate in the specific solution 30 is changed to a product using gas in the atmosphere, and the by-products such as the above-described hydrogen peroxide are removed. An enzyme that converts the by-product into a target gas can be used as the enzyme immobilized on the second enzyme-immobilized membrane 22 using the generated enzyme.
As the enzyme immobilized on the first enzyme-immobilized member or the second enzyme-immobilized member, a single enzyme may be used, or a plurality of enzymes may be used in combination.
When a plurality of enzymes are used in combination as the enzyme immobilized on the first enzyme immobilization member, for example, from the viewpoint of improving the gas supply ability, a specific enzyme such as a combination of glucose oxidase and pyranose oxidase is used. It is preferable to use an enzyme that uses a compound in the solution 30 (glucose solution) as a substrate and an enzyme that uses a product generated by a reaction between the enzyme and the substrate as a substrate.
The optimum pH, the optimum pH, and the like of the enzyme immobilized on the first enzyme-immobilized member or the second enzyme-immobilized member are not particularly limited, and may be determined according to the application and design.

<First enzyme immobilization member>
The first enzyme immobilization member contains an enzyme.
FIG. 1 shows an embodiment in which a first enzyme-immobilized membrane 12 as a first enzyme-immobilized member is formed on a first support 14. May be formed, for example, as a resin film containing an enzyme, and the first support 14 may be omitted, or the first support 14 may be impregnated with the enzyme of the first enzyme-immobilized membrane 12, At least a part of the one support 14 and the first enzyme-immobilized membrane 12 may be integrated.
The first enzyme-immobilized membrane 12 preferably contains an enzyme and a binder.
The enzyme contained in the first enzyme-immobilized membrane 12 may be selected from the above combinations according to the application.
The binder contained in the first enzyme-immobilized membrane 12 is not particularly limited, but for example, a curable resin can be used.
In the present disclosure, the curable resin refers to a resin obtained by curing a curable resin, and the curable resin refers to a resin having a property of being cured by irradiation with ultraviolet light, heating, or the like.
The curable resin in the first enzyme-immobilized film 12 is preferably an ultraviolet curable resin cured by ultraviolet light from the viewpoint of easy handling.
The curable resin in the first enzyme-immobilized membrane 12 is preferably a resin that becomes water-insoluble at least after curing.
Specifically, the ultraviolet curable resin preferably has a solubility in water of 100 parts by mass at 25 ° C. of less than 0.1 part by mass, and more preferably less than 0.05 part by mass. The lower limit of the dissolution amount is not particularly limited, and may be 0 parts by mass or more.

  The first enzyme-immobilized membrane 12 may further include other components. Other components include known protective materials and surfactants used for protecting enzymes, binders, the first support, and the like.

The first enzyme-immobilized film 12 is produced, for example, by applying a composition containing an enzyme and a curable resin to the first support 14 and curing the composition by irradiating ultraviolet rays or the like.
The curable resin used for forming the first enzyme-immobilized film 12 is not particularly limited, and a known ultraviolet curable resin can be used. For example, PVA-SbQ (ultraviolet curable resin) can be used. And polyvinyl alcohol to which a styrylpyridinium salt is added).

(First support)
As the first support 14, a known material can be used without any particular limitation, but it is preferable to use a dialysis membrane.
As the dialysis membrane, a dialysis membrane permeable to the enzyme substrate immobilized on the first enzyme immobilization membrane 12 is preferably used, and examples thereof include cellulose, hydrophilic polytetrafluoroethylene, and hydrophobic cotton. In addition to the dialysis membrane, various soft and hard porous bodies can be used.
The positional relationship between the first support 14 and the first enzyme-immobilized membrane 12 is not particularly limited, but when a gas is used in the enzyme reaction in the first enzyme-immobilized membrane 12, the first enzyme-immobilized membrane 12 may be used. It is preferable that the first support 14 exists between the conversion film 12 and the specific solution 30. With the above positional relationship, in the enzyme reaction in the first enzyme-immobilized membrane 12, not the gas in the specific solution 30, but the gas outside the device can be easily used. It is thought that it is easier to improve.

  Further, the thickness and area of the first enzyme-immobilized membrane 12 and the first support 14 and the amount of the enzyme contained therein may be appropriately set according to the use.

<Second enzyme immobilization member>
The second enzyme-immobilized member can have the same configuration as the first enzyme-immobilized member. That is, the second enzyme-immobilized membrane 22 as the second enzyme-immobilized member and the second support 24 are the same as the first enzyme-immobilized membrane 12 except that the type of the contained enzyme is different. A configuration can be adopted, and preferred embodiments are also the same.
Although the positional relationship between the second support 24 and the second enzyme-immobilized membrane 22 is not particularly limited, the second support 24 is located between the second enzyme-immobilized membrane 22 and the specific solution 30. Preferably it is present. With the above positional relationship, the gas generated by the enzyme reaction in the second enzyme-immobilized membrane 22 is suppressed from dissolving in the specific solution 30, so that the gas supply capability in the gas supply device 10 is more likely to be improved. it is conceivable that.

<Specific solution>
The specific solution 30 is not particularly limited, but is preferably an aqueous solution from the viewpoint of safety and the like.
The specific solution 30 contains at least one substrate selected from the group consisting of an enzyme substrate immobilized on the first enzyme-immobilized membrane and an enzyme substrate immobilized on the second enzyme-immobilized membrane. And a solution containing at least the enzyme substrate immobilized on the first enzyme-immobilized membrane.
The concentration of the substrate in the specific solution 30 is not particularly limited, and may be set according to the application. It is conceivable that the higher the concentration, the easier the enzymatic reaction proceeds, and the higher the gas supply ability. It is also conceivable to lower the concentration in consideration of the precipitation and denaturation of the substrate compound due to a change in temperature, depending on the application.
The pH of the specific solution 30 is not particularly limited, and may be, for example, an optimal pH of the enzyme immobilized on the first enzyme-immobilized membrane and / or the enzyme immobilized on the second enzyme-immobilized membrane. Alternatively, the pH is preferably set to a value close to this.

The specific solution 30 may further contain a compound other than the above substrate.
Compounds other than the substrate include known preservatives, wetting agents, defoamers, stabilizers, organic solvents, and the like. Further, when a body fluid or the like is used, it is conceivable that proteins, ions, and the like in biological components are also contained. The content of these compounds can be set as appropriate.

<Application>
The opening 42, the opening 46, and the opening 44 in the gas supply device 10 illustrated in FIG. 1 may be in contact with a gas, or one of them may be in contact with a liquid such as water. And both may be in contact with a liquid such as water.
When both are in contact with gas, for example, there is a mode in which the opening 42 is open and is in contact with the atmosphere, and the openings 46 and 44 are in contact with gas in a closed space. According to such an embodiment, a specific gas is supplied to the closed space through the gas supply device 10, and the concentration of the specific gas in the closed space increases.
When the opening 42 is in contact with the liquid and the openings 46 and 44 are in contact with the gas in the closed space, the gas supply device 10 causes the specific gas in the liquid to be removed from the gas in the closed space. Can be supplied to
When the opening 42 is in contact with a gas and the openings 46 and 44 are in contact with a liquid, the gas supply device 10 can supply a specific gas in the gas to the liquid.
When both are in contact with the liquid, for example, the gas in the liquid that is in contact with the opening 42 can be supplied to the liquid that is in contact with the openings 44 and 46.

  The gas supply device according to the present disclosure is capable of breathing in water by using a battery described later, an artificial lung for extracting oxygen from water (for example, using the gas supply device as a mask covering a respiratory unit such as a mouth and a nose). Can be used for applications such as an oxygen supply device for aquariums for breeding and cultivating fish and shellfish, an oxygen supply device to be mounted at the time of climbing and exercising, and an aerial art display such as a liquid column. it is conceivable that.

(Energy generator)
A gas release device according to the present disclosure includes, for example, a gas release device according to the present disclosure, and an energy generation unit (for example, a battery unit) that generates energy using gas released from the gas release device. It can be used as an energy generating device (for example, a battery).
Here, as an energy generating unit (battery unit) that generates (generates) energy using gas, a known air battery such as a known air zinc battery and an air lithium battery, or an air bio battery described later is used. No.
The gas used in the energy generating section (battery section) is not particularly limited, but is preferably oxygen from the viewpoint of being practically used at the present time.
It is conceivable that the power generation output of these energy generation units (battery units) increases as the gas concentration increases. Therefore, by using the gas release device according to the present disclosure and increasing the gas concentration of the atmosphere including the means for using gas in the energy generation unit (battery unit), the power generation output in the energy generation unit (battery unit) is improved. It is thought that it can be done.

Conventionally, for example, in an air battery, an attempt has been made to improve the power generation output by selecting an electrode, an electrolyte, and the like.However, it is often difficult to improve the gas concentration, and an attempt is made on the assumption that air is used. There were many. Therefore, the low concentration of the gas in the air (for example, the oxygen concentration) may be dominant to the power generation reaction speed in the battery.
Here, according to the gas discharge device according to the present disclosure, it is possible to discharge gas by a simple method, and therefore, the gas discharge device according to the present disclosure further improves the power generation output of the energy generation unit (battery unit). It is considered to be very useful from the viewpoint of doing.
The gas release device according to the present disclosure includes a power device (an example of an energy generation device) including a power generation unit (an example of an energy generation unit) that generates power using a released gas, in addition to the battery. It is also possible to use as. For example, a gas pressure actuator that operates a piston with the pressure of released gas, a rubber artificial muscle that uses deformation due to pressurization of a rubber tube, a diaphragm pump that combines a diaphragm and a check valve, an internal combustion engine that uses combustion of released gas Various applications are conceivable.
In particular, when blood is used as the specific solution 30 and an insulin pump is configured with a diaphragm as a drug injecting mechanism, oxygen is released in accordance with the glucose concentration (blood sugar level) in the blood and connected to the insulin reservoir, Insulin can be injected by a drug injection mechanism driven by the pressure of the released oxygen. Thereby, autonomous and continuous blood sugar level control using blood sugar as a driving force source becomes possible, and an artificial pancreas system can be realized.

The energy generation unit according to the present disclosure includes a positive electrode that is a first enzyme-immobilized electrode, a negative electrode that is a second enzyme-immobilized electrode, and an electrolyte including an enzyme substrate immobilized on the negative electrode. It is preferable that the enzyme immobilized on the positive electrode is in contact with the electrolyte, and the enzyme immobilized on the negative electrode is an energy generating unit that generates electric power by contacting the electrolyte.
When the energy generating unit in the present disclosure is a battery unit, a positive electrode that is a first enzyme-immobilized electrode, a negative electrode that is a second enzyme-immobilized electrode, and a substrate of the enzyme immobilized on the negative electrode are included. An electrolyte (hereinafter, also simply referred to as an “electrolyte”), and a battery unit in which the positive electrode is in contact with the electrolyte, and the negative electrode is in contact with the electrolyte. Bio battery ").
Hereinafter, an example of an air bio battery as an energy generation device according to the present disclosure will be described with reference to FIG. 2, but the air bio battery is not limited to the embodiment illustrated in FIG.

FIG. 2 is a schematic cross-sectional view illustrating an example of the air bio battery 100 that is an embodiment of the energy generation device according to the present disclosure.
The air bio battery 100 shown in FIG. 2 includes, in a case 140, a positive electrode 110 serving as a first enzyme-immobilized electrode, a negative electrode 120 serving as a second enzyme-immobilized electrode, and an electrolyte 130, The positive electrode is in contact with the electrolyte, and the negative electrode is in contact with the electrolyte. The positive electrode 110 is in contact with outside air via an electrolyte supply member 118 (an example of a gas-liquid separation member).
The positive electrode 110 has a structure in which the enzyme 112 is immobilized on the surface of the electrode 114. Further, it is preferable that an electron mediator is further fixed to the positive electrode 110.
The negative electrode 120 has a structure in which the enzyme 122 is immobilized on the surface of the electrode 124. Further, it is preferable that an electron mediator is further fixed to the negative electrode 120.
The substrate of the enzyme immobilized on the electrode may be further immobilized on at least one electrode selected from the group consisting of the positive electrode 110 and the negative electrode 120.
In the air bio battery 100 shown in FIG. 2, the positive electrode 110 (negative electrode 120) is fixed to the first electrode support 116 (second electrode support 126), but the first electrode support 116 ( The second electrode support 126) may be omitted.
Further, the shape of case 140 is not particularly limited.
In the air bio battery 100 shown in FIG. 2, the positive electrode 110 and the negative electrode 120 are in contact with the same electrolyte 130, but may be in contact with different electrolytes 130. Specifically, for example, there is an embodiment in which the electrolyte in contact with the positive electrode 110 and the electrolyte in contact with the negative electrode 120 are different electrolytes, and the electrolytes are in contact with each other via a semipermeable membrane (dialysis membrane).
The electrolyte supply member 118 is capable of supplying gas (outside air) without supplying the electrolyte 130 to the positive electrode 110 and leaking the electrolyte 130 out of the case 140.

<Combination of enzyme and substrate>
In the air bio battery 100 shown in FIG. 2, the enzyme 112 immobilized on the positive electrode 110, the enzyme 122 immobilized on the negative electrode 120, the substrate included in the electrolyte 130, and the like can be appropriately changed.
For example, an electrode on which bilirubin oxidase (BOD) is immobilized on the positive electrode 110 as the enzyme 112 and K ₃ [Fe (CN) ₆ ] is immobilized on the electron mediator, and glucose dehydrogenase ( An embodiment in which an electrode in which glucose dehydrogenase (GDH) and diaphorase (Dp) are immobilized, NADH is immobilized as a substrate, and vitamin K3 is immobilized as an electron mediator, and the electrolyte 130 contains D-glucose. Hereinafter, the air bio battery of the above embodiment is also referred to as “air bio battery A”.
In the present specification, NADH indicates reduced nicotinamide dinucleotide, and NAD ⁺ indicates oxidized nicotinamide dinucleotide.
The details of the reaction at the positive electrode or the negative electrode of the air bio battery A are shown below.

In the air bio battery A, the enzyme reactions (1) and (2) occur at the negative electrode 120 to generate electrons. In the positive electrode 110, the above reaction (3) occurs, and electrons are used. As described above, in the air biobattery A, it is possible to extract electric energy from glucose using an enzymatic reaction. Here, in the air bio battery 100 shown in FIG. 2, the enzyme 112 immobilized on the positive electrode 110 is in close contact with the electrolyte supply member 118. Thus, not only the electrolyte 130 but also oxygen can be taken in from the outside air, so that the above-mentioned reaction (3) is promoted and the power density is improved.
The optimal pH, optimal pH, and the like of the enzyme immobilized on the positive electrode 110 or the negative electrode 120 are not particularly limited, and may be determined according to the application and design.

  The combination of an enzyme, a substrate, and an electron mediator (Med) immobilized on the positive electrode 110, an enzyme, a coenzyme, an electron mediator, and a substrate included in the electrolyte 130 immobilized on the negative electrode 120 are as described in Table 2 below. However, the present invention is not limited to this.

<Positive electrode>
The positive electrode 110 can have a structure in which the enzyme 112 is immobilized on the surface of the electrode 114.
As long as the enzyme 112 is in contact with the electrolyte 130, it may be immobilized inside the electrode 114, for example.

〔electrode〕
The electrode 110 is not particularly limited, and a known electrode in the field of batteries can be used. A carbon electrode such as glassy carbon, a metal electrode such as a platinum electrode, or the like is used, and the enzyme is immobilized by various methods. Platinum electrodes are preferred from the viewpoint of easy conversion.
In the air bio battery 100 shown in FIG. 2, a platinum electrode deposited on the first electrode support 116 is used.
The size and shape of the electrode are not particularly limited, and may be designed according to the use or the like.

(First electrode support)
The first electrode support is not particularly limited, but is preferably a porous material from the viewpoint that the enzymatic reaction proceeds efficiently when a metal electrode is deposited and used.
The first electrode support is not particularly limited, but is preferably a hydrophobic material, and is preferably a plastic such as hydrophobic PTFE (polytetrafluoroethylene) or PET.
The first electrode support is not particularly limited, but from the viewpoint of ease of handling, for example, a film-like support is preferably used.

(Enzyme immobilization method)
The method for immobilizing the enzyme on the electrode is not particularly limited, and a known method may be used, and examples thereof include an LbL (Layer-by-Layer) method, a comprehensive immobilization method, and a crosslinking method.
The amount of the immobilized enzyme and the like are not particularly limited, and may be appropriately set depending on the use.
When the enzyme is immobilized, a substrate, a mediator, and other compounds may be immobilized together.

-LbL method-
The LbL method is a method in which a substrate is alternately immersed in a solution containing a cation and a solution containing an anion, thereby adsorbing ions by electrostatic interaction to form a thin film.
As the LbL method, a method of forming a self-assembled monolayer (SAM) (SAM / LbL method) is also preferably used.
For example, using a solution containing poly (diallyldimethyl ammonium chloride) (PDDA) that is positively charged and a solution containing poly (sodium 4-stylenesulfonate) (PSS) that is negatively charged, a PDDA solution and an enzyme are used as necessary. A method of forming the solution by applying the solution containing the substrate and / or the mediator and the solution containing the PSS to the electrode in this order is mentioned.
In addition, as the SAM / LbL method, it may be formed by a known method using a solution containing 2-mercaptoethanesulfonic acid sodium salt.

−Comprehensive immobilization method−
The entrapment immobilization method is a method in which an enzyme or the like is fixed to an electrode by solidifying the enzyme or the like in a gel or solid state with a polymer material and forming a layer of the polymer material containing the enzyme.
Examples of the polymer material used in the entrapping immobilization method include the above-described ultraviolet curable resin, and for example, PVA-SbQ (ultraviolet curable resin, polyvinyl alcohol to which a styrylpyridinium salt is added) is preferable.
For example, there is a method in which a solution containing PVA-SbQ, an enzyme, and if necessary, a solution containing a substrate and / or a mediator, and a solution containing PSS are prepared, applied to an electrode, and then subjected to ultraviolet irradiation.

-Cross-linking method-
The cross-linking method is a method in which a molecule containing an enzyme is insolubilized by reacting a polyfunctional reagent, an enzyme, and if necessary, a compound such as another protein to form a large molecule.
Examples of the polyfunctional reagent include glutaraldehyde.
Examples of other proteins include, but are not particularly limited to, BSA (bovine serum albumin, Bovine Serum Albumin) and the like.
For example, a method in which a solution in which an enzyme, BSA, an enzyme, and a mediator and / or a substrate are mixed as necessary is applied onto the electrode, and then a glutaraldehyde solution is applied and dried.

<Negative electrode>
The negative electrode 120 (the immobilized enzyme 122 and the electrode 124) and the second electrode support 126 are the same as the positive electrode 110 except that the enzyme, the substrate provided as needed, and / or the compound used as a mediator are different. The same electrodes as described above are used.
From the viewpoint of suppressing damage to the positive electrode or the negative electrode due to a difference in ionization tendency between the positive electrode and the negative electrode, the positive electrode and the negative electrode are preferably made of the same material except for the enzyme, the substrate, and the mediator.

<Electrolyte>
The electrolyte 130 is not particularly limited, but is preferably an aqueous solution from the viewpoint of safety and the like.
The electrolyte 130 is an electrolyte containing an enzyme substrate immobilized on the negative electrode 120.
The concentration of the substrate in the electrolyte 130 is not particularly limited, and may be set according to the application. It is conceivable that the higher the concentration, the easier the enzymatic reaction proceeds, and the higher the power generation output. It is also conceivable to lower the concentration in consideration of the deposition of the substrate compound due to a temperature change, etc., depending on the application.
The pH of the electrolyte 130 is not particularly limited, and is preferably, for example, an optimum pH of the enzyme immobilized on the negative electrode, or a pH close thereto.

The electrolyte 130 may further include a compound other than the above substrate.
Compounds other than the substrate include known preservatives, wetting agents, defoamers, stabilizers, organic solvents, and the like. The content of these compounds can be set as appropriate.

<Gas-liquid separation member (electrolyte supply member)>
The air bio battery 100 illustrated in FIG. 2 includes an electrolyte supply member 118 (an example of a gas-liquid separation member).
The electrolyte supply member 118 is in contact with the electrolyte 130, is in contact with the positive electrode 110, and preferably has an open side opposite to the positive electrode 110, and allows gas (outside air) to permeate without leaking the electrolyte 130 out of the case 140. Is possible.
As the electrolyte supply member, for example, by using a member having a void such as a film made of cotton mesh or PTFE, at least a part of the positive electrode 110 is in contact with the outside air, or the electrolyte existing between the outside air and the positive electrode 110 Since the amount is smaller than when the electrolyte supply member is not present, it is considered that a gas such as oxygen is easily used in the reaction at the positive electrode 110, and the power generation output is likely to be improved.
As the electrolyte supply member, any member having a gap can be used without particular limitation. For example, a water-absorbing member is preferably used from the viewpoint of easily supplying the electrolyte.

<Position relation>
The positional relationship between the positive electrode 110, the negative electrode 120, the electrolyte 130, and the like is not particularly limited.
When the enzymatic reaction in the positive electrode 110 is a reaction using a gas, the positive electrode 110 may be present near the interface between the electrolyte 130 and the outside air (particularly, in close contact with the electrolyte supply member 118) from the viewpoint of facilitating the use of the gas. preferable.

<Combined use with gas release device>
FIG. 3 shows a gas supply device, which is an embodiment of a gas release device according to the present disclosure, as an energy generation device according to the present disclosure, and a battery unit that generates energy using gas supplied by the gas supply device. FIG. 3 is a schematic cross-sectional view illustrating an example of a battery including (corresponding to a combination of the gas supply device 10 illustrated in FIG. 1 and the air bio battery 100 illustrated in FIG. 2).
The battery 200 shown in FIG.
A case 40 (a lower case), a first enzyme-immobilized membrane 12, a second enzyme-immobilized membrane 22, a first support 14, a second support 24, a specific solution 30, and an upper case 48 are provided. A gas supply device;
A case 140, a positive electrode 110, an electrolyte supply member 118, a first electrode support 116, a negative electrode 120, a second electrode support 126, and a battery unit (air bio battery) including the electrolyte 130 are provided.
Note that the specific solution 30 and the electrolyte 130 may be the same solution or different solutions. However, it is preferable that the specific solution 30 and the electrolyte 130 be the same so that the battery 200 can be constituted by one solution.
3, 52a and 52b, 54a and 54b, and 56a and 56b each represent a cross section of one O-ring. As shown in FIG. 3, it is possible to configure the gas supply device and the battery unit by fixing each member using a member such as an O-ring. Examples of the O-ring include those made of rubber and silicon.
In the battery 200 shown in FIG. 3, it is considered that the gas is supplied from the gas supply device to the space 142 including the opening 46 and the opening 44, and the enzyme reaction using the gas in the positive electrode 110 of the battery unit proceeds easily.
For example, when the battery unit is the air bio battery A, it is preferable to use the gas supply device A as the gas supply device.
By using the air bio-battery A and the gas supply device A in combination, a high concentration of oxygen is supplied to the positive electrode of the air bio-battery A, and the reaction using the oxygen easily proceeds. It is considered that the output increases.
As described above, a battery including the gas supply device according to the present disclosure and the battery unit that generates energy using the gas supplied by the gas supply device is considered to have excellent power generation output.

(Energy generator)
The energy generating apparatus according to the present disclosure includes a positive electrode that is a first enzyme-immobilized electrode, a negative electrode that is a second enzyme-immobilized electrode, an electrolyte including an enzyme substrate immobilized on the negative electrode, and the electrolyte. A gas-liquid separating member capable of transmitting gas without leaking, and an enzyme fixed to the positive electrode is in contact with the electrolyte and the gas-liquid separating member, and the negative electrode is in contact with the electrolyte to supply power. appear.
The energy generating device according to the present disclosure includes the above-described gas-liquid separating member as an essential component, and is the same as the above-described energy generating portion except that the enzyme fixed to the positive electrode also contacts the gas-liquid separating member. The preferred embodiment is also the same.
In addition, a preferable example of the energy generation device according to the present disclosure includes the above-described air bio battery.
In addition, the battery (an example of an energy generation device) according to the present disclosure includes a positive electrode that is a first enzyme-immobilized electrode, a negative electrode that is a second enzyme-immobilized electrode, and a substrate of the enzyme immobilized on the negative electrode. Wherein the positive electrode and the electrolyte are in contact with each other, and the negative electrode and the electrolyte are in contact with each other.
The battery according to the present disclosure is the same as the above-described air bio battery, and the preferred embodiment is also the same.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to the following examples unless it exceeds the gist of the present invention.

(Production of gas supply device)
<Preparation of the first enzyme-immobilized membrane>
Glucose oxidase (Sigma-Aldrich, 10 units / cm ² ), pyranose oxidase (Sigma-Aldrich, 2 units / cm ² ), and PVA-SbQ (Toyo Gosei Co., 6.25 mg / cm) ² ) A mixed solution was prepared using a phosphate buffer (pH 7, 50 mmol / l, 3 μl / cm ² ) as a solvent.
The above mixed solution was applied to a dialysis membrane (manufactured by Ieda Trading Co., Ltd.) as a first support, and was comprehensively immobilized by irradiation with ultraviolet rays to produce a first enzyme-immobilized membrane.

<Preparation of second enzyme-immobilized membrane>
Catalase (manufactured by Sigma-Aldrich, 18.3 units / cm ² ) and PVA-SbQ (manufactured by Toyo Gosei Co., Ltd., 6.25 mg / cm ² ), and a phosphate buffer (pH 7, 50 mmol / l, 3 μl / cm ² ) to prepare a solvent mixture.
The above mixed solution was applied to a dialysis membrane (manufactured by Ieda Trading Co., Ltd.) as a second support, and was comprehensively immobilized by irradiation with ultraviolet rays to prepare a second enzyme-immobilized membrane.

<Production of gas supply device>
Using the first and second enzyme-immobilized membranes produced, a gas supply device was produced.
FIG. 4 is a photograph of the gas supply device manufactured in the example.
As shown in FIGS. 4A and 4B, a first enzyme-immobilized membrane 12 applied to one surface of a transparent first support 14 and a transparent second support are provided on an upper case 210 and a lower case 212. 24, and the second enzyme-immobilized membrane 22 applied to one surface of each of them. The first enzyme-immobilized membrane 12 was immobilized using a translucent O-ring 52, and the second enzyme-immobilized membrane 22 was immobilized using a translucent O-ring 54.
Subsequently, as shown in FIG. C, a gas supply device was manufactured by fitting the upper case 210 and the lower case 212 together.
The space between the first enzyme-immobilized membrane 12 and the second enzyme-immobilized membrane 22 was filled with a 5 mmol / l D-glucose aqueous solution 214 (see FIG. 5). The D-glucose aqueous solution 214 corresponds to the specific solution 30 described above.
FIG. 5 is a schematic cross-sectional view showing a mounting direction of the first enzyme-immobilized membrane 12 and the second enzyme-immobilized membrane 22 in the gas supply device shown in FIG.
5, the direction indicated by arrow A is on the upper case side in FIG. 4, and the direction indicated by arrow B is on the lower case side in FIG.
As shown in FIG. 5, in the gas supply device manufactured in the example, the first support 14 exists between the first enzyme-immobilized membrane 12 and the D-glucose aqueous solution 214, and A second support 24 exists between the second enzyme-immobilized membrane 22 and the D-glucose aqueous solution 214.
In the obtained gas supply device, the area where the first support 14 and the D-glucose aqueous solution 214 are in contact is slightly larger than the area where the first oxygen-fixed membrane 12 and the air are in contact with 7.1 cm ² . The area where the second support 24 is in contact with the aqueous D-glucose solution 214 is slightly larger than the area where the second oxygen immobilization membrane 12 is in contact with air, which is 7.1 cm ² .

<Evaluation of gas supply device>
The oxygen supply capability of the obtained gas supply device was measured, and the gas supply capability of the gas supply device was evaluated.
Specifically, a tube was connected to the opening 42 in FIG. 4D, and an oxygen concentration meter (manufactured by Able) was connected to the end of the tube. The inside of the tube connected to the opening 42 and the oximeter was a closed space so that outside air did not enter the tube. The inner diameter of the tube was 1 mm, and the length of the tube was 5 cm.
The oxygen concentration was measured every second, and the measurement was continued for a total of 40 minutes. The measurement environment was room temperature (25 ° C.).
At the time of measurement, pure water was converted into a 5 mmol / L D-glucose solution (pH = 7.0) at 2 minutes after the start of measurement using a gas supply device in which the D-glucose solution was changed to pure water. Was changed.
FIG. 6 shows the measurement results.
The vertical axis of FIG. 6 represents the oxygen concentration (vol%), and the horizontal axis represents the time (minute).
From FIG. 6, it can be seen that, under the present implementation conditions, the oxygen concentration (about) 20.9% (vol%) in the atmosphere increased to about 29% (vol%) in about 20 minutes.
That is, it is understood that the gas supply device in the present embodiment can supply gas using an enzyme reaction. As a modified example of FIG. 5, a configuration in which the first enzyme-immobilized membrane 12 is present between the first support 14 and the D-glucose aqueous solution 214, a second support 24, A configuration in which the second enzyme-immobilized membrane 22 is present between the glucose aqueous solution 214 and the first support 14 is arranged on the arrow A side and / or the second support 24 is arranged on the arrow B side. When they are arranged, the increase in the oxygen concentration is slowed down. Therefore, the mounting direction is preferably as shown in FIG.
The 5 mmol / L D-glucose solution is an amount corresponding to the blood glucose level in the blood of a healthy individual. That is, it can be said that it is suggested that the gas supply device in the present embodiment can supply gas by using blood or the like.

(Production of battery)
<Preparation of positive electrode>
The positive electrode was formed by a method of forming a single-layer self-assembled monolayer on a glassy carbon (glassy carbon, GC) electrode or a platinum (Pt) electrode by a SAM / LbL method.

-Preparation of electrodes-
As the GC electrode, an electrode whose end face was polished using a polishing apparatus (PK-3 Electrode Polishing kit, manufactured by ALS) was used.
The Pt electrode was formed by forming Pt (thickness: 200 nm) on a hydrophobic porous PTFE film (diameter: 0.22 μm, thickness: 150 μm, Millipore) by a sputtering method.
Next, in order to increase the affinity between the surface of the Pt electrode and the solution, a hydrophilic treatment was performed using an atmospheric pressure plasma device (Aiplasma, Panasonic Electric). Finally, parts other than the electrode sensitive part and the terminal part were insulated and coated with polydimethylsiloxane (PDMS).

-Immobilization of enzyme-
The above-mentioned GC electrode includes a PDDA solution, a solution containing BOD (manufactured by Amano Enzyme Co., Ltd.), a solution containing K3 [Fe (CN) ₆ ] (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and a solution containing PSS. Immobilization of the enzyme was performed by sequentially applying.
Further, the Pt electrode coated with the hydrophilic treatment and insulated was immersed in a 2-mercaptoethanesulfonic acid sodium salt (MESNA, manufactured by Tokyo Kasei Kogyo) solution (1 mM, solvent: ethanol) for 1 hour, and the self-organized MESNA was placed on the surface of the Pt electrode. A monomolecular film was formed. Thereafter, the enzyme was immobilized in the same procedure as in the GC electrode.

<Preparation of negative electrode>
The negative electrode was formed on a GC electrode or a Pt electrode by the LbL method.
As the GC electrode or the Pt electrode, the same electrode as that used for the positive electrode was used.

-Immobilization of enzyme-
A PDDA solution, GDH (manufactured by Toyobo Co., Ltd.), Dp (manufactured by Oriental Yeast Co., Ltd.), NADH (manufactured by Oriental Yeast Co., Ltd.) and vitamin The enzyme was immobilized by applying a solution containing K3 (manufactured by Nacalai Tesque, Inc.) and a solution containing PSS in this order.

<Production of battery>
The above-described positive electrode and negative electrode were combined as shown in FIG.
Batteries 3 to 4 were manufactured by combining the gas supply device manufactured in the above-described example and the above-described positive electrode and negative electrode as shown in FIG.
As the electrolyte supply member 118, a mesh made of cotton (diameter: 700 μm, thickness: 100 μm) was used.
In the battery 5, the positive electrode and the negative electrode described above are combined as shown in FIG. 2, and the oxygen concentration in the outside air existing in the positive electrode portion is set to 26.8% using a commercially available air charger (MS-X1, Panasonic). (% By volume).
Table 3 shows the combinations of the positive electrode and the negative electrode used in Battery 1 to Battery 5.
As the electrolyte 130, a 5 mmol / L D-glucose solution was used.
In Table 3, the description of% in parentheses in the column of the gas supply device indicates the concentration (% by volume) of oxygen contained in the outside air existing in the positive electrode portion.

<Measurement of power generation output>
A bio battery was constructed, and the power generation characteristics in a 5 mmol / l glucose solution were examined. After connecting the produced enzyme-immobilized electrode to an electrometer (8240, ADCMT), a variable resistor (0 to 1000 kΩ) is connected between both electrodes. The output when the resistance value of the variable resistor was changed was measured with an electrometer, and the current density and the power density by power generation were calculated from the resistance value and the recorded voltage value, and the power generation characteristics were evaluated.

Table 3 shows the maximum power density obtained by the above method as an index of the power generation output.

FIG. 7 shows the evaluation results of the power generation characteristics of Battery 2 (2) and Battery 4 (4).
In FIG. 7, the vertical axis indicates the power density (μW / cm ² ) and the horizontal axis indicates the voltage (mV), and the maximum value in the vertical axis direction in each graph is the maximum power density.

The results of Battery 1 to Battery 5 indicate that the battery according to the present disclosure has power generation characteristics and can be used as a battery.
The amount of the 5 mmol / L D-glucose solution used as the electrolyte in the battery was equivalent to the blood glucose level in the blood of a healthy individual. In other words, it is suggested that the battery according to the present embodiment also enables power generation by using blood or the like.For example, in a biological implantable electronic device such as a pacemaker, the battery can be replaced semipermanently. No longer needed. Further, power generation by lactic acid or the like in sweat is also conceivable, and it can be said that application to a self-driven wearable health care device for athlete fatigue management and heat stroke monitoring is possible.
From the results of Battery 1 and Battery 2, the power generation output was approximately 2.3 times higher when the Pt electrode was used as the positive electrode and the negative electrode than when the Pt electrode was used as the positive electrode and the negative electrode. You can see that.
Also, from the results of Battery 2 and Battery 4, it can be seen that when the gas supply device according to the present disclosure is used, approximately 1.3 times the power generation output is obtained.

Reference Signs List 10 gas supply device 12 first enzyme-immobilized membrane 14 first support 22 second enzyme-immobilized membrane 24 second support 30 specific solution 40 case 42, 44, 46 opening 48 upper case 52, 52a , 52b, 54, 54a, 54b, 56a, 56b O-ring 100 Air bio battery 110 Positive electrode 112 Enzyme 114 Electrode 116 First electrode support 118 Electrolyte supply member 120 Negative electrode 122 Enzyme 124 Electrode 126 Second electrode support 130 Electrolyte 140 Case 142 Space 200 Battery 210 Upper case 212 Lower case 214 D-glucose aqueous solution A Upper case side B Lower case side

Claims (5)

A first enzyme-immobilized member, a second enzyme-immobilized member, and a substrate of the enzyme immobilized on the first enzyme-immobilized member and the enzyme immobilized on the second enzyme-immobilized member. A solution containing at least one substrate selected from the group consisting of substrates,
The first enzyme-immobilized member and the second enzyme-immobilized member are in contact with the solution,
Releasing gas to the opposite side of the second enzyme immobilization member from the side in contact with the solution,
Outgassing device.   The enzymes immobilized on the first enzyme immobilization member are glucose oxidase and pyranose oxidase, and the enzymes immobilized on the second enzyme immobilization member are catalase, and the substrate contained in the solution. 2. The gas release device according to claim 1, wherein D is glucose and the gas is oxygen. An energy generating device comprising: the gas releasing device according to claim 1 or 2; and an energy generating unit that generates energy by using gas released by the gas releasing device.   The energy generating unit includes a positive electrode that is a first enzyme-immobilized electrode, a negative electrode that is a second enzyme-immobilized electrode, and an electrolyte including a substrate of an enzyme immobilized on the negative electrode. The energy generator according to claim 3, wherein the enzyme immobilized on the negative electrode contacts the electrolyte, and the enzyme immobilized on the negative electrode contacts the electrolyte to generate power. A positive electrode, which is a first enzyme-immobilized electrode,
A negative electrode that is a second enzyme-immobilized electrode;
An electrolyte containing an enzyme substrate immobilized on the negative electrode,
A gas-liquid separator capable of transmitting gas without allowing the electrolyte to leak out,
An energy generating device, wherein an enzyme immobilized on the positive electrode contacts the electrolyte and the gas-liquid separating member, and the negative electrode contacts the electrolyte to generate electric power.