JP2571459B2 - Enzyme-containing paper - Google Patents
Enzyme-containing paperInfo
- Publication number
- JP2571459B2 JP2571459B2 JP2202279A JP20227990A JP2571459B2 JP 2571459 B2 JP2571459 B2 JP 2571459B2 JP 2202279 A JP2202279 A JP 2202279A JP 20227990 A JP20227990 A JP 20227990A JP 2571459 B2 JP2571459 B2 JP 2571459B2
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- paper
- glycerin
- containing paper
- present
- 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.)
- Expired - Fee Related
Links
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は、酵素含有紙に関するものであり、詳しく
は、反応基質を含む液体と接触させて使用する際に、含
有する酵素をコントロールして溶出せしめ、しかも、該
酵素の活性を高度に発揮せしめるように改良された酵素
含有紙に関するものである。Description: TECHNICAL FIELD The present invention relates to an enzyme-containing paper. More specifically, the present invention relates to an enzyme-containing paper which is used in contact with a liquid containing a reaction substrate to control the enzyme contained therein. The present invention relates to an enzyme-containing paper which is eluted and improved so as to exhibit the activity of the enzyme to a high degree.
近年、酵素は、各分野で利用されるようになり、その
使用方法も多岐に亘り、従って、酵素の形態も、液体、
粉末状だけではなく、種々の媒体に固定または付着させ
た状態で実用に供されている。特に、固定化酵素の技術
の進歩は顕著である。In recent years, enzymes have been used in various fields, and the methods of use have been diverse.
It is practically used not only in powder form but also in a state of being fixed or adhered to various media. In particular, advances in immobilized enzyme technology have been remarkable.
しかしながら、固定化酵素の技術は、酵素を如何にし
て媒体内に固定して溶出させないようにするかの点に重
点がおかれており、従って、反応基質を含む液体と接触
させ、必要量の酵素を溶出させるような使用態様には適
用できない。However, the technique of immobilized enzymes focuses on how to immobilize the enzyme in the medium so that it is not eluted, and therefore, is brought into contact with the liquid containing the reaction substrate and the required amount of It cannot be applied to the use mode in which the enzyme is eluted.
すなわち、固定化酵素の技術は、例えば、特開昭59−
2302049号公報に提案されたカタラーゼを有効成分とす
るコーヒーの突然変異原不活性剤のように、コーヒーに
カタラーゼを添加するような使用態様には適用できな
い。That is, the technique of immobilized enzymes is disclosed in, for example,
It cannot be applied to a use mode in which catalase is added to coffee, such as a coffee mutagenic inactivator containing catalase as an active ingredient proposed in 2302049.
一方、上記の公開公報には、カタラーゼの使用態様の
一つとして、フィルターペーパー又はクロスにカタラー
ゼを含浸付着させる方法が開示されている。On the other hand, the above-mentioned publication discloses a method of impregnating and attaching catalase to filter paper or cloth as one of the modes of use of catalase.
しかしながら、フィルターペーパー等に単に含浸附着
しただけでは、酵素が直ちに溶出すると共にその活性を
十分に発揮し得ないという問題がある。However, there is a problem in that simply impregnating the filter paper or the like implies that the enzyme elutes immediately and does not exhibit its activity sufficiently.
本発明は、反応基質を含む液体と接触させて使用する
際に、含有する酵素をコントロールして溶出せしめ、し
かも、該酵素の活性を高度に発揮せしめるように改良さ
れた酵素含有紙の提供を目的とするものである。The present invention provides an enzyme-containing paper improved so that when used in contact with a liquid containing a reaction substrate, the enzyme contained therein can be controlled and eluted, and the enzyme can exhibit its activity to a high degree. It is the purpose.
本発明者等は、上記目的を達成するために鋭意検討を
重ねた結果、酵素を特定の状態で紙に含有させるなら
ば、上記の目的を容易に達成し得るとの知見を得、本発
明の完成に至った。The present inventors have conducted intensive studies to achieve the above object, and as a result, obtained a finding that the above object can be easily achieved if the enzyme is contained in paper in a specific state. Was completed.
すなわち、本発明の要旨は、酵素をグリセリンとの混
合状態で含有することを特徴とする酵素含有紙に存す
る。That is, the gist of the present invention resides in an enzyme-containing paper containing an enzyme in a mixed state with glycerin.
以下、本発明を詳細に説明する。 Hereinafter, the present invention will be described in detail.
本発明の酵素含有紙は、例えば、グリセリン水溶液に
酵素を溶解させ、得られた水溶液を紙に含浸または塗布
し、次いで、乾燥する方法により得ることができる。The enzyme-containing paper of the present invention can be obtained, for example, by dissolving an enzyme in an aqueous glycerin solution, impregnating or applying the obtained aqueous solution to paper, and then drying the paper.
本発明においては、酵素は、特に制限されず如何なる
酵素をも使用し得るが、上記方法により酵素含有紙を得
る場合には、グリセリン水溶液に可溶な酵素が用いら
れ、このような酵素の具体例としては、ペロオキシダー
ゼ、β−ガラクトシダーゼ、リパーゼ、インベルター
ゼ、カタラーゼ、アスコルビン酸オキシダーゼ、アミラ
ーゼ、グリコアミラーゼ、トリプシン、キモトリプシ
ン、ペプシン、パパイン、パンクレアチン、アミノアシ
ラーゼ、ヌクレアーゼ、リボヌクレアーゼ、ATPデアミ
ナーゼ、ホスファターゼ、ストレプトキナーゼ、アピラ
ーゼ（ATP−ジキスファターゼ）、ATPクレアチンリン酸
転移酵素、ペクチナーゼ、マルターゼ、ラクターゼ、ウ
レアーゼ、クリコースイソメラーゼ、メリビアーゼ、ア
ルドラーゼ、セルラーゼ、アントシナーゼ、ナリンジナ
ーゼ、グリコースオキシダーゼ、アスパラキシダーゼ等
を挙げることができる。In the present invention, the enzyme is not particularly limited, and any enzyme can be used.When the enzyme-containing paper is obtained by the above method, an enzyme soluble in an aqueous glycerin solution is used. Examples include peroxidase, β-galactosidase, lipase, invertase, catalase, ascorbate oxidase, amylase, glycoamylase, trypsin, chymotrypsin, pepsin, papain, pancreatin, aminoacylase, nuclease, ribonuclease, ATP deaminase, phosphatase, phosphatase, streptase. Kinase, apyrase (ATP-dixphatase), ATP creatine phosphotransferase, pectinase, maltase, lactase, urease, glycose isomerase, melibiase, aldolase, cellulase, annase Tyrosinase, mention may be made of Narinjinaze, glyceryl course oxidase, the asparagus Kishida over zero, or the like.
一方、グリセンは、通常の市販品を使用することがで
き、また、紙は、天然パルプ又は合成パルプより得られ
る各種の紙あるいは紙製品（フィルターペーパー又はク
ロス等）を使用することができる。特に、本発明におけ
るグリセリンの効果は、乾燥によってパルプが収縮する
ような紙などを使用した場合に顕著である。On the other hand, for glycene, a normal commercial product can be used, and for paper, various papers or paper products (such as filter paper or cloth) obtained from natural pulp or synthetic pulp can be used. In particular, the effect of glycerin in the present invention is remarkable when paper or the like whose pulp shrinks upon drying is used.
本発明の酵素含有紙は、酵素を溶解したグリセリン水
溶液を含浸または塗布させた後、乾燥することより得ら
れる。乾燥温度は、酵素の種類より耐熱性が異なるた
め、その上限温度は異なるが、一般的には、通風乾燥機
を用いて20〜50℃とするのがよい。必要ならば、凍結乾
燥を行うこともできる。The enzyme-containing paper of the present invention is obtained by impregnating or applying an aqueous solution of glycerin in which an enzyme is dissolved, and then drying. The drying temperature has a different heat resistance depending on the type of the enzyme, and thus the upper limit temperature is different. However, in general, the drying temperature is preferably set to 20 to 50 ° C. using a ventilation dryer. If necessary, freeze-drying can be performed.
そして、本発明の酵素含有紙においては、酵素の濃度
は、酵素含有紙の使用目的に応じて任意に選択される
が、グリセリンの濃度は、これが余りにも低い場合は酵
素の高活性維持が期待できず、一方、余りにも高い場合
は取扱い上不便であるので、通常は１〜30重量％の範囲
から酵素の溶出速度等を勘案して適宜決定される。In the enzyme-containing paper of the present invention, the concentration of the enzyme is arbitrarily selected according to the purpose of use of the enzyme-containing paper. However, if the concentration of glycerin is too low, high enzyme activity is expected to be maintained. On the other hand, if it is too high, it is inconvenient to handle. Therefore, it is usually determined appropriately from the range of 1 to 30% by weight in consideration of the enzyme elution rate and the like.
なお、酵素濃度は、酵素含有紙の使用目的に応じ、予
め、単位紙当りの必要な酵素量を求め、これにより、紙
に含浸、塗布される酵素・グリセリン水溶液中の酵素量
を調整することにより、容易にコントロールすることが
できる。The enzyme concentration is determined in advance according to the intended use of the enzyme-containing paper by determining the required amount of enzyme per unit paper, and thereby adjusting the amount of enzyme in the aqueous solution of the enzyme / glycerin to be impregnated and applied to the paper. Can be easily controlled.
本発明の酵素含有紙において、酵素をグリセリンとの
混合状態で紙に含有させたことにより、酵素の高い活性
が維持される理由は、必ずしも明らかにされていない
が、次のように推定される。In the enzyme-containing paper of the present invention, the reason why the high activity of the enzyme is maintained by incorporating the enzyme into the paper in a mixed state with glycerin is not necessarily elucidated, but is presumed as follows. .
すなわち、酵素は、数万から数十分の分子量を有する
高分子量の蛋白質であり、フィルターペーパー等に単に
含浸附着した場合は、乾燥時におけるペーパー（セルロ
ース）の収縮に伴い酵素が一緒に収縮して変形するが、
グリセリンの共存状態においては、セルロースの収縮が
防止され、その結果、酵素の上記変形も阻止されて酵素
の基質特異性が維持されることによるものと推定され
る。That is, the enzyme is a high molecular weight protein having a molecular weight of tens of thousands to tens of minutes, and when simply impregnated on filter paper or the like, the enzyme shrinks together with the shrinkage of the paper (cellulose) during drying. Deform
It is presumed that, in the presence of glycerin, the contraction of cellulose is prevented, and as a result, the above-mentioned deformation of the enzyme is prevented, and the substrate specificity of the enzyme is maintained.
また、本発明の酵素含有紙においては、酵素は、グリ
セリン濃度に依存するものの容易に溶出されるが、その
理由は、酵素のグリセリンに対する親和力が大きく、そ
のために、酵素のセルロースに対する附着力が低下する
と同時に、グリセリンが紙の乾燥による収縮を防止して
酵素の溶出を容易にするためと推定される。In the enzyme-containing paper of the present invention, the enzyme is easily eluted although it depends on the glycerin concentration, because the enzyme has a large affinity for glycerin, and therefore, the adhesion of the enzyme to cellulose decreases. At the same time, it is presumed that glycerin prevents shrinkage due to drying of the paper and facilitates elution of the enzyme.
本発明の酵素含有紙は、コーヒーや紅茶等のフィルタ
ーペーパーやバッグ等の材料に使用し、コーヒー中の突
然変異原の不活性や有害物質（過酸化水素）の除去に使
用する他、酵素反応を利用した試験紙、防臭用包装紙な
どの各種の分野に応用可能である。The enzyme-containing paper of the present invention is used for materials such as filter papers and bags for coffee and tea, and is used for inactivating mutagen in coffee and removing harmful substances (hydrogen peroxide). It can be applied to various fields such as test papers and odor-proof wrapping papers that use the same.
以下、本発明を実施例により更に詳細に説明するが、
本発明は、その要旨を超えない限り、以下の実施例に限
定されるものではない。Hereinafter, the present invention will be described in more detail with reference to Examples.
The present invention is not limited to the following examples unless it exceeds the gist.
実施例１ 10重量％濃度のグリセリン水溶液99gに酵素活性162u/
mgのペロオキシダーゼ（天野製薬製）100mgを添加して
溶解させた。Example 1 An enzyme activity of 162 u /
100 mg of peroxidase (manufactured by Amano Pharmaceutical) was added and dissolved.
直径15cmの濾紙（東洋製、No.2濾紙）を上記の酵素溶
液に浸漬した後、油圧プレス機により脱水した。使用し
た濾紙の酵素溶液含浸前後の重量差から、保持酵素液量
を求めたところ、3.3gであった。A filter paper having a diameter of 15 cm (No. 2 filter paper, manufactured by Toyo) was immersed in the above enzyme solution, and then dehydrated with a hydraulic press. The amount of the retained enzyme solution was determined from the difference in weight of the used filter paper before and after the impregnation with the enzyme solution, and was 3.3 g.
次いで、プレス処理した濾紙を通風乾燥機にて30℃で
５時間乾燥して酵素含有紙を得た。これを試料１とす
る。Next, the pressed filter paper was dried at 30 ° C. for 5 hours with a ventilation dryer to obtain an enzyme-containing paper. This is designated as Sample 1.
上記操作において、10重量％濃度のグリセリン水溶液
の代りに、２重量％濃度のグリセリン水溶液と蒸留水を
それぞれ使用した他は、同様の操作により、酵素含有紙
を得た。それぞれの酵素含有紙を試料２（２重量％濃度
のグリセリン使用）、試料３（蒸留水使用）とする。In the above operation, an enzyme-containing paper was obtained by the same operation except that a 2% by weight aqueous glycerin solution and distilled water were used instead of the 10% by weight aqueous glycerin solution. Each of the enzyme-containing papers is designated as Sample 2 (using 2% by weight of glycerin) and Sample 3 (using distilled water).
なお、各試料の乾燥後の重量は、いずれも、3.2gであ
った。The weight of each sample after drying was 3.2 g.
上記の各試料から、0.05gの試験片を切取り、それぞ
れ、0.1Mのリン酸緩衝液10mlと共に50mlの三角フラスコ
に入れて振盪させ、振盪時間と溶出液の酵素活性の関係
を求めた。From each of the above samples, a test piece of 0.05 g was cut out, placed in a 50 ml Erlenmeyer flask together with 10 ml of a 0.1 M phosphate buffer and shaken, and the relationship between the shaking time and the enzyme activity of the eluate was determined.
結果を表−１に示す。 The results are shown in Table 1.
なお、酵素活性の測定は、次の方法により行った。 In addition, the measurement of the enzyme activity was performed by the following method.
＜酵素活性の測定方法＞ 50mlのネスラー管に0.1Mのリン酸緩衝液（pH6.0）2m
l、0.3％の過酸化水素溶液2mlを加えて、20℃で５分間
保持した後、0.667％のPyrogallorを15ml加えてよく混
和し、５分間静値する。<Method for measuring enzyme activity> 2 m of 0.1 M phosphate buffer (pH 6.0) in a 50 ml Nessler tube
1) Add 2 ml of 0.3% hydrogen peroxide solution, keep at 20 ° C. for 5 minutes, add 15 ml of 0.667% Pyrogallor, mix well, and let stand for 5 minutes.
次いで、試料を加えて振盪反応を行なう。20秒後に2N
の硫酸2mlを添加して混和し、15分〜30分放置する。Next, a sample is added and a shaking reaction is performed. 2N after 20 seconds
Add 2 ml of sulfuric acid and mix for 15-30 minutes.
次いで、ジエチルエーテル15mlを加えて振盪抽出を行
ない、上層を分取する。Subsequently, 15 ml of diethyl ether is added, and the mixture is extracted by shaking, and the upper layer is separated.
上記の抽出操作を５〜６回くり返して抽出液を100ml
にメスアップする。この液について420nmの波長での吸
光度（As）を測定する。Repeat the above extraction procedure 5-6 times to make 100 ml of extract
To mess up. The absorbance (As) of this solution at a wavelength of 420 nm is measured.
盲検として、試料の添加時期を2Nの硫酸の添加後にす
る以外は、同様の方法で吸光度（Ab）を測定する。As a blind test, the absorbance (Ab) is measured in the same manner except that the sample is added after the addition of 2N sulfuric acid.
上記条件下で20秒間に1mgのPurprogallinを生成する
のに要する酵素量を１単位とし、得られた吸光度から、
下記の式により試料の酵素活性を求める。Under the above conditions, the amount of enzyme required to produce 1 mg of Purprogallin in 20 seconds was defined as one unit, and from the obtained absorbance,
The enzyme activity of the sample is determined by the following equation.
酵素活性＝（As−Ab）×8.5×ｎ 但し、ｎは希釈倍率を表わす。Enzyme activity = (As-Ab) × 8.5 × n, where n represents a dilution factor.
〔発明の効果〕 以上説明した発明によれば、酵素をコントロールして
溶出せしめ、しかも、該酵素の活性を高度に発揮せしめ
るように改良された酵素含有紙が提供され、本発明の酵
素含有紙は、酵素を溶出して利用する各種分野において
効果的に応用し得る。 [Effects of the Invention] According to the invention described above, an enzyme-containing paper is provided which is improved so as to control and elute the enzyme, and to exhibit the activity of the enzyme to a high degree. Can be effectively applied in various fields where enzymes are eluted and used.
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭49−133605（ＪＰ，Ａ) 特開 昭63−160585（ＪＰ，Ａ) 特開 昭53−44687（ＪＰ，Ａ) 特開 昭61−6372（ＪＰ，Ａ) 特開 平２−23892（ＪＰ，Ａ) ──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-49-133605 (JP, A) JP-A-63-160585 (JP, A) JP-A-53-44687 (JP, A) 6372 (JP, A) JP-A-2-23892 (JP, A)
Claims (1)
ことを特徴とする酵素含有紙。1. An enzyme-containing paper containing an enzyme in a mixed state with glycerin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2202279A JP2571459B2 (en) | 1990-07-30 | 1990-07-30 | Enzyme-containing paper |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2202279A JP2571459B2 (en) | 1990-07-30 | 1990-07-30 | Enzyme-containing paper |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0491293A JPH0491293A (en) | 1992-03-24 |
| JP2571459B2 true JP2571459B2 (en) | 1997-01-16 |
Family
ID=16454909
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2202279A Expired - Fee Related JP2571459B2 (en) | 1990-07-30 | 1990-07-30 | Enzyme-containing paper |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2571459B2 (en) |
-
1990
- 1990-07-30 JP JP2202279A patent/JP2571459B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0491293A (en) | 1992-03-24 |
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