JP2990790B2 - Method for producing enzyme-immobilized carrier - Google Patents
Method for producing enzyme-immobilized carrierInfo
- Publication number
- JP2990790B2 JP2990790B2 JP2309944A JP30994490A JP2990790B2 JP 2990790 B2 JP2990790 B2 JP 2990790B2 JP 2309944 A JP2309944 A JP 2309944A JP 30994490 A JP30994490 A JP 30994490A JP 2990790 B2 JP2990790 B2 JP 2990790B2
- Authority
- JP
- Japan
- Prior art keywords
- carrier particles
- enzyme
- carrier
- granulation
- immobilized carrier
- 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 - Lifetime
Links
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Description
【発明の詳細な説明】 （産業上の利用分野） 本発明は表面に酵素を吸着或いは結合して保持する酵
素固定化担体の製造方法に関する。Description: TECHNICAL FIELD The present invention relates to a method for producing an enzyme-immobilized carrier that holds an enzyme by adsorbing or binding to the surface.
（従来の技術） 従来の一般的な酵素反応は、基質（原料）と酵素とを
反応槽に入れ、温度及びPHを調整しながら反応せしめて
目的の生成物を得るようにしてしていたが、これでは基
質を流しながら連続して反応物を得ることができないの
で、最近では担体に酵素を固定しこの担体を貫通して、
或いは担体に沿って基質を流すことで、基質に酵素を接
触させる方法が利用されている。(Prior art) In a conventional general enzymatic reaction, a substrate (raw material) and an enzyme are put into a reaction tank and reacted while adjusting the temperature and PH to obtain a desired product. However, in this case, it is not possible to obtain a reaction product continuously while flowing the substrate.
Alternatively, a method in which an enzyme is brought into contact with a substrate by flowing the substrate along a carrier is used.
斯かる担体としては有機材料が一般的であるが、有機
材料は機械的強度、耐熱性及び耐薬品性に問題があり、
このため無機材料を担体として用いることが提案されて
いる。Organic materials are generally used as such carriers, but organic materials have problems in mechanical strength, heat resistance and chemical resistance,
Therefore, it has been proposed to use an inorganic material as a carrier.
無機材料としては表面が疎水性を示す活性炭、骨炭、
多孔質炭素等の炭素材料と、表面が親水性を示すアルミ
ナ、シリカやベントナイト、ゼピオライト、アタパルジ
ャイト等の粘土鉱物がある。そして、セピオライトを用
いた酵素固定化担体の製造方法としては特開昭63−1906
37号に開示されるものがある。Activated carbon, bone char, whose surface is hydrophobic as inorganic materials,
There are carbon materials such as porous carbon and clay minerals such as alumina, silica, bentonite, zeolite and attapulgite whose surfaces are hydrophilic. A method for producing an enzyme-immobilized carrier using sepiolite is disclosed in JP-A-63-1906.
There is one disclosed in No. 37.
（発明が解決しようとする課題） 上述したように酵素固定化担体として無機材料は優れ
た特性を有するのであるが、炭素材料は酸性溶液或いは
アルカリ溶液に対する耐久性は高いが、酵素活性が低い
という欠点がある。一方、粘土鉱物は酵素活性は高い
が、通常酵素反応はPH4〜９程度の水溶液で行なうこと
を考えると、酸やアルカリに対する耐久性に問題があ
る。(Problems to be Solved by the Invention) As described above, an inorganic material has excellent properties as an enzyme-immobilized carrier, but a carbon material has high durability against an acidic solution or an alkaline solution, but has a low enzyme activity. There are drawbacks. On the other hand, clay minerals have a high enzyme activity, but have a problem in durability against acids and alkalis when considering that an enzyme reaction is usually performed in an aqueous solution having a pH of about 4 to 9.
（課題を解決するための手段） 上記課題を解決すべく本発明は、セピオライトやアパ
タルジャイト等の粘土鉱物に対し破砕、押出し造粒、転
動造粒等を施して担持粒子を造粒し、次いでこの担体粒
子を酸溶液で処理して可溶成分を除去した後に、担体粒
子を焼成するようにした。(Means for Solving the Problems) In order to solve the above-mentioned problems, the present invention provides a method of granulating supported particles by subjecting clay minerals such as sepiolite and apatalite to crushing, extrusion granulation, and tumbling granulation. Then, the carrier particles were treated with an acid solution to remove soluble components, and then the carrier particles were calcined.
（作用） 担体粒子を酸溶液で処理することで、酸に可溶な成分
が担体中から除去され、結果として酸に対する耐久性が
向上する。(Action) By treating the carrier particles with the acid solution, the components soluble in the acid are removed from the carrier, and as a result, the durability against the acid is improved.
（実施例） 以下に本発明の実施例を具体的数値を挙げて説明す
る。(Examples) Examples of the present invention will be described below with specific numerical values.
本発明は造粒工程、酸処理工程及び焼成工程からな
る。The present invention comprises a granulation step, an acid treatment step and a baking step.
［造粒工程］ 造粒工程はセピオライト原料のうち300メッシュ以下
のものを篩によって選定し、この原料に水を加えホモジ
ナイザーで分散させた後、110℃で乾燥固化させて粉砕
することで粒径300〜500μｍの担体粒子を造粒する。
又、造粒の仕方として、この破砕造粒の他、原料と水を
加えて粘土状にし、これを押し出し成形した後、裁断す
る押し出し造粒や、原料と小量の水とをミキサーで分
散、造粒させる転動造粒等各種の造粒方法が利用でき
る。[Granulation Step] In the granulation step, a sepiolite raw material having a mesh size of 300 mesh or less is selected by a sieve, water is added to the raw material, and the raw material is dispersed by a homogenizer. Granulate 300-500 μm carrier particles.
As a method of granulation, in addition to this crushing granulation, raw materials and water are added to make a clay, which is extruded, then extruded and granulated, and the raw materials and a small amount of water are dispersed with a mixer. Various granulation methods such as rolling granulation can be used.
尚、原料としてはセピオライトの他、表面が親水性を
示すアルミナ、シリカやアタパルジャイト、ベントナイ
ト等の粘土鉱物を用いてもよい。As a raw material, other than sepiolite, clay minerals such as alumina, silica, attapulgite, and bentonite whose surfaces are hydrophilic may be used.
［酸処理工程］ 上記の造粒工程で得られた担体粒子を約800℃で４時
間仮焼成する。この仮焼成で造粒状態を維持するととも
に後述するアルカリ金属の酸化物を溶出しやすくする。[Acid treatment step] The carrier particles obtained in the above granulation step are calcined at about 800 ° C for 4 hours. This calcining maintains the granulated state and facilitates the elution of an oxide of an alkali metal described later.
そして、仮焼成した担体粒子4gを塩酸溶液（PH≒２）
100mlに24時間浸漬し、担体中からMgO、K2O、Na2O等の
アルカリ金属の酸化物を溶出する。このときの処理温度
は30〜180℃とする。Then, 4 g of the calcined carrier particles are added to a hydrochloric acid solution (PH ≒ 2).
It is immersed in 100 ml for 24 hours to elute alkali metal oxides such as MgO, K 2 O and Na 2 O from the carrier. The processing temperature at this time is 30 to 180 ° C.
この後、塩酸溶液中から担体粒子を取り出して塩酸が
完全に除去されるまで繰返し洗浄して乾燥させる。Thereafter, the carrier particles are taken out from the hydrochloric acid solution, washed repeatedly until the hydrochloric acid is completely removed, and dried.
［焼成工程］ 上記の酸処理工程で得られた担体粒子を焼成温度800
℃,900℃及び1000℃で４時間保持して目的とする担体粒
子を得る。[Firing step] The carrier particles obtained in the above acid treatment step are baked at a firing temperature of 800.
C., 900.degree. C. and 1000.degree. C. for 4 hours to obtain target carrier particles.
尚、実施例では担体を粒子状のものとしたがバインダ
を用いて所定形状に成形してもよい。In the embodiment, the carrier is in the form of particles. However, the carrier may be formed into a predetermined shape using a binder.
以上によって得られた担体粒子と、酸処理工程を省略
し造粒工程から直接焼成工程に移行して得られた担体粒
子とを水銀圧入法による細孔評価、PH5水溶液24時間浸
漬による耐食性評価、酵素（プルラナーゼ）の固定化に
よる活性評価において比較した結果を第１図乃至第３図
のグラフに示す。The carrier particles obtained by the above, the carrier particles obtained by directly moving from the granulation step to the baking step omitting the acid treatment step and pore evaluation by mercury intrusion method, corrosion resistance evaluation by immersion in PH5 aqueous solution for 24 hours, The results of comparison in activity evaluation by immobilization of an enzyme (pullulanase) are shown in the graphs of FIG. 1 to FIG.
（効果） 以上の説明及びグラフから明らかなように本発明によ
れば、セピオライトやアパタルジャイト等の粘土鉱物を
造粒してなる担体粒子を酸処理し、この酸処理で担体中
の可溶成分を除去した後に、担体粒子を焼成するように
したので、酵素固定化担体の耐酸性が大幅に向上し、更
に比表面積及び酵素活性についても酸処理しないものよ
り向上した。(Effects) As is clear from the above description and the graph, according to the present invention, carrier particles obtained by granulating clay minerals such as sepiolite and apatalite are subjected to an acid treatment, and the acid treatment causes the carrier particles to become soluble in the carrier. Since the carrier particles were calcined after the removal of the components, the acid resistance of the enzyme-immobilized carrier was greatly improved, and the specific surface area and the enzyme activity were further improved as compared with those without acid treatment.
第１図は酵素固定化担体粒子の焼成温度と酸性溶液に対
する耐久性の関係を酸処理したものと酸処理しないもの
について比較したグラフ、第２図は酵素固定化担体粒子
の焼成温度と比表面積との関係を酸処理したものと酸処
理しないものについて比較したグラフ、第３図は酵素固
定化担体粒子の焼成温度と酵素活性との関係を酸処理し
たものと酸処理しないものについて比較したグラフであ
る。FIG. 1 is a graph comparing the relationship between the calcination temperature of the enzyme-immobilized carrier particles and the durability against an acidic solution of the enzyme-treated and non-acid-treated ones, and FIG. And FIG. 3 is a graph comparing the relationship between the calcination temperature of the enzyme-immobilized carrier particles and the enzyme activity between the acid-treated and non-acid-treated ones. It is.
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭62−36193（ＪＰ，Ａ) 特開 昭63−91083（ＪＰ，Ａ) 特開 昭63−190637（ＪＰ，Ａ) 特開 平２−252669（ＪＰ，Ａ) (58)調査した分野(Int.Cl.6，ＤＢ名) C12N 11/14 ──────────────────────────────────────────────────続 き Continuation of front page (56) References JP-A-62-36193 (JP, A) JP-A-63-91083 (JP, A) JP-A-63-190637 (JP, A) JP-A-2- 252669 (JP, A) (58) Field surveyed (Int. Cl. 6 , DB name) C12N 11/14
Claims (2)
が親水性を示す粘土鉱物またはこの粘土鉱物を仮焼した
ものに対し、粉砕、押出し造粒、転造造粒を施して担体
粒子を造粒し、次いでこの担体粒子を酸溶液で処理する
ことで担体粒子中の可溶成分を除去し、この後担体粒子
を焼成するようにしたことを特徴とする酵素固定化担体
の製造方法。1. A clay mineral whose surface is hydrophilic, such as sepiolite or apatalite, or a calcined product of this clay mineral, is subjected to pulverization, extrusion granulation, and roll granulation to granulate carrier particles. Then, the carrier particles are treated with an acid solution to remove soluble components in the carrier particles, and thereafter, the carrier particles are calcined.
法において、前記酸処理前の粘土鉱物を仮焼する際の温
度を1000℃以下とし、酸処理後の焼成温度を800〜1000
℃とすることを特徴とする酵素固定化担体の製造方法。2. The method for producing an enzyme-immobilized carrier according to claim 1, wherein the temperature at which the clay mineral before the acid treatment is calcined is 1000 ° C. or less, and the firing temperature after the acid treatment is 800 to 1000 ° C.
C., a method for producing an enzyme-immobilized carrier.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2309944A JP2990790B2 (en) | 1990-11-15 | 1990-11-15 | Method for producing enzyme-immobilized carrier |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2309944A JP2990790B2 (en) | 1990-11-15 | 1990-11-15 | Method for producing enzyme-immobilized carrier |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH04183393A JPH04183393A (en) | 1992-06-30 |
| JP2990790B2 true JP2990790B2 (en) | 1999-12-13 |
Family
ID=17999232
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2309944A Expired - Lifetime JP2990790B2 (en) | 1990-11-15 | 1990-11-15 | Method for producing enzyme-immobilized carrier |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2990790B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0735736A (en) * | 1993-07-19 | 1995-02-07 | Snow Brand Milk Prod Co Ltd | Separation agent for liquid chromatography and separation/analytic method employing same |
| ATE316651T1 (en) * | 1999-11-15 | 2006-02-15 | Arkray Inc | BIOSENSOR |
-
1990
- 1990-11-15 JP JP2309944A patent/JP2990790B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPH04183393A (en) | 1992-06-30 |
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