JPS58194748A - Glass or ceramic for immobilizing enzymes and vital substances - Google Patents

Abstract

PURPOSE:A glass or ceramic for immobilizing enzymes or vital substances that is prepared by the sol-gel method utilizing hydrolysis of metal alkoxide, thus showing high purity, stability and porosity with increased volume efficiency as a support. CONSTITUTION:A metal alkoxide such as ethyl silicate is hydrolyzed by mixing it with water and ethanol under stirring. After completion of hydrolysis, the mixture is placed in a mold for standing to convert the sol into the gel of silicic acid. Then, the resultant dried gel is placed in a crucible and calcined at about 900 deg.C to give the objective microporous silica glass. When aluminum isopropoxide or titanium butoxide is used as a metal alkoxide instead of ethyl silicate, Al2O3 or TiO2 ceramic is obtained.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to glass KII for life sciences, and more specifically, a novel miterporous glass produced by the sol-gel method using hydrolysis of a metal alkoxide compound.
! ! This is a glass related to YFi Cera 1 Tutus for fixing enzymes and biological substances.

Enzymes are proteins produced by living organisms and are 41I
It has a catalytic effect that causes a constant chemical reaction in tF substances, and its reaction efficiency is high, so it can be used for brewing, fermentation, and food.

@Medicine, etc. [plays a very important role in 1 yen.

but,#! The I reaction is a patch method in which the enzyme and substrate are mixed in water S* each time, and the enzyme is discarded after each reaction.

We have begun to try immobilizing (insolubilizing) the enzyme and using it repeatedly at medium speed, precise control and automation of reaction 1, IIA for separation of biological products, and improvement of enzyme stability. Porous silica glass has been taken up as one of the carriers by researchers.

Thin porous silica glass? I! 1 side KN11 element tlii
For example, as shown in Fig. 1, a method of bonding glass and glass through one molecule of a horizontal substance is used to determine the temperature.

The carrier glass used for such enzymes and biological materials is mostly porous glass made by utilizing the so-called phase separation phenomenon of glass.

For example, Nn*O(8w+t'jg), %OB(
24%r)t), 8(0@(see 68vnt)) is melted and molded, and then heated at 500-650℃ to form N(L*0-T3*O
Separation (b) of the sIIJwi1 and p<o, m1izy2 phases into glass progresses, and the two phases become continuous, forming the glass of the copper wire (Fig. 2). About 5% of this glass
When immersed in hydrochloric acid to remove the Ha, O-BIO, and Si
o, 964g with remaining phase and countless continuous 9 pores
A porous glass ω) of 1o and 1g is obtained.

Production of porous glass by such a method The elution rate tK of the KFi soluble component causes distortion and separation between the two phases, and there is a problem with using only a completely stable phase as the framework component. Since the treatment is performed using a solution, there are problems such as impure eyelids tending to remain in the holes.

In contrast, the present invention uses a novel microporous glass made by the sol-gel method using hydrolysis of a gold-woven alkoxide compound, which is highly impurity-free and highly suitable for guests. It presents enzyme and biological material-fixed glass with high porous friction and high volumetric efficiency.

The preparation of a glass carrier by a sol-gel method using hydrolysis of a gold jI flukoxide compound will be described below.

For example, silica glass uses a silicon alkoxide compound as a raw material, but if ethyl silicate is used as silicon alkoxide, ethyl silicate (tti(o a
, i, l, ), water and ethyl alcohol? The mixture is stirred to hydrolyze the ethyl silicate and convert the silicic acid sol into a gel. This interest 1! Although it can be formed into any shape depending on the requirements, water and alcohol can be expelled while heating this fixed-shaped object to obtain silica glass with a three-dimensional structure that is highly pure charcoal.

In this case, the dry gel formed by the sol-gel method is gradually heated to about 1000-1100"C and fired.
'This results in a complete three-dimensional structure of quartz with no porous properties.

However, depending on the conditions for hydrolysis and the conditions for changing from a sol to a gel, the removal of 1 wl of gas in the adsorbed water cannot be controlled, resulting in the formation of a microporous structure with a pore size ranging from several tens of angstroms to several hundred angstroms. In the case of silica glass, this microporous structure is stably maintained at 900'C@It.

Inside this microporous, free silanol groups (-
day i-oH) remains.

Based on past experimental analysis, the temperature ranges from approximately 900℃ to 1000℃.
It has been found that the microporous disappears when the structure with a volume of annealed copper is heated to about 20°C.

In the present invention, by processing in the upper layer 19 where microporous does not disappear, it is possible to obtain enzyme and biomaterial-fixed glass with high density and stability, which cannot be obtained by conventional methods. This made it possible.

Specific embodiments of the present invention will be explained below.

In one example, ethyl silicate was hydrolyzed by adding 5.4 mj of ethanol and 4 ml of ethyl silicate for 2 seconds.

Put the 4j1 mixture l1i11 into a Teflon-coated petri dish with a diameter of 50, cover tightly with a polyethylene chain plate with pinholes as a lid, and place in a constant temperature bath at 70°C.
The temperature was raised to 95° C. and left to stand for 4 days to form a dry gel.

Next, this IF-dried gel was placed in a crucible furnace and heated to 900° C. by heating and shutting off at a rate of 20° C./hour to obtain microporous silica glass with a fM diameter of 30 m.

This porous silica glass has a pore diameter of 150 to 200
The dust is distributed over five seconds, and the volume ratio is approximately 2
It had a pore ratio of 0.01s. Enzymes such as lactase, pepsin, and trypsin can be immobilized on the silanol groups inside this porous via other suitable organic molecules shown in Figure 1.

Microporous silica glass has a disc-like shape, but it can be divided into small blocks of appropriate size depending on the pK and used.

Above, Kinoto's example was explained using 8I Os silica glass, and Hashimoto explained that the microporous glass and 7U1Tx made by the sol-gel method using metal alkoxide compound V hydrolysis drops are basic KFi. This book is applicable to all gold II compounds.

aluminum isopropoxide (
Mu1JoscerajV using Al(OOMtS)s)
clear alumina or titanium butoxide (RI).
(o (14H9)4) in 9TjOt ceramics and these funds 4#1 parabolic etc. It is possible to create microporous glasses and ceramics in the same manner as in the above examples! ! Galasne shows an example of an immobilized enzyme using Seratax 1 as a carrier.

Part 1!1 Microporous glass ceramics are made by the sol-gel method using hydrolysis of compounds.
In addition to important points such as being highly pure W and free of impurities due to the characteristics of the manufacturing method, and eliminating the need for high temperature LI fisting and large amount of energy dissipation that are required in the production of ordinary glass and ceramics, * Since a large amount of free 10H@f, which is necessary for fixing * and biological substances, is retained in the microporous, it is predicted that the identification efficiency per one step of the body 11j will be extremely high.

For this reason, it is possible to increase the catalytic efficiency considerably compared to porous glass in which the soluble portion is S★ using glass phase separation, which has been the main method in the past. Ta.

Novel microporous skin glass according to the present invention
The cerami-cus is fermented due to the many times it has been used.
There is an incalculable 4 in the range of T and ten as l and bio-quality recitation materials.

[Brief explanation of the drawing]

Figure 1 schematically shows a system for stabilizing enzyme 1 on a plus carrier. No. 211 is a schematic continuation of the conventional method for producing porous glass using phase separation. One person, Kamiide, owns the stock as a company, Suiwa Seikokin (a)
(b) No. 21 “C +1

Claims (2)

[Claims] (1) Microporous monolithic glass or ceramics for immobilizing biological materials prepared by Kei sol-gel method using hydrochloric acid of metal alkoxide. (2) Ethyl silicate (5s
Silica glass or ceramics for fixing enzymes and biological substances according to claim 1, which is prepared by a sol-gel method using (oc, H,)4).