JP2001252073A - Method of preparing granular formed body for immobilizing enzyme or microorganism cell - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for improving the mechanical strengths of a polyurethane resin used as a hydrophilic resin for immobilizing an enzyme or microorganism cells and a method of manufacturing granular formed bodies having enough strength. SOLUTION: This method of preparing granular formed bodies for immobilizing an enzyme or microorganism cells comprises the dropwise addition of an aqueous liquid composition composed of (a) a hydrophilic urethane resin, which has at least two ethylenically unsaturated bonds in a molecule, obtained by reacting an adduct of a diepoxy compound and an ethylenically unsaturated carboxylic acid with a polyisocyanate compound, (b) a polymerization initiator, (c) a water-soluble polymeric polysaccharide having an activity to gel by the contact with an alkali metal ion or a polyvalent metal ion, to an aqueous solvent co containing an alkali metal ion or a polyvalent metal ion to gelatinize the composition into a granular state, or the continuous pouring of the aqueous composition on the surface of the aqueous solvent over a specified period to grow the liquid drops to a desired particle size and subsequently depositing the liquid drops to convert the drops to gel. Subsequently, the obtained granular gel is subjected to photopolymerization and/or thermal polymerization to harden the hydrophilic resin in the granular gel.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a method for producing a granular molded product for immobilizing enzymes or microorganisms.

[0002]

2. Description of the Related Art As methods for immobilizing enzymes or microorganisms, many methods such as an entrapment method, a physical adsorption method, and a covalent bonding method have been known. When used in a microbial reaction or an enzymatic reaction, the solid or sheet-like immobilized product obtained by these methods is usually cut into small pieces or ground, and then packed into a column. However, in this case, the immobilized substance often comes into close contact with each other, resulting in poor microbial reaction and enzymatic reaction efficiency, and often has a drawback that a channeling phenomenon occurs and the column is clogged.

[0003] For this reason, recently, by immobilizing enzymes or microbial cells as a granular molded product, it is easy to flow, it is easy to pack into a column, and the contact area between particles is reduced. It has been proposed to increase the efficiency of a microbial reaction or an enzymatic reaction (for example, Japanese Patent Publication No. 62-19837, Japanese Patent Application Laid-Open No. 10-21096).
No. 9).

On the other hand, as a material for immobilizing enzymes or microbial cells, various hydrophilic photocurable resins and thermosetting resins have been conventionally used. Among them, polyurethane resins have been put to practical use because they have good adhesion to microbial cells and have sufficient biological activity, but polyurethane resins that have been used conventionally have photopolymerization or thermal polymerization reactivity. Since the group is present only at the molecular end, when the molecular weight is increased, the concentration of the unsaturated group is reduced, so that the number of crosslinking points is reduced. As a result, there is a problem that a granular molded product having sufficient strength cannot be obtained.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to improve the mechanical strength of a polyurethane resin used as a hydrophilic resin for immobilizing enzymes or microbial cells to produce a granular molded product having sufficient strength. Is to provide a way to

[0006]

Means for Solving the Problems The inventors of the present invention have conducted intensive studies in order to achieve the above-mentioned object. As a result, the use of a specific hydrophilic urethane resin having a high unsaturated group concentration provides sufficient strength. It has been found that a granular molded product can be obtained, and the present invention has been completed.

Thus, according to the present invention, (a) a polyisocyanate compound obtained by reacting an adduct of a diepoxy compound with an ethylenically unsaturated carboxylic acid with a polyisocyanate compound is obtained.
A hydrophilic urethane resin having at least two ethylenically unsaturated bonds in the molecule, (b) a polymerization initiator, and (c) a water-soluble agent capable of gelling by contact with an alkali metal ion or a polyvalent metal ion. An aqueous liquid composition comprising a high molecular weight polysaccharide is dropped into an aqueous medium containing an alkali metal ion or a polyvalent metal ion to gel the composition into particles, or on the surface of the aqueous medium. After continuously pouring for a predetermined time and growing the droplets to a desired particle size, the droplets are allowed to settle and gel, and then the resulting granular gel is subjected to photopolymerization and / or thermal polymerization. And curing the hydrophilic resin in the granular gel, thereby providing a method for producing a granular molded product for immobilizing enzymes or microorganisms.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail. (A) Hydrophilic urethane resin In the present invention, the hydrophilic urethane resin (a) having at least two ethylenically unsaturated bonds in one molecule used for the production of the granular carrier for immobilizing enzymes or microbial cells includes: Generally 400 to 50,000, preferably 500 to 2,000, obtained by reacting a polyisocyanate compound with an unsaturated group-containing diol compound obtained by performing an addition reaction of a diepoxy compound and an ethylenically unsaturated carboxylic acid.
Number average molecular weight in the range of 0 and 0.04 to 5 mol / kg
A resin, preferably having an unsaturated group content in the range of 0.1 to 4 mol / kg resin, and having sufficient ionic or nonionic hydrophilic groups, such as hydroxyl groups, to be uniformly dispersed in the aqueous medium. Those containing a carboxyl group, an ether group, an amino group and the like can be mentioned.

Examples of the diepoxy compound include diglycidyl ether compounds of bisphenol compounds such as bisphenol A, bisphenol F, and bisphenol S; Celloxide 2021 and 2080, trade names of alicyclic diepoxy compounds manufactured by Daicel Chemical Industries, Ltd. 3
000, general formula

[0010]

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(Wherein R ₁ represents an alkylene group or the like)
T228 and CDM; a general formula manufactured by Nagase Kasei Co., Ltd.

[0012]

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[0013] The product name Denacol EX-20
1, 203, 211, 212, 701, 71
1 and 721, general formula

[0014]

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Denacol EX-810, 811, 811, 830, 832, 841, 850, 851, and 861, represented by the following general formula:

[0016]

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, 921, 921, 931 and 941; and the like, but are not limited thereto. These compounds can be used alone or in combination of two or more. Among them, those particularly preferably used in the present invention are Denacol E
X-830, 832, 841 and 861.

Examples of the ethylenically unsaturated carboxylic acid include (meth) acrylic acid; trade name ARONIX M-5300 (ω-carboxy-) manufactured by Toagosei Co., Ltd.
Polycaprolactone monoacrylate), 5400
(Monohydroxyethyl phthalate phthalate) and 5600 (Acrylic acid dimer); Viscoat # 2000 (Monohydroxyethyl phthalate phthalate), # 2100 (Monohydroxypropyl phthalate phthalate), # 200 manufactured by Osaka Organic Chemical Industry Co., Ltd. 2150 (hexahydrophthalic acid monohydroxypropyl acrylate)
And # 2180 (monohydroxypropyl acrylate of tetrahydrophthalic acid); and the like, but are not limited thereto. These compounds can be used alone or in combination of two or more. Among them, (meth) acrylic acid is particularly preferably used in the present invention.

Examples of the polyisocyanate compound to be reacted with an unsaturated group-containing diol compound obtained by an addition reaction between a diepoxy compound and an unsaturated carboxylic acid include methylene diisocyanate, ethylene diisocyanate, propylene diisocyanate, butylene diisocyanate, and octadecylene diisocyanate. , 1,4-
Aliphatic polyisocyanate compounds such as tetramethylene diisocyanate, 1,6-hexamethylene diisocyanate, 1,10-decamethylene diisocyanate, trimethylhexamethylene diisocyanate, dimer acid diisocyanate, lysine diisocyanate; xylylene diisocyanate, tolylene diisocyanate, phenylene Diisocyanate, 4,4'-diphenylmethane diisocyanate, triphenylmethane triisocyanate, 3-phenyl-2-ethylene diisocyanate, 4-methoxy-1,3-phenylene diisocyanate, 2,4-diisocyanate diphenyl ether,
4,4-diisocyanate diphenyl ether, 2,
Aromatic polyisocyanate compounds such as 4,6-tolylene triisocyanate, 2,4,4-triisocyanate diphenyl ether, naphthalene diisocyanate, 1,4-anthracene diisocyanate, cumene-2,4-diisocyanate; isophorone diisocyanate; 4,4'-methylenebis (cyclohexyl isocyanate), methylcyclohexane 2,4- (or 2,
Alicyclic polyisocyanate compounds such as 6) -diisocyanate and 4,4'-methylenebis (cyclohexyl isocyanate); and polymers (prepolymers) of these polyisocyanate compounds
Can also be used. These polyisocyanate compounds can be used alone or in combination of two or more. Among these, those preferably used in the present invention include tolylene diisocyanate,
4,4'-diphenylmethane diisocyanate and isophorone diisocyanate.

When producing the hydrophilic urethane resin (a),
If necessary, a hydroxyl group may be introduced by using a polyol compound, or an ether bond may be introduced by using polyethylene glycol (molecular weight: 6,000 or less), polypropylene glycol (molecular weight: 6,000 or less) to further improve hydrophilicity. it can.

Examples of the polyol compound include ethylene glycol, diethylene glycol, propylene glycol, dipropylene glycol, 1,2-butylene glycol, 1,3-butanediol, 1,10-decanediol, neopentyl glycol, 1,6 -Hexanediol, cyclohexanediol, 2-butene1,
4-diol, 3-cyclohexene-1,1-dimethanol, 3-methylene-1,5-pentanediol, (2
-Hydroxyethoxy) -1-propanool, hydrogenated bisphenol A, glycerin, polycaprolactone,
1,2,6-hexanetriol, trimethylolpropane, pentanetriol, trishydroxymethylaminomethane, 3- (2-hydroxyethoxy) -1,2-
Examples thereof include propanediol, pentaerythritol, dipentaerythritol, mannitol, and glucose.

Further, if necessary, an unsaturated group can be introduced into the molecular chain terminal of the hydrophilic urethane resin (a) to improve the curability. Therefore, the hydroxyl group and the ethylenically unsaturated group are added to the polyurethane resin. It is also possible to react a compound having a group or a compound having an ethylenically unsaturated group with an isocyanate group.

Examples of the compound having a hydroxyl group and an ethylenically unsaturated group include, for example, 2-hydroxyethyl (meth)
Acrylate, hydroxypropyl (meth) acrylate, allyl alcohol, butanediol mono (meth) acrylate, N-methylol (meth) acrylamide,
Propylene glycol mono (meth) acrylate, adducts of glycidyl (meth) acrylate and carboxyl group-containing compounds (eg, adipic acid, sebacic acid, azelaic acid, etc.), adducts of (meth) acrylic acid and epoxy compounds, etc. Can be mentioned.

Examples of the compound having an isocyanate group and an ethylenically unsaturated group include, for example, ethyl isocyanate methacrylate and diisocyanate (for example,
Monoadduct of isophorone diisocyanate) and hydroxyethyl (meth) acrylate.

In producing the hydrophilic urethane resin (a),
The reaction ratio between the diepoxy compound and the unsaturated carboxylic acid is generally from 1 / 1.8 to an equivalent ratio of epoxy group / carboxy group.
2.2, preferably in the range of 1/2 to 2.1. The reaction between the diepoxy compound and the unsaturated carboxylic acid can be carried out by a method known per se using, for example, an amine such as triethylamine, a quaternary ammonium salt such as tetrabutylammonium bromide as a catalyst.

The reaction ratio between the unsaturated group-containing diol compound and the polyisocyanate compound obtained by the above reaction is generally 1/3 to 3/1, preferably 1/2, in terms of the equivalent ratio of hydroxyl group to isocyanate group. ２2/1. The reaction between the unsaturated group-containing diol compound and the polyisocyanate compound can be performed by a method known per se. For example, both of these components are dissolved or dispersed in an inert organic solvent that does not inhibit the reaction, and a reaction catalyst such as tetrabutyltin or triphenylaluminum is used at about 20 to about 250 ° C. for about 10 minutes to about 24 hours.
It can be performed by reacting for a time. (B) Polymerization initiator Examples of the polymerization initiator used in the present invention include a photopolymerization initiator and a redox-based thermal polymerization initiator.

As the photopolymerization initiator, those conventionally known as those useful for the polymerization of ethylenically unsaturated compounds can be used without any particular limitation. Specifically, for example, benzoin, benzoin methyl ether,
Benzoin ethyl ether, benzoin butyl ether, diethoxyacetophenone, 2-hydroxy-2-
Methyl-1-phenylpropan-1-one, benzyldimethylketal, 1-hydroxycyclohexyl-phenylketone, 2-methyl-2-morpholino (4-thiomethylphenyl) propan-1-one, 2-benzyl-2
-Dimethylamino-1- (4-morpholinophenyl)-
Butanone, 2,4,6-trimethylbenzoyldiphenylphosphine oxide, 2,4,6-trimethylbenzoylphenylethoxyphosphine oxide, benzophenone, methyl O-benzoylbenzoate, hydroxybenzophenone, 2-isopropylthioxanthone,
2,4-dimethylthioxanthone, 2,4-diethylthioxanthone, 2,4-dichlorothioxanthone, 2,
4,6-tris (trichloromethyl) -S-triazine, 2-methyl-4,6-bis (trichloro) -S-triazine, 2- (4-methoxyphenyl) -4,6-bis (trichloromethyl)- S-triazine and the like can be mentioned. These photopolymerization initiators can be used alone or in combination of two or more.

In order to accelerate the photopolymerization reaction of these photopolymerization initiators, a photosensitization accelerator may be used in combination with the photopolymerization initiator. As a photosensitizer that can be used in combination,
For example, triethylamine, triethanolamine, methyldiethanolamine, methyl 4-dimethylaminobenzoate, ethyl 4-dimethylaminobenzoate, isoamyl 4-dimethylaminobenzoate, (2-dimethylamino) ethyl benzoate, Michler's ketone, 4,4 Tertiary amines such as'-diethylaminobenzophenone; alkyl phosphines such as triphenylphosphine; thioethers such as β-thiodiglycol. These photosensitizers can be used alone or in combination of two or more.

Further, as the redox-based thermal polymerization initiator, conventionally known ones can be used. For example, an oxidizing agent and a reducing agent capable of performing radical polymerization at a relatively low temperature of about -10 ° C. to 50 ° C. A polymerization initiator composed of a combination of agents is suitably used.

Examples of the oxidizing agent include organic peroxides such as benzoyl peroxide, methyl ethyl ketone peroxide, dicumyl peroxide, t-butyl perbenzoate and cumene hydroperoxide; and peroxodioxides such as ammonium peroxodisulfate and potassium peroxodisulfate. Sulfates; hydrogen peroxide and the like. Examples of the reducing agent include bisulfites such as sodium bisulfite; divalent iron salts such as ferrous sulfate and ferrous chloride; amines such as N, N-dimethylaniline and phenylmorpholine; Metal naphthenates such as cobalt naphthenate, manganese naphthenate, and copper naphthenate can be exemplified.

Among these redox-based thermal polymerization initiators, those which can be used particularly advantageously in the present invention are those in which the oxidizing agent is a peroxodisulfate or hydrogen peroxide, and the reducing agent is a bisulfite or a divalent salt. It is a combination of iron salts. (C) Water-soluble polymer polysaccharide The water-soluble polymer polysaccharide used in the present invention is water-soluble and insoluble or hardly soluble in water when contacted with an alkali metal ion or a polyvalent metal ion in an aqueous medium. A high molecular weight polysaccharide capable of transforming into a soluble gel, generally having a number average molecular weight in the range of about 3000 to about 2,000,000, and a water-soluble prior to contacting with alkali metal ions or polyvalent metal ions. Sexual condition, usually at least about 1
Those exhibiting a solubility of 0 g / l (25 ° C.) are preferably used.

Specific examples of the water-soluble polysaccharide having such properties include alkali metal salts of alginic acid and carrageenan.

In the case of carrageenan, these water-soluble high molecular weight polysaccharides are dissolved in an aqueous medium, and then contacted with an alkali metal ion such as potassium ion, sodium ion or the like. In the case, alkaline earth metal ions such as magnesium ion, calcium ion, strontium ion and barium ion; or at least one polyvalent metal ion among other polyvalent metal ions such as aluminum ion, cerium ion and nickel ion It can be gelled by contact with The concentration of the alkali metal ion or polyvalent metal ion at which gelation occurs varies depending on the type of the water-soluble polymer polysaccharide and the like, but is generally 0.01 to 5 mol / mol.
1, especially in the range of 0.01 to 3 mol / l.

The mutual use ratio of each of the above components (a), (b) and (c) is not strictly limited, and can be varied over a wide range according to the type of each component. Is the hydrophilic urethane resin 100 of the component (a).
It is appropriate that the components (b) and (c) be used in the following proportions with respect to parts by weight (the preferred range is in parentheses).

(B) Polymerization initiator: 0.1 to 10 parts by weight (0.3 to 5 parts by weight) (c) Water-soluble high molecular polysaccharide: 0.5 to 15 parts by weight (1
In addition, among the polymerization initiators (b), the redox-based thermal polymerization initiator is used in combination with an oxidizing agent and a reducing agent, and the mixing ratio of both is generally 5: 1-1:
5, suitably in the range of 2.5: 1 to 1: 2.5.

The granular molded product produced according to the present invention comprises:
Generally, it has a specific gravity in the range of 1.0 to 1.03, which is almost the same as water, but when it is used in a fluidized-bed large reactor or a reactor having a complicated structure, it is easy to move downward. It may be difficult to make it flow uniformly. In such a case, it is necessary to reduce the specific gravity of the granular molded product, and the specific gravity of the granular molded product can be reduced by adding inorganic minute hollow beads to the composition of the granular molded product.

The inorganic fine hollow beads used for adjusting the specific gravity of the granular molded product preferably contain silicon as a main component, particularly silicate or alumina silica as a main component, and have a hollow structure in the particles. Has a specific gravity of 0.3 to 0.7
Can be used. Specific examples include hollow glass beads and hollow celite.

Micro hollow beads generally have a size of 1 to 200 μm.
m, preferably can have an average particle size in the range of 3 to 50 [mu] m, also hollow size varies depending desired specific gravity, typically of a diameter of the bead _^1/2 or less, preferably it is advantageous is _^1/3 or less.

The ones hollow diameter is greater than _^1/2, there is a possibility that or disrupted by immobilizing molded particles in the resulting compression strength decreases.

The minute hollow beads are used in an amount of 0.1 to 10 parts by weight based on 100 parts by weight of the component (a) in the aqueous liquid composition comprising the components (a), (b) and (c). By adding, the specific gravity of the obtained granular molded product is 0.9.
It can be adjusted to fall within the range of 0 to 1.00.

On the other hand, when it is desired to increase the specific gravity of the granular molded product, the specific gravity of glass beads, fine barium sulfate, etc. is 1
The specific gravity of the granular composition is from 1 to 1 by adding the inorganic powder to the composition of the granular molded product in a ratio of 0.1 to 30 parts by weight based on 100 parts by weight of the component (a). .2
The specific gravity can be adjusted to fall within the range of 4.

The components (a) to (c) described above and the above-mentioned additives for adjusting specific gravity, which are added as necessary, are dissolved or dispersed in an aqueous medium to form an aqueous liquid composition. Prepared. Generally, the solid content concentration of the liquid composition is suitably in the range of 5 to 30% by weight. In addition,
When a redox-based thermal polymerization initiator is used as the component (c), one of an oxidizing agent and a reducing agent may be used, if necessary,
In an aqueous medium containing an alkali metal ion or a polyvalent metal ion, for example, 0.05 to 5% by weight, preferably 0.1 to 5% by weight.
You may make it contain in the density | concentration within the range of 1-2 weight%.

The aqueous liquid composition thus prepared is then added dropwise to an aqueous medium containing an alkali metal ion or a polyvalent metal ion of the type described above,
Or when obtaining a granular material having an average particle size of 5 mm or more,
The liquid composition is continuously poured for a predetermined time on the surface of the aqueous medium to grow droplets to a desired particle size, and then the droplets are allowed to settle, thereby causing the liquid composition to gel in a granular form. Can be

The aqueous liquid composition can be dropped into an aqueous medium containing an alkali metal ion or a polyvalent metal ion by, for example, a method of dropping the liquid composition from the tip of a syringe, or a method using centrifugal force. A method of scattering the composition into particles,
It can be carried out by a method such as a method of atomizing the liquid composition from the tip of the spray nozzle to make it into granules and dropping it. The addition of the aqueous liquid composition to the surface of the aqueous medium,
It can be performed by continuously supplying a thin liquid stream from a nozzle opening having a desired hole diameter. The size of the droplet is
It can be freely changed according to the particle size desired for the final granular immobilized product.
To about 5 mm, preferably about 0.5 to about 4 mm.
Conveniently, the droplets are in the range of ３3 cm.

The granular gel produced as described above is
The hydrophilic resin in the granular gel is cured by photopolymerization or thermal polymerization in a state of being dispersed in the aqueous medium as it is or after being separated from the aqueous medium. Thereby, the granular gel is substantially insoluble in water, and a granular molded product for immobilizing enzymes or microorganisms having high mechanical strength can be obtained.

When the above-mentioned curing is carried out by photopolymerization, the wavelength of the actinic ray that can be used varies depending on the kind of the hydrophilic resin contained in the granular gel, but generally about 25%.
Advantageously, a light source emitting light of a wavelength in the range from 0 to about 600 nm is used for irradiation. 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 according to the light intensity of the light source, the distance from the light source, and the like, but can be generally in the range of about 0.5 to about 10 minutes.

When the hydrophilic resin is cured by thermal polymerization, the granular gel contains a redox-type thermal polymerization initiator. The resin is cured before the target strength is obtained, but may be cured in a constant temperature atmosphere, if necessary. The temperature of the constant temperature atmosphere is generally 0 ° C to 50 ° C, particularly 20 ° C to 4 ° C.
A temperature in the range of 0 ° C. is preferred. In order to obtain the required mechanical strength, it is desirable to take at least 10 to 30 minutes for heat curing.

The curing of the hydrophilic resin can be carried out by a combination of photopolymerization and thermal polymerization.

The granular gel which has been thus cured by photopolymerization and / or thermal polymerization can be washed with water or an aqueous buffer solution and stored as it is or by freeze-drying.

The granular molded product for immobilizing enzymes or microorganisms produced by the present invention has a surface structure particularly suitable for the adhesion of enzymes or microorganisms, and allows the enzyme or microorganisms to adhere in large amounts. Microorganisms that can be attached to the carrier can be any of anaerobic microorganisms and aerobic microorganisms, and examples of the types of microorganisms include molds such as Aspargilus, Penicillium, and Fusarium;
Yeasts such as Saccharomyces, Phaffia, and Candida; and bacteria such as Zymomonas, Pseudomonas, Nitrosomonas, Nitrobacter, Paracoccus, Vibrio, Methanosarcina, and Bacillus.

When the curing temperature of the hydrophilic resin is low, such as room temperature, the enzymes and microorganisms can be prepared in advance by (a)
It may be mixed and immobilized in an aqueous liquid comprising the components (b) and (c).

Thus, according to the method for producing a particulate molded product for immobilization of the present invention, the particulate adhesive product has the same adhesion to enzymes or microbial cells as a granular molded product produced using a conventional hydrophilic urethane resin. In addition, a granular molded product in which the strength of the granular molded product is greatly improved can be obtained. Moreover, the granular molded product provided by the method of the present invention is excellent in fluidity in water, and therefore is most suitable for use in a fluidized-bed bioreactor or a stirred fermenter, etc. It is also possible to apply to a floor type bioreactor, a fermenter, and the like.

[0053]

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to the following examples. In the production examples, examples and comparative examples, “parts” and “%” are based on weight. Production example of hydrophilic urethane Production example 1 1500 parts of toluene in a four-necked flask, Denacol EX
1821 parts of -821 (manufactured by Nagase Kasei Kogyo Co., Ltd.), 0.1 part of hydroquinone and 1 part of tetraethylammonium bromide are stirred in, 720 parts of acrylic acid are added dropwise at 110 ° C. for 1 hour while blowing air, and 8 parts are added.
The reaction was carried out while maintaining the temperature for a time to obtain a solution of an unsaturated group-containing diol. Further, while blowing air, 888 parts of isophorone diisocyanate is added to the solution, stirred, and kept at 100 ° C. for 8 hours to react. After confirming that almost no residual isocyanate group was observed, 5400 parts of deionized water was added, and the pressure in the reaction vessel was reduced by a vacuum pump at 70 ° C. for 1 hour to remove toluene, and the ethylenically unsaturated group was removed. Containing hydrophilic urethane resin (unsaturation degree 2.
8 mol / kg; an average molecular weight of 3560) was obtained as a 40% aqueous solution (A). Production Example 2 1600 parts of toluene in a four-necked flask, Denacol EX
2300 parts of -841 (manufactured by Nagase Kasei Kogyo Co., Ltd.), 0.1 part of hydroquinone and 1 part of tetraethylammonium bromide are added and stirred, and while blowing air, 432 parts of acrylic acid are added dropwise at 110 ° C. over 1 hour.
The reaction was carried out while maintaining the temperature for a time to obtain a solution of an unsaturated group-containing diol. Further, while blowing air, 888 parts of isophorone diisocyanate was added to the solution, and the mixture was stirred and reacted at 100 ° C. for 8 hours. Further acrylic acid 2
-232 parts of hydroxyethyl was placed in a reaction vessel and reacted at 80 ° C for 3 hours while blowing air. After confirming that almost no isocyanate group remained, 5800 parts of deionized water was added, and 70 ° C was added. The pressure in the reaction vessel was reduced by a vacuum pump for 1 hour to remove toluene, and the ethylenically unsaturated group-containing hydrophilic urethane resin (unsaturation degree 2.
1 mol / kg; an average molecular weight of 3850) (about 40% aqueous solution (B)) was obtained. Production Example 3 1000 parts of toluene and celoxide 2 in a four-necked flask
081 (manufactured by Daicel Chemical Industries, Ltd.), 0.1 part of hydroquinone and 1 part of tetraethylammonium bromide are stirred and added, and while blowing air, 144 parts of acrylic acid are added dropwise at 110 ° C. over 1 hour, and further 8 hours. The reaction was performed while maintaining the temperature to obtain a solution of an unsaturated group-containing diol. Further, while blowing air, polyethylene glycol (average molecular weight: about 600)
And 444 parts of isophorone diisocyanate were added, stirred, and reacted at 100 ° C. for 8 hours. Further, 310 parts of ethyl isocyanate methacrylate is put into the reaction vessel,
The reaction was carried out at 80 ° C. for 3 hours while blowing air. After confirming that almost no isocyanate group remained, 3800 parts of deionized water was added, and the pressure inside the reaction vessel was reduced by a vacuum pump at 70 ° C. for 1 hour. To remove the ethylenically unsaturated group-containing hydrophilic urethane resin (unsaturation degree 1.6 mol / kg; average molecular weight 2500).
% Aqueous solution (C) was obtained. Production Example 4 (for comparison) polyethylene glycol (average molecular weight: about 2000) 20
00 parts and 444 parts of isophorone diisocyanate were placed in a reaction vessel and reacted at 80 ° C. for 2 hours. Further, 232 parts of 2-hydroxyethyl acrylate and 2 parts of hydroquinone are put in a reaction vessel, and while blowing air,
C. for 3 hours. After confirming that almost no isocyanate group remained, deionized water 40
Of a hydrophilic urethane resin containing an ethylenically unsaturated group (unsaturation degree: 0.75 mol / kg; average molecular weight: about 2
680) was obtained. Production Example 5 (for comparison) Polyethylene glycol (average molecular weight: about 600) 120
0 parts and 666 parts of isophorone diisocyanate were placed in a reaction vessel and reacted at 80 ° C. for 2 hours. Further, 232 parts of 2-hydroxyethyl acrylate and 2 parts of hydroquinone are put in a reaction vessel, and the mixture is blown at 80 ° C. while blowing air.
For 3 hours, and after confirming that almost no isocyanate group remains, deionized water 310
0 parts by weight, and an ethylenically unsaturated group-containing hydrophilic urethane resin (unsaturation degree: 0.95 mol / kg; average molecular weight: about 21
00) of about 40% aqueous solution (E). Example 1 100 parts of the urethane resin solution (A) of Production Example 1, 2 parts of benzoin isobutyl ether, 100 parts of distilled water and 2%
100 parts of an aqueous sodium alginate solution was mixed well, and the resulting aqueous liquid composition was dropped into a 5% aqueous solution of calcium chloride from a liquid level of about 10 cm from the tip of a syringe. As a result, granules having a particle size of about 2 mm were obtained. Was. The granules were placed on a flat petri dish, and irradiated with ultraviolet light from the upper and lower surfaces of the petri dish with a 1 kW high-pressure mercury lamp for 30 seconds to obtain a granulated product. Example 2 A granular molded product was obtained in the same manner as in Example 1 except that the urethane resin solution (B) of Production Example 2 was used instead of the urethane resin solution (A) used in Example 1. Example 3 A granular molded product was obtained in the same manner as in Example 1 except that the urethane resin solution (C) of Production Example 3 was used instead of the urethane resin solution (A) used in Example 1. Example 4 100 parts of the urethane resin solution (A) of Production Example 1, 1 part of ammonium peroxodisulfate, 1 part of sodium bisulfite
Parts, 100 parts of distilled water and 100 parts of a 2% aqueous sodium alginate solution were mixed well, and the resulting aqueous liquid composition was dropped into a 5% aqueous calcium chloride solution from a tip of a syringe at a liquid level of about 10 cm. Granules having a diameter of about 2 mm were obtained. This granular material is kept at 30 ° C. for 30 minutes.
This was left for a minute to obtain a granular molded product. Example 5 100 parts of the urethane resin solution (A) of Production Example 1, 2 parts of benzoin isobutyl ether, 1 part of ammonium peroxodisulfate, 1 part of sodium bisulfite, 100 parts of distilled water and 100 parts of a 2% aqueous sodium alginate solution were mixed well. The resulting aqueous liquid composition was dropped into a 5% aqueous solution of calcium chloride from the tip of a syringe at a liquid level of about 10 cm, and a granular material having a particle size of about 2 mm was obtained. The granular material was irradiated with ultraviolet light from the upper surface and the lower surface with a 1 kW high-pressure mercury lamp for 30 seconds, and further left at 30 ° C. for 30 minutes to obtain a granular molded product. Comparative Example 1 A granular molded product was obtained in the same manner as in Example 1 except that the urethane resin solution (D) of Production Example 4 was used instead of the urethane resin solution (A) used in Example 1. Comparative Example 2 A granular molded product was obtained in the same manner as in Example 1 except that the urethane resin solution (E) of Production Example 5 was used instead of the urethane resin solution (A) used in Example 1. Comparative Example 3 A granular molded product was obtained in the same manner as in Example 4 except that the urethane resin solution (D) of Production Example 4 was used instead of the urethane resin solution (A) used in Example 4.

Examples 1 to 5 and Comparative Examples 1 to 3 obtained above
Table 1 shows the compressive strength and bacterial cell adhesion of the granular molded product of Example 1. The evaluation of bacterial cell adhesion was performed as follows.

The granules were placed in a 500 ml Erlenmeyer flask, and then each of them was GY-10 medium (yeast extract 1).
g / L, glucose 100 g / L)
L was added. To this was added Zymomonas mobilis IFO 13756 at a concentration of 2%, and 30
Activated culture was carried out at 24 ° C. for 24 hours. After the activation, the surface of each of the immobilized granular formed products was washed with distilled water, and then the activated fermentation solution was replaced with a fresh medium, followed by standing culture for 24 hours, and the ethanol concentration was measured.

[0056]

[Table 1]

[0057]

According to the method for producing a granular molded product for immobilizing enzymes or microorganisms of the present invention, since a hydrophilic urethane resin having a high unsaturated group concentration is used, the compressive strength of the obtained granular molded product is extremely high. In addition, it is possible to obtain a granular molded product having excellent adhesion of enzymes or microbial cells.

Claims (3)

[Claims] 1. A hydrophilic urethane having at least two ethylenically unsaturated bonds in one molecule obtained by reacting (a) an adduct of a diepoxy compound and an ethylenically unsaturated carboxylic acid with a polyisocyanate compound. resin,
An aqueous liquid composition comprising (b) a polymerization initiator, and (c) a water-soluble polymer polysaccharide capable of gelling by contact with an alkali metal ion or a polyvalent metal ion,
The composition is gelled by dropping into an aqueous medium containing an alkali metal ion or a polyvalent metal ion, or is continuously poured onto the surface of the aqueous medium for a predetermined time to form a desired droplet. After growing to a particle size of, the droplets are allowed to settle and gel, and then the resulting granular gel is photopolymerized and / or thermally polymerized to cure the hydrophilic resin in the granular gel. A method for producing a granular molded product for immobilizing an enzyme or a microbial cell, characterized by the following. 2. The hydrophilic urethane resin (a) has a number-average molecular weight of 400 to 50,000 and an unsaturated group content of 0.
The method according to claim 1, wherein the amount of the resin is from 04 to 5 mol / kg. 3. The method according to claim 1, wherein the polymerization initiator (b) is a photopolymerization initiator or a redox-based thermal polymerization initiator.