JP6990906B2 - Porous molded body of mesoporous fine particles, carrier for supporting enzymes, enzyme complexes thereof, and methods for producing them. - Google Patents

Description

The present invention relates to a porous molded body of mesoporous fine particles, a carrier for carrying an enzyme, an enzyme complex thereof, and a method for producing these.

As an inorganic highly functional material, fine particles having natural micro or mesopores such as clay, zeolite, diatomaceous earth, and activated carbon have been known for a long time. These fine particles are not only used in industrial processes as molecular sieves, adsorbents, reaction catalysts, etc. due to their characteristics such as regular pores on the order of molecules, adsorptivity to specific molecules and ions, and catalytic ability resulting from the structure. , Has been used in various ways in ordinary life.

As an example of the use of zeolite, Patent Document 1 discloses a separation membrane in which zeolite is fixed on a breathable support as a separation membrane of a liquid mixture. Further, it is also disclosed that the support is made of a woven fabric, a knitted fabric, a braid, or a continuous foam made of a ceramic, a metal, or a synthetic resin.

Further, Patent Document 2 discloses a filter filter medium for an organic solvent or an organic detergent, which comprises an ultra-high molecular weight polyethylene resin and a fine filler having heat resistance. Further, it is also disclosed that the fine filler is one or more selected from zeolite, potassium titanate, titanium oxide, sepiolite, and activated carbon.

Further, Patent Document 3 discloses a hollow fiber membrane made of a polyamide resin. Further, it is also disclosed that the polyamide resin contains 5 to 100% by mass of filler particles, that is, about 4.7 to 50% by mass in the hollow fiber membrane.

Further, the present inventors have reported in Patent Document 4, a natural zeolite hollow fiber porous body, a zeolite membrane composite porous body, and a method for producing the same. This natural zeolite hollow fiber porous body is a wet-spun hollow fiber porous body composed of natural zeolite powder and 3-50% by weight of an organic polymer, and has a pore diameter of 0.01-100 microns. It is a multi-dimensional porous body in which pores of multiple diameters are distributed in a range, has water resistance that allows water to enter, and is characterized by exhibiting reversibly flexibility performance by keeping it in a water-containing state even after drying treatment. do.

As synthetic fine particles having mesopores, mesoporous substances such as mesoporous silica are known. Examples of the usage of this mesoporous substance include a method of immobilizing an enzyme or protein as a biocatalyst in these regular mesopores and using it as an immobilized support. For example, Non-Patent Document 1 describes mesoporous silica fine particles as an enzyme-immobilized support for separation of products and enzymes, reuse of enzymes, etc. in the production process of functional chemicals such as pharmaceuticals using enzymes. Disclose the usage.

Further, in recent years, a method of processing mesoporous silica into a molded body has been reported in order to make it easier to use. For example, Non-Patent Document 2 discloses a hollow filament molding method using mesoporous silica fine particle powder. Further, Non-Patent Document 3 discloses a pellet-shaped molding method.

Japanese Unexamined Patent Publication No. 2009-61369 Japanese Unexamined Patent Publication No. 2004-275845 JP-A-2010-104983 Japanese Unexamined Patent Publication No. 2012-72534

Nils Carlsson et al., "Enzymes Immobilized on Mesoporous Silica: Physical-Chemical Perspectives silica: A Physical-Chemical Perceptive, Vol. 20 Incecenc", Ad. , P. 339-360 Aria. Ali A. Rownaghi et al., "In situ Formation of a Monodispersed Sforicial Mesoporous Microge-Rosi-Rosi-Rosia-Toronica-Toronica-Torlon Hollow Fiber Complex , ChemSusChem, 2015, Volume 8, p. 3439-3450 Harsh Maheshwari et al., "Robust mesoporous silica compacts: multi-scale silica ceramics characterization". ) ”, Journal of Materials Science, 2016, Vol. 51, p. 4470-4480

Zeolites are aluminosilicates that have regularly arranged micropores and a part of Si, which is the main component in the skeleton structure, is replaced with Al, and this Si / Al ratio serves as a catalyst for zeolite. Not only does it affect the strength and function, but it also affects the affinity of the zeolite itself with the adsorption molecules, reaction molecules, and reaction solvents. The pores in zeolite are three-dimensional ordered pores of about 3 to 10 Å, whereas clay has a two-dimensional layered structure and diatomaceous earth has relatively large pores ranging in size from nano-sized to several micron. In addition to those having a skeleton mainly made of silica, activated carbon is mainly composed of carbon and has pores having a size of about 2 nm to 10 nm.

In addition to such naturally occurring porous materials, mesoporous silica, which is a silicate having ordered pores of 1 nm or more, which is difficult to obtain naturally, and mesoporous titanium oxide and carbon having titania as a skeleton instead of silica. In recent years, various functional materials having functions and pore sizes that cannot be obtained in nature, such as mesoporous carbon and hydrotalcite, which is a layered compound capable of adsorbing anions, have been developed. Since the functions of these fine particles are exhibited by contacting the reaction target on the surface of the particles, fine particles having a higher surface area have been promoted with the aim of improving the functions.

Mesoporous substances such as mesoporous silica are a group of porous particles having regular pores of about 1 to 50 nm, whereas zeolite has regular pores of 1 nm or less. Mesoporous silica was synthesized using layered silicate as a raw material using a quaternary alkylamine as a template, aluminosilicate in which a part of silica was replaced with alumina, and alumina or titania was used as a raw material instead of silica. , Mesoporous aluminum oxide, mesoporous titanium oxide, mesoporous carbon made of carbon, etc. have been synthesized.

All of these mesoporous substances are synthetic products using organic substances as templates, and since they can immobilize various organic substances such as proteins and enzymes in the regular pores, they are expected to be used in various fields. However, it is very expensive, and there are many submicron-sized fine particles, which makes separation and purification difficult, which is an issue of industrial usage.

On the other hand, most of the newly synthesized fine particles have a drawback that the dispersibility is low as they are as primary particles, and it becomes difficult to separate and recover them after the dispersibility is improved. In order to industrially utilize these functional fine particles, it is indispensable to form / member or separate them while maintaining the original surface area as much as possible. For example, since zeolite has poor self-sintering properties, pellet molding is generally performed via an inorganic or organic binder, but in either method, if the amount of the binder is small, the strength and water resistance are reduced. If the amount of the binder is large, the binder crushes the surface of the fine particles, which causes a problem that the original function is not sufficiently exhibited, and this problem is particularly large when an attempt is made to form a thin film.

As a molding method, Patent Document 1 describes a molding method such as secondary growth on a substrate from the stage of synthesis such as hydrothermal synthesis as one of the membrane forming methods of a separation membrane in which zeolite is fixed. As described, this method tends to be less plastic, although it is obtained, and can only be adopted in the case of specific fine particles capable of hydrothermal synthesis.

On the other hand, by compounding the inorganic fine particles as a filler in the molded body of the organic polymer, it is possible to improve the strength of the organic polymer film and impart functions such as optical properties, heat resistance and fire resistance. In these cases, it is considered that the affinity between the organic polymer and the filler and the orientation of the filler particles have a great influence on the properties of the composite, and surface modification of the filler particles and the resin side is actively performed to enhance the affinity. It has been.

Patent Document 2 has through holes having an average pore size of 0.01 to 5 μm, which are obtained by blending a high molecular weight polyethylene resin with a fibrous inorganic filler having heat resistance as a filter filter medium for an organic solvent for cleaning. However, a porous membrane having a film thickness of 25 to 300 μm is disclosed. However, since the method of melt molding of an organic polymer film described in Patent Document 2 performs high temperature heating melting, kneading, and extrusion molding of an organic polymer, it is used except for fine particles having resistance to these molding processes. Can not do it. In addition, since the surface of the inorganic particles is crushed during melt-kneading, a large amount of mineral oil or di-2-ethylhexyl phthalate must be used as a plasticizer and a pore-forming agent, and this plasticizer is further extracted. As an extractant for removal, a large amount of saturated hydrocarbon-based organic solvents such as hexane, heptane, octane, nonane, and decane, and halogenated hydrocarbon-based organic solvents such as trichlorethylene and tetrachloroethylene, which are highly carcinogenic, must be used.

In addition to such a melt molding method, a phase separation method can be considered as a molding method for organic polymers. The phase separation method is a method in which an organic polymer is dissolved in a solvent to form a slurry, which is then aggregated by invasion of a non-solvent or a poor solvent or a temperature change to form a slurry. Patent Document 3 discloses a polyamide hollow fiber membrane and a film forming method by a heat-induced phase separation method as a method for producing the same. However, in this method, the organic polymer slurry must be prepared by heating to 170 ° C. or higher, the film is formed by lowering the temperature to 100 ° C. or lower, and the filler is up to 100% by weight with respect to the resin, that is, the composition. If 50% by weight or more of the object is supported, the strength may not be obtained. As described above, this phase separation method is used as a film-forming method focusing on the properties of the organic polymer film, and due to the above-mentioned strength problem, the functional fine particles are used as the main raw material to form a porous body. It is not suitable as a film forming method.

In Patent Document 4, a composite of an inorganic adsorbent and an organic polymer is formed by utilizing phase separation in a poor solvent. According to this method, it is possible to produce a porous molded product containing 50% by weight or more of an inorganic adsorbent as a main raw material, and this molded product is stable in an aqueous solution for several years without powder dropping. It has strength that can be used as a filtration membrane.

However, Patent Document 4 is characterized in that the inorganic particles are dispersed while being pulverized in a good solvent for the slurry in order to highly disperse the inorganic fine particles in producing the slurry for molding. However, according to the method of Patent Document 4, when polysulfone is first mixed as an organic polymer in a state where synthetic zeolite having a high silica / alumina ratio is dispersed in a solvent, the solvent of polysulfone is determined by the interaction between the solvent and the fine particles and the balance of affinity. In some cases, dissolution into the solvent was hindered, and a slurry in which the inorganic particles were uniformly dispersed could not be obtained, or in some cases, powder was often dropped from the molded body. Further, in order to avoid such a problem, when an organic polymer having a high affinity with inorganic particles is used, the organic polymer penetrates not only on the surface of the inorganic particles but also inside the pores, and the inorganic particles are molded during molding. It was found that the micropores were easily blocked and crushed. As described above, in the prior art, when forming porous silicate fine particles having pores of various sizes and having a surface having a surface different from that of natural zeolite, the structure is dense and liquid permeation can be prevented from falling off. There is a problem that it is difficult to achieve both the porosity and the porosity that retains the pore function.

Further, in the basic research on the conventional immobilization enzyme using mesoporous silica fine particles as proposed in Non-Patent Document 1, "mesoporous silica fine particle powder" in the form of powder is mainly used, and generally, fine particle powder is used. Although it is often used in the form of a mesoporous silica fine particle powder as a part of a device or instrument such as a bioreactor, there are problems such as pulverization of the fine particle powder and powder removal.

Further, the molded product obtained by using the organic polymer disclosed in Non-Patent Document 2 as a binder may have a problem that it may be difficult to maintain the original pore diameter and surface area of mesoporous silica due to the influence of the organic polymer. was there.

Further, the molded product obtained by sintering disclosed in Non-Patent Document 3 has a problem that it may be difficult to maintain the original pore diameter and surface area of mesoporous silica due to the influence of the sintering temperature. ..

Therefore, the subject of the present invention is to retain the characteristics of porous silicates having mesopores such as mesoporous silica, mesoporous aluminum oxide, and mesoporous titanium oxide, and functional fine particles having similar surface properties. At the same time, it is an object of the present invention to provide a multi-dimensional porous molded body having high water resistance, filterability and reactivity, and a novel method capable of producing the same. Further, by this method, good results can be obtained even if a hydrophilic resin such as EVO H , which is cheaper, is used as a binder, not limited to the polyether sulfone, depending on the application.

Further, the subject of the present invention is that even if fine particle powder of mesoporous substance such as mesoporous silica is molded into an appropriate shape such as hollow filament or pellet shape according to the application, it is stable, has high mechanical strength, and is in powder form. To provide a porous molded body of mesoporous fine particles, an enzyme complex thereof, and a method for producing them, which can maintain the same physical properties as the mesoporous silica fine particles and can exert the same effect as powder as an immobilized support for the enzyme. Is.

As a result of diligent studies to achieve the above problems, the present inventors have conducted a method of phase-separating a mixed slurry with an organic polymer in a poor solvent as a method for forming porous particles, in which the poor solvent infiltrates. It is preferable from the viewpoint that the path becomes a through hole to enhance the reactivity of the molded body and partially expose the surface of the porous fine particles. , The organic polymer dissolved in a good solvent invades the pores of the porous particles and polymerizes, or completely covers the surface of the porous particles and shields the surface of the fine particles from the through holes of the molded body. I came to the view that it is important to keep in mind to avoid.

From that point of view
1. 1. To prepare a slurry for forming porous particles, first put the organic polymer in a good solvent and dissolve it at a temperature at which it can be sufficiently dissolved, then the fine particles do not react and the organic polymer can maintain a viscous slurry form. It is preferable to carry out the procedure such as adding fine particles little by little while stirring at high speed at a temperature.
2. 2. Further, in order to prevent powder falling, it is effective to use a hydrophilic organic polymer having a functional group having a high affinity with silanol formed on the silicate surface in the structure as a binder, but hydrophilic organic. It was found that it is preferable to avoid macromolecules that are in a small molecule state or that have low viscosity and easily penetrate into the pores. Further, since these hydrophilic organic polymers are more likely to have a dense aggregated structure in water than the hydrophobic organic polymers, they are likely to enter between submicron-sized fine particles and become dense in the molded body. In order to prepare a mixed slurry with fine particles, the nano-sized particles generated by high-speed stirring of the gas containing the fine particles and the viscous slurry when the fine particles are added to the viscous slurry in which the organic polymer is first dissolved are not defective. It has been found that extremely minute bubbles can be intentionally mixed, voids are likely to be generated between the molded particles, and the filterability and flexibility are improved. Furthermore, when the hydrophilic organic polymer has an aggregated structure that completely embeds the fine particles due to the balance of affinity between the poor solvent and the fine particles, the post-treatment after the formation of the molded body greatly destroys the molded body structure. It was found that the accessibility to fine particles can be improved without doing so and without causing a decrease in strength.

The present inventors have completed the present invention based on these findings.

That is, the present invention includes the following aspects.
(1) Functional fine particles having mesopores with a pore diameter of 2 nm to 50 nm, mainly composed of silicate, aluminosilicate, alumina or titania, and a particle size of 0.01 μm to 200 μm, and an organic polymer. It is a porous molded body containing
The organic polymer is a polyether salphon or an EVOH resin having a polyethylene molar ratio of 44% or more.
It has macropores with a pore diameter of 50 nm to 100 μm.
It is a porous body with a porosity of 34% or more.
The content of the functional fine particles in the molded product is 46% by mass or more, and the content is 46% by mass or more.
At least a part of the surface of the functional fine particles is exposed from the molded product.
The pore volume of the mesopores of the functional fine particles is maintained at 40% or more of the raw material functional fine particles.
A porous molded body of mesoporous fine particles.

(2) The porosity of the mesoporous fine particles according to (1) above, characterized in that the weight change of the porous molded body before and after stirring in water for 24 hours or more is 1% or less. Molded body.

(3) The porous molded body of mesoporous fine particles according to (1) or (2) above, wherein the shape of the porous molded body is granular, hollow particle-like or hollow thread-like.

(4) The porosity of the mesoporous fine particles according to any one of (1) to (3) above, wherein the functional fine particles and / or the organic polymer are a mixture of a plurality of types. Molded body.

(5) Raw Material of the Functional Fine Particles The function of the functional fine particles is adsorption performance, and the characteristics of the functional fine particles are maintained after molding of the porous molded body. 4) The porous molded body of mesoporous fine particles according to any one of.

(6) The porous molded body of mesoporous fine particles according to any one of (1) to (5) above, wherein particles having a size of several microns can be pressure-filtered from the dispersion liquid.

( 7 ) The gas contained in the functional fine particles is intentionally mixed by dispersing the functional fine particles in a dried state in an organic solvent in which the organic polymer is previously dissolved in a stirred state. And the process of obtaining a raw material slurry containing nanobubbles
The production of a porous molded body of mesoporous fine particles according to any one of (1) to ( 6 ) above, which comprises a step of molding by injecting the raw material slurry into a non-solvent. Method.

( 8 ) The method for producing a porous molded body of mesoporous fine particles according to ( 7 ) above, wherein the organic polymer has a hydrophilic group.

( 9 ) The above-mentioned (7) or (8) further comprises a step of subjecting the porous molded body of the mesoporous fine particles to a low-temperature heat treatment in ethanol under reduced pressure or in a low-concentration aqueous solution of a good solvent. ) . The method for producing a porous molded body of mesoporous fine particles.

( 10 ) Raw material of the functional fine particles The function of the functional fine particles includes the ability to immobilize one or more kinds of enzymes and / or proteins, and the content of the functional fine particles in the molded body is 46 mass. % Or more, the enzyme-supporting carrier which is the porous molded body according to any one of (1) to ( 6 ) above.

( 11 ) The complex of the enzyme-supporting carrier and the enzyme according to ( 10 ) above.

( 12 ) The complex according to ( 11 ) above, wherein the enzyme is an oxidoreductase, a hydrolase, a transfer enzyme, a desorption enzyme, an isomerase, and / or a synthase.

( 13 ) The complex according to ( 11 ) or ( 12 ) above, wherein the enzymatic reaction of the enzyme is an oxidation-reduction reaction, a hydrolysis reaction, a transfer reaction, an elimination reaction, an isomerization reaction, and / or a synthetic reaction. ..

( 14 ) Any of the above ( 11 ) to ( 13 ) in which one or more enzymes and / or proteins involved in the enzymatic reaction of the enzyme are immobilized in mesopores having a pore diameter of 2 nm to 50 nm. A method for producing the complex according to one.
A production method comprising an immobilization step of immobilizing the enzyme and / or protein on a porous molded body of mesoporous fine particles in a buffer solution adjusted to pH 3-11.

( 15 ) The production method according to ( 14 ) above, further comprising a washing step of washing the porous molded body of mesoporous fine particles after immobilization with a buffer solution adjusted to pH 3 to 11 multiple times.

( 16 ) Any one of ( 11 ) to ( 13 ) above, wherein each of one or more kinds of enzymes and / or proteins involved in the enzymatic reaction of the enzyme is immobilized on a porous molded body of mesoporous fine particles. It is an enzymatic reaction method using the complex described in the above.
Immobilization step of immobilizing the enzyme and / or protein in a porous molded body of mesoporous fine particles in a buffer solution adjusted to pH 3 to 11.
A reaction substrate is added to a buffer solution having a pH of 3 to 11 containing a complex of a porous molded body of mesoporous fine particles obtained in the immobilization step and an enzyme, or the mesoporous fine particles obtained in the immobilization step. The enzyme reaction method, which comprises an enzyme reaction step of adding a complex of the porous molded product and the enzyme to a buffer solution having a pH of 3 to 11 containing a reaction substrate to carry out an enzyme reaction involving the enzyme and / or protein.

( 17 ) Any one of ( 11 ) to ( 13 ) above, wherein each of one or more kinds of enzymes and / or proteins involved in the enzymatic reaction of the enzyme is immobilized on a porous molded body of mesoporous fine particles. An enzymatic reaction method using the complex described in the section.
Immobilization step of immobilizing the enzyme and / or protein in a porous molded body of mesoporous fine particles in a buffer solution adjusted to pH 3 to 11.
A washing step of washing a composite of a porous molded body of mesoporous fine particles obtained in the immobilization step and an enzyme with a buffer solution having a pH of 3 to 11.
An enzymatic reaction step in which a complex of a porous molded body of mesoporous fine particles after washing obtained in the washing step and an enzyme is subjected to an enzymatic reaction involving the enzyme and / or a protein in a reaction solution containing a reaction substrate. Enzyme reaction method including.
( 18 ) The enzymatic reaction method according to ( 16 ) or ( 17 ) above, wherein the enzymatic reaction is a method for producing a functional useful substance from a reaction substrate.
( 19 ) The enzymatic reaction method according to ( 16 ) or ( 17 ) above, wherein the enzymatic reaction is a method for decomposing an environmental pollutant that is a reaction substrate existing in the environment.
( 20 ) The enzyme reaction method according to ( 16 ) or ( 17 ) above, wherein the enzyme reaction method is a method for detecting or quantifying a reaction substrate that is or may be present in a test sample.
( 21 ) A kit, sensor or apparatus for an enzyme reaction involving the enzyme and / or protein, which comprises the complex according to any one of ( 11 ) to ( 13 ).

According to the present invention, porous silicates having mesopores such as mesoporous silica, mesoporous aluminum oxide, and mesoporous titanium oxide, and functional fine particles having similar surface textures are maintained at the same time. A multi-dimensional porous molded body having water resistance, filterability, and reactivity, and a novel method capable of producing the same are provided. Further, by this method, good results can be obtained even if a hydrophilic resin such as EVO H , which is cheaper, is used as a binder, not limited to the polyether sulfone, depending on the application.

Further, according to the present invention, even if fine particle powder of mesoporous substance such as mesoporous silica is molded into an appropriate shape such as hollow filament or pellet shape according to the application, it is stable and has high mechanical strength, and powdered mesoporous silica. Provided are a porous molded body of mesoporous fine particles, an enzyme complex thereof, and a method for producing the same, which can maintain the same physical properties as the fine particles and can exert the same effect as the powder as an immobilized support for the enzyme.

An SEM image of a cross section orthogonal to the longitudinal direction of the molded product of Example 1. An SEM image of a longitudinal cross section of the molded product of Example 1. The pore size distribution curve figure by the mercury porosimetry of the molded article of Example 1. FIG. The figure which showed the nitrogen adsorption isotherm of the molded article of Example 1 and the raw material particle A at room temperature. The figure which showed the nitrogen adsorption isotherm of the reference example 13 and the reference example 14 and the raw material particle F at room temperature. External view of the molded product of Example 18. Appearance image of (a) mesoporous silica powder, (b) molded product, and (c) enzyme complex of Example 23. The figure schematically showing the procedure for preparing the complex of the molded product of Example 18 and the azo reductase, the decomposition reaction of the azo dye, and the reuse of the immobilized enzyme. The figure which showed the immobilization amount and the immobilization rate of AzoR in Example 30. The figure which showed the change in the absorbance of Example 30 and the decomposition rate of methyl red. The figure which showed the immobilization amount of AzoR with respect to various mesoporous fine particles of Example 31. The figure which showed the decomposition rate and durability of methyl red by various AzoR-mesoporous fine particle complex of Example 31. 31 is an external view of (a) the molded product of Example 23, (b) the AzoR complex thereof, (c) the molded product of Comparative Example 8, and (d) the AzoR complex thereof. The figure which showed the influence of the free AzoR concentration on the decolorization rate. The figure which showed the immobilization amount of glucose dehydrogenase with respect to various mesoporous fine particles of Example 32. The figure which showed the density | concentration of the generated NADH by the various GDH-mesoporous fine particle complex of Example 32, and the durability of the immobilized enzyme. The figure which showed the influence of the unfixed free GDH concentration on the produced NADH concentration. The figure which showed the immobilization amount of the lipase with respect to the various mesoporous fine particles of Example 33. The figure which showed the free pyrene concentration and the durability of the immobilized enzyme in the triglyceride decomposition by the various lipase-mesoporous fine particle complex of Example 33. The figure which showed the influence of the unfixed free lipase concentration on the concentration of fluorescent substance (pyrene) released by triglyceride decomposition. The figure which showed the immobilization amount of protease with various mesoporous fine particles and activated carbon of Example 34. The figure which showed the degrading activity of gelatin and the durability of the immobilized enzyme by various protease-mesoporous fine particle complex and protease-activated carbon complex of Example 34.

Next, a preferred embodiment of the present invention will be described. In the present invention, the description of the numerical range includes not only both-end values but also all arbitrary intermediate values contained therein.

The porous molded body of micro or mesoporous fine particles of the present invention is a porous molded body containing functional fine particles and an organic polymer (hereinafter, the porous molded body of micro or mesoporous fine particles of the present invention is referred to as the present invention. It may be referred to as the porous molded body of the present invention).

The functional fine particles according to the present invention are mainly composed of silicate, aluminosilicate, alumina or titania having micro or mesopores having a pore diameter of 0.3 nm to 50 nm.

Here, the micropores are also called micropores or micropores, and refer to pores having a pore diameter of less than 1 nm. Mesopores are pores that are larger than micropores and have a pore diameter of 1 nm or more and 50 nm or less. Therefore, having micro or mesopores having a pore diameter of 0.3 nm to 50 nm means having pores having a pore diameter of 0.3 nm to 50 nm.

The pore diameter of the functional particles according to the present invention is 0.3 nm to 50 nm, preferably 0.3 nm to 10 nm, and more preferably 0.5 nm to 5 nm.

In addition, silicates, aluminosilicates, aluminas or titanias are pure silicates, aluminosilicates, aluminas or titanias, as well as other metallic elements introduced into defects in the skeleton of silicates, aluminosilicates or titania structures, silicates. And some of the silicon and aluminum elements in the structure of aluminosilicate are replaced with other metals (metal substituents).

Further, when the main component is silicate, aluminosilicate, alumina or titania, the functional fine particles are those in a pure phase thereof, or the mass ratio thereof is usually 50% or more, preferably 90% or more. To say.

The function of the functional fine particles is not particularly limited, and examples thereof include adsorption performance and removal ability. It is preferable that these functions retain their characteristics after molding the porous molded product.

The type, structure, and crystallinity of silicate, alumina, aluminosilicate, alumina, and titania according to the present invention are not particularly limited.

The particle size of the functional fine particles according to the present invention is 0.01 μm to 200 μm, preferably 0.01 μm to 10 μm, and more preferably 0.1 μm to 5 μm. Here, the particle size means the size of the agglomerated particles. Particles having a crystal particle size smaller than this range may form agglomerates and are not separated. When preparing the raw material slurry described later, the functional fine particles having a particle size in the above range are dispersed with the same particle size, or the aggregated particles are loosened and dispersed with a finer particle size. When the porous molded body of the present invention is hollow filamentous or finely granular, the functional fine particles are preferably dispersed in the porous molded body, the raw material slurry, or the solvent in the range of particle size of 0.01 μm to 10 μm. More preferably, the dispersion is in the range of 0.1 μm to 5 μm. The particle size can be determined, for example, as an average value of the diameters of 20 to 30 particles observed by transmission electron microscopy.

The functional fine particles according to the present invention can be modified functional fine particles chemically modified or treated so that they can be stably dispersed together with an organic polymer in a slurry solvent. Examples of the chemical modification or treatment of the functional fine particles include modification of the organic chain at the terminal of the silanol group.

The functional fine particles according to the present invention may be used alone or in a plurality of types, but when a mixture of a plurality of types is used, a single molded product can selectively adsorb or remove a plurality of reaction targets at the same time. It is preferable in that multiple functions can be added.

The content of the functional particles of the porous molded product of the present invention is 46% by mass or more, preferably 50% by mass or more, more preferably 60% by mass or more, still more preferably 67% by mass or more.

The organic polymer according to the present invention is not particularly limited, but has functionality of silicate, alumina, aluminosilicate or titania as a main component, which is a porous body in the porous molded body of the present invention and has micro or mesopores. It is preferable to have a functional group having a high affinity with fine particles (for example, a hydrophilic group such as a hydroxyl group, a sulfonyl group, or a carboxyl group), and it is suitable for a general-purpose amphipathic organic solvent such as dimethylsulfoxide at a relatively low temperature. In terms of weight, it can be dissolved in an amount of about 7 to 20% by weight or more, a uniform slurry can be prepared when these are dispersed, and the slurry can be further aggregated and molded in a non-solvent or a poor solvent. And more preferable. More specifically, among the thermoplastic resins, polyether sulfone, polysulfone having relatively high hydrophilicity, polyethylene-polyvinyl alcohol copolymer resin (EVOH resin) having an OH group, and polyethylene-vinyl acetate copolymer resin. Kenzoku, acrylic resin (PMMA resin), polyamideimide resin and the like are preferable. More preferably, it is a polyether sulfone, a polyethylene-polyvinyl alcohol copolymer resin having a polyethylene molar ratio of 44% or more, or a polyamide-imide resin.

The organic polymer according to the present invention may be used alone or in a plurality of types, but the physical properties such as the strength of the molded body and the chemical properties such as chemical resistance and heat resistance are the properties of the organic polymer. Since it depends largely on the above, it is possible to control the properties of these molded bodies by using a mixture of a plurality of types.

The organic polymer according to the present invention is preferably chemically modified or treated modified functional fine particles. Examples of the chemical modification or treatment of the organic polymer include the addition of a hydroxyl group to the polyether sulfone and the saponification treatment of the polyethylene-vinyl acetate copolymer resin.

The content ratio of the hydrophilic group of the organic polymer according to the present invention is not particularly limited, but is preferably less than 70 mol%, more preferably less than 56 mol%. When the content ratio of the hydrophilic group is low, the hydrophilicity does not become too high, and it tends to be easy to form an arbitrary shape without spreading like a film on the surface of a poor solvent for coagulation.

The polyethylene content of the organic polymer according to the present invention is not particularly limited, but is preferably less than 70 mol%, more preferably 48 mol% or less. When the polyethylene content is low, it dissolves in a solvent and tends to increase the affinity with fine particles.

The content of the organic polymer in the porous molded product of the present invention is appropriately selected depending on the shape of the molded product and the type of functional fine particles, but is usually 5 to 50 weight with respect to the total weight of the porous molded product. %, Preferably around 15-40% by weight. When the content of the organic polymer is in this range, the molded product is mechanically stable, and the micro or mesopores tend to function without closing the particle gaps of the functional fine particles.

The porous molded article of the present invention is not limited depending on the production method thereof, but for example, the main component is silicate, aluminosilicate, alumina or titania having micro or mesopores in an organic solvent in which an organic polymer is previously dissolved. It can be produced by a method including a step of dispersing the functional fine particles to obtain a raw material slurry and a step of molding by injecting the raw material slurry into a non-solvent.

When the organic polymer of the porous molded product of the present invention is a PMMA resin or a polyamide-imide resin, it can be produced by a method further including a step of treating with a polymerization accelerator or a curing accelerator after molding.

This production method is a method for obtaining a molded product by a phase separation method from a raw material slurry containing functional fine particles and an organic polymer.

In addition to functioning as a binder for functional fine particles, the organic polymer has a role of retaining the gaps between the functional fine particles so as not to be crushed even during use after forming the porous body by forming macropores. The organic polymer is not particularly limited, and examples thereof include those exemplified as the organic polymer related to the above-mentioned molded product. However, it is soluble in a kind of organic solvent that is difficult to volatilize at room temperature at a relatively low temperature, and is functional fine particles. Those having a high affinity with are appropriately selected. Those having a hydrophilic group are preferable because of their affinity with silicates and aluminosilicates. Further, from the viewpoint of preventing infiltration into micro or mesopores in the raw material slurry, from the viewpoint of obtaining sufficient viscosity and moldability at a low concentration during molding, and from the viewpoint of the structure and strength of the molded product after molding, polystyrene conversion A relatively high molecular weight having a weight average molecular weight (Mw) of 50,000 or more is preferable. The amount of the organic polymer added to the raw material slurry is appropriately selected depending on the shape of the molded product and the type of functional fine particles, but is usually 5 to 50% by weight based on the total weight of the obtained dried molded product. It is preferably around 15 to 40% by weight. If the addition rate of the organic polymer is too small, it becomes difficult to form a stable molded body, and if it is too large, the particle gaps of the raw material functional fine particles are closed, and the micro or mesopores tend to not function. Further, when PMMA resin or the like is used, the strength of the molded product can be increased by performing the repolymerization treatment after molding.

As the solvent for dissolving the organic polymer, as described above, the organic polymer can be dissolved at a relatively low temperature in an amount of about 6 to 20% by weight or more based on the weight of the solvent, and is a non-solvent for coagulating the resin. Functional fine particles that are compatible with a poor solvent and have micro or mesopores that are porous and are mainly composed of silicate, aluminosilicate, alumina or titania, cause a chemical or physical reaction with it. It is preferable that the solvent can be uniformly dispersed without any problem. Examples of such a solvent include dimethyl sulfoxide, dimethylformamide, dimethylacetamide, and n-methylpyrrolidone.

In the present invention, the preparation of the raw material slurry in which the functional fine particles are dispersed in an organic solvent in which an organic polymer is dissolved is prepared by using silicates having micro or mesopores, aluminosilicates or functional fine particles containing titania as a main component. This can be done by adding the above-mentioned solvent together with the above-mentioned organic polymer serving as a binder and stirring the mixture.

When the functional fine particles of the raw material are sufficiently dried before addition, it is preferable in that the local polymerization of the organic polymer in the raw material slurry before molding is prevented and the strength of the composite after molding is prevented from being lowered. .. The drying temperature is not particularly limited, but is preferably a temperature at which dehumidification does not occur within a range in which the properties of the raw material fine particles do not change. The concentration of the functional fine particles in the raw material slurry is not particularly limited, but is appropriately selected depending on the type and size of the functional fine particles, the difference in viscosity, and the type of the molded product or the organic polymer. For hollow fiber membrane molding, the concentration of functional fine particles in the raw material slurry is preferably 6 to 40% by weight (mass%). When this concentration is low, molding becomes difficult, or the porosity in the molded body becomes low, and when the concentration is high, the distribution with the organic substance that becomes the binder in the molded body tends to be uneven, and the flexibility and pressure resistance are high. May not be obtained.

The order of adding the functional fine particles and the organic polymer at the time of preparing the raw material slurry is not particularly limited, but the viscous slurry in which the organic polymer is sufficiently dissolved in a preheated solvent is dried at high speed. A method in which the raw material powder in the state is added little by little and dispersed is preferable because it is easy to obtain a uniform and highly viscous slurry at a relatively low temperature in a short time, and more effectively, nanobubbles can be generated. In the method of adding the organic polymer later, the dissolution of the organic polymer is inhibited depending on the concentration of addition due to the balance of the affinity between the fine particles and the solvent, and as a result, it may be difficult to obtain a stable slurry.

The stirring speed at the time of preparing the raw material slurry is not particularly limited, but is preferably 200 rpm or more in the gas phase. When the stirring speed is in this range, macropores are likely to be formed.

In the method for producing a porous molded body of micro or mesoporous fine particles of the present invention, the raw material slurry prepared as described above can be solidified and molded by injecting it into a non-solvent, but the raw material slurry can be molded in a poor solvent. It can also be solidified and molded. This makes it possible to obtain a self-supporting water-resistant composite porous material having a pore diameter distribution of several angstroms to several tens of microns.
Examples of the non-solvent include water alcohol and the like. Examples of the poor solvent include an aqueous solution of the solvent used for the raw material slurry.

In the method for producing a porous molded body of micro or mesoporous fine particles of the present invention, a step of subjecting the porous molded body of micro or mesoporous fine particles to a low-temperature heat treatment in ethanol under reduced pressure or in a low-concentration aqueous solution of a good solvent. Can be further included.

As the molding method, a mold suitable for the porous molded body can be used depending on the type of the porous molded body. For example, when the shape of the molded body is a hollow thread, molding is performed by an existing method using a nozzle or the like. Can be done.

The porous molded body of the present invention has macropores in the range of 50 nm to 100 μm and has a porosity of 34% or more, and at least a part of the surface of the functional fine particles is exposed from the molded body. The pore volume of the micro or mesopores of the functional fine particles is maintained at 40% or more of the raw material functional fine particles.

In the present specification, the fact that the porous molded product has macropores in the range of 50 nm to 100 μm means that the poor solvent, which is a coagulating liquid, is an approach path to the raw material slurry side, that is, a contact interface when the porous molded product is produced. This means that the coagulation of the raw material causes the formation of macroscopic pores leading from the outside to the inside of the porous molded product. The size of these macropores can be confirmed by having a peak in this range of the pore size distribution curve when measured with a porosimeter, etc., and the fine pores, which are the characteristics of the raw material particles, and the particles are the organic polymer of the binder. It means that the macroscopic pores to be formed coexist. Further, since the entry path of the poor solvent corresponds to the gap between the fine particles in the raw material slurry at the same time, the surface of the raw material particles is easily exposed in the macropores.

Further, in the porous molded body of the present invention, micropores are provided in the dense layer formed on the wall of the macropores by degassing under reduced pressure for a short time in ethanol or a good solvent at a low temperature after molding. It is possible to modify the macropores so that the surface of the raw material particles is exposed.

Since the porous molded product of the present invention has such characteristics, a substance to be adsorbed or removed or reacted is diffused in the molded product as compared with a normal molded product, and comes into contact with the surface of the raw material particles contained therein. It is easy to carry out, and as a result, it is excellent in that the reaction speed and the reaction efficiency are increased.

Further, the porous molded body of the present invention has properties of micro or mesoporous fine particles such as ion exchange property and adsorptive property in addition to heat resistance, water resistance and humidity control.

The porous molded article of the present invention preferably has a weight change of 1% or less before and after stirring continuously in water for 24 hours or more. Here, the fact that there is a change in weight means that the fine particles are removed from the porous molded body and fall into the water, or the properties of the fine particles and the organic polymer are changed. When the weight change is in this range, it is preferable from the viewpoint that when the molded product is used in water, it is considered that almost no fine particles are washed away due to this deterioration.

The porous molded product of the present invention can regain its flexibility by being immersed in alcohols after being dried at a low temperature of 60 ° C. or lower. Since heating at 150 ° C. or higher tends to lose the flexibility of the porous molded product, it is preferable to store or process the porous molded product at a temperature lower than 150 ° C.

The shape of the porous molded product of the present invention is not particularly limited, but it is preferably granular, hollow particles, or hollow threads. In particular, when the shape is hollow particles or hollow threads, the reaction target substances such as the substances to be adsorbed and the substances to be removed are more likely to diffuse into the porous molded body than in the case where there is no hollow structure, and the reaction rate and reaction are increased. There is no raw material particles or hollow structure, such as the point that efficiency can be improved, the point that it is easy to scale up to modules, and the fact that substances are supplied from the inside and outside of the porous molded body and reacted on the surface. It can be expected to be superior in various points such as widening the application to reactions that cannot be achieved.

Further, the porous molded body of the present invention can be used as a molded body such as a hollow fiber membrane or a pellet, as a microfiltration membrane that can be used for filtration and purification of water, a carrier of a reactant in an aqueous solution, and the like. Is. Although this microfiltration membrane is a water resistant microfiltration membrane, it also has the pore characteristics and reactivity of micro or mesoporous fine particles.

The composite membrane produced from the porous molded body of the present invention has an action of adsorbing and removing harmful substances and an ion exchange action of an aqueous solution containing cations, and can be a water-resistant and self-supporting composite membrane. This composite membrane can be suitably used, for example, as a treatment agent for wastewater for the food industry.

The porous molded body of the present invention is capable of pressure-filtering particles having a size of several microns from the dispersion liquid. When used as a membrane material, neutral particles having a large size that cannot be removed by raw material particles such as microorganisms. Is preferable in that it can be removed at the same time.

Regarding the storage of the molded product of the present invention after molding, if it is stored in water or alcohol as it is without being dried, it can be used while maintaining its flexibility. Further, the molded body of the present invention can maintain the pore characteristics, adsorption characteristics, reactivity and the like of the functional porous fine particles before molding even after molding, while being a stable molded body even in the liquid phase.

The molded product of the present invention can be used as a carrier for carrying an enzyme. The molded product of the present invention can form a complex with an enzyme as a carrier for carrying an enzyme. In particular, among the molded bodies of the present invention, the functional fine particles are mesoporous fine particles having pore diameters of 2 nm to 50 nm and particle sizes of 0.01 μm to 200 μm, and are functional fine particles in the molded body. A molded product having a content of 46% by mass or more is suitable as a carrier for carrying an enzyme. Further, when the function of the functional fine particles, which is the raw material of the functional fine particles constituting the molded product of the present invention, contains the ability to immobilize one or more kinds of enzymes and / or proteins, it is suitable as a carrier for carrying an enzyme. Is.

This enzyme is not particularly limited, but is preferably an oxidative-reducing enzyme, a hydrolase, a transferase, a desorption enzyme, an isomerase, and / or a synthase. Specific examples of the enzyme include azoreductase, lipase, glucose dehydrogenase, and protease.
This enzyme can be used for enzymatic reactions of oxidation-reduction reaction, hydrolysis reaction, transfer reaction, elimination reaction, isomerization reaction, and / or synthetic reaction.

The method for producing the complex is not particularly limited, but for example, one or more enzymes and / or proteins involved in the enzymatic reaction of the above enzymes are porous with mesoporous fine particles in a buffer solution adjusted to pH 3 to 11. It can include an immobilization step of immobilizing the quality molded body. Further, it is more preferable to include a washing step of washing the porous molded body of the mesoporous fine particles after the completion of immobilization with a buffer solution adjusted to pH 3 to 11 multiple times.

The enzyme reaction method using the complex of the present invention is not particularly limited, and for example, 1) immobilization of an enzyme and / or a protein in a porous molded body of mesoporous fine particles in a buffer solution adjusted to pH 3 to 11. Step 2) Add the reaction substrate to the buffer solution having a pH of 3 to 11 containing the complex of the porous molded body of the mesoporous fine particles obtained in the immobilization step and the enzyme, or 3) the immobilization step. An enzymatic reaction step in which a complex of a porous molded body of mesoporous fine particles obtained in 1 above and an enzyme is added to a buffer solution having a pH of 3 to 11 containing a reaction substrate to carry out an enzymatic reaction involving the above-mentioned enzyme and / or protein. , The method of including. Further, as another enzyme reaction method, 1) an immobilization step of immobilizing an enzyme and / or a protein in a buffer solution adjusted to pH 3 to 11 in a porous molded body of mesoporous fine particles, and 2) the immobilization step. Cleaning step of washing the complex of the obtained porous molded body of mesoporous fine particles with the enzyme with a buffer solution of pH 3 to 11 3) The porous molded body of the obtained mesoporous fine particles and the enzyme after washing obtained in the washing step. Examples thereof include an enzymatic reaction step of carrying out an enzymatic reaction involving the above-mentioned enzyme and / or protein in a reaction solution containing a reaction substrate, and a method of containing the complex with.

These enzymatic reactions are not particularly limited, but are, for example, a method for producing a functional useful substance from a reaction substrate, a method for decomposing an environmental pollutant that is a reaction substrate existing in the environment, and a method present in a test sample. Examples include methods for detecting or quantifying reaction substrates that may or may be present.

The complex of the porous molded body and the enzyme of the present invention can be used for an enzyme reaction kit, a sensor or an apparatus involving the above-mentioned enzyme and / or protein. The use of this enzyme reaction kit, sensor or device is not particularly limited, but it is for producing a functional useful substance from a reaction substrate, or for decomposing an environmental pollutant which is a reaction substrate existing in the environment. , Can be used to detect or quantify the reaction substrate or the product after the enzymatic reaction that may be present in the test sample.

(Example)
Next, the present invention will be described in more detail based on Examples, but the present invention is not limited to the following Examples and the like. The porosity and pore size distribution of the porous molded product according to the present invention can be evaluated by a measuring method such as mercury porosity. Further, the retention rate of the pore volume of the micro or mesopores of the raw material fine particles can be evaluated by comparing the gas adsorption or steam adsorption measurement results with the measured values of the raw material particles.

The physical characteristics of the fine particles used in the examples are shown in Tables 1 and 6, and the physical characteristics of the obtained molded product are shown in Tables 2 to 4 and 7 as a list.

(Example 1)
9.98 g of an ethylene-vinyl alcohol copolymer resin (EVAL G156B, Claret Co., Ltd.) having an ethylene content (molar ratio) of 48 mol% as an organic polymer is added to dimethyl sulfoxide (Wako Pure Chemical Industries, Ltd., special grade) 102. It was mixed with 88 g and stirred at 200 rpm at 40 ° C. overnight to dissolve it. While heating and stirring this, 19.99 g of MCM41 type synthetic mesoporous silica (SiO ₂ , Sigma-Aldrich No. 643653) (raw material particles A) having a particle diameter of 0.2 μm to 2 μm and a pore diameter of 2 nm to 3 nm was added. , Prepared a slurry for forming a porous body. At this time, the concentration of the particles in the slurry was 15% by weight, the amount of the ethylene-vinyl alcohol copolymer resin added was 7.5% by weight, and the amount of MCM particles charged into the molded product was 67% by weight. After stirring overnight, use a double-ended nozzle with a slurry discharge port diameter of 3 mmφ and a core liquid discharge port diameter of 0.7 mmφ in a dry-wet spinning device, and put the obtained slurry into a cylinder pump at 20.5 ml / min. Then, while injecting into a hot water bath at 37 ° C., hot water at 29 ° C. was flowed as a core liquid at 7 ml / min to form a film in the form of a hollow thread.

The produced hollow fiber membrane has an outer diameter of 1.2 mm and an inner diameter of 0.7 mm (see FIG. 1), and after being dried overnight at 60 ° C., the porosity is about 62% when measured with a mercury porosity, according to the pore size distribution curve. It was a multi-dimensional porous molded body having nanopores with a diameter of 4 nm to 10 nm and macropores of 0.1 μm-1 μm derived from MCM41 (see FIG. 2). 0.0299 g of a 60 ° C. dried product of this hollow filamentous molded product was placed in 3 ml of distilled water, continuously infiltrated at 100 rpm for 36 hours or more, dried at 60 ° C. again, and weighed to be 0.0297 g. The weight change is within 1%, and it is considered that there is almost no powder loss. According to thermal analysis, the organic polymer decomposes at 200 ° C or higher when heated to 600 ° C, and the weight loss due to this decomposition is 34% by weight. Therefore, the content of MCM41 particles in the molded body is 66% by weight. Conceivable. Further, according to the nitrogen adsorption isotherm, the relative pressure (P / Po) range of the hysteresis of the isotherm of the molded body and the raw material particles is the same, and it is considered that the size of the mesopores of the raw material particles is almost maintained. Further, since the pore volume in the molded body was 0.28 cm ³ / g and the mesopore volume of the raw material MCM41 powder particles was 0.59 cm ³ / g, the MCM containing the pore volume of the molded body was contained. When converted in terms of the amount of particles, it is considered that 71% of the pore volume of the powder sample is retained (see FIG. 3). Further, FIG. 4 shows the nitrogen adsorption isotherm of the molded product of Example 1 and the raw material particles A at room temperature. In addition, the pretreatment was performed under the reduced pressure condition of 60 ° C. for 24 hours, and converted by the particle content.

(Example 2)
Completely 0.54 g of polyether sulfoxide (Veradel PES 3000P, Solvay Advanced Polymers Co., Ltd., weight average molecular weight (Mw) = 57,000) as an organic polymer to 5.50 g of dimethyl sulfoxide (Wako special grade) at 50 ° C. To this, 0.55 g of the synthetic mesoporous silica (raw material particles A) used in Example 1 was mixed to prepare a slurry. At this time, the concentration of the particles in the slurry was 8.3% by weight, the amount of the polyether salphon added was 8.2% by weight, and the amount of the MCM particles charged to the molded product was 50% by weight. This slurry was dropped into a beaker containing hot water at about 32 ° C. and solidified for 2 hours or more, dried at 60 ° C., and further stirred and washed at room temperature for 25 hours or more until the white turbidity disappeared to form MCM-PES pellets. ..

After stirring and washing until the white turbidity disappeared, 0.0288 g of the pellet was stirred in 3 mL of water at room temperature for 72 hours or more, and the dry weight was measured. As a result, almost no change in weight was observed. When this was heated to 600 ° C by thermal analysis, the organic polymer decomposed at 450 ° C or higher, and the weight loss due to decomposition was 47%. Therefore, the content of MCM particles in the pellet is about 53%. it is conceivable that. (Reversible endothermic peaks and weight loss not found in the raw material particles and organic polymers were observed around 120 ° C, and it is considered that new adsorption sites derived from molding were generated.) The void ratio was 78% as measured by a mercury porosimeter. According to the pore size distribution curve, it was a multidimensional porous body having nanopores with a diameter of 4 nm to 10 nm and macropores with a diameter of 0.1 μm to 1 μm derived from MCM41. According to the nitrogen adsorption isotherm, the pore volume of the molded body was 0.50 cm ³ / g, and the pore volume of the powder sample measured under the same conditions was 0.59 cm ³ / g. When converted into the amount of MCM41 particles contained in, it is considered that almost all the mesopore volume of MCM41 in the molded body is maintained.

( Reference example 3)
As an organic polymer, 0.60 g of methyl methacrylate resin (PMMA polymer CASNo.9011-14-7, Tokyo Chemical Industry Co., Ltd., M788, Mw = 135,000 to 140,000) is dimethylformamide (Wako special grade) 7. It was completely dissolved in 0 g at 50 ° C., and 0.60 g of the synthetic mesoporous silica (raw material particles A) used in Example 1 was mixed with this to prepare a slurry. This slurry was dropped into a beaker containing hot water at about 32 ° C., solidified for 2 hours or more, and dried at 60 ° C. overnight to form MCM-acrylic pellets. Since this pellet had a very low mechanical strength, 0.20 g of this pellet was added to 4.98 g of ethanol (Wako special grade) together with 0.051 g of benzoyl peroxide (Biorad), and the temperature was slowly raised to finally. It was taken out after heating at 60 ° C. for 4 hours. The pellets taken out had changed to soft and hard-to-break properties like rubber. Further, when this was dried at 60 ° C., pellets having a compressive strength of about 1.4 MPa were obtained.

When the pellets after polymerization and curing were heated to 600 ° C by thermal analysis, the organic polymer decomposed at around 400 ° C, and the weight loss due to decomposition was 50%. Therefore, the MCM particles in the pellets. The content of is considered to be about 50%. According to the nitrogen adsorption isotherm, the pore volume in the molded body was 0.30 cm ³ / g, and the mesopore volume of the raw material MCM41 powder particles was 0.96 cm ³ / g. When converted into the amount of MCM particles contained in the pore volume, it is considered that 63% of the pore volume of the powder sample is retained.

(Example 4)
0.10 g of the organic polymer used in Example 1 was placed in 0.7 mL of dimethyl sulfoxide (Wako special grade), heated and stirred to dissolve it, and the mesoporous titanium oxide having a particle diameter of 10 to 100 nm and an average pore diameter of 4.2 nm was dissolved therein. (Alfa Aesar No. 43250) (raw material particles B) 0.20 g was mixed to prepare a slurry. The slurry was dropped into a beaker containing room temperature water to solidify it, and dried at 60 ° C. to form mesoporous TiO ₂ -EVOH pellets.

According to thermal analysis, the weight loss due to decomposition of the organic polymer when heated to 600 ° C. is 47%, and the weight loss when the raw material particles are heated is 18%. Therefore, the content of the particles in the pellet is 65. It is considered to be about%. Further, when the pore volume in the molded body calculated by pretreatment at 100 ° C. and the measurement result of nitrogen adsorption is converted into the amount of the mesoporous titanium oxide particles contained, the mesoporous volume of the mesoporous titanium oxide in the molded body is 64%. It is considered that the degree is maintained.

(Example 5)
The same as in Example 2 except that MSU-X type synthetic mesoporous aluminum oxide (Al ₂ O ₃ , Sigma-Aldrich No. 517747) (raw material particles C) having an average pore diameter of 5.7 nm is used as the raw material particles. Mesoporous alumina-PES pellets were formed. According to thermal analysis, the content of particles in the pellet was about 50%.

(Example 6)
FSM22-PES pellets were formed in the same manner as in Example 2 except that FSM22 type synthetic mesoporous silica (raw material particles D) having a pore diameter of 8 nm was used as the raw material particles. According to thermal analysis, the content of particles in the pellet was about 55%.

( Reference example 7)
An ethylene-vinyl alcohol copolymer resin (Soanol A4412, Nippon Synthetic Chemical Industry Co., Ltd.) having an ethylene content of 44 mol% was dissolved in the dimethyl sulfoxide used in Example 1 as an organic polymer, and the particle size was 2- as raw material particles. High silica in the same manner as in Example 2 except that synthetic high silica zeolite (SiO ₂ / Al ₂ O ₃ = 100) (raw material particle E) having a crystal particle diameter of 1 μm or less and a pore diameter of 0.90 nm is used at 3 μm. Zeolite-EVOH pellets were formed. After drying overnight at 60 ° C., the porosity was 62% as measured by a mercury porosity meter. According to the thermal analysis of the pellets after drying at 60 ° C., the weight loss when heated to 600 ° C. is considered to be 50% by weight, and the content of the zeolite particles in the molded body is considered to be 50% by weight. Further, according to the nitrogen adsorption isotherm, the total pore volume in the molded body was 80% or more of the powder sample in terms of the amount of zeolite particles.

( Reference example 8)
34.5 g of the ethylene-vinyl alcohol copolymer resin having an ethylene content of 48 mol% used in Example 1 as an organic polymer was mixed with 181.71 g of dimethyl sulfoxide used in Example 1 and mixed at 48 ° C. at 200 rpm overnight. It was stirred and dissolved. While the mixture was heated and stirred, 68.01 g of the synthetic high silica zeolite (raw material particles E) used in Reference Example 7 was added to prepare a slurry for forming a porous body. At this time, the concentration of the zeolite particles in the slurry was 22.0% by weight, and the amount of the zeolite particles charged into the molded product was 67% by weight. After stirring overnight, use a double-ended nozzle with a slurry discharge port diameter of 6 mmφ and a core liquid discharge port diameter of 2.5 mmφ, put the obtained slurry in a cylinder pump, and put it in a water bath at 34 ° C. at 40.0 ml / min. While injecting, water at 25 ° C. was flowed as a core liquid at 70.0 ml / min to form a film in the form of a hollow thread.

The produced hollow fiber membrane had an outer diameter of 4.8 mm and an inner diameter of 3.0 mm. According to thermal analysis, the weight loss when heated to 600 ° C. is 36% by weight, and the content of zeolite particles in the molded product is considered to be 64% by weight. According to mercury porosity, the porosity after drying overnight at 60 ° C. is 40%, and after further heating at 150 ° C. for 2 hours to crystallize EVOH, the porosity is 37%, and the pore size distribution curve. According to the report, it was a porous body having a macropore of 0.28 μm. According to the nitrogen adsorption measurement, the total pore volume in the molded body after the drying treatment at 60 ° C. was 89% or more of the powder sample in terms of the amount of zeolite particles.

( Reference example 9)
35.03 g of the resin used in Example 1 as an organic polymer was mixed with 181.29 g of dimethyl sulfoxide used in Example 1 and dissolved by stirring overnight at 202 rpm at 50 ° C. While heating and stirring this, 60.20 g of synthetic high silica zeolite (SiO ₂ / Al ₂ O ₃ = 40) (raw material particles F) having a particle diameter of 10 μm, a crystal particle diameter of 2 μm-4 μm, and a pore diameter of 0.58 nm was added. Then, a slurry for forming a porous body was prepared. At this time, the concentration of the particles in the slurry was 21.8% by weight, and the amount of the zeolite particles charged into the molded product was 63% by weight. After stirring overnight, a dry-wet spinning device was used to put the obtained slurry into a cylinder pump at 22.3 ml / min and inject it into a water bath at 25 ° C. as a core liquid. A film was formed into a hollow thread by flowing water at 11.7 ml / min at 11.7 ml / min.

The produced hollow fiber membrane had an outer diameter of 2.4 mm and an inner diameter of 1.4 mm. According to thermal analysis, the weight loss when heated to 600 ° C. is 36% by weight, and the content of zeolite particles in the molded product is considered to be 64% by weight. After drying overnight at 60 ° C. and further heating at 150 ° C. for 2 hours, the porosity was 40% as measured by a mercury porosity, and according to the pore size distribution curve, it was a porous body having macropores of 0.43 μm. According to the nitrogen adsorption measurement, the total pore volume in the molded body after the drying treatment at 60 ° C. is 100% or more of the powder sample in terms of the amount of zeolite particles, of which the low pressure portion due to the zeolite pores (relative pressure P / Po < The amount of nitrogen adsorbed in 0.12) was 64% or more of the powder sample.

( Reference example 10)
Polysulfone (Udel PSF p-1700, Solvay Advanced Polymers Co., Ltd.) is used as the organic polymer, and dimethylformamide of Reference Example 3 is used as a good solvent for the slurry . The raw material particles F were added while stirring the previously dissolved solvent to prepare a molding slurry, and the hollow yarn was spun under the same conditions using the same nozzle as in Reference Example 9.

The produced hollow fiber membrane had an outer diameter of 1.8 mm and an inner diameter of 1.0 mm. The content of the zeolite particles in the molded product by thermal analysis was 59% by weight, and it is considered that there was a loss of 4-5% of fine particles with respect to the charging ratio at the time of spinning. After drying overnight at 60 ° C., the porosity was 66% as measured by a mercury porosity, and according to the pore size distribution curve, it was a multidimensional porous body having macropores of 0.22 μm and 1.43 μm. According to the nitrogen adsorption measurement, the total pore volume in the molded body after the drying treatment at 60 ° C. is 85% of the powder sample in terms of the amount of zeolite particles, of which the low pressure portion due to the zeolite pores (relative pressure P / Po <0). The amount of nitrogen adsorbed in 12.12) was 77% or more of that of the powder sample.

( Reference example 11)
While 203.50 g of dimethylformamide used in Reference Example 3 was heated and stirred, 20.24 g of the methyl methacrylate resin used in Reference Example 3 was gradually mixed as an organic polymer, and the mixture was stirred at 50 ° C. and 330 rpm overnight. And dissolved. While this was heated and stirred, 40.22 g of the synthetic high silica zeolite (raw material particles F) used in Reference Example 9 was added, and stirring was further continued for 4-5 hours to prepare a slurry for forming a porous body. .. At this time, the concentration of the zeolite particles in the slurry was 15.0% by weight, and the amount of the zeolite particles charged into the molded product was 67% by weight. After stirring for 5 hours, the obtained slurry was put into a cylinder pump using the double-mouthed nozzle used in Example 1 and injected into a hot water bath at 30 ° C. at 20.6 ml / min to form a core liquid of 25. A film was formed into a hollow thread by flowing water at ℃ at 7.6 ml / min.

The produced hollow fiber membrane had an outer diameter of 2.2 mm and an inner diameter of 1.0 mm. According to thermal analysis, the weight loss when heated to 600 ° C. is 39% by weight, and the content of zeolite particles in the molded product is considered to be 61% by weight. According to mercury porosity, the void ratio after drying at 60 ° C. overnight was 55%. According to the pore size distribution curve, it was a multi-dimensional porous body having macropores with a diameter of 2.5 μm and a diameter of 6 μm. According to the nitrogen adsorption measurement, the total pore volume in the molded body after drying at 60 ° C. is 82% of the powder sample in terms of the amount of zeolite particles, of which the low pressure part due to the zeolite pores (relative pressure P / Po <0.12). ) Was 64% or more of the powder sample.

( Reference example 12)
In order to improve the strength and flexibility of the hollow fiber obtained in Reference Example 11, 4 g of benzoyl peroxide was dissolved in 396.2 g of ethanol (Wako special grade 99.5%) in the same manner as in Reference Example 3. Hollow fibers after drying overnight at room temperature were placed in a container and heated at 40-50 ° C. for 2 hours for repolymerization. After further washing and drying at room temperature, heat treatment was performed again in ethanol under reduced pressure for 2 hours. According to the mercury porosity, the void ratio was 40% due to the polymerization treatment, which was smaller than that before the treatment, and macropores were formed at 0.43, 1.3, and 2.5 μm diameters according to the pore size distribution curve. According to the nitrogen adsorption measurement, the total pore volume in the molded body after vacuum drying treatment at 150 ° C. for 24 hours is 97% of the powder sample in terms of fine particle content, of which the low pressure part due to the micropores (relative pressure P / Po). The amount of nitrogen adsorbed in <0.12) was 77% or more of the powder sample, and the pore retention rate was also increased as compared with the hollow yarn before the treatment. Further, 0.3087 g of a 60 ° C. dried product of the hollow fiber was placed in 30 ml of distilled water, continuously infiltrated at 120 rpm for 30 hours or more, dried again at 60 ° C. overnight, and weighed. No weight loss was observed.

( Reference example 13)
Reference Example 9-12 in a state where 110.95 g of a normal methylpyrrolidone solution (Vilomax HR-16NN, Toyobo Co., Ltd.) containing 14% by weight of a polyamide-imide resin as an organic polymer was heated and stirred at 45 ° C. and 335 rpm. 31.19 g of the synthetic high-silica zeolite (raw material particles F) used in the above was added, and stirring was continued overnight to prepare a slurry for forming a porous body. At this time, the concentration of the zeolite particles in the slurry was 22% by weight, and the amount of the zeolite particles charged into the molded product was 67% by weight. After stirring for 5 hours, the obtained slurry was put into a cylinder pump using the double-mouthed nozzle used in Example 1 and injected into a hot water bath at 30 ° C. at 21.4 ml / min to form a core liquid of 25. A film was formed into a hollow thread by flowing water at ° C. at 10.4 ml / min.

The produced hollow fiber membrane had an outer diameter of 2.6 mm and an inner diameter of 1.0 mm, and had a bending strength of about 4-5 MPa. According to thermal analysis, the content of zeolite particles in the molded product was around 65% by weight. According to mercury porosity, the void ratio after drying at 60 ° C. overnight was 50%. According to the pore size distribution curve, it was a multi-dimensional porous body having mesopores with a diameter of 20 nm or less and macropores with a diameter of about 0.2 μm. According to the nitrogen adsorption measurement, the total pore volume in the molded body after the drying treatment at 60 ° C. was 98% or more of the powder sample in terms of the amount of zeolite particles, but the low pressure part (relative pressure P / Po) due to the zeolite pores. The amount of nitrogen adsorbed in <0.12) was 24% or less of that of the powder sample, and the pore volume of the mesopores was large, but the micropores were considerably crushed.

( Reference example 14)
In order to improve the micropore function of the polyamide-imide hollow fiber obtained in Reference Example 13, the hollow fiber after drying at room temperature was heat-treated in ethanol under reduced pressure at 40-50 ° C. as in Reference Example 12. It was done for 2 hours. According to the thermal analysis, the zeolite content was 65% by weight, and no change was observed. According to the mercury porosity, the void ratio was 59%, which was higher than that before the treatment, and according to the pore size distribution curve, macropores were formed in the diameter of 0.43 μm instead of the diameter of 0.2 μm. According to the nitrogen adsorption measurement, the total pore volume in the molded body after vacuum drying at 150 ° C for 24 hours was 0.17 cm ³ / g, which was 1.4 times that of the raw material particles, of which the low pressure part due to the micropores (relative). The amount of nitrogen adsorbed at a pressure of P / Po <0.12) was 84% or more of that of the powder sample, and the pore retention rate was significantly increased as compared with the hollow yarn before the treatment. Moreover, when the strength of this hollow fiber was measured, the bending strength was 4-6 MPa, and no decrease in strength was observed as compared with that before the treatment. FIG. 5 shows a comparison of nitrogen adsorption isotherms with respect to the raw material particles F of Reference Example 13 and Reference Example 14 (modified product of Reference Example 13) at room temperature. It should be noted that the pretreatment was performed under a reduced pressure condition of 150 ° C. for 24 hours, and the conversion was based on the content of the particles F.

( Reference example 15)
As the raw material particles, 20.10 g of synthetic high silica zeolite (SiO ₂ / Al ₂ O ₃ = 52) (raw material particles G) having a particle diameter of 0.5-1 μm, a crystal particle diameter of 50 nm, and a pore diameter of 0.58 nm was used in Example 2. It was mixed in advance with 50.10 g of the dimethyl sulfoxide used, and 9.98 g of the resin used in Example 2 was added as an organic polymer while heating and stirring at 47 ° C. at 200 rpm. After stirring at 200 rpm-300 rpm for 4 hours, 20.07 g of dimethyl sulfoxide heated to 50 ° C. was added while continuing the heating and stirring, and the stirring was further continued for 3 hours to prepare a slurry for forming a porous body. At this time, the concentration of the particles in the slurry was 20% by weight, and the amount of the zeolite particles charged into the molded product was 67% by weight. Using the double-mouthed nozzle used in Example 1, an attempt was made to spin a hollow fiber while flowing a slurry at 19 to 22.5 mL / min under the condition of a free running distance of 5 cm and a hot water bath at 27 ° C., but the core liquid was 0. When -2 mL / min was flowed, the injection of the slurry was interrupted and became intermittent, and hollow fiber molding was not possible, and a pellet-shaped molded body was obtained instead.

When the pellet-shaped molded product after washing and drying was measured with a mercury porosity, the void ratio was around 69%. The particle content estimated from the thermal analysis was 67%, which was in line with the charging ratio. According to the nitrogen adsorption measurement, the total pore volume in the molded body is 56% of the powder sample in terms of the zeolite particle amount, and the nitrogen adsorption amount in the low pressure part (relative pressure P / Po <0.12) due to the zeolite pores is powder. It was 57% of the sample.

(Example 16)
1.68 g of the polyether sulfoxide used in Example 2 as an organic polymer was dissolved in 11.70 g of dimethyl sulfoxide at 50 ° C., and 0.85 g of raw material particles A and 1.73 g of raw material particles F were added thereto. Pellets were prepared in the same manner as in Example 2 except that the molding slurry was prepared. At this time, the concentration of the fine particles in the slurry was 16% by weight, and the charging ratio of the fine particles to the molded body was 20% by weight for the raw material particles A and 41% by weight for the raw material particles F. When the obtained pellets were subjected to thermal analysis, the weight loss due to the decomposition of organic matter was about 20%, and the content of fine particles was 80%, which was higher than the charging ratio. According to the mercury porosity, the void ratio of the molded body was 69%, and according to the pore size distribution curve, it was a multidimensional porous body having mesopores in a region of about 5 nm in addition to a plurality of macropores. Further, according to the nitrogen adsorption measurement, the total pore volume in the molded product was about 50% of that of the powder sample in terms of the fine particle content.

( Reference example 17)
As shown in Table 2, the molded product of Comparative Example 7, which was molded using PMMA resin as an organic polymer and further subjected to repolymerization treatment with benzoyl peroxide, has a low retention rate of micropores, but in ethanol under reduced pressure. When the boiling treatment at 40-55 ° C. was carried out for 1-2 hours, as shown in Table 3, a remarkable improvement was obtained in the adsorption performance test results of the micropores. (Table 3 shows the results of the cobalt adsorption test from the aqueous solution conducted using Reference Example 11 and Comparative Example 7 and their modified products.)

(Comparative Example 1)
In Example 2, pellets containing only polyether sulfone were prepared in the same manner as in Example 2 except that the raw material particles were not added and a double amount of polyether sulfone, which is an organic polymer, was added. According to mercury porosity, the pellet was a porous body with a void ratio of 42%, but the total pore volume of the micro to mesopores due to nitrogen adsorption was 0.01 mL / g or less.

(Comparative Example 2)
1.15 g of polysulfone used in Reference Example 10 as an organic polymer was added to 1.15 g of raw material particles A and 20.36 g of dimethylformamide used in Reference Examples 3, 10 and 11 at the same time, and the mixture was stirred to prepare a molding slurry. Other than the prepared ones, pellet molding was attempted in the same manner as in Example 2, but molding was not possible.

(Comparative Example 3)
An attempt was made to prepare pellets in the same manner as in Reference Example 7, except that an ethylene-vinyl alcohol copolymer resin (Soanol D2908, Nippon Synthetic Chemical Industry Co., Ltd.) having an ethylene content of 29 mol% was used as the organic polymer. The prepared slurry spread in the form of a film on the surface of the poor solvent for coagulation, and pellet-like molding could not be performed.

(Comparative Example 4)
As the solvent for the slurry, a mixed solvent of 95.5 g of dimethylformamide used in Reference Examples 3, 10 and 11 and 54.83 g of dimethylacetamide (Wako special grade) was used, and 60.12 g of raw material particles F was dispersed therein in advance. An attempt was made to prepare a slurry for forming a porous body in the same manner as in Reference Example 10, except that polysulfone as an organic polymer was added while heating and stirring, but polysulfone did not mix in the slurry and the stirrer was used. When stopped, precipitation occurred at the bottom and a uniform slurry could not be obtained.

(Comparative Example 5)
After dissolving 0.54 g of an amorphous polyester resin (Byron 200, Toyobo Co., Ltd.) as an organic polymer in the dimethylformamide used in Reference Examples 3, 10 and 11, the raw material particles F0. An attempt was made to prepare a molding slurry by adding 54 g, but a uniform slurry could not be obtained.

(Comparative Example 6)
As described in Reference Example 15, a slurry for molding was prepared by a method of premixing the raw material particles G, but the injection of the slurry was not stable and the hollow fiber could not be molded.

(Comparative Example 7)
16.23 g of the methyl methacrylic resin used in Reference Examples 3 and 11 as an organic polymer was dissolved in dimethyl sulfoxide 160.69 g by heating at 45 ° C. and then dissolved, and then the natural mordenite powder from Kamiayashi (Shin-Tohoku) was dissolved while stirring. Chemical Industry Co., Ltd., raw material particles H) 40.0 g were added while sieving with a 30-mesh SUS wire mesh to prepare a molding slurry. At this time, the concentration of the fine particles in the slurry was 18% by weight, and the amount of the zeolite particles charged into the molded product was 71% by weight. After stirring overnight, the obtained slurry was put into a cylinder pump using the double-mouthed nozzle used in Example 1 in a dry-wet spinning apparatus and injected into a hot water bath at 40 ° C. at 22.8 ml / min. At the same time, water at 25 ° C. was flowed as a core liquid at 10.1 ml / min to form a hollow thread-like film.

The produced hollow fiber had an outer diameter of 2.2 mm and an inner diameter of 1.0 mm. Since it is very brittle as it is, the hollow fiber after being dried overnight at room temperature in a container in which 8 g of benzoyl peroxide is dissolved in ethanol (Wako special grade 99.5%) 1L 791.82 g) as in Reference Example 3. Was added and heated at 40-60 ° C. for 2 hours for repolymerization. When the hollow yarn after the treatment was washed and dried and subjected to thermal analysis, the weight loss due to decomposition of organic substances was 40% by weight, and the content of zeolite particles in the molded product was 60% by weight, but the molding slurry was higher than that at the time of molding. It is probable that the loss when adding the raw material to the product was large. According to mercury porosity, the void ratio was 46%. According to the nitrogen adsorption measurement, the total pore volume in the molded body after vacuum drying treatment at 150 ° C. for 24 hours is 24% of the powder sample in terms of fine particle content, especially the low pressure part due to the micropores (relative pressure P / Po). The amount of nitrogen adsorbed in <0.12) was 5% or less of the powder sample, and the same was true even when the hollow yarn before the polymerization treatment was measured.

Table 1 shows the physical characteristics of the functional fine particles used in Examples , Reference Examples and Comparative Examples.

Tables 2 to 4 show the physical characteristics of the (porous) molded product obtained in Examples , Reference Examples or Comparative Examples.

The cobalt (CoCl ₂ ) adsorption performance of the molded articles after ethanol treatment of Reference Example 11, Comparative Example 7 and Comparative Example 7 was compared with the respective raw material particles by a batch test at room temperature for 24 hours. The results are shown in Table 5. 0.3 g of each adsorbent was added to 50 ppm of a 30 mL cobalt aqueous solution. The Si / Al ratio in the zeolite is different between the raw materials F and H. In the zeolite particles, the substitution of Si in the structure with Al creates an ion exchange site, so that the Co adsorption capacity of the raw material H is considerably larger than that of F. In Reference Example 11, the amount of cobalt adsorbed by the PMMA resin on the molded product is higher than that of the raw material F, but the amount of cobalt adsorbed on the PMMA molded product of Comparative Example 7 is considerably lower than that of the raw material H. When Comparative Example 7 was heated in ethanol under reduced pressure as in Reference Examples 12 and 14, the adsorption capacity of cobalt equivalent to that of the raw material particles was observed as shown in Table 5.

(Example 18)
9.98 g of an ethylene-vinyl alcohol copolymer resin (EVAL G156B, Kuraray Co., Ltd.) having an ethylene content (molar ratio) of 48 mol% as an organic polymer is mixed with 102.88 g of dimethyl sulfoxide (Wako special grade) at 40 ° C. It was dissolved by stirring overnight at 200 rpm. While heating and stirring this, 19.99 g of MCM-41 type synthetic mesoporous silica (SiO ₂ , Sigma-Aldrich No. 643653) (raw material particles I) having a particle diameter of 0.2 μm to 2 μm and a pore diameter of 2 nm to 3 nm was added. Then, a slurry for forming a porous body was prepared. At this time, the concentration of the particles in the slurry was 15% by weight, the amount of the ethylene-vinyl alcohol copolymer resin added was 7.5% by weight, and the amount of MCM particles charged into the molded product was 67% by weight. After stirring overnight, use a double-ended nozzle with a slurry discharge port diameter of 3 mmφ and a core liquid discharge port diameter of 0.7 mmφ in a dry-wet spinning device, and put the obtained slurry into a cylinder pump at 20.5 ml / min. Then, while injecting into a hot water bath at 37 ° C., hot water at 29 ° C. was flowed as a core liquid at 7 ml / min to form a hollow fiber-like film to obtain "MCM-41-EVOH hollow fiber". FIG. 6 shows an external image of the obtained molded product.

The produced hollow fiber membrane has an outer diameter of 1.2 mm and an inner diameter of 0.7 mm, and after being dried overnight at 60 ° C., the porosity is around 62% when measured with a mercury porosity. According to the pore size distribution curve, it is derived from MCM-41. It was a multi-dimensional porous molded body having nanopores having a diameter of 4 nm to 10 nm and macropores of 0.1 μm-1 μm. 0.0299 g of a 60 ° C. dried product of this hollow filamentous molded product was placed in 3 ml of distilled water, continuously infiltrated at 100 rpm for 36 hours or more, dried at 60 ° C. again, and weighed to be 0.0297 g. The weight change is within 1%, and it is considered that there is almost no powder loss. According to thermal analysis, the organic polymer decomposes at 200 ° C or higher when heated to 600 ° C, and the weight loss due to this decomposition is 34% by weight. Therefore, the content of MCM-41 particles in the molded product is 66% by weight. It is believed that there is. Further, according to the nitrogen adsorption isotherm, the relative pressure (P / Po) range of the hysteresis of the isotherm of the molded body and the raw material particles is the same, and it is considered that the size of the mesopores of the raw material particles is almost maintained. Further, since the pore volume in the molded body was 0.28 cm ³ / g and the mesopore volume of the raw material MCM-41 powder particles was 0.59 cm ³ / g, the pore volume of the molded body was included. When converted into the amount of MCM particles, it is considered that 71% of the pore volume of the powder sample is retained.

(Example 19)
Completely 0.54 g of polyether sulfoxide (Veradel PES 3000P Solvay Advanced Polymer Co., Ltd., weight average molecular weight (Mw) = 57,000) as an organic polymer to 5.50 g of dimethyl sulfoxide (Wako special grade) at 50 ° C. It was dissolved and 0.55 g of the synthetic mesoporous silica (raw material particles I) used in Example 18 was mixed thereto to prepare a slurry. At this time, the concentration of the particles in the slurry was 8.3% by weight, the amount of the polyether salphon added was 8.2% by weight, and the amount of the MCM particles charged to the molded product was 50% by weight. This slurry is dropped into a beaker containing hot water at about 32 ° C., solidified for 2 hours or more, dried at 60 ° C., and further stirred and washed at room temperature for 25 hours or more until the white turbidity disappears. Was molded.

After stirring and washing until the white turbidity disappeared, 0.0288 g of the pellet was stirred in 3 mL of water at room temperature for 72 hours or more, and the dry weight was measured. As a result, almost no change in weight was observed. When this was heated to 600 ° C by thermal analysis, the organic polymer decomposed at 450 ° C or higher, and the weight loss due to decomposition was 47%. Therefore, the content of MCM particles in the pellet is about 53%. it is conceivable that. (Reversible endothermic peaks and weight loss not found in the raw material particles and organic polymers were observed around 120 ° C, and it is considered that new adsorption sites derived from molding were generated.) The void ratio was 78% as measured by a mercury porosimeter. According to the pore size distribution curve, it was a multidimensional porous body having nanopores with a diameter of 4 nm to 10 nm and macropores with a diameter of 0.1 μm to 1 μm derived from MCM-41. According to the nitrogen adsorption isotherm, the pore volume of the molded body was 0.50 cm ³ / g, and the pore volume of the powder sample measured under the same conditions was 0.59 cm ³ / g. When converted into the amount of MCM-41 particles contained in, it is considered that almost all the mesopore volume of MCM-41 in the molded body is retained.

(Example 20)
"FSM-22-4.5nm-PES pellets" were prepared in the same manner as in Example 19 except that FSM-22 type synthetic mesoporous silica (raw material particles J) having a pore diameter of 4.5 nm was used as the raw material particles. Molded. According to thermal analysis, the content of particles in the pellet was about 50%.

(Example 21)
"FSM-22-6nm-PES pellets" were formed in the same manner as in Example 19 except that FSM-22 type synthetic mesoporous silica (raw material particles K) having a pore diameter of 6 nm was used as the raw material particles. According to thermal analysis, the content of particles in the pellet was about 52%.

(Example 22)
"FSM-22-8nm-PES pellets" were formed in the same manner as in Example 19 except that FSM-22 type synthetic mesoporous silica (raw material particles L) having a pore diameter of 8 nm was used as the raw material particles. According to thermal analysis, the content of particles in the pellet was about 55%.

(Example 23)
"SBA-15-4nm-PES pellets" were formed in the same manner as in Example 19 except that SBA-15 type synthetic mesoporous silica (raw material particles M) having a pore diameter of 4 nm was used as the raw material particles. According to thermal analysis, the content of particles in the pellet was about 50%. Further, FIG. 7 shows an external image of the SBA-15 type synthetic mesoporous silica (raw material particles M) used in Example 23, the obtained molded product, and the composite of the molded product and the enzyme (lipase). (A) is a powder (10 mg) of typical mesoporous silica (SBA-15 type, raw material particles M), (b) is a molded product (20 mg), and (c) is the molded product. It is a complex with an enzyme (lipase). (A) and (b) are white, and (c) is brown.

(Example 24)
"SBA-15-6nm-PES pellets" were formed in the same manner as in Example 19 except that SBA-15 type synthetic mesoporous silica (raw material particles N) having a pore diameter of 6 nm was used as the raw material particles. According to thermal analysis, the content of particles in the pellet was about 50%.

(Example 25)
"SBA-15-8nm-PES pellets" were formed in the same manner as in Example 19 except that SBA-15 type synthetic mesoporous silica (raw material particles O) having a pore diameter of 8 nm was used as the raw material particles. According to thermal analysis, the content of particles in the pellet was about 50%.

(Example 26)
"SBA-16-5nm-PES pellets" were formed in the same manner as in Example 19 except that SBA-16 type synthetic mesoporous silica (raw material particles P) having a pore diameter of 5 nm was used as the raw material particles. According to thermal analysis, the content of particles in the pellet was about 54%.

(Example 27)
0.53 g of the organic polymer used in Example 18 was placed in 5.55 mL of dimethyl sulfoxide (Wako special grade), heated and stirred to dissolve it, and MSU-X type synthetic mesoporous material having an average pore diameter of 5.8 nm was added thereto. A slurry was prepared by mixing 0.54 g of aluminum oxide (Al ₂ O ₃ , Sigma-Aldrich No. 517747) (raw material particles Q). This slurry was dropped into a beaker containing room temperature water to solidify it, and dried at 60 ° C. to form "mesoporous Al ₂ O ₃ -EVOH pellets".

According to thermal analysis, the weight loss due to decomposition of the organic polymer when heated to 600 ° C. is 54%, and the content of particles in the pellet is considered to be about 46%.

(Example 28)
The same as in Example 19 except that MSU-X type synthetic mesoporous aluminum oxide (Al ₂ O ₃ , Sigma-Aldrich No. 517747) (raw material particles Q) having an average pore diameter of 5.8 nm was used as the raw material particles. "Mesoporous Al ₂ O ₃ -PES pellets" were formed. According to thermal analysis, the content of particles in the pellet was about 50%.

(Example 29)
0.10 g of the organic polymer used in Example 18 was placed in 0.7 mL of dimethyl sulfoxide (Wako special grade), heated and stirred to dissolve it, and the mesoporous titanium oxide having a particle diameter of 10 to 100 nm and an average pore diameter of 4.2 nm was dissolved therein. (Alfa Aesar No. 43250) (raw material particles R) 0.20 g was mixed to prepare a slurry. This slurry was dropped into a beaker containing room temperature water to solidify it, and dried at 60 ° C. to form "mesoporous TiO ₂ -EVOH pellets".

According to thermal analysis, the weight loss due to decomposition of the organic polymer when heated to 600 ° C. is 47%, and the content of particles in the pellet is considered to be about 65%. Further, the specific surface area in the molded body calculated from the results of steam adsorption measurement after pretreatment at 100 ° C. was 127 m ² / g, the pore volume of the raw material powder was 0.29 cm ³ / g, and the specific surface area was 235 m ² / g. Therefore, it is considered that almost all the mesoporous volume of the mesoporous titanium oxide in the molded body was maintained when converted into the amount of the mesoporous titanium oxide particles contained.

(Comparative Example 8)
In Example 19, "PES pellets", which are pellets containing only polyether sulfone, were produced in the same manner as in Example 19 except that the raw material particles were not added and a double amount of polyether sulfone, which is an organic polymer, was added. did. According to the mercury porosity, the pellet was a porous body with a void ratio of 42%, but according to the nitrogen adsorption measurement, the total pore volume of the molded body was 0.01 mL / g or less.

Table 6 shows the physical properties of various mesoporous particles used in the production of porous molded bodies of mesoporous particles.

Table 7 shows the porous molded body of the mesoporous fine particles obtained in Examples 18 to 29, and the molded body produced in Example 19 using only the organic polymer polyether sulfone without the raw material particles. Shows the physical properties of.

(Example 30: Production of AzoR-MCM-41-EVOH hollow fiber complex and enzymatic reaction)
In this example, the immobilization of azo reductase (AzoR) on the molded product (MCM-41-EVOH hollow yarn) obtained in Example 18 and the evaluation of the enzyme activity were performed. FIG. 8 shows an example of an experimental procedure for producing a complex of MCM-41-EVOH hollow yarn and AzoR, a reduction decomposition reaction of an azo dye (methyl red), and reuse of an immobilized enzyme.

(1-1) Method for Producing AzoR-MCM-41-EVOH Hollow Thread Complex The MCM-41-EVOH hollow obtained in Example 18 was used as a porous molded body of mesoporous fine particles in the enzyme-immobilized support. Threads (average pore size: 2.66 nm) were used. In addition, MCM-41 type synthetic mesoporous silica (raw material particles I) was used as the mesoporous silica fine particles before molding for comparison. As the azo reductase, AzoR (dimer, number of amino acid residues: 201, molecular weight (monomer): about 23 kD) derived from Escherichia coli was used.

First, a circular plasmid DNA for AzoR expression was prepared by inserting the AzoR gene amplified from the chromosomal DNA of Escherichia coli into the pET100 / D-TOPO vector. Next, the cyclic plasmid DNA was introduced into recombinant Escherichia coli, and AzoR was produced using an Escherichia coli protein expression system.

When immobilizing the enzyme, it was cut into 1 mL of a buffer solution (25 mM Tris-HCl (pH 7.5)) containing AzoR (0.387 mg) in advance to a length of 5 mm according to the procedure shown in FIG. MCM-41-EVOH hollow yarn (15 or 30 mg) weighed into a microtube and equilibrated with the same buffer was compounded by gently mixing at room temperature for 20 hours using a rotator. AzoR-MCM-41-EVOH hollow yarn composite was obtained by performing the centrifugation operation and the washing operation using the same buffer solution twice. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used as it is, but in this example and the following Examples 31, 32, and 33, the molded body is used. In order to evaluate the binding stability of the enzyme to, a washing step is performed together with the following centrifugation step.

Specifically, the dispersion liquid of the AzoR-MCM-41-EVOH hollow fiber complex (unwashed) was centrifuged (4 ° C., 18,000 G, 10 minutes), and all the supernatant was recovered. .. Subsequently, 1 mL of a washing buffer solution (using the buffer solution selected at the time of immobilization) was added, and the mixture was stirred with a Vortex Mixer at room temperature for about 5 seconds to resuspend the complex of AzoR and the mesoporous silica molded product. After that, centrifugation (4 ° C., 18,000 G, 10 minutes) was performed, and a washing operation was performed to collect all the supernatant. The washing operation was repeated again using 1 mL of the washing buffer solution, and finally, an AzoR-MCM-41-EVOH hollow fiber complex (washed) was obtained. Hereinafter, it is referred to as "AzoR-MCM-41-EVOH hollow fiber".
In addition, MCM-41 type synthetic mesoporous silica (raw material particles I) (10 mg) was used for comparison, and a complex was obtained by the same procedure. Hereinafter, it is referred to as "AzoR-MCM-41-powder".

The amount of enzyme immobilized on the molded product and the immobilization rate are based on the amount of enzyme before immobilization (0.387 mg in 1 mL), and the amount of free enzyme contained in the recovered liquid in the above-mentioned centrifugation step and washing step. Evaluated by subtraction and division of. FIG. 9 shows the amount and rate of AzoR immobilized on MCM-41-powder (typical mesoporous silica (MCM-41 type, raw material particles I) or MCM-41-EVOH hollow fiber of Example 18). show.

From FIG. 9, it was observed that the immobilization amount and immobilization rate of AzoR with respect to the MCM-41-EVOH hollow fiber tended to increase in proportion to the amount of the molded product. From Example 18, considering that the content of MCM-41 particles in the molded product is 66% by weight, the MCM-41 particles contained in 15 mg and 30 mg of the MCM-41-EVOH hollow fiber are 10 mg and 20 mg, respectively. It is considered to be equivalent. Here, comparing the MCM-41-powder (10 mg) and the MCM-41-EVOH hollow yarn (15 mg), although the amount of MCM-41 particles is the same, the hollow yarn molded body is more than the powder. Although the amount of enzyme immobilization and the immobilization rate were lower in the case, it was found that the molded product also retained the enzyme immobilization ability equivalent to 82.5% of the powder.

(1-2) Evaluation of Degradation Activity of Methyl Red The enzyme activity of each of the AzoR-MCM-41-EVOH hollow yarn and AzoR-MCM-41-powder obtained in (1-1) is used as a reaction substrate for an azo dye. It was evaluated by the decomposition rate (bleaching rate) of (methyl red).
Specifically, as shown in FIG. 8, each of the complexes is subjected to the following enzymatic reaction, and the change in absorbance of various reaction substrates (methyl red and coenzyme (NADH)) is examined to examine the enzyme. The activity was evaluated.

The enzymatic reaction was carried out with respect to the AzoR-MCM-41-EVOH hollow yarn and AzoR-MCM-41-powder obtained in (1-1) in a microtube, and a reaction substrate containing methyl red, NADH, etc. It was started by adding 1 mL).
At that time, the reaction composition of the reaction substrate was adjusted to be "25 mM Tris-HCl (pH 7.5), 0.05 mM methyl red, 0.3 mM NADH, 1 μM FMN, reaction solution volume: 1 mL". For comparison, free AzoR without an immobilized support was also reacted by mixing with a similar reaction substrate.
The reaction conditions were to maintain the heated state at 37 ° C. for 30 minutes while gently mixing using a rotator. At the end of the reaction, the supernatant was collected by centrifugation (4 ° C., 18,000 G, 5 minutes), and the absorbance of the recovered solution at 340 nm or 430 nm was measured using a microplate reader (SpectraMax M2e; manufactured by Molecular Devices). By doing so, the consumption rate of NADH 30 minutes after the reaction in the methyl red decomposition reaction or the decomposition rate (bleaching rate) of methyl red was evaluated. 10 and 8 show changes in the absorbance of NADH and methyl red in the reduction decomposition of methyl red by AzoR-MCM-41-EVOH hollow yarn and AzoR-MCM-41-powder, and the decomposition rate of methyl red. FIG. 10 shows the change in absorbance of (a) coenzyme (NADH) and (b) methyl red in the reductive decomposition of the azo dye (methyl red) by the AzoR-mesoporous silica complex (powder or molded product). The decomposition rate of methyl red is shown in (c) of FIG.

From FIGS. 10 (a) and 10 (b), in the case of free AzoR, AzoR-MCM-41-powder, or AzoR-MCM-41-EVOH hollow fiber without an immobilized support, NADH and methyl are used. There was a tendency for the absorbance of red to decrease significantly. This indicates that the decomposition reaction of methyl red with the consumption of NADH is efficiently proceeding even in the immobilized enzyme.

Interestingly, focusing on the change in absorbance after the enzymatic reaction in FIG. 10 (a), a larger decrease in absorbance was shown in the case of the immobilized enzyme as compared with the case of the free enzyme. From this, it is inferred that NADH was consumed more efficiently, or that the immobilized enzyme and NADH form a complex.

Furthermore, the absorbances of NADH and methyl red in the absence of the enzyme showed the same values when both MCM-41-powder and MCM-41-EVOH hollow yarn were used as compared with the reaction solution itself. From this, the reaction substrate itself, that is, NADH and methyl red molecules are hardly adsorbed on the mesopores of the MCM-41-powder, the mesopores of the MCM-41-EVOH hollow yarn, and the organic polymer portion. It was suggested.

Further, from FIG. 10 (c), the decomposition rate of methyl red by the enzyme immobilized on the MCM-41-powder and the MCM-41-EVOH hollow fiber was the same as that in the case of the unfixed free enzyme. It was found that the original enzyme activity was maintained even in the immobilized state.

(Example 31: Production of complex of AzoR and various porous molded products and enzymatic reaction)
In this example, the porous molded body of the mesoporous fine particles obtained in Examples 18 to 28 and the molded product produced using only the organic polymer polyether sulfone in Comparative Example 8 without the raw material particles. Immobilization of azo reductase (AzoR) on the body and evaluation of enzyme activity were performed.

(1-1) Method for Producing Composites of AzoR and Various Porous Molds Eleven types of moldings obtained in Examples 18 to 28 were used as porous moldings of mesoporous fine particles on the immobilized support of AzoR. Used the body. Further, the "PES pellet" obtained in Comparative Example 8 was used as a molded product containing no raw material particles for comparison. Further, nine kinds of synthetic mesoporous silica (raw material particles I to Q) were used as the mesoporous silica fine particles before molding for comparison.

When immobilizing the enzyme, weigh 1 mL of a buffer solution (25 mM MES-NaOH (pH 6)) containing AzoR (0.328 mg) and each well of a 96-well deep-bottom type plate in advance, and use the same buffer. Eleven types of molded bodies and "PES pellets" (equivalent to 20 mg) equilibrated using the liquid were mixed (25) using a stirrer (TAITEC, Deep Well Maximizer, BioShaker M.BR-022UP). Complex was obtained by performing a centrifugation operation at ° C., 1,200 rpm, 24 hours) twice, and performing a centrifugation operation and a washing operation using the same buffer solution twice to obtain a composite of AzoR and various porous molded bodies. .. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used as it is, but in this embodiment, in order to evaluate the binding stability of the enzyme to the molded body. In addition, a cleaning step is performed together with the following centrifugation step.

Specifically, the dispersion liquid of the complex (unwashed) of the AzoR and various porous molded products was centrifuged (25 ° C., 4,700 rpm, 10 minutes), and all the supernatant was recovered. .. Subsequently, 1 mL of a washing buffer solution (using the buffer solution selected at the time of immobilization) was added, and the composite of AzoR and the mesoporous silica molded product was stirred at 25 ° C. for 1 hour using the above-mentioned stirring device. , Centrifugation (25 ° C, 4,700 rpm, 10 minutes) was performed, and a washing operation was performed to collect all the supernatant. The cleaning operation was repeated again using 1 mL of the cleaning buffer solution, and finally, a complex (cleaned) of AzoR and various porous molded products was obtained. Hereinafter, it is referred to as "AzoR-mesoporous fine particle molded product".

Also, for comparison, 9 types of synthetic mesoporous silica (raw material particles I to Q) (equivalent to 10 mg) were used, and a centrifuge for microtubes, rotators, and microtubes (25 ° C, 20,000 G, 10). The composite was obtained by the same procedure as that of the molded product except that the minutes were used. Hereinafter, it is referred to as "AzoR-mesoporous fine particle powder".

The amount of the enzyme immobilized on the molded body and powder of various mesoporous fine particles is the amount of the enzyme before immobilization (0.328 mg in 1 mL), and the amount of the free enzyme contained in the recovered liquid in the above-mentioned centrifugation step and washing step. Calculated by subtracting the amount. FIG. 11 shows the amount of AzoR immobilized on the mesoporous fine particle molded product, the molded product containing no raw material particles, or the mesoporous fine particle powder. (A) is the amount of AzoR immobilized on various molded bodies of mesoporous fine particles. (B) is the amount of AzoR immobilized on various powders of mesoporous fine particles.

From FIGS. 11 (a) and 11 (b), in the immobilized support other than the mesoporous Al ₂ O ₃ -EVOH pellet, the SBA-16-5 nm-powder, or the mesoporous Al ₂ O ₃ powder, the AzoR is generally equivalent. The amount of immobilization was shown. From Table 7, considering that the content of the mesoporous fine particles in the 11 types of molded bodies obtained in Examples 18 to 28 is 52% by weight on average, the mesoporous fine particles contained in 20 mg of the molded body correspond to 10 mg. The immobilization amount of AzoR shown in FIG. 11 (a) was equivalent to the immobilization amount of AzoR with respect to the mesoporous fine particle powder (10 mg) in FIG. 11 (b). It suggests that it retains the same enzyme immobilization ability as powder. Here, SBA-16-5nm-powder or mesoporous Al ₂ O3 powder and SBA-16-5nm-PES pellets, mesoporous Al ₂ O _{3 -EVOH} pellets or mesoporous Al containing these powders. Comparing with _{2O3 -} PES pellets, although almost no enzyme was immobilized on the above two types of powder, among the above three types of molded products, especially as an organic polymer, polyether monkeys were used. In the case of SBA _- 16-5nm-PES pellets and mesoporous _Al2O3 -PES pellets molded using a phone (PES), the same enzyme fixation as when using PES pellets containing no raw material particles The amount of chemical conversion was shown. From this, it was found that the enzyme (AzoR) was non-specifically adsorbed on the PES surface portion of the molded product, and when a molded product using polyethylene polyvinyl alcohol copolymer resin (EVOH) was used as the organic polymer. Suggested that it may be specifically immobilized on the mesopores contained in the molded product. Therefore, from the results of Example 30 and FIG. 11, it was clarified that a molded product using EVOH, such as MCM-41-EVOH hollow fiber, is suitable for immobilization of AzoR.

(1-2) Evaluation of Degradation Activity of Methyl Red 11 types of AzoR-mesoporous fine particle molded product obtained in (1-1), AzoR-PES pellets (without raw material particles), and 9 types of AzoR-mesoporous fine particles. The enzymatic activity of each powder was evaluated by the decomposition rate (bleaching rate) of the azo dye (methyl red) as a reaction substrate. At this time, the durability (reaction 5 times) of the complex in repeated use was also evaluated.
Specifically, the following enzymatic reaction was caused in each of the complexes, and the decolorization rate was determined by examining the change in the absorbance of methyl red.

Prior to the enzymatic reaction, the various AzoR-mesoporous microparticle powders were placed from microtubes into each well of a 96-well deep-bottomed plate containing AzoR-mesoporous microparticle moldings and AzoR-PES pellets (without raw material particles). Transferred. The enzymatic reaction contained methyl red for the AzoR-mesoporous fine particle compact, AzoR-PES pellets (without raw material particles), and AzoR-mesoporous fine particle powder in each well of the 96-well deep type plate. Instead, a Tris buffer solution (0.966 mL) containing only NADH and FMN was added, and the mixture was stirred using the above-mentioned stirring device (30 ° C., 1,200 rpm, 10 minutes), and then a methyl red solution (0. It was started by adding (034 mL) (first reaction).
At that time, the reaction composition of the reaction substrate was adjusted to be "25 mM Tris-HCl (pH 7.5), 0.05 mM methyl red, 0.2 mM NADH, 1 μM FMN, reaction solution volume: 1 mL". From the second reaction onward, the reaction was started by adding a buffer solution, a substrate, or the like to the AzoR complex separated from the supernatant after the reaction after the following centrifugation operation in the same procedure as described above.
For comparison, free AzoR without an immobilized support was also reacted by mixing with a similar reaction substrate.
The reaction conditions were to maintain the heated state at 30 ° C. for 10 minutes while mixing (1,200 rpm) using the above-mentioned stirring device. Subsequently, after 10 minutes have passed, the entire supernatant is recovered by centrifugation (30 ° C., 4,700 rpm, 5 minutes) in a heated state, and the absorbance of the recovered solution at 430 nm is measured using the above-mentioned microplate reader. By doing so, the decomposition rate (bleaching rate) of methyl red was evaluated. FIG. 12 shows the decolorization rate of methyl red during repeated use of various AzoR complexes (reaction 5 times). The decomposition rate (bleaching rate) of methyl red by the AzoR-mesoporous fine particle complex (a: molded body, b: powder) and the durability (reaction 5 times) in repeated use of the immobilized enzyme were shown.

13 (a) shows the molded product of Example 23, and (b) shows the complex (AzoR-SBA-15-4nm-PES pellet) of the molded product and AzoR after the completion of the methyl red decomposition reaction. C) shows the molded product of Comparative Example 8, and (d) shows a photograph of the appearance of the composite (AzoR-PES pellets (without raw material particles)) of the molded product and AzoR after the completion of the methyl red decomposition reaction.

From FIGS. 12 (a) and 12 (b), a bleaching rate of about 90% of methyl red is shown as in FIG. 10 (c) except for SBA-15-6 nm-powder, and high durability in repeated use is shown. Was recognized.

On the other hand, FIG. 14 shows the results of evaluating the enzymatic activity of free AzoR (enzyme amount: 0, 1, 10, 100 μg). FIG. 14 shows the effect of the unfixed free AzoR concentration on the decomposition rate (bleaching rate) of methyl red. From FIG. 14, it was found that the optimum amount of enzyme that enables the bleaching rate of methyl red of 90% or more is 1 to 10 μg, and the enzyme activity is not expressed at 100 μg. From FIG. 11, considering that about 100 to 200 μg of an enzyme is immobilized on various AzoR complexes, a part of the immobilized enzyme, that is, an enzyme corresponding to 1 to 10 μg is involved in the reaction. It is inferred that they are doing it. This suggests that the activity of the free enzyme is significantly reduced when the high-concentration enzyme is used, but the original enzyme activity can be stably expressed in the immobilized state.

Interestingly, when the appearances of the AzoR-mesoporous fine particle molded product and the AzoR-PES pellets (without raw material particles) after the reaction in FIG. 12A were confirmed, the mesoporous fine particle molded product exhibited a pink color. , The PES pellet (without raw material particles) exhibited the same white color as before the reaction (FIG. 13). This suggests that in the AzoR-mesoporous fine particle molded product, a complex of AzoR-NADH-methyl red is formed in the mesopores in the molded product. On the other hand, in the AzoR-PES pellets (without raw material particles), it is considered from FIG. 11 that a considerable amount of AzoR is non-specifically adsorbed on the PES surface portion of the molded product, but the adsorbed AzoR is a reaction substrate. It is inferred that they do not form a complex. That is, it is considered that the decomposition reaction of highly active methyl red by the AzoR-PES pellets (without raw material particles) shown in FIG. 12 (a) is due to the contribution of a small amount of free AzoR desorbed from the molded product. Therefore, it was suggested that AzoR immobilized on the mesopores in the molded product greatly contributed to the enzymatic reaction by the AzoR-mesoporous fine particle molded product.

(Example 32: Production of a complex of GDH and various porous molded products and enzymatic reaction)
In this example, the porous molded body of the mesoporous fine particles obtained in Examples 18 to 28 and the molded product produced using only the organic polymer polyether sulfone in Comparative Example 8 without the raw material particles. Immobilization of glucose dehydrogenase (GDH: derived from Bacillus sp., Wako Pure Chemical Industries, Ltd.) on the body and evaluation of enzyme activity were performed.

(1-1) Method for Producing Composites of GDH and Various Porous Molds Eleven types of moldings obtained in Examples 18 to 28 were used as porous moldings of mesoporous fine particles on the immobilized support of GDH. Used the body. Further, the "PES pellet" obtained in Comparative Example 8 was used as a molded product containing no raw material particles for comparison. Further, nine kinds of synthetic mesoporous silica (raw material particles I to Q) were used as the mesoporous silica fine particles before molding for comparison.

When immobilizing the enzyme, weigh 1 mL of a buffer solution (25 mM MES-NaOH (pH 6)) containing GDH (0.363 mg) and each well of a 96-well deep type plate in advance, and use the same buffer. 11 kinds of molded bodies equilibrated with the liquid and "PES pellets" (equivalent to 20 mg) are combined by mixing (25 ° C., 1,200 rpm, 24 hours) using the above-mentioned stirring device. Then, the centrifugation operation and the washing operation using the same buffer were performed twice to obtain a composite of GDH and various porous molded bodies. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used as it is, but in this embodiment, in order to evaluate the binding stability of the enzyme to the molded body. In addition, a cleaning step is performed together with the following centrifugation step.

Specifically, the dispersion liquid of the complex (unwashed) of the GDH and various porous molded products was centrifuged (25 ° C., 4,700 rpm, 10 minutes), and all the supernatant was recovered. .. Subsequently, 1 mL of a washing buffer solution (using the buffer solution selected at the time of immobilization) was added, and the composite of GDH and the mesoporous silica molded product was stirred at 25 ° C. for 1 hour using the above-mentioned stirring device. , Centrifugation (25 ° C, 4,700 rpm, 10 minutes) was performed, and a washing operation was performed to collect all the supernatant. The cleaning operation was repeated again using 1 mL of the cleaning buffer solution, and finally, a complex (cleaned) of GDH and various porous molded products was obtained. Hereinafter, it is referred to as "GDH-mesoporous fine particle molded product".

Also, for comparison, 9 types of synthetic mesoporous silica (raw material particles I to Q) (equivalent to 10 mg) were used, and a centrifuge for microtubes, rotators, and microtubes (25 ° C, 20,000 G, 10). The composite was obtained by the same procedure as that of the molded product except that the minutes were used. Hereinafter, it is referred to as "GDH-mesoporous fine particle powder".

The amount of enzyme immobilized on the molded body and powder of various mesoporous fine particles is the amount of free enzyme contained in the recovered liquid of the above-mentioned centrifugation step and washing step from the amount of enzyme before immobilization (0.363 mg in 1 mL). Calculated by subtracting the amount. FIG. 15 shows the amount of GDH immobilized on the mesoporous fine particle molded product, the molded product containing no raw material particles, or the mesoporous fine particle powder. FIG. 15A shows the amount of glucose dehydrogenase (GDH) immobilized on the molded article of various mesoporous fine particles. (B) shows the amount of glucose dehydrogenase (GDH) immobilized on the powder of various mesoporous fine particles.

From FIGS. 15 (a) and 15 (b), as a whole, it is compared with the molded body and powder of SBA-16-5 nm, the molded body and powder of mesoporous Al ₂ O3 _, or the PES pellet containing no raw material particles. A larger amount of GDH immobilization was shown when MCM-41, FSM-22, or SBA-15 type synthetic mesoporous silica moldings and powders were used. Comparing the powder with MCM-41, FSM-22, or SBA-15 type synthetic mesoporous silica molded product, the amount of immobilization on the fine particle powder was slightly higher, but FSM-22 or SBA- Focusing on 15 types of synthetic mesoporous silica, it was found that the amount of GDH immobilization tended to increase in proportion to the pore size as a whole for both the molded product and the powder. It is considered that this is because the amount of immobilization depends on the pore size because GDH having a larger molecular size is not saturated and adsorbed as compared with the saturated adsorption of AzoR in Example 31. That is, it is suggested that the mesoporous fine particles contained in the molded product have the same enzyme immobilization ability as the powder. Here, considering from the amount of GDH immobilized on the molded body and powder of SBA-16-5 nm, the molded body and powder of mesoporous Al ₂ O3 _, or the PES pellet containing no raw material particles, EVOH as an organic polymer is considered. It is presumed that more GDH was adsorbed non-specifically on the PES surface portion of the molded body using PES than that of the molded body using PES. Nevertheless, in the molded product of MCM-41, FSM-22, or SBA-15 type synthetic mesoporous silica, the amount of immobilization is more than twice that in the case of using PES pellets (without raw material particles). It was found that a considerable amount of GDH was immobilized on the mesoporous portion of the molded product.

(1-2) Evaluation of NADH production activity 11 types of GDH-mesoporous fine particle molded product obtained in (1-1), GDH-PES pellets (without raw material particles), and 9 types of GDH-mesoporous fine particle powder The activity of each enzyme was evaluated by the molar concentration of reduced NADH produced from the coenzyme (oxidized NAD ⁺ ) as a reaction substrate. At this time, the durability (reaction 5 times) of the complex in repeated use was also evaluated.
Specifically, the following enzymatic reaction was caused in each of the complexes, and the produced NADH concentration was determined by examining the change in the absorbance of NADH.

The various GDH-mesoporous particulate powders are dispensed from the microtubes into each well of a 96-well deep-bottomed plate that already contains the GDH-mesoporous particulate compact and GDH-PES pellets (without raw material particles) prior to the enzymatic reaction. Transferred. The enzymatic reaction was glucose-free with respect to the GDH-mesoporous fine particle compact, GDH-PES pellets (without raw material particles), and GDH-mesoporous fine particle powder in each well of the 96-well deep-bottomed type plate. , Tris buffer (0.967 mL) containing only NAD ⁺ is added, and the mixture is stirred using the above-mentioned stirring device (30 ° C., 1,200 rpm, 10 minutes), and then a glucose solution (0.033 mL) is added. It was started by adding (first reaction).

At that time, the reaction composition of the reaction substrate was adjusted to be "25 mM Tris-HCl (pH 7.5), 0.5 mM NAD ⁺ , 10 mM glucose, reaction solution volume: 1 mL". From the second reaction onward, the reaction was started by adding a buffer solution, a substrate, or the like to the GDH complex separated from the supernatant after the reaction after the following centrifugation operation in the same procedure as described above. For comparison, free GDH without an immobilized support was also reacted by mixing with a similar reaction substrate.

The reaction conditions were to maintain the heated state at 30 ° C. for 10 minutes while mixing (1,200 rpm) using the above-mentioned stirring device. Subsequently, after 10 minutes have passed, the entire supernatant is recovered by centrifugation (30 ° C., 4,700 rpm, 5 minutes) in a heated state, and the absorbance of the recovered solution at 340 nm is measured using the above-mentioned microplate reader. By doing so, the production efficiency of NADH (NADH concentration) was evaluated. FIG. 16 shows the generated NADH concentration during repeated use of various GDH complexes (reaction 5 times). FIG. 16 (a) shows the concentration of NADH produced by the molded body of the GDH-mesoporous fine particle complex and the durability (reaction 5 times) in repeated use of the immobilized enzyme. (B) shows the concentration of NADH produced by the powder of the GDH-mesoporous fine particle complex and the durability (reaction 5 times) in repeated use of the immobilized enzyme.

From FIGS. 16 (a) and 16 (b), as a whole, GDH immobilized on the molded body and powder of MCM-41 type synthetic mesoporous silica remained low in activity, but FSM-22 or SBA- GDH immobilized on a molded body of 15 types of synthetic mesoporous silica and powder showed a tendency for the enzyme activity to increase in proportion to the pore size. Further, when the activities of the GDH-mesoporous fine particle molded product and the GDH-PES pellet (without raw material particles) are compared from FIG. 16 (a), most of the GDH-mesoporous fine particle molded products are obtained from the GDH-PES pellet (without raw material particles). Also clearly expresses high activity, which suggests the high stability of the enzyme immobilized on the mesopores of the molded product. In the GDH-PES pellets (without raw material particles), it is considered from FIG. 15 that a considerable amount of GDH is non-specifically adsorbed on the PES surface portion of the molded product, but from FIG. 16 (a), it is in an adsorbed state. It was found that the activity of GDH was extremely low. From the above, it was suggested that GDH immobilized on the mesopores in the molded product greatly contributed to the reaction of producing highly active NADH by the GDH-mesoporous fine particle molded product.
Further, when the molded product and the powder were compared, in the case of GDH immobilized on the molded product, although the activity of the first reaction was low, the activity tended to gradually increase with the number of repeated uses. This means that in the initial stage of the reaction, the diffusion of the reaction substrate in the macropores and mesopores in the molded product is rate-determining, and the contact frequency between the enzyme immobilized in the mesopores and the reaction substrate is low. It is inferred that there is.

On the other hand, FIG. 17 shows the results of evaluating the enzyme activity of free GDH (enzyme amount: 0, 1, 10, 100 μg). FIG. 17 shows the effect of the unfixed free GDH concentration on the produced NADH concentration. From FIG. 17, the amount of enzyme that gives a reaction yield of 100% (~ 0.5 mM NADH) is 10 to 100 μg, and the amount of enzyme required to produce 0.1 to 0.5 mM NADH is 1 to ~. It was found to be in the range of 10 μg. From FIG. 16, the concentration of NADH produced by various GDH complexes is about 0 to 0.4 mM, and from FIG. 15, it is considered that about 100 to 300 μg of enzyme is immobilized on the GDH complex. It is presumed that a part of the immobilized enzyme, that is, an enzyme equivalent to 1 to 10 μg is involved in the reaction.

(Example 33: Production of complex of lipase and various porous molded products and enzymatic reaction)
In this example, the porous molded body of the mesoporous fine particles obtained in Examples 18 to 28 and the molded product produced using only the organic polymer polyether sulfone in Comparative Example 8 without the raw material particles. Immobilization of lipase (derived from Phycomyces nitens, Wako Pure Chemical Industries, Ltd.) on the body and evaluation of enzyme activity were performed.

(1-1) Method for Producing Composites of Lipase and Various Porous Molds Eleven types of moldings obtained in Examples 18 to 28 were used as porous moldings of mesoporous fine particles on the immobilized support of lipase. Used the body. Further, the "PES pellet" obtained in Comparative Example 8 was used as a molded product containing no raw material particles for comparison. Further, nine kinds of synthetic mesoporous silica (raw material particles I to Q) were used as the mesoporous silica fine particles before molding for comparison.

When immobilizing the enzyme, weigh 1 mL of a buffer solution (25 mM MES-NaOH (pH 6)) containing lipase (0.456 mg) and each well of a 96-well deep type plate in advance, and use the same buffer. 11 kinds of molded bodies equilibrated with the liquid and "PES pellets" (equivalent to 20 mg) are combined by mixing (25 ° C., 1,200 rpm, 24 hours) using the above-mentioned stirring device. Then, the centrifugation operation and the washing operation using the same buffer were performed twice to obtain a composite of lipase and various porous molded bodies. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used as it is, but in this embodiment, in order to evaluate the binding stability of the enzyme to the molded body. In addition, a cleaning step is performed together with the following centrifugation step.

Specifically, the dispersion liquid of the complex (unwashed) of the above-mentioned lipase and various porous molded products was centrifuged (25 ° C., 4,700 rpm, 10 minutes), and all the supernatant was recovered. .. Subsequently, 1 mL of a washing buffer solution (using the buffer solution selected at the time of immobilization) was added, and the complex of the lipase and the mesoporous silica molded product was stirred at 25 ° C. for 1 hour using the above-mentioned stirring device. , Centrifugation (25 ° C, 4,700 rpm, 10 minutes) was performed, and a washing operation was performed to collect all the supernatant. The cleaning operation was repeated again using 1 mL of the cleaning buffer solution, and finally, a complex (cleaned) of lipase and various porous molded products was obtained. Hereinafter, it is referred to as "lipase-mesoporous fine particle molded product".

Also, for comparison, 9 types of synthetic mesoporous silica (raw material particles I to Q) (equivalent to 10 mg) were used, and a centrifuge for microtubes, rotators, and microtubes (25 ° C, 20,000 G, 10). The composite was obtained by the same procedure as that of the molded product except that the minutes were used. Hereinafter, it is referred to as "lipase-mesoporous fine particle powder".

The amount of the enzyme immobilized on the molded body and powder of various mesoporous fine particles is the amount of the enzyme before immobilization (0.456 mg in 1 mL), and the amount of the free enzyme contained in the recovered liquid in the above-mentioned centrifugation step and washing step. Calculated by subtracting the amount. FIG. 18 shows the amount of lipase immobilized on the mesoporous fine particle molded product, the molded product containing no raw material particles, or the mesoporous fine particle powder. FIG. 18A shows the amount of lipase immobilized on the molded product of various mesoporous fine particles. (B) shows the amount of lipase immobilized on the powder of various mesoporous fine particles.

Synthetic mesoporous of MCM-41, FSM-22, SBA-15 type, SBA-16 type, or mesoporous aluminum oxide type, excluding mesoporous _{Al2O3 -} EVOH pellets from FIGS. 18 (a) and 18 (b). Comparing the silica molded body and the powder, the amount of lipase immobilized on the molded body and the powder is the same. Focusing on FSM-22 or SBA-15 type synthetic mesoporous silica, both the molded body and the powder Overall, the amount of lipase immobilized tended to increase in proportion to the pore size. This is similar to the tendency of GDH immobilization in Example 32, and it is considered that the amount of immobilization depends on the pore diameter because lipase is not saturated and adsorbed to the mesopores. That is, it is suggested that the mesoporous fine particles contained in the molded product have the same enzyme immobilization ability as the powder.

Further, from FIG. 18A, only the mesoporous _{Al2O3 -} EVOH pellets had a low lipase immobilization amount, but other than that, MCM-41, FSM-22, SBA-15 type, and SBA. When a molded body of synthetic mesoporous fine particles of -16 type or mesoporous aluminum oxide type was used, the amount of lipase immobilized was about 2 to 3 times that of PES pellets containing no raw material particles. rice field. From this, it was found that a considerable amount of lipase was immobilized on the mesopore portion of the molded product.

(1-2) Evaluation of Hydrolytic Activity of Triglyceride 11 types of lipase-mesoporous fine particle molded product, lipase-PES pellets (without raw material particles), and 9 types of lipase-mesoporous fine particles obtained in (1-1). The enzymatic activity of each of the powders was evaluated by the molar concentration of the fluorescent substance (pyrene) bound to the fatty acid produced by the hydrolysis of the fluorescent triglyceride as a reaction substrate. At this time, the durability (reaction 5 times) of the complex in repeated use was also evaluated.
Specifically, the following enzymatic reaction was caused in each of the complexes, and the fluorescence intensity of pyrene was examined to determine the free pyrene concentration.

Prior to the enzymatic reaction, the various lipase-mesoporous particulate powders were dispensed from the microtubes into each well of a 96-well deep-bottomed plate already containing the lipase-mesoporous particulate molding and lipase-PES pellets (without raw material particles). Transferred. The enzymatic reaction was carried out against the lipase-mesoporous fine particle compact, lipase-PES pellets (without raw material particles), and lipase-mesoporous fine particle powder in each well of the 96-well deep type plate. (0.9 mL) was added, and the mixture was stirred using the above-mentioned stirring device (30 ° C., 1,200 rpm, 10 minutes), and then started by adding a fluorescent triglyceride solution (0.1 mL) (Reaction 1). The second time).

At that time, the reaction composition of the reaction substrate was adjusted to be "150 mM PBS (pH 8.2), 1 μM fluorescent triglyceride, reaction solution volume: 1 mL". From the second reaction onward, the reaction was started by adding a buffer solution, a substrate, or the like to the lipase complex separated from the supernatant after the reaction after the following centrifugation operation in the same procedure as described above. For comparison, free lipase without an immobilized support was also reacted by mixing with a similar reaction substrate.

The reaction conditions were to maintain the heated state at 30 ° C. for 10 minutes while mixing (1,200 rpm) using the above-mentioned stirring device. Subsequently, after 10 minutes have passed, all the supernatant is recovered by centrifugation (30 ° C., 4,700 rpm, 5 minutes) in a heated state, and the fluorescence intensity of the recovered liquid is measured using the above-mentioned microplate reader (). Measurement wavelength: Excitation 342 nm / Fluorescence 400 nm) was used to evaluate the fatty acid production efficiency (pyrene concentration). FIG. 19 shows the free pyrene concentration at the time of repeated use of various lipase complexes (reaction 5 times). FIG. 19 (a) shows the concentration of free pyrene in the decomposition of triglyceride by the molded product of the lipase-mesoporous fine particle complex and the durability (reaction 5 times) in repeated use of the immobilized enzyme. FIG. 19 (b) shows the concentration of free pyrene in the decomposition of triglyceride by the powder of the lipase-mesoporous fine particle complex and the durability (reaction 5 times) in the repeated use of the immobilized enzyme.

From FIGS. 19 (a) and 19 (b), it was observed that, as a whole, the concentration of free pyrene tended to be lower when the lipase-mesoporous fine particle compact was used as compared with the lipase-mesoporous fine particle powder. , No widespread increase or decrease in pyrene concentration was shown as in the case of using mesoporous fine particle powder. Specifically, the lipase-mesoporous fine particle molded product excluding the MCM-41-EVOH hollow fiber showed a constant pyrene concentration (20 to 40 nM) even after 5 repeated reactions. Further, from FIG. 19 (a), the lipase-PES pellet (without raw material particles) gradually decreased in activity with the number of repeated uses, and hardly exhibited the activity in the fifth reaction. In the lipase-PES pellets (without raw material particles), it is considered from FIG. 18 that a considerable amount of lipase is non-specifically adsorbed on the PES surface portion of the molded body, but the stability of the lipase in the adsorbed state is extremely low. It has been found. On the other hand, most lipase-mesoporous fine particle compacts express sustained enzyme activity in 1 to 5 reactions, which means that the enzyme immobilized in the mesopores of the compact has high stability. It suggests. From the above, it was suggested that the lipase immobilized on the mesopores in the molded body greatly contributed to the hydrolysis reaction of the highly active triglyceride by the lipase-mesoporous fine particle molded body.
In addition, FIG. 20 shows the results of evaluating the enzyme activity of free lipase (enzyme amount: 0, 1, 10, 100 μg). FIG. 20 shows the effect of unfixed free lipase concentration on the concentration of fluorescent substance (pyrene) released by triglyceride decomposition. FIG. 20 shows that the pyrene concentration tends to increase in proportion to the enzyme concentration. From FIG. 20, the amount of the enzyme that gives the produced pyrene concentration (20 to 40 nM) by the lipase-mesoporous fine particle molded product shown in FIG. 19 (a) is considered to be in the range of 50 to 100 μg. From FIG. 18, considering that about 100 to 250 μg of the enzyme is immobilized on the lipase complex, it is inferred that about half of the immobilized enzymes are involved in the reaction. To.

(Example 34: Production of complex of protease and various porous molded products and enzymatic reaction)
In this example, the porous molded body of the mesoporous fine particles obtained in Examples 22 and 25, and the protease N (PN: Streptomyces genus, Nagasechem) for the activated carbon pellets (derived from rice husks, Dexerials Co., Ltd.) for comparison. Tex Co., Ltd.) was immobilized and the enzyme activity was evaluated.

(1-1) Method for Producing Complexes of Proteases and Various Porous Molds Two types of moldings obtained in Examples 22 and 25 were used as porous moldings of mesoporous fine particles on the immobilized support of protease. Used the body. In addition, as a porous body having a different surface affinity for comparison, "activated carbon pellets" having a larger mesopore volume than conventional activated carbon were used. Further, two kinds of synthetic mesoporous silica (raw material particles L and O) and "activated carbon powder" were used as the mesoporous silica fine particles before molding for comparison.

When immobilizing the enzyme, 1 mL of buffer solution (100 mM Tris-HCl (pH 7)) containing protease (0.271 mg), two types of compacts pre-weighed in microtubes, and "activated carbon pellets" (Equivalent to 20 mg) is compounded by mixing (4 ° C., 15 hours) using a rotator, and the protease and various porosities are performed by performing the centrifugation operation and the washing operation using the same buffer solution twice. A composite with the quality molded product was obtained. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used as it is, but in this embodiment, in order to evaluate the binding stability of the enzyme to the molded body. In addition, a cleaning step is performed together with the following centrifugation step.

Specifically, the dispersion liquid of the complex (unwashed) of the above-mentioned protease and various porous molded bodies (or activated carbon pellets) is centrifuged (4 ° C., 20,000 G, 10 minutes). All supernatants were collected. Subsequently, 1 mL of a washing buffer solution (using the buffer solution selected at the time of immobilization) was added, and a complex of the protease and the mesoporous silica molded product (or activated carbon pellet) was added using a Vortex Mixer at room temperature for about 5 seconds. After stirring, centrifugation (4 ° C., 20,000 G, 10 minutes) was performed, and a washing operation was performed to collect all the supernatant. The washing operation was repeated again using 1 mL of the washing buffer solution, and finally, a complex (washed) of the protease and various porous molded bodies (or activated carbon pellets) was obtained. Hereinafter, it is referred to as "protease-mesoporous fine particle molded product" or "protease-activated carbon pellet".

Further, for comparison, two kinds of synthetic mesoporous silica (raw material particles L and O) and "activated carbon powder" (equivalent to 10 mg) were used to obtain a complex by the same procedure as the molded product. Hereinafter, it is referred to as "protease-mesoporous fine particle powder" or "protease-activated carbon powder".

The amount of the enzyme immobilized on the molded body and powder of various mesoporous fine particles is the amount of the enzyme before immobilization (0.271 mg in 1 mL), and the amount of the free enzyme contained in the recovered liquid in the above-mentioned centrifugation step and washing step. Calculated by subtracting the amount. FIG. 21 shows the amount of protease immobilized on the mesoporous fine particle molded product or the mesoporous fine particle powder. FIG. 21 (a) shows the amount of protease immobilized on the molded product of various mesoporous fine particles. FIG. 21 (b) shows the amount of protease immobilized on the powder of various mesoporous fine particles.

Comparing FSM-22 (raw material particles L), SBA-15 (raw material particles O) type synthetic mesoporous silica and activated carbon from FIGS. 21 (a) and 21 (b), both the molded product and the powder have "activated carbon> FSM-". A tendency was observed that the amount of protease immobilized increased in the order of "22> SBA-15". Comparing the molded product and the powder, the amount of protease immobilization remained low when the molded product was used, but the powder was used for the protease immobilization behavior due to the difference in the immobilization support. Similar to the case, this suggests that the mesoporous fine particles contained in the compact retain approximately the same enzyme immobilization ability as the powder.

(1-2) Evaluation of Hydrolytic Activity of Gelatin The protease complex obtained in (1-1) (two types of protease-mesoporous fine particle molded product, protease-activated carbon pellet, two types of protease-mesoporous fine particle powder, and , Protease-activated carbon powder) was evaluated for enzymatic activity. The evaluation was performed by relative activity based on the fluorescence intensity of the unfixed free enzyme, using the fluorescence intensity that increases with the hydrolysis of fluorescent gelatin as a reaction substrate as an index. At this time, the durability (reaction 10 times) of the complex in repeated use was also evaluated.
Specifically, the following enzymatic reactions were caused in each of the complexes to determine the degrading activity of gelatin.

The enzymatic reaction was carried out against the protease complex (protease-mesoporous fine particle molded product, protease-activated carbon pellet, protease-mesoporous fine particle powder, and protease-activated carbon powder) in each microtube obtained in (1-1). , Fluorescent gelatin, sodium chloride, or Tris buffer containing calcium chloride (0.4 mL) was added (first reaction).

At that time, the reaction composition of the reaction substrate was adjusted to be "45 mM Tris-HCl (pH 7.6), 0.1 mg / mL fluorescent gelatin, 135 mM sodium chloride, 4.5 mM calcium chloride". From the second reaction onward, the reaction was started by adding a reaction substrate to the protease complex separated from the supernatant after the reaction after the following centrifugation operation in the same procedure as described above. In addition, for comparison, a free protease that did not use an immobilized support was also reacted by mixing with a similar reaction substrate.

The reaction conditions were to maintain the heated state at 37 ° C. for 30 minutes while mixing using a rotator. Subsequently, the supernatant was completely recovered by centrifugation (4 ° C., 20,000 G, 5 minutes), and the fluorescence intensity of the recovered solution was measured using the above-mentioned microplate reader (measurement wavelength: excitation 495 nm / fluorescence 520 nm). By doing so, the decomposition efficiency of gelatin was evaluated. FIG. 22 shows the relative activity of various protease complexes during repeated use (reaction 10 times) based on the efficiency of gelatin decomposition by free protease. FIG. 22 (a) shows the degrading activity of gelatin by the molded product of the protease-mesoporous fine particle complex and the durability (reaction 10 times) in repeated use of the immobilized enzyme. FIG. 22 (b) shows the degrading activity of gelatin by the powder of the protease-mesoporous fine particle complex and the durability (reaction 10 times) in repeated use of the immobilized enzyme.

From FIGS. 22 (a) and 22 (b), it was observed that, as a whole, the gelatin decomposition activity tended to be lower when the protease-mesoporous fine particle molded product was used as compared with the protease-mesopaphoric fine particle powder. Both the molded product and the powder showed remarkably excellent expression rate of enzyme activity and repeated durability as compared with activated carbon (pellets and powder).
Specifically, the gelatin decomposition activity at the 10th reaction was 3.9% when the protease-activated carbon powder was used, whereas it was about 100% when the protease-mesoporous fine particle powder was used. (FSM type: 100%, SBA type: 98.3%), showing extremely high repeatability (FIG. 22 (b)).
On the other hand, when the protease-activated carbon pellets were used, the gelatin decomposition activity at the 10th reaction was 41.1%, which was higher than that of the activated carbon powder, but when the protease-mesoporous fine particle molded product was used. Was found to have higher activity (FSM type: 92.4%, SBA type: 84.6%) (FIG. 22 (a)).

From the results shown in FIGS. 21 and 22, activated carbon (pellets and powders) having high surface hydrophobicity has higher enzyme activity than mesoporous silica (molds and powders) even though the protease immobilization rate is higher. The extremely low values showed the superiority of the mesoporous silica powder and its molded product in this reaction system.
Comparing FSM-type and SBA-type mesoporous silica, it was found that the decomposition activity of gelatin tended to increase in the order of "FSM-22>SBA-15" in both the molded product and the powder. , It is considered that the result depends on the amount of the protease immobilized (FSM-22> SBA-15) shown in FIG. 21.
Further, from the results of FIG. 21, in view of the fact that the amount of protease immobilization remained at a lower value (70 to 80% in the case of powder) when the molded product was used than in the case of powder, molding was performed. It is inferred that the mesoporous fine particles contained in the body retain the activity-expressing ability of the enzyme equivalent to that of the powder.

As described in detail above, the porous molded body of micro or mesoporous fine particles of the present invention is a porous body having good handling by a phase separation method of a mixed slurry of porous fine particle powder having adsorptivity and the like with an organic polymer resin. It is obtained by molding into. Further, according to the present invention, it is possible to provide a microfiltration membrane composed of the above-mentioned porous molded body of micro or mesoporous fine particles. The porous molded article of the present invention can be suitably used as a treatment agent for, for example, industrial wastewater.

Claims (21)

Porous containing functional fine particles having a pore diameter of 2 nm to 50 nm, which are mainly composed of silicate, aluminosilicate, alumina or titania and having a particle size of 0.01 μm to 200 μm, and an organic polymer. It is a quality molded product
The organic polymer is a polyether salphon or an EVOH resin having a polyethylene molar ratio of 44% or more.
It has macropores with a pore diameter of 50 nm to 100 μm.
It is a porous body with a porosity of 34% or more.
The content of the functional fine particles in the molded product is 46% by mass or more, and the content is 46% by mass or more.
At least a part of the surface of the functional fine particles is exposed from the molded product.
The pore volume of the mesopores of the functional fine particles is maintained at 40% or more of the raw material functional fine particles.
A porous molded body of mesoporous fine particles. The porous molded body of mesoporous fine particles according to claim 1, wherein the weight change of the porous molded body before and after stirring in water for 24 hours or more is 1% or less. The porous molded body of mesoporous fine particles according to claim 1 or 2, wherein the shape of the porous molded body is granular, hollow particle-like or hollow thread-like. The porous molded body of mesoporous fine particles according to any one of claims 1 to 3, wherein the functional fine particles and / or the organic polymer are a mixture of a plurality of types. Raw material of the functional fine particles Any one of claims 1 to 4, wherein the function of the functional fine particles is adsorption performance, and the characteristic expression of the function is maintained after molding of the porous molded body. The porous molded body of the mesoporous fine particles according to the section. The porous molded body of mesoporous fine particles according to any one of claims 1 to 5, wherein particles having a size of several microns can be pressure-filtered from the dispersion liquid. By dispersing the functional fine particles in a dried state in an organic solvent in which the organic polymer is previously dissolved , the gas contained in the functional fine particles is intentionally mixed to form nanobubbles. The process of obtaining the raw material slurry containing
The method for producing a porous molded body of mesoporous fine particles according to any one of claims 1 to 6 , further comprising a step of molding by injecting the raw material slurry into a non-solvent. The method for producing a porous molded body of mesoporous fine particles according to claim 7 , wherein the organic polymer has a hydrophilic group. The mesoporous according to claim 7 or 8 , further comprising a step of subjecting the porous molded body of the mesoporous fine particles to a low-temperature heat treatment in ethanol under reduced pressure or in a low-concentration aqueous solution of a good solvent. A method for producing a porous molded body of fine particles. Raw material for the functional fine particles The function of the functional fine particles includes the ability to immobilize one or more types of enzymes and / or proteins, and the content of the functional fine particles in the molded product is 46% by mass or more. The enzyme-supporting carrier which is the porous molded body according to any one of claims 1 to 6 . The complex of the enzyme-supporting carrier according to claim 10 and the enzyme. The complex according to claim 11 , wherein the enzyme is an oxidoreductase, a hydrolase, a transferase, a desorption enzyme, an isomerase, and / or a synthase. The complex according to claim 11 or 12 , wherein the enzymatic reaction of the enzyme is an oxidation-reduction reaction, a hydrolysis reaction, a transfer reaction, an elimination reaction, an isomerization reaction, and / or a synthetic reaction. The complex according to any one of claims 11 to 13 , wherein each of one or more kinds of enzymes and / or proteins involved in the enzymatic reaction of the enzyme is immobilized in a mesopore having a pore diameter of 2 nm to 50 nm. It ’s a way to make a body,
A production method comprising an immobilization step of immobilizing the enzyme and / or protein on a porous molded body of mesoporous fine particles in a buffer solution adjusted to pH 3-11. The production method according to claim 14 , further comprising a washing step of washing the porous molded body of mesoporous fine particles after immobilization with a buffer solution adjusted to pH 3 to 11 multiple times. The complex according to any one of claims 11 to 13 , wherein each of one or more kinds of enzymes and / or proteins involved in the enzymatic reaction of the enzyme is immobilized on a porous molded body of mesoporous fine particles. It is an enzymatic reaction method using
Immobilization step of immobilizing the enzyme and / or protein in a porous molded body of mesoporous fine particles in a buffer solution adjusted to pH 3 to 11.
A reaction substrate is added to a buffer solution having a pH of 3 to 11 containing a complex of a porous molded body of mesoporous fine particles obtained in the immobilization step and an enzyme, or the mesoporous fine particles obtained in the immobilization step. An enzymatic reaction step in which a complex of the porous molded product of No. 1 and an enzyme is added to a buffer solution having a pH of 3 to 11 containing a reaction substrate to carry out an enzymatic reaction involving the enzyme and / or a protein, and an enzymatic reaction method including the above. .. The complex according to any one of claims 11 to 13 , wherein each of one or more kinds of enzymes and / or proteins involved in the enzymatic reaction of the enzyme is immobilized on a porous molded body of mesoporous fine particles. It is an enzymatic reaction method using
Immobilization step of immobilizing the enzyme and / or protein in a porous molded body of mesoporous fine particles in a buffer solution adjusted to pH 3 to 11.
A washing step of washing a composite of a porous molded body of mesoporous fine particles obtained in the immobilization step and an enzyme with a buffer solution having a pH of 3 to 11.
An enzymatic reaction step in which a complex of a porous molded body of mesoporous fine particles after washing obtained in the washing step and an enzyme is subjected to an enzymatic reaction involving the enzyme and / or a protein in a reaction solution containing a reaction substrate. Enzyme reaction method including. The enzymatic reaction method according to claim 16 or 17 , wherein the enzymatic reaction is a method for producing a functional useful substance from a reaction substrate. The enzymatic reaction method according to claim 16 or 17 , wherein the enzymatic reaction is a method for decomposing an environmental pollutant which is a reaction substrate existing in the environment. The enzyme reaction method according to claim 16 or 17 , wherein the enzyme reaction method is a method for detecting or quantifying a reaction substrate that is or may be present in a test sample. A kit, sensor or apparatus for an enzyme reaction involving the enzyme and / or protein, which comprises the complex according to any one of claims 11 to 13 .

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