JPH0365644A - Enzyme electrode - Google Patents
Enzyme electrodeInfo
- Publication number
- JPH0365644A JPH0365644A JP1201208A JP20120889A JPH0365644A JP H0365644 A JPH0365644 A JP H0365644A JP 1201208 A JP1201208 A JP 1201208A JP 20120889 A JP20120889 A JP 20120889A JP H0365644 A JPH0365644 A JP H0365644A
- Authority
- JP
- Japan
- Prior art keywords
- electrode
- enzyme
- coenzyme
- membrane
- inner membrane
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 61
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 61
- 239000012528 membrane Substances 0.000 claims abstract description 53
- 239000005515 coenzyme Substances 0.000 claims abstract description 51
- 239000000758 substrate Substances 0.000 claims abstract description 14
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 5
- 230000027756 respiratory electron transport chain Effects 0.000 claims description 5
- 230000000295 complement effect Effects 0.000 claims description 4
- 238000009792 diffusion process Methods 0.000 claims description 4
- 230000035945 sensitivity Effects 0.000 abstract description 9
- 238000006243 chemical reaction Methods 0.000 abstract description 4
- 230000015556 catabolic process Effects 0.000 abstract 1
- 238000006731 degradation reaction Methods 0.000 abstract 1
- 230000036632 reaction speed Effects 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 49
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 24
- 239000000243 solution Substances 0.000 description 14
- 238000005259 measurement Methods 0.000 description 9
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 7
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 7
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 7
- 229940098773 bovine serum albumin Drugs 0.000 description 7
- 229950006238 nadide Drugs 0.000 description 7
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 6
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- -1 poly(tetrafluoroethylene) Polymers 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 238000004528 spin coating Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 108010093096 Immobilized Enzymes Proteins 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 238000004401 flow injection analysis Methods 0.000 description 2
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108020005199 Dehydrogenases Proteins 0.000 description 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 1
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 229910021607 Silver chloride Inorganic materials 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000003411 electrode reaction Methods 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 229920002635 polyurethane Polymers 0.000 description 1
- 239000004814 polyurethane Substances 0.000 description 1
- 229910052594 sapphire Inorganic materials 0.000 description 1
- 239000010980 sapphire Substances 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000006276 transfer reaction Methods 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
[産業上の利用分野]
本発明は酵素電極に関し、ざらに詳しくは酵素と補酵素
とを固定化した酵素電極に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to an enzyme electrode, and more particularly to an enzyme electrode in which an enzyme and a coenzyme are immobilized.
[従来の技術]
溶液中の物質の濃度を測定するために、その物質を基質
とする酵素を電極表面に固定化した酵素電極の作製が試
みられている。その原理は、電極を溶液中に浸漬して電
極表面で酵素反応を起こさせ、生成物の量を電気化学的
に検出づることによって溶液中の物質濃度を測定するも
のである。これまでに、グルコースオキシダーゼなどの
酸化酵素についての酵素電極が作製されている。[Prior Art] In order to measure the concentration of a substance in a solution, attempts have been made to produce an enzyme electrode in which an enzyme using the substance as a substrate is immobilized on the electrode surface. The principle is to immerse an electrode in a solution, cause an enzyme reaction to occur on the electrode surface, and measure the concentration of a substance in the solution by electrochemically detecting the amount of product. Enzyme electrodes for oxidizing enzymes such as glucose oxidase have been produced so far.
一方、酸化還元酵素の中には、補酵素を必要とするもの
がある。例えば、工業的に有用なアルコールデヒドロゲ
ナーゼ、乳酸デヒドロゲナーゼなどの脱水素酵素は、補
酵素としてニコチンアミドアデニンジヌクレオチド(以
下、NADで表す〉を必要とする。これらの酵素を酵素
電極に応用する場合、酵素反応によって生成した補酵素
誘導体(例えば、アルコールデヒドロゲナーゼではNA
Dの還元体〉を電極反応によって検出して基質量を求め
る。On the other hand, some oxidoreductases require coenzymes. For example, industrially useful dehydrogenases such as alcohol dehydrogenase and lactate dehydrogenase require nicotinamide adenine dinucleotide (hereinafter referred to as NAD) as a coenzyme. When applying these enzymes to enzyme electrodes, Coenzyme derivatives produced by enzymatic reactions (for example, in alcohol dehydrogenase, NA
The amount of the substrate is determined by detecting the reduced form of D> by an electrode reaction.
従来、このような酵素電極のほとんどは、酵素のみを電
極上に固定化し、補酵素は固定化せずに溶液から供給す
る方法がとられている。例えば、補酵素溶液を用いるフ
ローインジエクションアノリンス法によるエタノール測
定系が報告されている( Analytica Ch
imica へcta 、 214. 161〜17
2(1988) )。また、酵素と同時に補酵素を固定
化した電極についての試みもなされており、カーボン上
に酵素と補酵素溶液を塗布した電極の例がある(特願昭
56−6961@)。Conventionally, most of these enzyme electrodes employ a method in which only the enzyme is immobilized on the electrode, and the coenzyme is supplied from a solution without being immobilized. For example, an ethanol measurement system using a flow injection anorinse method using a coenzyme solution has been reported (Analytica Ch
imica to cta, 214. 161-17
2 (1988)). Attempts have also been made to create an electrode in which a coenzyme is immobilized at the same time as an enzyme, and there is an example of an electrode in which an enzyme and a coenzyme solution are coated on carbon (Japanese Patent Application No. 56-6961@).
[発明が解決しようとする課題]
しかしながら、補酵素を溶液から供給する方法には、補
酵素溶液が不安定で長期間保存することができない、補
酵素溶液が高価であるなどの欠点がある。また、フロー
インジェクション法は、装置が大がかりであり、簡便な
測定法ではない。これらの点から、酵素と同時に補酵素
を固定化した電極が必要となる。しかし、酵素と補酵素
を塗布しただけの電極は、補酵素を多量に固定化できな
いので感度が低く、分子の小さな補酵素の溶液への散逸
が多いので、繰り返し使用に耐えないという問題点があ
る。[Problems to be Solved by the Invention] However, the method of supplying a coenzyme from a solution has drawbacks such as the coenzyme solution being unstable and cannot be stored for a long period of time, and the coenzyme solution being expensive. Furthermore, the flow injection method requires a large-scale apparatus and is not a simple measurement method. From these points, an electrode in which coenzymes are immobilized at the same time as enzymes is required. However, electrodes coated with just enzymes and coenzymes have low sensitivity because they cannot immobilize large amounts of coenzymes, and the coenzymes with small molecules often dissipate into the solution, so they cannot withstand repeated use. be.
本発明は以上述べたような従来の問題点を解決するため
になされたもので、酵素を固定化すると同時に、補酵素
を安定に、かつ多量に固定化した測定感度の優れた酵素
電極を提供することを目的とする。The present invention has been made in order to solve the conventional problems as described above, and provides an enzyme electrode with excellent measurement sensitivity that immobilizes enzymes and at the same time stably immobilizes coenzymes in large quantities. The purpose is to
[課題を解決するための手段]
本発明は、電極と、酵素および該酵素と相補的に使用さ
れる補酵素を含有する膜(内膜)と、前記補酵素の拡散
が遅い膜(外膜〉とが順次基板上に形成されてなること
を特徴とする酵素電極、および電極と、酵素、該酵素と
相補的に使用される補酵素および該補酵素と電子間の電
子授受を媒介する試薬を含有する膜(内膜〉と、前記補
酵素の拡散が遅い膜(外膜〉とが順次基板上に形成され
てなることを特徴とする酵素電極である。[Means for Solving the Problems] The present invention comprises an electrode, a membrane (inner membrane) containing an enzyme and a coenzyme used complementary to the enzyme, and a membrane (outer membrane) in which the coenzyme diffuses slowly. 〉 and 〉 are sequentially formed on a substrate, and an enzyme, an enzyme, a coenzyme used complementary to the enzyme, and a reagent that mediates electron transfer between the coenzyme and electrons. This enzyme electrode is characterized in that a membrane containing the coenzyme (inner membrane) and a membrane in which the coenzyme diffuses slowly (outer membrane) are successively formed on a substrate.
本発明で用いる外膜は、基質の拡散速度に対する補酵素
の拡散速度の比が小さいものが適しており、例えばグル
タルアルデヒドで架橋した牛血清アルブミン膜が挙げら
れる。The outer membrane used in the present invention is suitably one in which the ratio of the coenzyme diffusion rate to the substrate diffusion rate is small, such as a bovine serum albumin membrane crosslinked with glutaraldehyde.
[作用]
本発明においては、補酵素の拡散が遅い外膜で内膜を被
覆することによって、内膜にある補酵素が溶液中へ散逸
することを防ぎ、内膜中の補酵素11度を高濃度に保つ
ことができる。これによって、当該酵素電極の基質に対
する測定感度を向上させるとともに、繰り返し使用に耐
えるようになる。[Function] In the present invention, by coating the inner membrane with an outer membrane in which coenzyme diffusion is slow, the coenzyme in the inner membrane is prevented from dissipating into the solution, and the coenzyme 11 degree in the inner membrane is prevented from dissipating into the solution. Can be kept at high concentration. This improves the measurement sensitivity of the enzyme electrode to the substrate and makes it durable for repeated use.
ざらに、酵素電極の堅牢性も向上する。また、補酵素と
電極の電子移動反応速度が遅い場合、この反応を触媒す
る試薬(メゾイエイタ〉を内膜に加えることにより、測
定感度の低減化を防ぐことができる。Furthermore, the robustness of the enzyme electrode is also improved. Furthermore, when the electron transfer reaction rate between the coenzyme and the electrode is slow, a reduction in measurement sensitivity can be prevented by adding a reagent (mesoieter) that catalyzes this reaction to the inner membrane.
[実施例]
次に、本発明の実施例について図面を参照して詳細に説
明する。[Example] Next, an example of the present invention will be described in detail with reference to the drawings.
実施例1
酵素としてアルコールデヒドロゲナーゼ、補酵素として
NADを用いて本発明に基づく酵素電極を作製した。酵
素電極のベースには、特願昭63−282721@に示
した方法で作製した、サファイア基板上に金を蒸着した
電極を用いた。酵素膜の調製は、リフトオフ法(特願昭
59−209165号参照〉に使用したスピン塗布によ
った。2U/μlアルコールデヒドロゲナーゼ、100
IIIHNADを含んだ15%牛血清アルブミン水溶液
に、グルタルアルデヒドを0.5%になるように加えて
電極上に滴下し、2000 rplnで30秒間スピン
塗布した。室温で架橋反応を進めて、酵素・補酵素固定
化電極を得た。この酵素電極をAとする。Example 1 An enzyme electrode based on the present invention was produced using alcohol dehydrogenase as an enzyme and NAD as a coenzyme. The base of the enzyme electrode was an electrode prepared by the method shown in Japanese Patent Application No. 63-282721@, in which gold was deposited on a sapphire substrate. The enzyme membrane was prepared by spin coating using the lift-off method (see Japanese Patent Application No. 59-209165). 2U/μl alcohol dehydrogenase, 100%
Glutaraldehyde was added to a 15% bovine serum albumin aqueous solution containing IIIHNAD to a concentration of 0.5%, and the mixture was dropped onto the electrode and spin-coated at 2000 rpm for 30 seconds. The crosslinking reaction was carried out at room temperature to obtain an enzyme/coenzyme immobilized electrode. This enzyme electrode is designated as A.
電極Aの表面に外膜を被覆した。外膜の材質としては、
ポリ(テトラフルオロエチレン)(PTFE)製の膜、
ポリウレタン膜、グルタルアルデヒドで架橋した牛血清
アルブミン膜等が使用可能であった。特に、グルタルア
ルデヒドで架橋した牛血清アルブミン膜は、調製が容易
であること、膜の電極への付着性がよいこと、架橋度を
変化させることで透過性を制御できることなどから優れ
ていた。The surface of electrode A was coated with an outer membrane. The material of the adventitia is
membrane made of poly(tetrafluoroethylene) (PTFE),
Polyurethane membranes, bovine serum albumin membranes cross-linked with glutaraldehyde, etc. could be used. In particular, bovine serum albumin membranes crosslinked with glutaraldehyde were excellent because they were easy to prepare, had good adhesion of the membrane to electrodes, and the permeability could be controlled by changing the degree of crosslinking.
15%牛血清アルブミン水溶液にグルタルアルデヒドを
5%になるように加えて電極Aに滴下した後、電極を2
000 rpHlで30秒間スピン塗布して膜厚約11
JMの外膜を被覆した。この酵素電極をBとする。第1
図(a)は本実施例によって得られた酵素と補酵素を固
定化した酵素電極Bの断面図である。図中、1は基板、
2は電極、3は酵素と補酵素を固定化した内膜、4は補
酵素の散逸を防ぐ外膜である。Add 5% glutaraldehyde to a 15% bovine serum albumin aqueous solution and drop it onto electrode A.
Spin coating for 30 seconds at 000 rpm to obtain a film thickness of approximately 11
The adventitia of JM was coated. This enzyme electrode is designated as B. 1st
Figure (a) is a cross-sectional view of enzyme electrode B on which the enzyme and coenzyme obtained in this example are immobilized. In the figure, 1 is the substrate,
2 is an electrode, 3 is an inner membrane on which enzymes and coenzymes are immobilized, and 4 is an outer membrane that prevents dissipation of coenzymes.
上記、A、B電極について、エタノールに対する電流応
答を測定した。測定装置の構成を第2図に示す。図中、
6は酵素電極、7は対極、8は銀・塩化銀参照電極、9
はエタノールを含む緩衝溶液、10はポテンシオスタッ
ト、11は記録計である。The current response to ethanol was measured for the above A and B electrodes. The configuration of the measuring device is shown in Figure 2. In the figure,
6 is an enzyme electrode, 7 is a counter electrode, 8 is a silver/silver chloride reference electrode, 9
1 is a buffer solution containing ethanol, 10 is a potentiostat, and 11 is a recorder.
酵素反応で生成するNADの還元体の酸化に必要な電位
である0、75 VVS、AQ/AgC1を酵素電極6
に印加し、反応溶液にエタノールを加えて得られた電流
の変化を記録した。Enzyme electrode 6
was applied, and the change in current obtained by adding ethanol to the reaction solution was recorded.
第3図は、A、B電極で得られた500 mMエタノー
ルに対する電流応答である。外膜のない電極Aは、補酵
素の散逸によって電流応答が得られないのに対して、外
膜を備えた電極Bは、十分な電流応答を得ることができ
た。FIG. 3 shows the current response to 500 mM ethanol obtained at the A and B electrodes. Electrode A without an outer membrane could not obtain a current response due to dissipation of the coenzyme, whereas electrode B with an outer membrane could obtain a sufficient current response.
第4図は、500 mMエタノールの繰り返し測定に対
する電流応答の減少を示したもので、図中、口は電極A
の場合、士は電極Bの場合である。外膜のない電極Aで
は測定2回目以降の電流応答はほとんどなく、複数回の
測定は不可能であるのに対し、外膜を備えた電極Bは繰
り返し使用することができた。Figure 4 shows the decrease in current response to repeated measurements of 500 mM ethanol, where the mouth is electrode A.
In the case of , 2 is the case of electrode B. With electrode A without an outer membrane, there was almost no current response after the second measurement and multiple measurements were impossible, whereas electrode B with an outer membrane could be used repeatedly.
実施例2
実施例1の酵素電極作製に際して、アルコールデヒドロ
ゲナーゼ、NADと共に、NADの還元体と電極の間の
電子授受を媒介する試薬(メゾイエイタ)を固定化した
。メゾイエイタとしては、フェリシアン化カリウム、メ
ルトラブル−が使用可能であった。ここではフェリシア
ン化カリウムを用いた例を示す。Example 2 In producing the enzyme electrode of Example 1, a reagent (mesoieta) that mediates electron transfer between the reduced form of NAD and the electrode was immobilized along with alcohol dehydrogenase and NAD. As the mesoieta, potassium ferricyanide and melt rub could be used. Here, an example using potassium ferricyanide is shown.
2U/μlアルコールデヒドロゲナーゼ、100m)I
NADとIHmMフェリシアン化カリウムを含んだ15
%牛血清アルブミン水溶液に、グルタルアルデヒドを0
.5%になるように加えて平板金電極上に滴下し、20
00 rpmで30秒間スピン塗布して内膜を得た。こ
の電極に5%グルタルアルデヒドを含む15%牛血清ア
ルブミン水溶液を滴下し、200Orpmで5秒間回転
して膜厚約1脚の外膜を被覆した。メゾイエイタを含有
した酵素電極をCとする。第1図(b)は本実施例によ
って得られた酵素と補酵素とメゾイエイタを固定化した
酵素電極の断面図である。図中1は基板、2は電極、4
は補酵素の散逸を防ぐ外膜、5は酵素と補酵素とメゾイ
エイタを固定化した内膜である。2U/μl alcohol dehydrogenase, 100m)I
15 containing NAD and IHmM potassium ferricyanide
% bovine serum albumin aqueous solution with 0 glutaraldehyde
.. Add it to 5% and drop it onto the flat metal electrode, and add 20%.
The inner membrane was obtained by spin coating at 00 rpm for 30 seconds. A 15% bovine serum albumin aqueous solution containing 5% glutaraldehyde was dropped onto this electrode, and the electrode was rotated at 200 rpm for 5 seconds to coat the outer membrane with a thickness of about one leg. The enzyme electrode containing mesoieta is designated as C. FIG. 1(b) is a cross-sectional view of an enzyme electrode on which the enzyme, coenzyme, and mesoieta obtained in this example were immobilized. In the figure, 1 is the substrate, 2 is the electrode, 4
5 is an outer membrane that prevents the dissipation of coenzymes, and 5 is an inner membrane that immobilizes enzymes, coenzymes, and mesoieta.
第2図の測定装置で電極B、Cのエタノールに対する電
流応答を測定した。0.3Vvs、 A(J/AgCJ
!の電位を設定した各酵素電極の500 mMエタノー
ルに対する電流応答を第5図に示す。同図かられかるよ
うに、NADの還元体と電極の間の電子授受を媒介する
フェリシアン化カリウムが固定化されている電極Cは、
低い電位でもエタノールに対する十分な電流応答が得ら
れ、測定感度がさらに向上したものであった。The current response of electrodes B and C to ethanol was measured using the measuring device shown in FIG. 0.3V vs, A(J/AgCJ
! Figure 5 shows the current response of each enzyme electrode to 500 mM ethanol, with the potential set at . As can be seen from the figure, electrode C has immobilized potassium ferricyanide, which mediates electron transfer between the reduced form of NAD and the electrode.
A sufficient current response to ethanol was obtained even at a low potential, and the measurement sensitivity was further improved.
[発明の効果]
以上説明したように、本発明の酵素電極は補酵素の溶液
への逃げを防ぐ緻密な外膜を設けているので、測定感度
や再使用性の点から優れている。[Effects of the Invention] As explained above, the enzyme electrode of the present invention is provided with a dense outer membrane that prevents coenzymes from escaping into the solution, and is therefore excellent in terms of measurement sensitivity and reusability.
ざらに、この外膜は補酵素とともにメゾイエイタを固定
化することができ、これによってメゾイエイタの効果を
併せ持った酵素電極を提供することができる。In general, this outer membrane can immobilize mesoieta together with coenzymes, thereby making it possible to provide an enzyme electrode that also has the effects of mesoieta.
第1図は本発明による酵素電極の断面図、第2図は酵素
電極を用いてエタノール′a度を測定する装置の構成図
、第3図は本発明の一実施例のエタノールに対する電流
応答を示す特性図、第4図は本発明の酵素電極の再使用
性を従来例による場合と比較して示す図、第5図は本発
明の一実施例のエタノールに対する電流応答を示す特性
図である。
1・・・基板 2・・・電極3・・・
酵素と補酵素を固定化した内膜4・・・外膜
5・・・酵素と補酵素とメゾイエイタを固定化した内膜
6・・・酵素電極 7・・・対極8・・・
参照電極
9・・・緩衝溶液
10・・・ポテンシオスタット
11・・・記録計
代
理
人Fig. 1 is a cross-sectional view of an enzyme electrode according to the present invention, Fig. 2 is a block diagram of an apparatus for measuring ethanol concentration using an enzyme electrode, and Fig. 3 is a current response to ethanol in an embodiment of the present invention. FIG. 4 is a diagram showing the reusability of the enzyme electrode of the present invention in comparison with a conventional example, and FIG. 5 is a characteristic diagram showing the current response to ethanol of an embodiment of the present invention. . 1...Substrate 2...Electrode 3...
Inner membrane 4 with immobilized enzymes and coenzymes...Outer membrane 5...Inner membrane 6 with immobilized enzymes, coenzymes, and mesoieta...Enzyme electrode 7...Counter electrode 8...
Reference electrode 9... Buffer solution 10... Potentiostat 11... Recorder agent
Claims (2)
補酵素を含有する膜(内膜)と、前記補酵素の拡散が遅
い膜(外膜)とが順次基板上に形成されてなることを特
徴とする酵素電極。(1) An electrode, a membrane containing an enzyme and a coenzyme used complementary to the enzyme (inner membrane), and a membrane in which the coenzyme diffuses slowly (outer membrane) are sequentially formed on a substrate. An enzyme electrode characterized by:
素および該補酵素と電子間の電子授受を媒介する試薬を
含有する膜(内膜)と、前記補酵素の拡散が遅い膜(外
膜)とが順次基板上に形成されてなることを特徴とする
酵素電極。(2) an electrode, an enzyme, a coenzyme used complementary to the enzyme, and a membrane (inner membrane) containing a reagent that mediates electron transfer between the coenzyme and the electrons, and the coenzyme having slow diffusion; An enzyme electrode characterized in that a membrane (outer membrane) is sequentially formed on a substrate.
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1201208A JPH0365644A (en) | 1989-08-04 | 1989-08-04 | Enzyme electrode |
US07/559,685 US5196340A (en) | 1989-08-04 | 1990-07-30 | Enzyme electrode containing an enzyme and a coenzyme immobilized in separate layers of a membrane |
DE69023430T DE69023430T2 (en) | 1989-08-04 | 1990-08-03 | Enzyme electrode. |
EP90114973A EP0415124B1 (en) | 1989-08-04 | 1990-08-03 | An enzyme electrode |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1201208A JPH0365644A (en) | 1989-08-04 | 1989-08-04 | Enzyme electrode |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH0365644A true JPH0365644A (en) | 1991-03-20 |
Family
ID=16437144
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP1201208A Pending JPH0365644A (en) | 1989-08-04 | 1989-08-04 | Enzyme electrode |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH0365644A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005013210A (en) * | 2003-06-05 | 2005-01-20 | Sony Corp | Immobilizing carrier and method for producing the same, electrode and method for producing the same, and device for utilizing electrode reaction and method for producing the same |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5816695A (en) * | 1981-07-10 | 1983-01-31 | ジ−・デイ・サ−ル・アンド・コンパニ− | Production of teststerone |
JPS5853747A (en) * | 1981-09-28 | 1983-03-30 | Hitachi Kyowa Kogyo Kk | Automatic analyzer for fluorine in urine |
JPS6257939A (en) * | 1985-09-04 | 1987-03-13 | Kyowa Kikai Seisakusho:Kk | Sliver carriage traverse apparatus of mule spinning ravme |
-
1989
- 1989-08-04 JP JP1201208A patent/JPH0365644A/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5816695A (en) * | 1981-07-10 | 1983-01-31 | ジ−・デイ・サ−ル・アンド・コンパニ− | Production of teststerone |
JPS5853747A (en) * | 1981-09-28 | 1983-03-30 | Hitachi Kyowa Kogyo Kk | Automatic analyzer for fluorine in urine |
JPS6257939A (en) * | 1985-09-04 | 1987-03-13 | Kyowa Kikai Seisakusho:Kk | Sliver carriage traverse apparatus of mule spinning ravme |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005013210A (en) * | 2003-06-05 | 2005-01-20 | Sony Corp | Immobilizing carrier and method for producing the same, electrode and method for producing the same, and device for utilizing electrode reaction and method for producing the same |
US8236153B2 (en) | 2003-06-05 | 2012-08-07 | Sony Corporation | Immobilization support, process for producing the same, electrode, process for producing the same, electrode reaction utilizing apparatus and process for producing the same |
US9365882B2 (en) | 2003-06-05 | 2016-06-14 | Sony Corporation | Immobilization support, process for producing the same, electrode, process for producing the same, electrode reaction utilizing apparatus and process for producing the same |
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