JP2001190274A - Method for producing granular formed product for immoibilizing enzyme or microorganism cell - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing a granular formed product capable of improving properties for adhering enzymes or cells of microorganisms to the granular formed product for immobilizing the enzymes or cells of microorganisms. SOLUTION: This method for producing a granular formed product for immobilizing enzymes or cells of microorganisms is to drop a water-based liquid composition containing (a) a hydrophilic resin having at least two ethylenic unsaturated bonds in the molecule, (b) a polymerization initiator and (c) an acidic aqueous solution of chitosan into a basic water-based medium to bring the composition to granular gel or continuously pouring the composition on the surface of the water-based medium for a predetermined time to grow droplets to a predetermined granular size and precipitate and bring the droplets to gel, subsequently optically polymerize and/or thermally polymerize the obtained granular gel and cure 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). However, conventional photocurable resins for immobilizing enzymes or microbial microbial cells or granular molded products mainly composed of thermosetting resins are microbial cells having a negative charge, such as Pseudomonas spp., Paracoccus spp. There is a problem that adhesion to bacteria such as Nitrosomonas sp. Is poor and sufficient biological activity cannot be obtained, and improvement thereof is desired. In addition, the enzyme was hardly adsorbed on the granular molded article carrier, so that it was extremely difficult to fix and adhere the enzyme.

[0004]

SUMMARY OF THE INVENTION The main object of the present invention is to produce a granular molded article capable of improving the adhesion of an enzyme or a microorganism to a granular molded article on which an enzyme or a microorganism is immobilized. Is to provide a way.

[0005]

Means for Solving the Problems The present inventors have conducted intensive studies in order to achieve the above-mentioned object, and as a result, as a constituent material of a granular carrier for immobilizing enzymes or microorganisms using a hydrophilic resin, By using chitosan together, they found that the above problems could be solved at once,
The present invention has been completed.

Thus, according to the present invention, there are provided (a) a hydrophilic resin having at least two ethylenically unsaturated bonds in one molecule, (b) a polymerization initiator, and (c) an aqueous chitosan aqueous solution. The aqueous liquid composition is dropped into a basic aqueous medium to gel the composition into granules, or is continuously poured onto the surface of the aqueous medium for a predetermined time to form the droplets into the desired particles. After growing to a diameter, the droplets are settled and gelled, and then the resulting granular gel is photopolymerized and / or thermally polymerized to cure the hydrophilic resin in the granular gel. The present invention provides a method for producing a granular molded product for immobilizing an enzyme or a microbial cell.

The aqueous liquid composition may further contain, if necessary, (d) an amine compound having at least one ethylenically unsaturated bond in one molecule.

Hereinafter, the present invention will be described in more detail. (A) Hydrophilic resin In the present invention, the hydrophilic resin (a) having at least two ethylenically unsaturated bonds in one molecule used for the production of the granular carrier for immobilizing enzymes or microorganisms includes: , Generally from 300 to 30,000, preferably from 500 to 200
A ionic or nonionic hydrophilic group having a number average molecular weight in the range of 00 and sufficient to disperse evenly in the aqueous medium, such as hydroxyl, amino, carboxyl, phosphate, sulfonic acid groups And those containing an ether bond or the like are preferably used. As such a hydrophilic resin (a), those already known as a photocurable resin for encapsulation and immobilization can be similarly used (for example, Japanese Patent Publication No. 55-4).
No. 0, Japanese Patent Publication No. 55-20676, Japanese Patent Publication No. 62
19837). Representative examples include those described below. (I) A compound having a photopolymerizable or thermally polymerizable ethylenically unsaturated group at both ends of a polyalkylene glycol: For example, 1 mole of a polyethylene glycol having a molecular weight of 400 to 6000 is converted to 2 moles of both ends hydroxyl groups with 2 moles of (meth) acrylic acid. Esterified polyethylene glycol di (meth) acrylates.

[0009] Polypropylene glycol di (meth) acrylates having a molecular weight of 200 to 4000, wherein the hydroxyl groups at both ends of 1 mol of polypropylene glycol are esterified with 2 mol of (meth) acrylic acid.

The hydroxyl groups at both terminals of 1 mol of polyethylene glycol having a molecular weight of 400 to 6000 are urethanized with 2 mol of a diisocyanate compound such as tolylene diisocyanate, xylylene diisocyanate, or isophorone diisocyanate. An unsaturated polyethylene glycol urethane compound to which 2 mol of a saturated monohydroxyethyl compound has been added.

The hydroxyl groups at both terminals of 1 mole of polypropylene glycol having a molecular weight of 200 to 4000 are urethanized with 2 moles of a diisocyanate compound such as tolylene diisocyanate, xylylene diisocyanate, or isophorone diisocyanate. Unsaturated polypropylene glycol urethanates to which 2 moles of a saturated monohydroxy compound are added, and the like. (Ii) High acid value unsaturated polyester resin: for example, an acid value obtained by esterification of a polyhydric alcohol with a polyvalent carboxylic acid component containing an unsaturated polyvalent carboxylic acid is 40 to 20.
0 unsaturated polyester salts and the like. (Iii) High acid value unsaturated epoxy resin: For example, an acid value of 40 obtained by adding an acid anhydride to a hydroxyl group remaining in an addition reaction product of an epoxy resin and an unsaturated carboxyl compound such as (meth) acrylic acid. ~ 200 unsaturated epoxy resins and the like. (Iv) anionic unsaturated acrylic resin: for example, a carboxyl group or a phosphate group obtained by copolymerizing at least two kinds of (meth) acrylic monomers selected from (meth) acrylic acid and (meth) acrylic acid ester as well as/
Or a resin in which a thermally polymerizable ethylenically unsaturated group is introduced into a copolymer containing a sulfonic acid group. (V) Cationic unsaturated acrylic resin For example, obtained by subjecting a copolymer of a (meth) acrylate ester containing an unsaturated amino compound unit in an amount of more than 5% by weight to an addition reaction of glycidyl (meth) acrylate or the like. Such as unsaturated acrylic resin. (Vi) Unsaturated celluloses For example, an addition reaction product of a water-soluble cellulose and an unsaturated glycidyl compound such as glycidyl (meth) acrylate or an unsaturated acid anhydride. (Vii) Unsaturated polyamide: for example, an addition reaction of an adduct of a diisocyanate such as tolylene diisocyanate or xylylene diisocyanate with an ethylenically unsaturated hydroxy compound such as 2-hydroxyethyl acrylate to a water-soluble polyamide such as gelatin. And unsaturated polyamides obtained by

The hydrophilic resins as exemplified above can be used alone or in combination of two or more.

Among these hydrophilic resins, those which can be particularly advantageously used in the present invention are compounds having an ethylenically unsaturated group capable of photopolymerization or heat polymerization at both terminals of the polyalkylene glycol of the above (i). Typical examples are ENT-100 from Kansai Paint Co., Ltd.
0, ENT-2000, ENT-4000, ENTG-
2000, ENTG-3800, and the like. (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 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 by 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; 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 thermal polymerization initiators, those which can be particularly advantageously used 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) Chitosan acidic aqueous solution Chitosan is also called β-1,4-poly-D-glucosamine, and is generally represented by the following chemical formula obtained by deacetylating chitin with a concentrated alkaline solution. It is the white powdery substance shown.

[0019]

Embedded image

The chitosan used in the present invention is preferably of a low molecular weight type having an average molecular weight in the range of 10,000 to 230,000.
It is used as a chitosan acidic aqueous solution dissolved in an aqueous solution of an acid such as acetic acid or citric acid or an aqueous solution of a mixture of these acids.

The chitosan acidic aqueous solution can be gelled by dropping it into a basic aqueous medium. (D) Amine compound As the amine compound (d) having at least one ethylenically unsaturated bond in one molecule which can be used as required in the present invention, at least one amine compound in one molecule may be used.
And preferably a compound having one or two polymerizable double bonds and a secondary or tertiary amino group, specifically, for example, N, N-dimethylaminoethyl (meth) Nitrogen-containing alkyl (meth) acrylates such as acrylate, N, N-diethylaminoethyl (meth) acrylate and Nt-butylaminoethyl (meth) acrylate; N, N-dimethylaminoethyl (meth) acrylamide, N, N- Nitrogen-containing polymerizable unsaturated amides such as dimethylaminopropyl (meth) acrylamide; alkanolamines (eg, N-methyldiethanolamine, N, N-dimethylethanolamine, triethanolamine, etc.) and isocyanate group-containing ethylenically unsaturated monomers ( For example, ethyl isocyanate methacrylate, isophorone diiso Such adducts of a diisocyanate compound such as Aneto and (meth) monoadduct of 2-hydroxyethyl acrylate) can be cited.

Of these amine compounds (d), particularly preferred in the present invention are adducts of alkanolamines with isocyanate group-containing ethylenically unsaturated monomers.

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

(B) Polymerization initiator: 0.1 to 5 parts by weight (0.3 to 3 parts by weight) (c) Chitosan: 0.5 to 15 parts by weight (1 to 8 parts by weight) (d) Amine compound : 0 to 100 parts by weight (5 to 50 parts by weight) Among the polymerization initiators of the above (b), the redox-based thermal polymerization initiator is used in combination with an oxidizing agent and a reducing agent. The ratio is generally 5: 1 to 1: in molar ratio.
5, suitably within the range of 2.5: 1 to 1: 2.5.

The components (a) to (d) described above are dissolved or dispersed in an aqueous medium to prepare an aqueous liquid composition. Generally, the solid content concentration of the liquid composition is suitably in the range of 5 to 30% by weight.

The aqueous liquid composition thus prepared is then dropped into a basic aqueous medium, or when a granular material having an average particle size of 5 mm or more is obtained, the basic aqueous medium surface The liquid composition is continuously poured for a predetermined period of time to grow the droplets to a desired particle size, and then the droplets are allowed to settle, whereby the liquid composition is gelled in a granular form.

The basic aqueous medium can be prepared, for example, by adding a hydroxide such as sodium hydroxide or potassium hydroxide in water;
It can be carried out by dissolving carbonates such as potassium carbonate and calcium carbonate; bicarbonates such as sodium bicarbonate; water-soluble amines such as alkanolamine; and tris (hydroxymethyl) aminomethane.

The granular gel formed as described above is dispersed in a basic aqueous medium as it is, or after being separated from the basic aqueous medium, the granular gel is subjected to photopolymerization and / or
Alternatively, the hydrophilic resin in the granular gel is cured by thermal polymerization.

When the granular gel is cured by photopolymerization, the granular gel is irradiated with an actinic ray. The wavelength of the actinic ray used at this time varies depending on the type of the hydrophilic resin contained in the granular gel and the like.
It is advantageous to use a light source emitting light of a wavelength in the range of 00 nm for the irradiation.

Examples of such a light source include a low-pressure mercury lamp, a high-pressure mercury lamp, a fluorescent lamp, a xenon lamp, a carbon arc lamp, and sunlight. The irradiation time needs to be changed according to the light intensity of the light source, the distance from the light source, etc.
Generally, it can be 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 granular gel which has been thus cured by photopolymerization or thermal polymerization can be washed with water or a buffered aqueous solution and stored as it is or by freeze-drying.

The granular product for immobilizing enzymes or microbial cells produced by the present invention uses chitosan, which is a cationic high molecular polysaccharide, as a constituent component, so that enzymes or microorganisms having a negative charge can be used. Can be attached in large quantities. Examples of the enzyme that can be attached to the carrier include glucose oxidase, β-galactosidase, catalase, invertase, and lipase.The microorganism can be any of anaerobic microorganisms and aerobic microorganisms. For example, specifically, for example, molds such as Aspargillus, Penicillium, and Fusarium, yeasts such as Saccharomyces, Phaffia, and Candida; Zymomonas, Pseudomonas, Nitrosomonas, and Nitrobactor, Bacteria such as Paracoccus, Vibrio, Methanosarcina, and Bacillus can be mentioned. The granular molded product produced according to the present invention is particularly effective in improving the adhesion to microorganisms of the genera Pseudomonas, Nitromonas and Paracoccus.

Further, according to the method of the present invention, a granular molded product for immobilizing an enzyme or a microorganism can be produced by an extremely simple operation, and continuous production is also possible. Furthermore, the granular molded product produced by the method of the present invention has the advantage that the strength of the carrier is significantly improved over the carrier of chitosan alone, and that it can be used even in a plant requiring physical strength.

[0035]

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 Examples and Comparative Examples, “parts” and “%” are based on weight.

Example 1 Polyethylene glycol 20 having a number average molecular weight of about 4000
00 parts, 1 mol of isophorone diisocyanate (222
g) and 1 mol of 2-hydroxyethyl methacrylate (1
30 g) of the mixture, 100 parts of an unsaturated group-containing hydrophilic obtained by reacting the mixture, 100 parts of a 1.5% aqueous solution of chitosan acid and 2 parts of benzoin butyl ether are mixed well. This hydrophilic resin-chitosan mixture was poured into a 1 M aqueous solution of sodium hydroxide from a nozzle at the tip of a syringe at a liquid level of about 10 μm.
By dropping the powder from a cm, a granular material having a particle size of about 4 mm was obtained. Further, the granules are placed in a Petri dish, and a wavelength of 30
Irradiation with actinic light of 0 to 400 nm for 3 minutes resulted in a granular carrier having a compressive strength of 20 kg / cm ² .

Example 2 Polyethylene glycol 40 having a number average molecular weight of about 4000
00 parts and 444 parts of isophorone diisocyanate
It was put in a container and reacted at 80 ° C. for 2 hours. In addition,
132 parts of 2-hydroxyethyl lylate and hydroquino
2 parts into a reaction vessel, while blowing air at 80 ° C.
For 3 hours to obtain an unsaturated group-containing hydrophilic resin. This
100 parts of an unsaturated group-containing hydrophilic resin, N, N-dimethyl
Aminoethyl acrylate 30 parts, peroxodisulfuric acid
1.3 parts of ammonium, 1.3 parts of anhydrous sodium bisulfite
Parts and 100 parts of 1.5% chitosan acidic aqueous solution
Aqueous composition obtained by 1M sodium hydroxide
In the aqueous solution, the liquid level is about 10 cm from the tip of the syringe.
Upon dripping, a granular material having a particle size of about 4 mm was obtained.
Leave the granules at 30 ° C. for 30 minutes and compress
Strength 20kg / cm ^TwoWas obtained.

Comparative Example 1 100 parts of the same hydrophilic resin as used in Example 1, 1 part of 1% aqueous sodium alginate solution and 2 parts of benzoisobutyl ether are mixed well. The photocurable resin-alginic acid mixed solution was dropped into a 1M-calcium chloride solution from a nozzle at the tip of a syringe at a liquid level of 10 cm to obtain a granular material having a particle size of about 4 mm. Further, the granules were placed in a Petri dish and irradiated with actinic rays having a wavelength of 300 to 400 nm for 3 minutes. The compressive strength was 25 kg / c.
m ² of granular support was obtained.

Comparative Example 2 The same unsaturated group-containing hydrophilic resin 10 as used in Example 2
0 parts, N, N-dimethylaminoethyl acrylate 30
An aqueous liquid composition obtained by thoroughly mixing 1.3 parts of ammonium peroxodisulfate, 1.3 parts of anhydrous sodium bisulfite and 100 parts of a 1% aqueous sodium alginate solution was added to a 1M aqueous solution of calcium chloride using a syringe. When the solution was dropped from the tip from a liquid level of about 10 cm, a granular material having a particle size of about 4 mm was obtained. This granular material is directly used for 30
The mixture was left at 30 ° C. for 30 minutes to obtain a granular molded product having a compressive strength of 20 kg / cm ² . The carrier obtained in the above Example 1 and Comparative Example 1 was used for Paracoccus denitrificans IFO1.
The microorganisms were allowed to adhere to the surface of the carrier by allowing the suspension to stand at room temperature for 2 days in a 2442 denitrifying cell suspension, and then the denitrifying ability was measured.

In the measurement of the denitrification capacity, after washing 50 g of the microorganism-adhering carrier, the carrier was poured into 250 mL of artificial wastewater (nitrate nitrogen concentration: 400 mg / L) having the composition shown in Table 1 below.
The treatment was performed at 30 ° C. The processing capacity at that time is NO _x -N
(Nitrate nitrogen and nitrite nitrogen concentrations) were measured. As a result, as shown in FIG. 1, nitrogen was reduced to about 50 mg / L in 4 days in the case of using the granular molded product of Example 1, whereas in the case of using the granular molded product of Comparative Example 1, In four days, the nitrogen was reduced to only about 180 mg / L, and that of Example 1 was about 1.6 times higher in processing capacity. Furthermore, when the amount of bacterial cells attached to the carrier was compared by measuring the amount of ATP (adenosine triphosphate), the one of Example 1 showed about 7 times higher value.

In addition, 5 g of the carrier obtained in Example 2 and Comparative Example 2 was mixed with M / 1 containing 0.1% of invertase.
The solution was put into 100 mL of a 5-phosphate buffer (pH 5.5), adsorbed and immobilized at 5 ° C. for 1 hour, washed, and invertase activity was measured using sucrose as a substrate. as a result,
The specific activity with respect to the non-immobilized invertase was 50% in the granular molded product of Example 2, whereas it was 10% in the granular molded product of Comparative Example 2. Table 1: Composition of artificial wastewater 2.888 g KNO ₃ 0.184 g Na ₂ HPO ₄ .12H ₂ O 0.01 g NaCl 0.0047 g CaCl ₂ .2H ₂ O 0.0047 g MgSO ₄ .7H ₂ O 0.0167 g Tap Water 1L pH 7.0

[0042]

According to the method of the present invention for producing a granular molded article for immobilizing an enzyme or a microorganism, granulation is carried out using chitosan. Therefore, the carrier produced is a microorganism or enzyme having a negative charge due to the positive charge of chitosan. Has the property of being easily adsorbed. Therefore, the amount of microorganisms and enzymes adsorbed on the carrier can be increased, and therefore, by using the granular molded product produced by the method of the present invention, the speed of the microorganism reaction or enzyme reaction can be improved.

[Brief description of the drawings]

FIG. 1 is a graph showing the denitrifying activity of a carrier for immobilizing denitrifying bacteria using the granular molded products obtained in Example 1 and Comparative Example 1.

Continued on the front page F term (reference) 4B029 AA21 BB02 BB16 CC04 CC13 4B033 NA12 NA22 NB02 NB13 NB34 NB36 NB49 NB62 NC04 NC12 ND04 4J011 AA05 HA02 HB22 JB22 JB26 PA53 PA69 PA86 PA88 PA90 PA96 PB40 QA03 QA03 QA03 QA03 QA03 QA03 QA03 RA10 RA15 SA01 SA21 SA31 SA51 SA61 SA64 SA78 SA83 SA84 UA01 UA06 WA10 4J027 AA01 AB02 AB05 AB14 AB32 AC03 AC04 AC06 AC08 AC09 AD02 AE01 AE07 AG04 AG09 AG12 AG23 AG24 AG27 AH00 AH03 AJ01 AJ05 BA01 CBCB CB CB CB CB CB CB CB CB CB CB CB CB CB CB CB CB CB CB CB CB CD01

Claims (4)

[Claims] 1. An aqueous liquid composition comprising (a) a hydrophilic resin having at least two ethylenically unsaturated bonds in one molecule, (b) a polymerization initiator, and (c) an acidic aqueous solution of chitosan. The composition may be gelled into granules by dropping into a basic aqueous medium, or may be continuously poured onto the surface of the aqueous medium for a predetermined time to grow the droplets to a desired particle size. After that, the droplets are allowed to settle to form a gel, and then the obtained granular gel is subjected to photopolymerization and / or thermal polymerization to cure the hydrophilic resin in the granular gel, and is characterized by being an enzyme or a microbial cell. A method for producing a granular molded product for immobilization. 2. The method according to claim 1, wherein the aqueous liquid composition further comprises an amine compound (d) having at least one ethylenically unsaturated bond in one molecule. 3. The method according to claim 2, wherein the amine compound (d) is selected from nitrogen-containing alkyl (meth) acrylates, nitrogen-containing polymerizable unsaturated amides and adducts of alkanolamines and isocyanate group-containing ethylenically unsaturated monomers. Method. 4. The method according to claim 1, wherein the polymerization initiator (b) is a photopolymerization initiator or a redox thermal polymerization initiator.