JPS62175177A - Production of granular molded material containing immobilized enzyme or microbial cell - Google Patents

Abstract

PURPOSE:To obtain the titled molded material, obtained by dripping a liquid composition containing an unsaturated group-containing photo-setting PVA, photopolymerization initiator, polysaccharide, gelling with polyvalent metal ions, enzyme, etc., into an aqueous solution containing polyvalent metal ion and boron and gelatinizing the composition and having improved immobillization efficiency and strength. CONSTITUTION:A liquid composition containing (A) a photo-setting polyvinyl alcohol having ethylenically unsaturated bonds, e.g. methacryloylated polyvinyl alcohol, etc., (B) a photopolymerization initiator, e.g. benzoin, etc., (C) a water- soluble polysaccharide having the ability to gel on contact with at least one polyvalent metal ion, e.g. carrageenan, etc., and (D) an enzyme or microbial cells is dripped into an aqueous solution containing the polyvalent metal ion and boron and gelatinized to afford the aimed molded material.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the immobilization of enzymes or microbial cells, and more particularly to a method for immobilizing enzymes or microbial cells in the form of granular molded products.

Conventionally, methods for immobilizing enzymes or microorganisms include comprehensive method,
Many methods are known, such as physical adsorption methods and covalent bonding methods. When a lump or sheet-like immobilized product obtained by these methods is used for a microbial reaction or an enzymatic reaction, it is usually cut into small pieces or crushed and then packed into a rear ram. However, in this case, the immobilized substances often come into close contact with each other, which reduces the efficiency of microbial reactions and enzymatic reactions, and also has drawbacks such as frequent channeling phenomena that clog the column.

The present inventors believe that if enzymes or microbial cells can be immobilized as a granular molded product, it will be easier to fill the fluidized column and the contact area between particles will be reduced, increasing the efficiency of microbial reactions and enzymatic reactions. Considering that it is possible to do so, we have previously proposed a method for preparing a granular gel of enzymes or microorganisms using a photocurable resin and a water-soluble polymeric polysaccharide (see JP-A-59-11182).

However, since photocurable resins are generally expensive,
There has been a need for a photocurable resin that is inexpensive and forms defined granules with excellent mechanical strength. Therefore, the inventors conducted extensive research into a method for immobilizing enzymes or microbial cells in the form of granular moldings easily and at low cost, and found that polyvinyl alcohol with photocurable properties was used as the immobilization carrier. Using a combination of this material and a water-soluble polymeric polysaccharide capable of gelling upon contact with polyvalent metal ions, it is possible to remove boric acid from an aqueous medium containing polyvalent metal ions very easily and inexpensively. The present inventors have discovered that it is possible to produce a granular immobilized product with high mechanical strength without loss of enzymes or microbial cells, and have completed the present invention. Thus, according to the present invention, (a) a photocurable polyvinyl alcohol having ethylenically unsaturated bonds, (b) a photopolymerization initiator, and (c) an aqueous solution capable of gelling upon contact with polyvalent metal ions. A liquid composition comprising a polysaccharide, and (d) an enzyme or a microbial cell is dropped into an aqueous medium containing polyvalent metal ions and boric acid to gel the composition into particles. Provided is a method for producing a granular immobilized molded product of enzymes or microorganisms, which is characterized by the following. The method of the present invention will be explained in more detail below.

(a) Photocurable polyvinyl alcohol Polyvinyl alcohol is inexpensive, has little toxicity, and has excellent mechanical strength. The polyvinyl alcohol generally has a degree of saponification of 100 to 75 and a degree of polymerization of 300 to 3.0.
00 is used, but in consideration of handling viscosity and mechanical strength, it is preferably one with a degree of saponification of 90 to 80 and a degree of polymerization of 500 to 2,500. The photosensitive group having an ethylenically unsaturated bond introduced to impart photocurability does not necessarily have to be a wood-philic group since polyvinyl alcohol itself is hydrophilic. In other words, it is sufficient that enzymes or microorganisms can be uniformly dispersed in an aqueous medium in which they are suspended, and that when irradiated with active light having a wavelength in the range of about 250 to about 600 nm, it hardens and becomes substantially insoluble in water. Any material that can replace resin can be suitably used. In order to introduce ethylenically unsaturated bonds into polyvinyl alcohol, there are a method of focusing on the OH group of polyvinyl alcohol and reacting with the OH group to introduce a photosensitive group, and a method of utilizing a graft reaction. The former method is generally used, and reaction methods include esterification, urethanization, ketalization, acetalization, and etherification. Typical photosensitive groups with ethylenically unsaturated bonds to be introduced into polyvinyl alcohol include acryloyl groups,
Examples include a methacryloyl group, an acrylamide group, an allyl group, a vinyl ether group, a vinyl 1,000-ethyl group, and a vinylamine group. The above photosensitive groups may be introduced singly or in combination of two or more. Among these photocurable polyvinyl alcohols into which a photosensitive group has been introduced, those that are particularly advantageously used in the present invention include photocurable polyvinyl alcohols in which N-methylol acrylamide is introduced using an acetalization reaction. and polyvinyl alcohol.

(b) Photopolymerization initiator - For the purpose of promoting the photopolymerization reaction of the photocurable polyvinyl alcohol mentioned in (a), a photopolymerization initiator (photosensitizer) is added to the liquid composition according to the present invention. Include. The photopolymerization initiators that can be used are those that decompose by light irradiation to generate radicals, which act as polymerization initiation species to cause a cross-linking reaction between resins having polymerizable unsaturated groups.For example, benzoin, α-carbonyl alcohols such as acetoin; acyloin ethers such as benzoin methyl ether, benzoin ethyl ether, benzoin propyl ether, anituin ethyl ether, and piparoin ethyl ether; polycyclic aromatic compounds such as naphthol and hydroxyanthracene; α-substituted acyloins such as methylbenzoin and α-methoxybenzoin; 2-cyano-2
Examples include azoamide compounds such as butylazoformamide; metal salts such as launyl nitrate and ferric chloride; mercaptans; disulfides; halogen compounds; and dyes.

These photopolymerization initiators can be used alone or in combination of two or more, usually at a ratio of 0.01 to 10 PHR.

(c) Water-soluble polymeric polysaccharide In order to achieve gelation and granularity of an enzyme or microbial immobilization carrier in an aqueous medium, the method of the present invention uses the above-mentioned (a) as an immobilization carrier. There is one major feature in using a water-soluble polymeric polysaccharide in combination with the photocurable polyvinyl alcohol described in .

The water-soluble polymeric polysaccharide used in the present invention is a polymeric polysaccharide that is water-soluble and has the ability to change into a gel that is insoluble or poorly soluble in water when it comes into contact with polyvalent metal ions in an aqueous medium. , generally from about 3,000 to about 2,000,000
0 and typically at least about 10 g/1 (2
Those exhibiting a solubility of 5°C) are preferably used.

Specific examples of water-soluble polymeric polysaccharides having such characteristics include alkali metal salts of alginic acid, carrageenan, mannan, chitosan, and the like.

The metal salt of alginic acid, which is one of these water-soluble polymeric polysaccharides, is dissolved in an aqueous medium and contains polyvalent metal ions,
For example, magnesium ions, calcium ions,
Alkaline earth metal ions such as rontium ions and barium ions, aluminum ions, cerium ions,
It can gel when contacted with at least one other polyvalent metal ion such as nickel ion.

It should be noted that the water-soluble polymeric polysaccharide that can be used in the present invention does not need to have gelation ability upon contact with all polyvalent metal ions, but at least one type of polyvalent metal ion, preferably It is sufficient that it has the ability to gel when contacted with alkaline earth metal ions. The concentration of polyvalent metal ions at which gelation occurs varies depending on the type of water-soluble polymeric polysaccharide, but is generally at least 0.01 mo/9
It is ゜.

(d) Enzymes or microbial cells There are no particular restrictions on the types of enzymes or microbial cells that can be immobilized by the method of the present invention, and according to the method of the present invention, any type of enzyme or microbial cells can be immobilized. It can be immobilized without substantially deactivating its enzyme activity.

Representative examples of enzymes and microbial cells that can be immobilized by the method of the present invention are as follows.

(b) Examples of enzymes Lactate dehydrogenase (1, 1, 2, 3) Lactate oxidase (1, 1, 3, 2) Glucose oxidase (1, 1, 3, 4) Formate dehydrogenase (1, 2, 1, 2 ) aldehyde dehydrogenase (1,
2,1,3) Aldehyde oxidase (1,2,3,1
) xanthine oxidase (1,2,3,2) pyruvate oxidase (1,2,3,3) pyruvate reductase (1,2,4,1) L-amino acid oxidase (1
,4,3,2) D-amino acid oxidase (1,4,3
,3) Catalase (1,11,1,6) (2,6,1,lJ Hexokinase (2,7,11) Glucokinase (2,7,1,2) Fructokinase (2,7,1, 4) Phosphoglucokinase (2,7,1,10) Phosphofructokinase (2,7,1,11) Pyruvate kinase (2,7,1,40) Carboxylesterase (3,1
,1,1) Aryl esterase (3,1,1,2) Lipase (3,1,1,3) Phospholipase A (3,1,1,4) Acetyl esterase (3,1,1,6) Glucoamylase (3,2,1,3) Cellulase (3,2,1,4) Inulase (3,2,1,7) α-glucosidase (3,2,1,20) β-glucosidase (3,2,1 , 21) α-galactosidase (3
, 2, 1, 22) β-galactosidase (3, 2, 1,
23) Invertase (3,2,1,26) Pepsin (3,4,4,1) Trypsin (3,4,4,4) Chymotrypsin A (3,4,4,5) Cathepsin A (3,4) Papain (3,4,4,10) Thrombin (3,4,4,13) Amidase (3,5,1,4) Urease (3,5,1,5) Penicillin acidase (3,5,1, 11) Amine acylase (3,5,1,14) adenine deaminase (3
, 5, 4, 2) A, T, P,ase (3, 6, 1, 3)
Oxalate decarboxylase (4,1,1,1) Tryptophan decarboxylase (4,1,1,27) Aldolase (4,1,2,13 ) malate sudase (4,1,3,2) tryptophan synthase (4,2,1,20) aldolase (4,3
, 1, 1) Lysine racemase (5, 1, 1, 5) Glucose-6-phosphorus imerase (5, 3, 1, 9) Steroid delta-imerase (5, 3, 3, 1) Maxinyl-CoA synthetase (6 , 2, 1, 5) etc. (Note) Numbers in parentheses represent enzyme numbers.

(b) Example of microbial cell Lactobacillus bulgaricus
) Aerobacter aerogenes Bacillus subtilis
) Proteus vulgaris
aris) Arthrobacter simplex x,
7 Escharichia coli
li) Pseudomonas putida
Achromobacter Iiquidum) Curvularia Iun
ata) Corynebacterium glutamicum
um) Nocardia rhodocras (Nocardia
rhodcrous) Methanosarcina berkerii
Such.

Each of the components (a), (b), (c) and (d) described in ``2'' below can be made into a liquid composition by sufficiently mixing them with each other in an aqueous medium. . The aqueous medium that can be used is preferably water or an aqueous buffer solution, but in some cases, a mixture of water-soluble alcohols and water or an aqueous buffer solution, a mixture of water-soluble ketones and water or an aqueous buffer solution, or water. It is also possible to use an ester solvent solution that can be uniformly mixed with the aqueous buffer solution.

- The mutual usage ratio of each component (a), (b), (c) and (d) is not strictly limited and can be varied over a wide range depending on the type of each component, etc. However, in general, it is appropriate to use the following proportions to 100 parts by weight of photocurable polyvinyl alcohol (a) r& (within the range being suitable).

(b) Photopolymerization initiator = 0.5 to 5□ (1 to 3 parts by weight) (c) Water-soluble polymeric polysaccharide = 0.5 to 15 parts by weight (1 to 3 parts by weight)
(d) Enzymes or microbial cells in Yaju Town: O, OO1-50 heavy toughness (0
, 01 to 20 Shigemachi, and the aqueous medium is 1 to the total of (a) to (d) above.
It can be used in a range of 0 to 1.500 parts by weight (50 to 900 parts by weight).

The liquid composition prepared as described above is then dropped into an aqueous medium containing polyvalent metal ions and boric acid to form a gel into particles.

Both polyvalent metal ions and boric acid are essential components in order to gel the above-mentioned liquid composition into particles, and the gelation cannot be achieved even if either one is missing. That is, when polyvalent metal ions are lacking, the above-mentioned liquid composition gels but does not become granular. On the other hand, if boric acid is lacking, the liquid composition described in "-" becomes granular, but this gelatinized material has almost no mechanical strength and cannot be put to practical use. The above liquid composition is extremely easily gelled into granules due to the synergistic action of both the polyvalent metal ions and boric acid.

In addition, gelation with boric acid takes time even if it can be made into granules, so it is difficult to continuously produce granular fixed molded products. On the other hand, if polyvalent metal ions and boric acid are used at the same time, a granular fixed molded product is generated instantly, and it does not adsorb easily or change the granular state, so continuous production is possible. Become. This significantly reduces manufacturing costs.

The polyvalent metal ions that can be included in the aqueous medium include:
One is selected that has the ability to gel the water-soluble polymeric polysaccharide in the liquid composition.

Preparation of an aqueous medium containing selected polyvalent metal ions and boric acid involves adding water-soluble compounds of the polyvalent metal, such as halides, carbon salts, hydrogen carbonates, and sulfates of the polyvalent metal, to the aqueous medium. This can be carried out by dissolving boric acid after dissolving nitrate or the like. The concentration of polyvalent metal ions in the aqueous medium at this time can generally be in the range of 0.01 to 2 mou/u, preferably 0.1 to 2 mou/u. On the other hand, the concentration of boric acid is generally 0.01 to 10m0/b
sentence, preferably within the range of 0.1 to 4 mo views/sentence.

Further, the pH of the aqueous medium containing the selected polyvalent metal ion and boric acid is in the acidic range. If you want this to be in the neutral or alkaline range, use a monovalent alkali metal,
It may coexist with polyvalent metal ions and boric acid. The concentration of this alkali metal can be arbitrarily determined depending on the pH to be set.

The liquid composition may be dropped into the aqueous medium containing polyvalent metal ions and boric acid by, for example, dropping the liquid composition from the tip of a narrow tube such as the injection side, or by using centrifugal force. This can be carried out by methods such as a method in which the liquid composition is dispersed in the form of granules, or a method in which the liquid composition is atomized into granules and dropped from the tip of a spray nozzle. The size of the droplets to be dropped can be freely changed depending on the particle size desired for the final granular immobilized product, but usually the diameter is about 0.1 to about 5 mm, preferably about 0.5 to about 3 mm. It is convenient to apply it as droplets.

Gelation of the dropped liquid composition in an aqueous medium is thought to occur in two ways: by the water-soluble polymeric polysaccharide and polyvalent metal ions, and by photocurable polyvinyl alcohol and boric acid. Gelation between the water-soluble polymeric polysaccharide and the polyvalent metal ion occurs immediately, and although the mechanism of gelation at that time is not precisely known, the water-soluble polymeric polysaccharide aggregates with the halide or salt solution, and then It is presumed that the anion groups contained in the water-soluble polymeric polysaccharide form ionic bonds via polyvalent metal ions, thereby three-dimensionally crosslinking and gelling. - It is presumed that the gelation caused by force-curable polyvinyl alcohol and boric acid is caused by chemical bonding between the hydroxyl group of the photocurable polyvinyl alcohol and the boron of boric acid in a monodiol type. This gelation occurs from the surface to the inside of the granular molded product, but since internal crosslinking is achieved by photocuring, there is no need to gel the inside with boric acid.

In addition, it takes a long time, from 1 to 24 hours, depending on the particle size and residual hydroxyl groups, to gel the inside using boric acid alone, and moreover, it does not provide the same mechanical strength as that achieved by photocuring. Therefore, it is considered that about 1 to 30 minutes is sufficient for the gelation of photocurable modified polyvinyl alcohol.

Furthermore, a surfactant may be used for the purpose of assisting the granulation caused by gelation. This surfactant may be contained in the liquid composition to be dropped, or may be placed in an aqueous medium containing polyvalent metal ions and boric acid. The concentration of the surfactant may be about 0.001% to 5%. Furthermore, any type of surfactant can be used, but nonionic surfactants are preferred as they do not have to worry about reacting with the compound being used. Examples include sorbitan fatty acid ester, glycerin fatty acid ester, propylene glycol fatty acid ester, and sucrose fatty acid ester.

2) Room temperature is usually sufficient for gelation, but if necessary, gelation may be carried out under heating to a degree that does not inactivate the enzyme or microbial cells, or may be carried out under cooling. .

The granular gel produced as described above can be dispersed as it is in an aqueous medium, or after being separated from the aqueous medium and irradiated with actinic light, the photocurable polyvinyl in the granular gel can be cured. Hardens the alcohol. As a result, the granular gel can be obtained as a granular immobilized enzyme or microorganism that is substantially insoluble in water and has high mechanical strength.

The wavelength of the actinic light that can be used for photocuring as described above varies depending on the type of photocurable modified polyvinyl alcohol contained in the granular gel, but is generally about 250 to about 60 Or+m.
Advantageously, a light source is used for the irradiation which emits light with a wavelength in the range of . Examples of such light sources include low pressure mercury lamps, high pressure mercury lamps, fluorescent lamps, xenon lamps, carbon arc lamps, sunlight, and the like. The irradiation time needs to be changed depending on the light intensity of the light source, the distance from the light source, etc., but can generally be within the range of about 0.5 to about 10 minutes. Note that if the irradiation is performed in an inert gas atmosphere, the irradiation time may be shortened.

Irradiation should be done so that the actinic rays are distributed as evenly as possible over the entire granular gel that has been formed. For example, when irradiating active rays to a granular gel that has been separated from an aqueous medium, the separated granular gel should be irradiated with active rays. It is convenient to arrange the irradiation material in substantially a single layer in a suitable transparent glass plate or glass container and to irradiate the glass plate or glass container both from above and from below.

The granular gel thus produced can be washed with water or an aqueous buffer solution and stored as is, or the granular gel can be lyophilized and stored.

Thus, according to the present invention, granular immobilized enzymes or microorganisms having a particle size of about 0.5 to about 5 mm can be produced by extremely simple operations, and continuous production is also possible.

According to the method of the present invention, the granular immobilization of enzymes or microbial cells using a synthetic carrier is facilitated, and the use of water-soluble polymeric polysaccharides provides a protective effect on the activity of enzymes and microbial cells. Furthermore, by using polyvinyl alcohol, which is inexpensive and has excellent biocompatibility, as an immobilization carrier, advantages such as a wider range of applications can be obtained in terms of cost and long-term storage.

These granular immobilized products can be easily used particularly in reactors whose scale is not too large, fluidized bed type glue reactors, and the like.

Next, the present invention will be further explained by examples.

Example 1 Degree of polymerization 1000. To 100 parts by weight of a 20% photocurable prepolymer aqueous solution consisting of a mixture of 100 g of polyvinyl alcohol with a saponification degree of 87.0 to 89.0 and 0.3 mol (30 g) of N-methylolacrylamide, 20 parts by weight of a 3% sodium alginate aqueous solution was added. , 1 part by weight of benzoin isobutyl ether, 40 parts by weight of distilled water and 20 parts by weight of the enzyme invertase (0.1% concentration) were added and mixed well, and the resulting photocurable resin-enzyme mixture was diluted with 3% boric acid and
．． Aqueous solution containing 1M calcium chloride (pH=
When the solution was dropped from a liquid level of 10 cm from the injection needle at the tip of the syringe into the solution (prepared in Example 6), granules with a particle diameter of 2.0 mm were obtained.

When this granular material was placed in a Petri dish with a flat bottom and active light having a wavelength of 300 to 400 r+m was irradiated for 3 minutes from the top and bottom surfaces of the Petri dish, a granular immobilized enzyme with good mechanical strength was obtained.

The invertase activity of these particles was increased to 1 using sucrose as a substrate.
When 111 was added, the specific activity against non-immobilized invertase was 70%.

Example 2 Degree of polymerization 1500. 100g of polyvinyl alcohol with a saponification degree of 87-89 and 0.2g of N-methylolacrylamide
To 100 parts by weight of a 20% photocurable prepolymer aqueous solution consisting of mol (20 g), add 3% ginaan aqueous solution 2
0 parts by weight, 1 part by weight of benzoin ethyl ether, 60 parts by weight of distilled water, and 3 parts by weight of 2% glucose imerase bacterial enzyme solution (sodium bicarbonate buffer, pH = 8) to prepare a homogeneous mixed solution. did. This homogeneous mixture was poured from the tip of the syringe needle into an aqueous solution containing 3% boric acid and 5% potassium chloride (adjusted to pH 6 with NaOH) from the liquid level at 15c+n.
When it was dropped from the position, granules with a particle size of 2.5 mm were obtained.

When this granular material was transferred to a Petri dish with a flat bottom and irradiated with actinic light having a wavelength of 300 to 400 nm from both sides and the bottom surface, a granular immobilized enzyme with good mechanical strength was obtained.

The activity of this granular glucose isomerase was measured at 60°C in a pH 8 sodium bicarbonate buffer using glucose as a substrate.
As a result of measurement, the specific activity against non-immobilized glucose isomerase was 80%.

Example 3 Degree of polymerization 1100. To 100 parts by weight of a 20% photocurable prepolymer aqueous solution consisting of 100 g of polyvinyl alcohol with a saponification degree of 87.0 to 89.0 and 0.3 mol (30 g) of N-methylol acrylamide, 20 parts by weight of a 3% sodium alginate aqueous solution, A homogeneous liquid mixture was prepared by adding 1 part by weight of benzoin ethyl ether, 50 parts by weight of distilled water, and 10 parts by weight of Bacillus subtilis cell suspension.

The resulting mixture was injected from the tip of a syringe into an aqueous solution containing 3% boric acid and 0.1M calcium chloride (pH adjusted with NaOH).
When added dropwise into the solution (prepared at 6.0%), it gelled into a spherical shape. When the spherical gel was transferred to a Petri dish and irradiated with actinic light having a wavelength of 300 to 400 nm for 3 minutes from the -L and bottom surfaces of the Petri dish, spherical immobilized microorganisms with a diameter of 3 mm were obtained.

Example 4 Degree of polymerization 500. To 100 parts by weight of a 20% photocurable prepolymer aqueous solution consisting of 100 g of polyvinyl alcohol with a saponification degree of 87.0 to 89.0 and 0.3 mol (3 og) of N-methylol acrylamide, 20 parts by weight of a 3% sodium alginate aqueous solution, 0.5 parts by weight of α-hydroxyisobutylphenone, 40 parts by weight of distilled water, and 20 parts by weight of a 0.5% glucose oxidase aqueous solution dissolved in 0.1M acetate buffer (pH 5, 6) were uniformly mixed to obtain this product. Pour the mixture from the tip of the syringe into 3% boric acid and 0.1M
When it was dropped into an aqueous solution (adjusted to pH=6 with NaOH) containing calcium chloride and 0.01% sorbitan monolaurate, it gelled into a spherical shape. This was placed in a Petri dish and irradiated with light of 300 to 400 nm from both sides and the bottom for 3 minutes to obtain granular immobilized microorganisms with a particle size of 2 mm.

Example 5 Degree of polymerization 1000. To 100 parts by weight of a 20% photocurable prepolymer aqueous solution consisting of 100 g of polyvinyl alcohol with a saponification degree of 87.0 to 89.0 and 0.3 mol (30 g) of N-methylolacrylamide, 20 parts by weight of a 3% aqueous ginan solution was added. 0.5 parts by weight of α-hydroxyisobutylphenone, 40 parts by weight of distilled water, and
20 parts by weight of the coli cell suspension was uniformly mixed and dispersed.

The thus obtained photocurable resin-microbial cell mixture was supplied to a rotating disk, and the mixture was scattered by the centrifugal force, and the scattered particles contained 3% boric acid and 0.3M potassium. It was dropped into an aqueous solution (adjusted to pH=6 with NaOH) and gelled into particles. This was placed in a Petri dish and irradiated with light of 300 to 400 nm from the upper and lower surfaces for 3 minutes at the same time, making it possible to produce immobilized bacterial cells with a particle size of 3IIffl.

Example 6 Polymerization degree 1000 in argon gas atmosphere. Saponification degree 87
．． 100 g of polyvinyl alcohol of 0 to 89,0 and N-
To 100 parts by weight of a 20% photocurable prepolymer aqueous solution consisting of 0.3 mol (30 g) of methylol acrylamide, 20 parts of a 3% alginic acid aqueous solution, α
-0.5 parts by weight of hydroxyisobutylphenone, 50 parts by weight of distilled water, and 10 parts by weight of Mesanosarcina berkelii cell suspension were uniformly mixed. Obtained photocurable resin
When the bacterial cell mixture was dropped from the tip of a syringe into an aqueous solution containing 3% boric acid and 0.1M potassium chloride (adjusted to pH=8 with NaOH), it gelled into particles. Place this in a Petri dish and irradiate it with light of 300 to 400 r+m for 3 minutes from the - side and the bottom side at the same time to obtain a particle size of 2 m.
m of granular immobilized microorganisms were obtained.

Claims (1)

[Scope of Claims] (a) Photocurable polyvinyl alcohol having an ethylenically unsaturated bond (b) Photopolymerization initiator (c) Aqueous solution capable of gelling upon contact with at least one polyvalent metal ion A liquid composition comprising a polymeric polysaccharide and (d) an enzyme or a microbial cell is dropped into an aqueous medium containing a polyvalent metal ion and boric acid to gel the composition into particles. A method for producing a granular immobilized molded product of enzymes or microorganisms, characterized by: