JP2002262865A - Catalase-occluding microcapsule and method for producing the same - Google Patents
Catalase-occluding microcapsule and method for producing the sameInfo
- Publication number
- JP2002262865A JP2002262865A JP2001064449A JP2001064449A JP2002262865A JP 2002262865 A JP2002262865 A JP 2002262865A JP 2001064449 A JP2001064449 A JP 2001064449A JP 2001064449 A JP2001064449 A JP 2001064449A JP 2002262865 A JP2002262865 A JP 2002262865A
- Authority
- JP
- Japan
- Prior art keywords
- catalase
- aqueous solution
- divinylbenzene
- hydrogen peroxide
- occluding
- 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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- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Manufacturing Of Micro-Capsules (AREA)
- Polymerisation Methods In General (AREA)
- Graft Or Block Polymers (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明はカタラーゼ包括マイ
クロカプセル及びその製造方法、ならびに該カタラーゼ
包括マイクロカプセルを用いて過酸化水素を分解する方
法に関する。The present invention relates to a microcapsule encapsulating catalase and a method for producing the same, and a method for decomposing hydrogen peroxide using the microcapsule encapsulating catalase.
【０００２】[0002]
【従来の技術】カタラーゼは過酸化水素を水と酸素に分
解する酵素であるが、カタラーゼを担体に固定化しない
でそのままで使用した場合には熱や高濃度の過酸化水素
の酸化力等の外的要因により酵素活性が損失したり、酵
素の再使用が困難であるとの問題がある。2. Description of the Related Art Catalase is an enzyme that decomposes hydrogen peroxide into water and oxygen. There are problems that the enzyme activity is lost due to external factors, and that it is difficult to reuse the enzyme.
【０００３】[0003]
【発明が解決しようとする課題】本発明の目的は上記の
従来技術の欠点を克服し、外的要因に対して酵素活性が
安定であり容易に再使用ができる固定化カタラーゼを提
供することにある。SUMMARY OF THE INVENTION An object of the present invention is to provide an immobilized catalase which overcomes the above-mentioned disadvantages of the prior art and has a stable enzyme activity against external factors and can be easily reused. is there.
【０００４】[0004]
【発明を解決するための手段】前記課題を解決するため
発明者らが鋭意検討した結果、カタラーゼを含有する水
溶液からなる水相と、ポリジビニルベンゼン、ポリスチ
レン、又はジビニルベンゼンとスチレンの共重合体から
なり該水相を包括する有機相とからなるカタラーゼ包括
マイクロカプセルが本発明の目的を達成できることを見
いだし本発明に到達した。即ち本発明は、（１）カタラ
ーゼを含有する水溶液からなる水相と、ポリジビニルベ
ンゼン、ポリスチレン、又はジビニルベンゼンとスチレ
ンの共重合体からなり且つ該水相を包括する有機相とか
らなるカタラーゼ包括マイクロカプセル、（２）カタラ
ーゼを含有する水溶液と、ジビニルベンゼン及び／又は
スチレンからなるモノマー、界面活性剤及び重合開始剤
を含有する有機溶媒とを混合して一次乳化液を得た後、
該一次乳化液をアラビアゴム水溶液に分散させ二次乳化
液を得、次いで該二次乳化液を窒素雰囲気下に保ちなが
らモノマーの重合を行うカタラーゼ包括マイクロカプセ
ルの製造方法、（３）水溶液において（１）記載のカタ
ラーゼ包括マイクロカプセルと過酸化水素とを接触させ
る分解方法に関する。以下に本発明を詳細に説明する。Means for Solving the Problems As a result of intensive studies by the present inventors to solve the above-mentioned problems, an aqueous phase comprising an aqueous solution containing catalase and polydivinylbenzene, polystyrene, or a copolymer of divinylbenzene and styrene is used. The present inventors have found that a catalase-encapsulated microcapsule comprising an organic phase comprising the aqueous phase can achieve the object of the present invention, and reached the present invention. That is, the present invention provides (1) a catalase comprising an aqueous phase comprising an aqueous solution containing catalase and an organic phase comprising polydivinylbenzene, polystyrene, or a copolymer of divinylbenzene and styrene and including the aqueous phase. Microcapsules, (2) an aqueous solution containing catalase, and a monomer composed of divinylbenzene and / or styrene, a surfactant and an organic solvent containing a polymerization initiator are mixed to obtain a primary emulsion,
A method for producing a catalase-encapsulated microcapsule in which the primary emulsion is dispersed in an aqueous gum arabic solution to obtain a secondary emulsion, and then the monomer is polymerized while keeping the secondary emulsion under a nitrogen atmosphere; The present invention relates to a method for decomposing catalase-encapsulated microcapsules and hydrogen peroxide according to 1). Hereinafter, the present invention will be described in detail.
【０００５】[0005]
【発明の実施の形態】本発明により得られるカタラーゼ
包括マイクロカプセルは、表面が滑らかな平均粒子径１
３０〜１５０μｍの球状の単殻カプセルである。本発明
のカタラーゼ包括マイクロカプセルの製造方法は、
（１）一次乳化液の調製、（２）一次乳化液からの二次
乳化液の調製、及び（３）二次乳化液中のモノマーの重
合の３工程から成る。BEST MODE FOR CARRYING OUT THE INVENTION The catalase-encapsulated microcapsules obtained by the present invention have a smooth surface with an average particle diameter of 1
It is a spherical single-shell capsule of 30 to 150 μm. The method for producing the catalase-encapsulated microcapsules of the present invention comprises:
It comprises three steps: (1) preparation of a primary emulsion, (2) preparation of a secondary emulsion from the primary emulsion, and (3) polymerization of monomers in the secondary emulsion.
【０００６】（１）一次乳化液の調製 カタラーゼの水溶液を、ジビニルベンゼン及び／又はス
チレンからなるモノマー、界面活性剤及び重合開始剤を
含有する有機溶媒に、ホモジナイザーで攪拌速度１０，
０００〜２０，０００ｒｐｍで攪拌しながら徐々に加
え、加え終わってから５分間以上攪拌を続けて、一次乳
化液を調製する。この時使用するカタラーゼは、動物、
植物、微生物等の由来にこだわらないが、６０℃〜７５
℃で酵素活性を有する必要がある。カタラーゼは、市販
のカタラーゼを用いることも出来、例えば三菱ガス化学
（株）製アスクスーパーを使用することが出来る。界面
活性剤としてはＳｐａｎ８０及びＳｐａｎ６０が好適に
使用される。重合開始剤としては２，２’−アゾビス
（２，４−ジメチルバレロニトリル）が好適に使用され
る。有機溶媒としてはトルエン及びベンゼンが好適に使
用される。モノマー等を含有する有機溶媒中の各成分の
濃度は、モノマー６０〜８０重量％、重合開始剤４〜５
重量％、界面活性剤３〜７重量％、有機溶媒１０〜３０
重量％である。(1) Preparation of Primary Emulsion An aqueous solution of catalase is mixed with an organic solvent containing a monomer composed of divinylbenzene and / or styrene, a surfactant and a polymerization initiator with a homogenizer at a stirring speed of 10,
Add slowly while stirring at 2,000 to 20,000 rpm, and continue stirring for 5 minutes or more after the addition is completed to prepare a primary emulsion. Catalase used at this time is animal,
It does not care about the origin of plants, microorganisms, etc.
It must have enzyme activity at ℃. As the catalase, a commercially available catalase can also be used. For example, Ask Super manufactured by Mitsubishi Gas Chemical Company, Ltd. can be used. As the surfactant, Span80 and Span60 are preferably used. As the polymerization initiator, 2,2′-azobis (2,4-dimethylvaleronitrile) is preferably used. As the organic solvent, toluene and benzene are preferably used. The concentration of each component in the organic solvent containing the monomer and the like is such that the monomer is 60 to 80% by weight, the polymerization initiator 4 to 5
Weight%, surfactant 3-7 weight%, organic solvent 10-30
% By weight.
【０００７】（２）二次乳化液の調製 次に一次乳化液を、分散安定剤であるアラビアゴムと水
との混合物に、攪拌しながら注ぎいれる。攪拌速度は３
００〜５００ｒｐｍが好適である。アラビアゴムと水と
の混合比は１：９９〜５：９５で、一次乳化液に対し１
５倍〜２５倍量のアラビアゴムと水との混合物を加え、
二次乳化液を調製する。(2) Preparation of Secondary Emulsion Next, the primary emulsion is poured into a mixture of gum arabic, which is a dispersion stabilizer, and water with stirring. The stirring speed is 3
00-500 rpm is suitable. The mixing ratio of gum arabic and water is 1:99 to 5:95, and is 1 to primary emulsion.
Add a mixture of 5 to 25 times the amount of gum arabic and water,
Prepare a secondary emulsion.
【０００８】（３）モノマーの重合 次いで、攪拌と窒素置換を行いながら、二次乳化液を６
０℃〜７５℃に加温し、１〜４時間重合を行う。重合終
了後水洗を行い、カタラーゼ包括マイクロカプセルが得
られる。(3) Polymerization of Monomer Next, while stirring and purging with nitrogen, the secondary emulsified liquid
Heat to 0 ° C to 75 ° C and carry out polymerization for 1 to 4 hours. After the completion of the polymerization, washing is performed to obtain catalase-encapsulated microcapsules.
【０００９】カタラーゼ包括マイクロカプセルは、過酸
化水素水溶液や半導体工場排水等の過酸化水素含有排水
に投入するか、或いはカラムに充填し過酸化水素水溶液
または過酸化水素含有排水を通液することで、過酸化水
素を分解する。カタラーゼ包括マイクロカプセルを使用
する際の過酸化水素水溶液の温度は１℃〜８０℃、ｐＨ
は２〜１２が好ましい。The catalase-encapsulated microcapsules are charged into a hydrogen peroxide aqueous solution or into a wastewater containing hydrogen peroxide such as semiconductor factory wastewater, or filled in a column and passed through the aqueous hydrogen peroxide solution or the wastewater containing hydrogen peroxide. Decomposes hydrogen peroxide. The temperature of the aqueous hydrogen peroxide solution when using catalase-encapsulated microcapsules is 1 ° C to 80 ° C, pH
Is preferably 2 to 12.
【００１０】[0010]
【実施例】以下実施例によって本発明をさらに詳細に記
述するが、下記実施例は本発明を制限するものではな
い。 実施例１ カタラーゼ（三菱ガス化学（株）製アスクスーパー）２
０ｇを、ジビニルベンゼン１４ｇ、２，２’−アゾビス
（２，４−ジメチルバレロニトリル）１ｇ、Ｓｐａｎ８
０を１ｇ、トルエン４ｇの混合液に攪拌速度１７，００
０ｒｐｍで攪拌しつつ徐々に添加した。添加終了後、更
に５分間攪拌し一次乳化液を得た。次に、得られた一次
乳化液をアラビアゴム２２．８ｇと水７３７．２ｇの混
合液に、攪拌速度３００ｒｐｍで攪拌しつつ徐々に添加
した。添加終了後、窒素置換を行いながら、攪拌速度３
００ｒｐｍで攪拌しつつ、６５℃で１時間３０分間ジビ
ニルベンゼンの重合反応を行った。反応終了後カプセル
を水で洗浄し、減圧ろ過し、平均粒径１３０〜１５０μ
ｍのカタラーゼ包括マイクロカプセル１０ｇを得た。The present invention will be described in more detail with reference to the following Examples, which do not limit the present invention. Example 1 Catalase (Ask Super manufactured by Mitsubishi Gas Chemical Co., Ltd.) 2
0 g, divinylbenzene 14 g, 2,2′-azobis (2,4-dimethylvaleronitrile) 1 g, Span8
Was added to a mixed solution of 1 g of toluene and 4 g of toluene at a stirring speed of 17,00.
The solution was gradually added while stirring at 0 rpm. After completion of the addition, the mixture was further stirred for 5 minutes to obtain a primary emulsion. Next, the obtained primary emulsion was gradually added to a mixture of 22.8 g of gum arabic and 737.2 g of water while stirring at a stirring speed of 300 rpm. After completion of the addition, the stirring speed is set to 3 while purging with nitrogen.
The polymerization reaction of divinylbenzene was carried out at 65 ° C. for 1 hour and 30 minutes while stirring at 00 rpm. After completion of the reaction, the capsule was washed with water, filtered under reduced pressure, and the average particle size was 130 to 150 μm.
Thus, 10 g of catalase-encapsulated microcapsules of m were obtained.
【００１１】実施例２ ０．０５Ｍ、ｐＨ７のりん酸緩衝液を用いて０．０５ｇ
／ｍｌのカタラーゼ包括マイクロカプセル懸濁液を調製
した。これを２５℃及び６５℃で３０分間保温し、その
後それぞれの２５℃でのカタラーゼ活性を測定し、２５
℃保温カタラーゼの活性を１００％とした場合の６５℃
保温カタラーゼの相対活性を調べた。比較として、遊離
のカタラーゼの水溶液を２５℃及び６５℃で３０分間保
温し、その後それぞれの２５℃でのカタラーゼ活性を測
定し、２５℃保温カタラーゼの活性を１００％とした場
合の６５℃保温カタラーゼの相対活性を調べた。結果を
表１に示した。Example 2 0.05 g of a phosphate buffer of 0.05 M, pH 7 was used.
/ Ml catalase-encapsulated microcapsule suspension was prepared. This was kept at 25 ° C. and 65 ° C. for 30 minutes, and then the catalase activity at each 25 ° C. was measured.
65 ° C assuming 100% activity of heat-protected catalase
The relative activity of warm catalase was examined. For comparison, an aqueous solution of free catalase was kept at 25 ° C. and 65 ° C. for 30 minutes, and then the catalase activity at each 25 ° C. was measured. Was examined for relative activity. The results are shown in Table 1.
【００１２】 表１ ６５℃でのカタラーゼ相対活性 カタラーゼ包括マイクロカプセル １００％ 遊離カタラーゼ ８６％ Table 1 Catalase relative activity at 65 ° C Catalase inclusive microcapsules 100% Free catalase 86%
【００１３】[0013]
【発明の効果】本発明のカタラーゼ包括マイクロカプセ
ルは、外的要因に対して酵素活性が安定である。The catalase-encapsulated microcapsules of the present invention have a stable enzyme activity against external factors.
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ テーマコート゛(参考） Ｃ１２Ｎ 9/08 Ｃ１２Ｎ 9/96 9/96 Ｂ０１Ｊ 13/02 Ｂ Ｆターム(参考） 4B033 NA02 NA23 NB34 NB62 NC06 ND04 ND08 4B050 CC07 HH02 JJ06 KK20 LL10 4G005 AA01 AB14 BA02 BB06 BB08 BB12 BB17 DB16Y DB16Z DB25X DC09W DD15Y DD15Z DE02Y DE02Z EA06 4J011 JA08 JB08 JB26 JB29 PA55 PB35 PC07 4J026 AA07 BA05 BA07 BB01 BB03 DB03 DB08 DB12 DB22 ──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. 7 Identification symbol FI Theme coat ゛ (Reference) C12N 9/08 C12N 9/96 9/96 B01J 13/02 BF Term (Reference) 4B033 NA02 NA23 NB34 NB62 NC06 ND04 ND08 4B050 CC07 HH02 JJ06 KK20 LL10 4G005 AA01 AB14 BA02 BB06 BB08 BB12 BB17 DB16Y DB16Z DB25X DC09W DD15Y DD15Z DE02Y DE02Z EA06 4J011 JA08 JB08 JB26 JB29 PA55 PB35 DB07 BA07 DB07 BA07 DB07
Claims (5)
相と、ポリジビニルベンゼン、ポリスチレン、又はジビ
ニルベンゼンとスチレンの共重合体からなり且つ該水相
を包括する有機相とからなることを特徴とするカタラー
ゼ包括マイクロカプセル。1. An aqueous phase comprising an aqueous solution containing catalase and an organic phase comprising polydivinylbenzene, polystyrene, or a copolymer of divinylbenzene and styrene and including the aqueous phase. Catalase-encapsulated microcapsules.
ルベンゼン及び／又はスチレンからなるモノマー、界面
活性剤及び重合開始剤を含有する有機溶媒とを混合して
一次乳化液を得た後、該一次乳化液をアラビアゴム水溶
液に分散させ二次乳化液を得、次いで該二次乳化液を窒
素雰囲気下に保ちながらモノマーの重合を行うことを特
徴とするカタラーゼ包括マイクロカプセルの製造方法。2. A primary emulsion is obtained by mixing an aqueous solution containing catalase and an organic solvent containing a monomer composed of divinylbenzene and / or styrene, a surfactant and a polymerization initiator. A method for producing catalase-encapsulated microcapsules, comprising dispersing the liquid in an aqueous solution of gum arabic to obtain a secondary emulsion, and then polymerizing the monomer while maintaining the secondary emulsion in a nitrogen atmosphere.
請求項２記載の製造方法。3. The method according to claim 2, wherein the organic solvent is toluene or benzene.
で、反応時間が１〜４時間である請求項２記載の製造方
法。4. The reaction temperature in the polymerization is from 60 to 75 ° C.
The method according to claim 2, wherein the reaction time is 1 to 4 hours.
ゼ包括マイクロカプセルと過酸化水素とを接触させるこ
とを特徴とする過酸化水素の分解方法。5. A method for decomposing hydrogen peroxide, comprising bringing the catalase-encapsulated microcapsules according to claim 1 into contact with hydrogen peroxide in an aqueous solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2001064449A JP2002262865A (en) | 2001-03-08 | 2001-03-08 | Catalase-occluding microcapsule and method for producing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2001064449A JP2002262865A (en) | 2001-03-08 | 2001-03-08 | Catalase-occluding microcapsule and method for producing the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2002262865A true JP2002262865A (en) | 2002-09-17 |
Family
ID=18923265
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2001064449A Pending JP2002262865A (en) | 2001-03-08 | 2001-03-08 | Catalase-occluding microcapsule and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2002262865A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016510589A (en) * | 2013-02-26 | 2016-04-11 | アヴェント インコーポレイテッド | Preparation of stabilized catalase enzyme using polyvinyl alcohol |
-
2001
- 2001-03-08 JP JP2001064449A patent/JP2002262865A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016510589A (en) * | 2013-02-26 | 2016-04-11 | アヴェント インコーポレイテッド | Preparation of stabilized catalase enzyme using polyvinyl alcohol |
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