JPS60244281A - Sensor for dissolved hydrogen concentration - Google Patents
Sensor for dissolved hydrogen concentrationInfo
- Publication number
- JPS60244281A JPS60244281A JP59100618A JP10061884A JPS60244281A JP S60244281 A JPS60244281 A JP S60244281A JP 59100618 A JP59100618 A JP 59100618A JP 10061884 A JP10061884 A JP 10061884A JP S60244281 A JPS60244281 A JP S60244281A
- Authority
- JP
- Japan
- Prior art keywords
- dissolved hydrogen
- hydrogen concentration
- sensor
- ion
- dissolved
- 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
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
【発明の詳細な説明】
〔発明の技術分野〕
この発明は、水溶液中の溶存水素濃度を測定する溶存水
素濃度センサに関するものである。DETAILED DESCRIPTION OF THE INVENTION [Technical Field of the Invention] The present invention relates to a dissolved hydrogen concentration sensor that measures dissolved hydrogen concentration in an aqueous solution.
水素をエネルギ源として成育可能な微生物の生育条件を
調べるための微生物の培養過程におい℃微生物培養液中
の溶存水素濃度を測定することは非常に重要であるが、
微生物の培養過程で、直接培養液中の溶存水素濃度を測
定する手段は従来存在していなかった。In the process of culturing microorganisms to investigate the growth conditions for microorganisms that can grow using hydrogen as an energy source, it is very important to measure the dissolved hydrogen concentration in the microorganism culture solution.
Conventionally, there has been no means to directly measure the dissolved hydrogen concentration in the culture solution during the microbial culture process.
本発明は、上記のような問題を解決するためになされた
もので、溶存水素を取り込んで成育する微生物を固定化
微生物として、水素イオン感応性センサのイオン感応部
に装着することにＬす、溶存水素濃度と微生物の代謝産
物の量との相関ケ利用して、溶存水素濃度に対応する微
生物の代謝産物を水素イオン感応性センサに対する信号
とし。The present invention was made in order to solve the above-mentioned problems, and involves attaching microorganisms that take in dissolved hydrogen and grow as immobilized microorganisms to the ion-sensitive part of a hydrogen ion-sensitive sensor. By utilizing the correlation between the dissolved hydrogen concentration and the amount of microbial metabolites, the microbial metabolites corresponding to the dissolved hydrogen concentration are used as signals for the hydrogen ion-sensitive sensor.
養液中の溶存水素濃度を測定することかできる存水素濃
度センサン提供するものである、発明の実施例〕
以下、本発明の一実施例を図に基づいて説明す、。第１
図において、（１）はＰＨ−ＩＳＦ’ＢＴ ）ランジス
タ、１は溶存水素を取り込んで成育する微生物固定化（
３１を表面に装着したＰＨ−ＩＳＦＥＴ ）ランジスタ
、）は両Ｐ）Ｔ −ＩＳＦ’ＥＴ ）ランジスタ（１１
Ｊ２＋の電位を一定二保つための擬似参照電極、［５１
，（６１Ｊ力はリード線−４１はガラスまたはプラスチ
ック等でなるセンサ支を体、である。Embodiment of the Invention Providing a Hydrogen Concentration Sensor Capable of Measuring Dissolved Hydrogen Concentration in Nutrient Solution] Hereinafter, an embodiment of the present invention will be described with reference to the drawings. 1st
In the figure, (1) is a PH-ISF'BT) transistor, and 1 is a microorganism immobilization (PH-ISF'BT) transistor that grows by taking in dissolved hydrogen.
PH-ISF'ET) transistor with 31 mounted on the surface, ) is both P) T-ISF'ET) transistor (11
Pseudo reference electrode for keeping the potential of J2+ constant, [51
, (61J) The lead wire 41 is the sensor support made of glass or plastic.
上記の微生物を固定化した膜（３１は、 Ａｌｃａｌｉ
ｇｅｎｅｓ：ｕｔｒｏｐｈｕｓ ’ｘ自家栄養条件で、
常法に従って、純粋音養した菌体の懸濁液から菌体乞回
収したものを２−寒天溶液に、４Ｘ１０ ｃｅｌｌｓ、
４！の濃度で菌体ｔＷｍしてＰＩ（−ＩＳＦ’ＥＴ ）
ランジスタ（２＋に装着し固定［ヒ膜として形成される
。擬似参照電極４は白金線で構成される。また、上記Ｐ
Ｈ−ＩＳＦＥＴ ）ランジスタ＋１１．＋２１は、通常
の金属酸化物型電界効果型トランジスタの製造法に準拠
して製作さね、それぞれ単独で水素イオンに感応して、
水溶液のＰＨ（水素イオン濃度）を測定することができ
る。The membrane on which the above microorganisms are immobilized (31 is Alcali
genes: Utrophus 'x under autotrophic conditions,
According to a conventional method, the cells collected from a suspension of pure cultured cells were added to a 2-agar solution, 4×10 cells,
4! PI(-ISF'ET) with bacterial cells tWm at a concentration of
The pseudo reference electrode 4 is composed of a platinum wire.
H-ISFET) transistor +11. +21 was manufactured in accordance with the manufacturing method of ordinary metal oxide field effect transistors, and each one was individually sensitive to hydrogen ions.
The pH (hydrogen ion concentration) of an aqueous solution can be measured.
次に、上記のよ）に構成された溶存水素濃度センサの作
用について説明する。溶存水素濃度センサを試料溶液中
に浸すと、試料溶液中に水素があれば固定化した微生物
、Ａｌｃａｌｉｇｅｎｅｓ ｃｕｔｈｒｏｐｈｕａは溶
存水素濃度に応じて、式（１）および（２）で示される
代謝を連続的に行５．。Next, the operation of the dissolved hydrogen concentration sensor configured as above will be explained. When the dissolved hydrogen concentration sensor is immersed in the sample solution, if there is hydrogen in the sample solution, the immobilized microorganism, Alcaligenes cuthrophua, continuously performs the metabolism shown by equations (1) and (2) according to the dissolved hydrogen concentration. 5. .
ＣＯ，＋ 訃７−−「ＨＣＯＯ、−−（２１ＮＡＤＨＮ
ＡＤ
式（１）では、菌体内のヒドロゲナーゼが溶存水素ビ使
ってＮＡＤ ｗ ＮＡＤＨに還元し、式（２）では、溶
存している二酸化炭素Ｙ ＮＡＤ）Ｔ Ｏ） 遣元力を
使ってギ酸にするが、この時、溶液中の水素イオンを消
費するので、式（２）の反応が生じるとＰＨ値はアルカ
リ側に変化する。CO, + Death 7--"HCOO,--(21NADHN
AD In Equation (1), hydrogenase within the bacterial cell uses dissolved hydrogen to reduce it to NAD w NADH, and in Equation (2), hydrogenase reduces it to formic acid using dissolved carbon dioxide Y NAD)TO) However, since hydrogen ions in the solution are consumed at this time, when the reaction of formula (2) occurs, the pH value changes to the alkaline side.
従って、溶存水素濃度に応じて上記Ｑ〕ような代諸が行
われると、 Ａｌｃａｌ ｉｇｅｎｅｓ ｅｕｔｈｒｏ
ｐｈｕｓ ｋ固定化した膜（３）の周辺のＰＴ（値と、
試料溶液自体のＰＴ−Ｔ値との間に差が生じる。夫々の
ＰＴ（値は、第１図に示したＰ）Ｔ −ＩＳＦＥＴ ト
ランジスタ（１）および（２）によってモニタされ、両
者のＰＨ値の差をめることにより、溶存水素濃度ケ測定
することができる。Therefore, if the above procedures are performed according to the dissolved hydrogen concentration, Alcal igenes euthro
PT (value and
A difference occurs between the PT-T value of the sample solution itself and the PT-T value of the sample solution itself. The respective PT (values are monitored by the P)T-ISFET transistors (1) and (2) shown in Figure 1, and the dissolved hydrogen concentration can be measured by calculating the difference between the two PH values. can.
例えば、０．０２Ｍリン酸緩衝液（ＰＨ値７．０）を用
いて、その液中に水素を吹き込んで溶存水素濃度ｆ １
．５ｐｐｍとした場合、本発明による溶存水素濃度セン
サを用いてその変化を測定すると、第２図に示されるよ
うな応答曲線が得られた。For example, using a 0.02M phosphate buffer (PH value 7.0), hydrogen is blown into the solution to increase the dissolved hydrogen concentration f 1
．． When the dissolved hydrogen concentration sensor according to the present invention was used to measure changes in the dissolved hydrogen concentration at 5 ppm, a response curve as shown in FIG. 2 was obtained.
このよ５に構成され、たセンサを用いると、５０ｐｐｍ
までの溶存水素濃度を測定することかできたなお、上記
実施例においては、擬似参照電極として白金線を用いて
いるが、金や銀などの貴金属−あるいは銀・塩化銀電極
などの安全な参照電極ケ用いても良い。・
また、菌体の固定化に寒天を用いたが、に−カラギーナ
ン、あるいは感光性樹脂を用いても良０更にまた、水素
イオン感応性センサは、Ｐ）Ｔ−ＩＳ−ＦＥＴ ）ラン
ジスタに限ることなく、水素イオン感応性ガラス電極を
用いても同様な効果を得ることができる。If a sensor configured like this is used, 50 ppm
In the above example, a platinum wire was used as a pseudo reference electrode, but a safe reference electrode such as a precious metal such as gold or silver or a silver or silver chloride electrode could be used. Electrodes may also be used.・ In addition, although agar was used to immobilize the bacterial cells, carrageenan or photosensitive resin may also be used.Furthermore, the hydrogen ion sensitive sensor is limited to P) T-IS-FET) Transistor Similar effects can also be obtained by using a hydrogen ion-sensitive glass electrode.
以上のように、本発明によれば、溶存水素を取り込んで
成育することができる固定化された微生物と水素イオン
感応センサとを組合せて溶存水素濃度センサを構成した
ので、従来測定することかできなかった、微生物培養液
などの水溶液中の溶存水素濃度を簡易に、かつ実時間で
直接測定できるとい）効果が得られる。As described above, according to the present invention, a dissolved hydrogen concentration sensor is constructed by combining an immobilized microorganism that can take in and grow dissolved hydrogen and a hydrogen ion sensitive sensor, so that conventional measurement is not possible. It is possible to directly measure the dissolved hydrogen concentration in aqueous solutions such as microbial culture fluids easily and in real time.
第１図は本発明による一実施例の溶存水素濃度センサの
構成図、第２図は本発明の溶存水素濃度センサによる測
定応答曲線ケ示すグラフ。
（１１，＋２１・・・ＰＨ−ＩＳＦＥＴ ）ランジスタ
、（３）微生物固定化膜、（４１・・・擬似参照電極、
（６１，（７１・・・リード線、（８）・・・センサ支
持体。
代理人 弁理士 木 村 三 朗
手続補正書（自発）
昭和５９年 ７月３０日
特許庁長官殿
】、事件の表示 特１郭昭５９−１００６１８号２、発
明の名称
溶存水素濃度センサ
３、補正をする者
代表者片山仁へ部
４、代理人
５゜
参、補正の対象
６、補正の内容
（１）明細書の「特許請求の範囲」を別紙のとおシ補正
する。
（２）明細書第２頁第８行の「調べるための微生物の培
養過程において」を「調べるために」と補正する。
（３）次のとおり補正する。
以 上
別紙
特許請求の範囲（補止）
「１．溶存水素を堰り込んで成育する微生物を固定化微
生物として、水素イオン感応性センサのイオン感応部に
装着して構成したことを特徴とする溶存水素濃度センサ
。
２、上記水素イオン感応性センサとして水素イオン感応
性電界効果型トランジスタ（以後ｐ）ｌ−ＩＳＦＥＴと
称する）を用いたことを特徴とする特許請求の範囲第１
項記載の溶存水素濃度センサ。
６、上記固定化微生物に用いられ、溶存水素を取り込ん
で成育する微生物として、
を用φたことを特徴とする特許請求の範囲第１項記載の
溶存水素濃度センサ。」FIG. 1 is a block diagram of a dissolved hydrogen concentration sensor according to an embodiment of the present invention, and FIG. 2 is a graph showing a measurement response curve of the dissolved hydrogen concentration sensor of the present invention. (11, +21...PH-ISFET) transistor, (3) microorganism immobilization membrane, (41... pseudo reference electrode,
(61, (71...Lead wire, (8)...Sensor support body. Agent: Patent attorney Sanro Kimura Procedural amendment (spontaneous) July 30, 1980, Commissioner of the Japan Patent Office) Indication Special No. 1 Kakusho 59-100618 2, Name of the invention Dissolved hydrogen concentration sensor 3, Person making the amendment Representative: Hitoshi Katayama Department 4, Agent 5゜ Reference: Subject of the amendment 6, Contents of the amendment (1) Details (2) Amend "in the process of culturing microorganisms for investigation" to "for investigation" in line 8 of page 2 of the specification. (3 ) The above appended claims (supplementation) ``1. A microorganism that grows by immersing dissolved hydrogen is attached to the ion-sensing part of a hydrogen ion-sensitive sensor as an immobilized microorganism. 2. A dissolved hydrogen concentration sensor characterized by comprising: 2. A hydrogen ion sensitive field effect transistor (hereinafter referred to as p)l-ISFET) is used as the hydrogen ion sensitive sensor. Range 1
Dissolved hydrogen concentration sensor described in section. 6. The dissolved hydrogen concentration sensor according to claim 1, wherein φ is used as the microorganism that is used as the immobilized microorganism and grows by taking in dissolved hydrogen. ”
Claims (1)
物として、水素イオン感応性センサのイオン感応部に装
着して構成したことを特徴とする溶存水素濃度センサ２ ２、上記水素イオン感応性センサとして水素イオン感応
性電界効果型トランジスタ（以後ＰＴ（（ＳＦＷＩ’と
称する）１に：用いたことな特徴とする特許請求の範囲
第１項記載の溶存水素濃度センサ。 ３、上記固定化微生物に用いられ、溶存水素を取り込ん
で成育する微生物として、 Ａｌｃａｌｉｇｅｎｅｓ属 Ｄｅｓｕｌｆｏｖｉｂｒｉｏ 属 Ｄｅｓｕｌｆｏｔｏｍａｃｕｌｕｍ属 Ｍｅｔｈａｎｏｂａｃｔｅｒｉｕｍ属 Ｃ１ｏｓｔｏｒｉｄｉｕｍ属 な用いたことな特徴とする特許請求の範囲第１項記載の
溶存水素濃度センサ。[Scope of Claims] 1. Dissolved hydrogen concentration sensor 2, characterized in that a microorganism that takes in dissolved hydrogen and grows is attached to an ion-sensitive part of a hydrogen ion-sensitive sensor as an immobilized microorganism. 2. The above-mentioned Dissolved hydrogen concentration sensor according to claim 1, characterized in that the hydrogen ion sensitive field effect transistor (hereinafter referred to as PT (hereinafter referred to as SFWI') 1) is used as the hydrogen ion sensitive sensor. 3. 2. The dissolved hydrogen concentration sensor according to claim 1, wherein the microorganisms used as the immobilized microorganisms and which grow by taking in dissolved hydrogen include members of the genus Alcaligenes, genus Desulfovibrio, genus Desulfotomaculum, genus Methanobacterium, and genus C1ostridium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59100618A JPS60244281A (en) | 1984-05-21 | 1984-05-21 | Sensor for dissolved hydrogen concentration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59100618A JPS60244281A (en) | 1984-05-21 | 1984-05-21 | Sensor for dissolved hydrogen concentration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS60244281A true JPS60244281A (en) | 1985-12-04 |
Family
ID=14278823
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59100618A Pending JPS60244281A (en) | 1984-05-21 | 1984-05-21 | Sensor for dissolved hydrogen concentration |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS60244281A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0870823A1 (en) * | 1997-03-07 | 1998-10-14 | Istituto Trentino Di Cultura | System for monitoring the metabolic activity of living cells |
-
1984
- 1984-05-21 JP JP59100618A patent/JPS60244281A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0870823A1 (en) * | 1997-03-07 | 1998-10-14 | Istituto Trentino Di Cultura | System for monitoring the metabolic activity of living cells |
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