JP2018090767A - Micro- or mesoporous fine-particle porous molded body, enzyme-supporting carrier, enzyme complex thereof, and production method thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To maintain the characteristics of a porous silicate having micropores and micro or mesopores and functional fine particles having similar surface properties, and at the same time, to have high water resistance, filterability and reactivity. A molded product and a method for producing the same are provided. SOLUTION: A porous molded body of micro or mesoporous fine particles is mainly composed of silicate, aluminosilicate, alumina or titania having micro or meso pores having a pore diameter of 0.3 nm to 50 nm, and has a particle size of 0. A porous molded body containing functional fine particles having a size of 01 μm to 200 μm and an organic polymer, which has macropores having a pore diameter of 50 nm to 100 μm and a porosity of 34% or more. The content of the functional fine particles in the molded body is 46% by mass or more, at least a part of the surface of the functional fine particles is exposed from the molded body, and the pore volume of the micro or mesopores is the raw material functional fine particles. 40% or more of the particles are retained. [Selection diagram] Fig. 1

Description

  The present invention relates to a porous molded body of micro to mesoporous fine particles, an enzyme-supporting carrier, an enzyme complex thereof, and a production method thereof.

  As inorganic highly functional materials, fine particles having natural micro to mesopores such as clay, zeolite, diatomaceous earth, activated carbon and the like 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 in the molecular order, adsorptivity to specific molecules and ions, and catalytic ability resulting from the structure. It has been used in various ways in general life.

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

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

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

  In addition, in the patent document 4, the present inventors reported on a natural zeolite hollow fiber porous body, a zeolite membrane composite porous body, and a production method thereof. This natural zeolite hollow fiber porous body is a wet-spun hollow fiber porous body composed of natural zeolite powder and 3 to 50% by weight of an organic polymer, and has a pore diameter of 0.01 to 100 microns. It is a multi-element porous body in which pores with a plurality of diameters are distributed in the range, has water resistance that can be submerged, and exhibits reversible flexibility performance by making it water-containing even after drying treatment. To do.

  As synthetic fine particles having mesopores, mesoporous materials such as mesoporous silica are known. Examples of the method of using the mesoporous material include a method of using the mesoporous material as an immobilization support by immobilizing a biocatalyst enzyme or protein in these regular mesopores. For example, Non-Patent Document 1 discloses that in the production process of functional chemicals such as pharmaceuticals using an enzyme, mesoporous silica fine particles as a support for immobilizing an enzyme for separation of the product and the enzyme, reuse of the enzyme, etc. Disclose usage.

  In recent years, a method for processing mesoporous silica into a molded body has been reported for easy use. For example, Non-Patent Document 2 discloses a hollow fiber molding method using mesoporous silica fine particle powder. Non-Patent Document 3 discloses a pellet molding method.

JP 2009-61369 A JP 2004-275845 A JP 2010-104983 A JP 2012-72534 A

Nils Carlsson et al., “Enzymes immobilized on mesoporous silica: Physico-chemical perspective (A physical-chemical 20%, Ainical-chemical 20) , P. 339-360 Ali A. Al A. Rownahi et al., “In Situ Formation of a Monodispersed Carbon-Horsoporous Bio-Nanosilica-Torlo hollow fiber composite. ChemSusChem, 2015, 8, p. 3439-3450 Harsh Maheshwari et al., “Strong mesoporous silica compacts: Multi-scale charactor physicotactics: Robust mesoporous chemicals: ), Journal of Materials Science, 2016, 51, p. 4470-4480

  Zeolite is an aluminosilicate having regularly arranged micropores, and a part of Si as a main component in the skeleton structure is substituted with Al, and this Si / Al ratio is used as a catalyst of zeolite. Not only affects the strength and function, but also affects the affinity of the zeolite itself with adsorbed molecules, reactive molecules, and reaction solvents. The pores in zeolite are three-dimensional regular pores of about 3 to 10 mm, whereas clay has a two-dimensional layered structure, and diatomaceous earth has relatively large pores ranging from nano size to several microns. Further, in addition to those having a skeleton mainly composed of silica, activated carbon has carbon as a main component and 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 regular pores of 1 nm or more, which is not easily obtained in nature, mesoporous titanium oxide having a titania skeleton instead of silica, carbon In recent years, various functional materials having functions and pore sizes that cannot be obtained in nature have been developed, such as mesoporous carbon produced by Hf and hydrotalcite, which is a layered compound capable of adsorbing anions. Since the function of these fine particles is expressed 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 materials 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. Starting with the synthesis of mesoporous silica using quaternary alkylamine as a template with layered silicate as a raw material, aluminosilicate in which a part of the silica was replaced with alumina, and using alumina and titania instead of silica as raw materials Mesoporous aluminum oxide, mesoporous titanium oxide, mesoporous carbon using carbon, and the like have been synthesized.

  These mesoporous materials are all synthetic products using organic substances as templates, and various organic substances such as proteins and enzymes can be immobilized in the regular pores, so they are expected to be used in various fields. However, it is very expensive, and there are many submicron-sized fine particles, so that separation and purification are difficult, which is a problem in industrial use.

  On the other hand, many of the newly synthesized fine particles have the disadvantage that the primary particles remain as dispersible and the separation and recovery become difficult after the dispersibility is improved. In order to use these functional fine particles industrially, a molding / membering or separation method with the original surface area maintained as much as possible is essential. For example, since zeolite has poor self-sintering properties, pellet molding is generally performed via an inorganic or organic binder. However, in any method, if the amount of the binder is small, strength and water resistance are reduced. When the amount of the binder is large, the binder is crushed on the surface of the fine particles, so that the original function is not sufficiently developed, and this problem is particularly serious when trying to form a thin film.

  As a forming method, Patent Document 1 discloses a forming method such as secondary growth on a substrate or the like from a synthesis stage such as hydrothermal synthesis as one of methods for forming a separation membrane in which zeolite is fixed. Although described, this technique tends to be less plastic but can only be molded in the case of specific particulates capable of hydrothermal synthesis.

  On the other hand, by combining inorganic fine particles as fillers with an organic polymer molded body, the strength of the organic polymer film can be improved, and functions such as optical properties, heat resistance, and fire resistance can be imparted. In these, the affinity between the organic polymer and the filler and the orientation of the filler particles are thought to greatly affect the properties of the composite, and the surface of the filler particles and the resin side is modified to increase the affinity. It is.

  Patent Document 2 has through-holes having an average pore diameter of 0.01 to 5 μm obtained by blending a high-molecular-weight polyethylene resin with a fibrous inorganic filler having heat resistance as a filter medium for a cleaning organic solvent. And the porous membrane whose film thickness is 25-300 micrometers is disclosed. However, the method of melt molding of the organic polymer film described in Patent Document 2 performs high-temperature heating melting, kneading, and extrusion molding of the organic polymer, so that it can be used in addition to 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 pore-forming agent, and this plasticizer is further extracted. As an extractant for removal, a large amount of a saturated hydrocarbon-based organic solvent such as hexane, heptane, octane, nonane, decane or the like, or a halogenated hydrocarbon-based organic solvent such as trichloroethylene or tetrachloroethylene having high carcinogenicity must be used.

  In addition to such a melt molding method, a phase separation method can be considered as a method for molding an organic polymer. The phase separation method is a technique in which an organic polymer is dissolved in a solvent to form a slurry, which is aggregated by invasion of a non-solvent or a poor solvent, temperature change, or the like. Patent Document 3 discloses a polyamide hollow fiber membrane and a method for producing the membrane by a thermally induced phase separation method as its production method. However, in this method, the organic polymer slurry must be produced by heating to 170 ° C. or more, and the film is formed by lowering the temperature to 100 ° C. or less, and the filler is made up to 100% by weight with respect to the resin. If 50% by weight or more of the product is supported, the strength may not be obtained. As described above, this phase separation method is used as a film forming method with an emphasis on the properties of the organic polymer film. From the above-mentioned problems of strength, the functional fine particles are made porous as the main raw material. 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 using phase separation in a poor solvent. According to this method, it is possible to produce a porous molded body containing 50% by weight or more of an inorganic adsorbent as a main raw material, and this molded body is stable in an aqueous solution for several years without causing powder falling. It has the 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 forming the molding slurry. However, when polysulfone is mixed as an organic polymer in a state where a synthetic zeolite having a high silica / alumina ratio is dispersed in a solvent in accordance with the technique of Patent Document 4, the solvent of polysulfone is balanced by the interaction and affinity balance between the solvent and fine particles. There were cases where dissolution into the inside was hindered and a slurry in which inorganic particles were uniformly dispersed could not be obtained, and there were cases where powder fall off from the molded body increased. In addition, in order to avoid such problems, when an organic polymer having high affinity with inorganic particles is used, the organic polymer penetrates not only into the surface of the inorganic particles but also into the pores, and the inorganic particles It has been found that the micropores are easily clogged. As described above, in the prior art, when forming porous silicate microparticles having pores of various sizes and having a surface different in hydrophilicity / hydrophobicity from that of natural zeolite, the fineness and liquid permeability of the structure can prevent powder falling. There has been a problem that it is difficult to achieve both the porosity and the porosity maintaining the pore function.

  In addition, in basic research on immobilized enzymes using conventional mesoporous silica fine particles as proposed by Non-Patent Document 1, “mesoporous silica fine particle powder” in a powder state is mainly used, and generally fine particle powders are used. In particular, when mesoporous silica fine particle powder is used as a part of a device or instrument such as a bioreactor, there is a problem that the fine particle powder is pulverized and powdered off.

  Further, the molded body obtained using the organic polymer disclosed in Non-Patent Document 2 as a binder 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 body 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 problem of the present invention is not limited to natural zeolite particles, but the surface has low hydrophilicity such as synthetic high silica zeolite, and microparticles such as mesoporous silica, mesoporous aluminum oxide, and mesoporous titanium oxide. Or porous silicate with mesopores, and multi-component porous molded body having high water resistance, filterability and reactivity while maintaining the characteristics of functional fine particles with similar surface properties It is to provide a new technique that can be done. In addition, this method is not limited to polysulfone or polyethersulfone, and good results can be obtained even if polyamide imide with high weather resistance or cheaper hydrophilic resin such as EVOH or PMMA is used as a binder depending on the application. It is done.

  In addition, the problem of the present invention is that the fine powder of mesoporous material such as mesoporous silica is stable and has high mechanical strength even if it is molded into an appropriate shape such as a hollow fiber shape or a pellet shape according to the use, and the powdery 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 mesoporous silica fine particles and can further exhibit the same effect as powder as an enzyme immobilization support. It is.

  As a result of intensive studies to achieve the above-mentioned problems, the inventors of the present invention have adopted a method of phase separation of a mixed slurry with an organic polymer in a poor solvent as a method for forming porous particles. It is preferable from the viewpoint that the route becomes a through hole to increase the reactivity of the molded body and partially expose the surface of the porous fine particles. , Organic polymer dissolved in a good solvent penetrates into the pores of the porous particles, 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 It came to the view that it is important to keep in mind.

From that perspective,
1. Preparation of slurry for forming porous particles can be performed by first dissolving the organic polymer in a good solvent at a temperature at which it can be sufficiently dissolved, and then the fine particles do not react and the organic polymer can maintain a viscous slurry. It is preferable to carry out the procedure of adding fine particles little by little while stirring at high speed at a temperature,
2. In order to prevent powder falling, it is effective to use a hydrophilic organic polymer having a functional group having a high affinity for silanol formed on the silicate surface in the structure as a binder. It has been found that it is preferable to avoid polymers that are in a low molecular state or that have low viscosity and easily enter the pores. Furthermore, since these hydrophilic organic polymers tend to have a dense aggregate structure in water compared to hydrophobic organic polymers, the molded product tends to become dense by entering between fine particles of submicron size. In the preparation of slurry mixed with fine particles, when adding fine particles to a viscous slurry in which organic polymer is dissolved, the nano-size generated by high-speed stirring of the gas containing the fine particles and the viscous slurry does not become a defect. It has been found that extremely fine bubbles can be intentionally mixed to such a degree that voids are easily generated between the formed particles, and the filterability and flexibility are improved. Furthermore, if the hydrophilic organic polymer has an agglomeration structure that completely embeds the fine particles due to the balance of the affinity between the poor solvent and the fine particles, the post-treatment after the formation of the molded product will greatly destroy the molded product structure. It has been found that the accessibility to the fine particles can be increased without degrading and without causing a decrease in strength.

  Based on these findings, the present inventors have completed the present invention.

That is, the present invention includes the following aspects.
(1) Functional fine particles having a microparticle or mesopore with a pore diameter of 0.3 nm to 50 nm, mainly composed of silicate, aluminosilicate, alumina, or titania and having a particle diameter of 0.01 μm to 200 μm; A porous molded body containing molecules,
Macropores with a pore diameter of 50 nm to 100 μm,
A porous body having a porosity of 34% or more;
The content of the functional fine particles in the molded body is 46% by mass or more,
At least part of the surface of the functional fine particles is exposed from the molded body,
The micro volume or mesopore volume of the functional fine particles is maintained at 40% or more of the raw material functional fine particles,
A porous molded body of micro or mesoporous fine particles characterized by the above.

(2) The micro or mesoporous fine particles according to (1) above, wherein the weight change before and after stirring of the porous molded body when continuously stirred in water for 24 hours or more is 1% or less. Quality molded body.

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

(4) The micro or mesoporous fine particles according to any one of (1) to (3), wherein the functional fine particles and / or the organic polymer is a mixture of a plurality of types. Quality molded body.

(5) The function of the functional fine particles as a raw material functional fine particle is an adsorption performance, and the characteristic expression of the function is maintained after the formation of the porous molded body. 4) A porous molded body of micro or mesoporous fine particles according to any one of 4).

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

(7) The micro or thru | or as described in any one of said (1)-(6) characterized by the said organic polymer being polyether sulfone, polysulfone, EVOH resin, PMMA resin, or polyamide-imide resin. A porous molded body of mesoporous fine particles.

(8) A step of obtaining a raw material slurry by dispersing functional fine particles mainly composed of silicate, aluminosilicate, alumina or titania having micro to mesopores in an organic solvent in which the organic polymer is previously dissolved;
A step of molding by injecting the raw material slurry into a non-solvent, the porous molded body of micro to mesoporous fine particles according to any one of (1) to (7), Production method.

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

(10) The organic polymer is a PMMA resin or a polyamide-imide resin, and further comprises a step of treating with a polymerization accelerator or a curing accelerator after molding, according to (8) or (9), A method for producing a micro- or mesoporous fine-particle porous molded body.

(11) The step (8) to (10), further comprising a step of subjecting the porous molded body of micro to mesoporous fine particles to 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 micro or mesoporous fine particles according to any one of the above.

(12) The functional fine particles are mesoporous fine particles having mesopores having a pore diameter of 2 nm to 50 nm and a particle diameter of 0.01 μm to 200 μm. It contains the immobilization ability of more than one kind of enzyme and / or protein, and the content of the functional fine particles in the molded body is 46% by mass or more. An enzyme-supporting carrier, which is a porous molded body.

(13) A complex of the enzyme-supporting carrier according to (12) and an enzyme.

(14) The complex according to (13), wherein the enzyme is an oxidoreductase, a hydrolase, a transferase, a dehydrogenase, an isomerase, and / or a synthase.

(15) The complex according to (13) or (14), wherein the enzyme reaction of the enzyme is an oxidation-reduction reaction, hydrolysis reaction, transfer reaction, elimination reaction, isomerization reaction, and / or synthesis reaction .

(16) Any one of (13) to (15), wherein each of one or more enzymes and / or proteins involved in the enzyme reaction of the enzyme is immobilized in a mesopore having a pore diameter of 2 nm to 50 nm. A method for producing the composite 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 adjusted to pH 3-11.

(17) The production method according to (16), further including a washing step of washing the porous molded body of mesoporous fine particles after completion of immobilization multiple times with a buffer adjusted to pH 3-11.

(18) In any one of the above (13) to (15), each of one or more enzymes and / or proteins involved in the enzyme reaction is immobilized on a porous molded body of mesoporous fine particles. An enzyme reaction method using the described complex,
An immobilization step of immobilizing the enzyme and / or protein on a porous molded body of mesoporous fine particles in a buffer adjusted to pH 3-11;
A reaction substrate is added to a buffer solution having a pH of 3 to 11 containing a complex of a porous compact of mesoporous fine particles obtained in the immobilization step and an enzyme, or mesoporous fine particles obtained in the immobilization step An enzyme reaction method comprising an enzyme reaction step in which a complex of the porous molded body and the enzyme is added to a pH 3 to 11 buffer solution containing a reaction substrate to perform an enzyme reaction involving the enzyme and / or protein.

(19) In any one of (13) to (15) above, each of one or more enzymes and / or proteins involved in the enzyme reaction is immobilized on a porous molded body of mesoporous fine particles. An enzyme reaction method using the described complex,
An immobilization step of immobilizing the enzyme and / or protein on a porous molded body of mesoporous fine particles in a buffer adjusted to pH 3-11;
A washing step of washing a complex 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 enzyme reaction step of performing an enzyme reaction involving the enzyme and / or protein in a reaction solution containing a reaction substrate, a complex of a porous molded body of mesoporous fine particles obtained after the washing step and the enzyme obtained in the washing step; Enzymatic reaction method comprising.
(20) The enzyme reaction method according to (18) or (19), wherein the enzyme reaction is a method for producing a functional useful substance from a reaction substrate.
(21) The enzyme reaction method according to (18) or (19), wherein the enzyme reaction is a method for decomposing an environmental pollutant serving as a reaction substrate present in the environment.
(22) The enzyme reaction method according to (18) or (19), wherein the enzyme reaction method is a method for detecting or quantifying a reaction substrate present or possibly present in a test sample.
(23) An enzyme reaction kit, sensor, or apparatus involving the enzyme and / or protein, comprising the complex according to any one of (13) to (15).

  According to the present invention, not limited to natural zeolite particles, the surface has low surface hydrophilicity such as synthetic high silica zeolite and microparticles such as mesoporous silica, mesoporous aluminum oxide, and mesoporous titanium oxide. Multi-porous molded body having high water resistance, filterability, and reactivity while maintaining the characteristics of porous silicate having functional fine particles having surface properties similar to the above, and a novel method capable of producing the same Is provided. In addition, this method is not limited to polysulfone or polyethersulfone, and good results can be obtained even if polyamide imide with high weather resistance or cheaper hydrophilic resin such as EVOH or PMMA is used as a binder depending on the application. It is done.

  In addition, according to the present invention, powdered mesoporous silica is stable and has high mechanical strength even if fine particles of mesoporous material such as mesoporous silica are molded into an appropriate shape such as a hollow fiber shape or a pellet shape according to the application. Provided are 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 fine particles and can exhibit the same effect as a powder as an enzyme immobilization support.

2 is an SEM image of a cross section orthogonal to the longitudinal direction of the molded body of Example 1. FIG. 3 is an SEM image of a cross section in the longitudinal direction of the molded body of Example 1. FIG. The pore diameter distribution curve figure by mercury porosimetry of the molded object of Example 1. FIG. The figure which showed the nitrogen adsorption isotherm at the room temperature of the molded object of Example 1 and the raw material particle A. FIG. The figure which showed the nitrogen adsorption isotherm in Example 13 and Example 14 and the raw material particle F at room temperature. The external appearance image of the molded object of Example 18. FIG. (A) Mesoporous silica powder of Example 23, (b) Molded product, (c) Appearance image of enzyme complex. The figure which showed typically the procedure until the preparation method of the composite_body | complex of the molded object of Example 18, and the decomposition reaction of an azo dye, and reuse of an immobilized enzyme. The figure which showed the fixed amount and fixed rate of AzoR in Example 30. FIG. The figure which showed the light absorbency change of Example 30 and the decomposition rate of methyl red. The figure which showed the fixed amount of AzoR with respect to the various mesoporous microparticles of Example 31. The figure which showed the decomposition rate and durability of methyl red by the various AzoR-mesoporous fine particle composites of Example 31. (A) The molded body of Example 23, (b) the AzoR composite, (c) the molded body of Comparative Example 8, and (d) the appearance image of the AzoR composite in Example 31. The figure which showed the influence of the free AzoR density | concentration which gives to a decoloring rate. The figure which showed the fixed quantity of glucose dehydrogenase with respect to the various mesoporous microparticles | fine-particles of Example 32. The figure which showed the durability of the production | generation NADH density | concentration by the various GDH-mesoporous microparticle composite_body | complex of Example 32, and the immobilized enzyme. The figure which showed the influence of the unfixed free GDH density | concentration given to the production | generation NADH density | concentration. The figure which showed the fixed amount of the lipase with respect to the various mesoporous microparticles | fine-particles of Example 33. The figure which showed the free pyrene density | concentration in the triglyceride decomposition | disassembly by the various lipase-mesoporous fine particle complex of Example 33, and durability of the fixed enzyme. The figure which showed the influence of the unfixed free lipase density | concentration which gives to the fluorescent substance (pyrene) density | concentration liberated by triglyceride decomposition | disassembly. The figure which showed the fixed amount of the protease with respect to various mesoporous microparticles | fine-particles of Example 34 and activated carbon. The figure which showed the degradation activity of gelatin by the various protease-mesoporous microparticles | fine-particles complex of Example 34, and protease-activated carbon complex, and durability of the fixed enzyme.

  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 included therein.

  The porous molded body of micro to mesoporous 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 to mesoporous particles of the present invention is It may be referred to as a porous molded body of the invention).

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

  Here, the micropore is also called a micropore or a micropore, and refers to a pore having a pore diameter of less than 1 nm. Mesopores are pores larger than micropores and having a pore diameter of 1 nm to 50 nm. Therefore, having a micro or mesopore having a pore diameter of 0.3 nm to 50 nm means having a pore 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, more preferably 0.5 nm to 5 nm.

  In addition, silicate, aluminosilicate, alumina or titania is pure silicate, aluminosilicate, alumina or titania, and other metal elements introduced into defects in the skeleton of silicate, aluminosilicate or titania structure, silicate In addition, silicon (aluminum silicate) in which a part of silicon or aluminum element in the structure of aluminosilicate is replaced with another metal is included.

  In addition, silicate, aluminosilicate, alumina or titania is the main component, the functional fine particles are those of these pure phases, or the mass ratio thereof is usually 50% or more, preferably 90% or more. Say.

  Further, the function of the functional fine particles is not particularly limited, and examples thereof include adsorption performance and removal ability. These functions are preferably maintained after the formation of the porous molded body.

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

  The particle diameter of the functional fine particles according to the present invention is 0.01 μm to 200 μm, preferably 0.01 μm to 10 μm, more preferably 0.1 μm to 5 μm. Here, the particle size means the size of the aggregated particles. Particles having a crystal particle size smaller than this range may include agglomerates that are not separated. When adjusting the raw material slurry to be 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 to become a finer particle size. When the porous molded body of the present invention is in the form of hollow fibers or fine particles, the functional fine particles are preferably dispersed in a particle size range of 0.01 μm to 10 μm in the porous molded body, raw material slurry, or solvent. More preferably, it disperses in the range of 0.1 μm to 5 μm. The particle diameter can be obtained, for example, as an average value of the diameters of 20 to 30 particles by observation with a transmission electron microscope.

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

  The functional fine particles according to the present invention may be single or plural types, but when the functional fine particles are a mixture of plural types, a plurality of reaction targets can be selectively adsorbed or removed simultaneously with one molded body. It is preferable at the point which can provide a some function.

  The content of the functional particles in the porous molded body of the present invention is 46% by mass or more, preferably 50% by mass or more, more preferably 60% by mass or more, and further preferably 67% by mass or more.

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

  The organic polymer according to the present invention may be a single type or a plurality of types, but physical properties such as strength of the molded body, chemical properties such as chemical resistance and heat resistance are properties of the organic polymer. Therefore, it is also possible to control the properties of these molded products by using a mixture of a plurality of types.

  The organic polymer according to the present invention is preferably a modified functional fine particle that has been chemically modified or treated. Examples of the chemical modification or treatment of the organic polymer include imparting a hydroxyl group to polyethersulfone and saponification treatment of a 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%, and more preferably less than 56 mol%. If 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 into a film on the surface of the poor solvent for coagulation.

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

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

  The porous molded body of the present invention is not limited depending on the production method thereof.For example, the silicate, aluminosilicate, alumina or titania having micro to mesopores in an organic solvent in which an organic polymer is dissolved in advance is a main component. It can be manufactured by a method comprising a step of obtaining a raw material slurry by dispersing functional fine particles and a step of forming the raw material slurry by injecting the raw material slurry into a non-solvent.

  Moreover, when the organic polymer of the porous molded body of the present invention is a PMMA resin or a polyamideimide resin, it can be produced by a method further comprising a step of treating with a polymerization accelerator or a curing accelerator after molding.

  This manufacturing method is a method of obtaining a molded body from a raw material slurry containing functional fine particles and an organic polymer by a phase separation method.

  In addition to functioning as a binder for the functional fine particles, the organic polymer plays a role of maintaining macroscopic pores so that the gaps between the functional fine particles are not crushed during use after the formation of the porous body. The organic polymer is not particularly limited, and examples thereof include those exemplified as the organic polymer according to the above-mentioned molded body. However, the organic polymer is soluble at a relatively low temperature in a kind of organic solvent that hardly volatilizes at room temperature, and functional fine particles. Are selected as appropriate. Those having a hydrophilic group are preferred because of their affinity with silicates and aluminosilicates. From the viewpoint of preventing entry into micro or mesopores in the raw slurry, from the viewpoint of obtaining sufficient viscosity and moldability at a low concentration during molding, from the viewpoint of the structure and strength of the molded body after molding, in terms of polystyrene Those having a relatively high molecular weight and a weight average molecular weight (Mw) of 50,000 or more are preferred. The amount of the organic polymer added to the raw slurry is appropriately selected depending on the shape of the molded body and the type of functional fine particles, but is usually 5 to 50% by weight with respect to the total weight of the molded body after drying, Preferably, it is 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 product, and if it is too much, the particle gaps of the raw material functional fine particles are blocked and the micro or mesopores tend not to function. Moreover, when PMMA resin etc. are used, the intensity | strength of a molded object can be raised by performing a repolymerization process after shaping | molding.

  As described above, the solvent for dissolving the organic polymer can dissolve about 6 to 20% by weight or more of the organic polymer with respect to the weight of the solvent at a relatively low temperature. Functional fine particles that are compatible with poor solvents and have micro or mesopores as porous materials, mainly composed of silicates, aluminosilicates, alumina, or titania, cause chemical or physical reaction with them. It is preferable that the solvent is capable of being uniformly dispersed. Examples of such a solvent include dimethyl sulfoxide, dimethylformamide, dimethylacetamide, and n-methylpyrrolidone.

  In the present invention, the functional fine particles are dispersed in an organic solvent in which an organic polymer is dissolved, and the preparation of the raw material slurry is performed using functional fine particles mainly composed of silicate, aluminosilicate or titania having micro to mesopores. It can carry out by stirring in addition to the above-mentioned solvent with the organic polymer used as the above-mentioned binder.

  It is preferable that the functional fine particles of the raw material are sufficiently dried before the addition in order to prevent local polymerization before molding of the organic polymer in the raw material slurry and to prevent a decrease in strength of the composite after molding. . The drying temperature is not particularly limited, but is preferably a temperature at which dehumidification is performed within a range where the property change of the raw material fine particles does not occur. The concentration of the functional fine particles in the raw slurry is not particularly limited, but is appropriately selected depending on the difference in viscosity depending on the type and size of the functional fine particles, and the type of the molded body and organic polymer. Regarding hollow fiber membrane molding, the concentration of functional fine particles in the raw 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 serving as the binder in the molded body tends to be uneven, and flexibility and pressure resistance May not be obtained.

  The order of addition of the functional fine particles and the organic polymer at the time of preparing the raw slurry is not particularly limited, but the viscous slurry in which the organic polymer is sufficiently dissolved in a preheated solvent is rapidly stirred and dried. A method of adding and dispersing small amounts of the raw material powder in a state is preferable because it is easy to obtain a slurry having a uniform and high viscosity at a relatively low temperature in a short time, and more effectively generating nanobubbles. In the method of adding the organic polymer later, 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 slurry is not particularly limited, but is preferably 200 rpm or more in the gas phase. When the stirring speed is within this range, macropores are easily formed.

In the method for producing a porous molded body of micro to mesoporous fine particles of the present invention, the raw material slurry prepared as described above can be solidified by injection into a non-solvent, and the raw material slurry can be formed in a poor solvent. It can also be solidified and molded. Thereby, a self-supporting water-resistant composite porous material having a pore size distribution of several angstroms to several tens of microns can be obtained.
Examples of the non-solvent include hydroalcohol. Examples of the poor solvent include an aqueous solution of a solvent used for the raw material slurry.

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

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

  The porous molded body of the present invention is a porous body having macropores in a diameter range of 50 nm to 100 μm and 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. In addition, 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 porous molded body has macropores in the diameter range of 50 nm to 100 μm means that the poor solvent, which is a coagulating liquid, enters the raw material slurry side, that is, the contact interface when the porous molded body is produced. Thus, the solidification of the raw material means that macroscopic pores leading from the outside to the inside of the porous molded body are formed. The size of the macropores can be confirmed by having a peak in this range of the pore diameter distribution curve when measured with a porosimeter or the like, and the micropores that are the characteristics of the raw material particles and the organic polymer as the binder It means that the macroscopic pores to be formed coexist. Moreover, since the poor solvent entry path corresponds to a gap between the fine particles in the raw material slurry at the same time, the surface of the raw material particles is easily exposed to the macropores.

  In addition, the porous molded body of the present invention provides micropores in a dense layer formed on the wall of the macropores by performing vacuum degassing for a short time in ethanol or in a low-temperature good solvent after molding. Thus, the surface of the raw material particles can be modified so as to be exposed in the macropores.

  Since the porous molded body of the present invention has such characteristics, the substance to be adsorbed or removed or reacted in the molded body diffuses in contact with the surface of the encapsulated raw material particles as compared with a normal molded body. It is excellent in that the reaction rate and reaction efficiency are increased as a result.

  In addition to heat resistance, water resistance and humidity control properties, the porous molded body of the present invention has characteristics of micro to mesoporous fine particles such as ion exchange properties and adsorbability.

  The porous molded body of the present invention preferably has a weight change of 1% or less before and after stirring when continuously stirred in water for 24 hours or more. Here, the weight change means that the fine particles are removed from the porous molded body and fall into water, or the properties of the fine particles and the organic polymer are changed. When the weight change is within this range, it is preferable from the viewpoint that when the molded body is used in water, the loss of fine particles due to this deterioration hardly occurs.

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

  The shape of the porous molded body of the present invention is not particularly limited, but is preferably in the form of particles, hollow particles, or hollow fibers. In particular, when the shape is a hollow particle shape or hollow fiber shape, the reaction target substance such as the adsorption target or the removal target substance is easily diffused into the porous molded body, compared with the case where there is no hollow structure. A compact that does not have raw material particles or a hollow structure, such as improved efficiency, easy scale-up to modules, etc., and a reaction by supplying substances from the inside and outside of the porous molded body It can be expected to be superior in various respects such as the application to reactions that cannot be achieved.

  In addition, 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 water filtration or purification, as a carrier of a reactant in an aqueous solution, etc. It is. Although this microfiltration membrane is a water-resistant microfiltration membrane, it has the pore characteristics and reactivity of micro to 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, an ion exchange effect of an aqueous solution containing cations, and is water-resistant and can be a self-supporting composite membrane. This composite membrane can be suitably used, for example, as a treatment agent for wastewater for food industry.

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

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

  The molded product of the present invention can be used as an enzyme-supporting carrier. The molded body of the present invention can form a complex with an enzyme as an enzyme-supporting carrier. In particular, among the molded articles of the present invention, the functional fine particles are mesoporous fine particles having mesopores having a pore diameter of 2 nm to 50 nm and a particle diameter of 0.01 μm to 200 μm, and the functional fine particles in the molded article. A molded product having a content of 46% by mass or more is suitable as an enzyme-supporting carrier. Moreover, when the function of the functional fine particles which are the raw materials of the functional fine particles constituting the molded article of the present invention includes the ability to immobilize one or more kinds of enzymes and / or proteins, it is suitable as an enzyme-supporting carrier. It is.

Although this enzyme is not specifically limited, It is preferable in it being an oxidoreductase, a hydrolase, a transferase, a detachment enzyme, an isomerase, and / or a synthetic enzyme. Specific examples of the enzyme include azo reductase, lipase, glucose dehydrogenase, and protease.
This enzyme can be used for enzyme reaction of oxidation-reduction reaction, hydrolysis reaction, transfer reaction, elimination reaction, isomerization reaction, and / or synthesis reaction.

  The method for producing the complex is not particularly limited. For example, the porous mesoporous microparticles in a buffer solution in which one or two or more enzymes and / or proteins involved in the enzyme reaction of the enzyme are adjusted to pH 3-11. An immobilization step of immobilizing the molded product may be included. Furthermore, it is further preferable to include a washing step of washing the porous molded body of mesoporous fine particles after completion of immobilization multiple times with a buffer adjusted to pH 3-11.

  The enzyme reaction method using the complex of the present invention is not particularly limited. For example, 1) Immobilization in which an enzyme and / or protein is immobilized on a porous molded body of mesoporous fine particles in a buffer adjusted to pH 3-11. Step 2) 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 3) the immobilization step. Enzyme reaction step in which the complex of mesoporous fine particles obtained in step 1 and a complex of an enzyme is added to a pH 3 to 11 buffer containing a reaction substrate to perform an enzyme reaction involving the enzyme and / or protein. The method of including is mentioned. Other enzyme reaction methods include 1) an immobilization step in which the enzyme and / or protein is immobilized on a porous molded body of mesoporous fine particles in a buffer adjusted to pH 3 to 11, and 2) in the immobilization step. Washing step of washing the obtained mesoporous fine particle porous compact and enzyme complex with a buffer solution having a pH of 3 to 11, and 3) the washed mesoporous fine particle porous compact and enzyme obtained in the washing step. And an enzyme reaction step in which an enzyme reaction involving the enzyme and / or protein is performed in a reaction solution containing a reaction substrate.

  These enzyme reactions are not particularly limited. For example, a method for producing a functional useful substance from a reaction substrate, a method for decomposing an environmental pollutant serving as a reaction substrate present in the environment, or a sample present in a test sample. Or a method for detecting or quantifying a reaction substrate that may be present.

  The composite of the porous molded body and the enzyme of the present invention can be used in an enzyme reaction kit, sensor, or apparatus involving the enzyme and / or protein. The use of the enzyme reaction kit, sensor, or device is not particularly limited, but for producing functional useful substances from reaction substrates, and for decomposing environmental pollutants that become reaction substrates present in the environment. The reaction substrate or the product after the enzymatic reaction that may be present in the test sample may be used for detection or quantification.

(Example)
EXAMPLES Next, although this invention is demonstrated further in detail based on an Example, this invention is not limited at all by the following Examples etc. The porosity and pore size distribution of the porous molded body according to the present invention can be evaluated by a measuring technique such as mercury porosimetry. Further, the retention rate of the micro volume or the mesopore volume of the raw material fine particles can be evaluated by comparing the gas adsorption and vapor adsorption measurement results with the measured values of the raw material particles.

  The physical properties of the fine particles used in the examples are shown in Tables 1 and 6, and the physical properties of the obtained molded products are listed in Tables 2 to 4 and Table 7.

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

The produced hollow fiber membrane has an outer diameter of 1.2 mm, an inner diameter of 0.7 mm (see FIG. 1), and after drying overnight at 60 ° C., the porosity is around 62% as measured by a mercury porosimeter. And MCM41-derived multi-porous material having nanopores with a diameter of 4 nm to 10 nm and macropores of 0.1 μm to 1 μm (see FIG. 2). When 0.0299 g of a 60 ° C. dry product of this hollow fiber-shaped molded product is placed in 3 ml of distilled water, continuously infiltrated at 100 rpm for 36 hours or more, dried again at 60 ° C. and weighed, it is 0.0297 g. The weight change is within 1%, and it is considered that there is almost no powder falling. According to thermal analysis, when heated to 600 ° C., the organic polymer is decomposed at 200 ° C. or more, 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, it is considered that the relative pressure (P / Po) region of the hysteresis of the isotherm of the compact and the raw material particles coincides, and the size of the mesopores of the raw material particles is almost maintained. Furthermore, since the pore volume in the compact was 0.28 cm ³ / g and the mesopore volume of the raw material MCM41 powder particles was 0.59 cm ³ / g, MCM containing the pore volume of the compact was included. 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). FIG. 4 shows a nitrogen adsorption isotherm at room temperature between the molded body of Example 1 and the raw material particles A. In addition, it pre-processed on 60 degreeC 24 hours pressure reduction conditions, and converted by particle content.

(Example 2)
Polyethersulfone as organic polymer (Veradel PES 3000P, Solvay Advanced Polymers Co., Ltd.) 0.54 g of weight average molecular weight (Mw) = 57,000 was completely added to 5.50 g of dimethyl sulfoxide (Wako special grade) at 50 ° C. The slurry was mixed with 0.55 g of synthetic mesoporous silica (raw material particles A) used in Example 1 to prepare a slurry. At this time, the concentration of particles in the slurry was 8.3% by weight, the amount of polyethersulfone added was 8.2% by weight, and the amount of MCM particles charged to the compact was 50% by weight. The slurry was dropped into a beaker containing hot water of 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 no turbidity was observed, to form MCM-PES pellets. .

When 0.0288 g of the pellet after stirring and washing until the cloudiness disappeared was stirred in 3 mL of water at room temperature for 72 hours or more and the dry weight was measured, almost no change in weight was observed. When this was heated to 600 ° C. by thermal analysis, the organic polymer was 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. (It is considered that a reversible endothermic peak and weight loss not found in the raw material particles and the organic polymer were observed around 120 ° C., and that a new adsorption site derived from molding was generated.) When measured with a mercury porosimeter, the porosity was 78%. According to the pore size distribution curve, it was a multi-element porous body having 4 nm to 10 nm diameter nanopores derived from MCM41 and 0.1 μm to 1 μm macropores. Further, according to the nitrogen adsorption isotherm, the pore volume of the compact 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. It is considered that almost all the mesopore volume of MCM41 in the molded body is retained in terms of the amount of MCM41 particles contained in the compact.

(Example 3)
6. 0.60 g of methyl methacrylate resin (PMMA polymer CAS No. 9011-14-7, Tokyo Chemical Industry Co., Ltd., M0088, Mw = 135,000 to 140,000) as an organic polymer was added to dimethylformamide (Wako Special Grade). The resultant 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 therewith to prepare a slurry. This slurry was dropped into a beaker containing hot water of about 32 ° C., solidified for 2 hours or more, and dried at 60 ° C. overnight to form MCM-acrylic pellets. Since the mechanical strength of this pellet was very low, 0.20 g of this pellet was put together with 0.051 g of benzoyl peroxide (Bio-Rad) in 4.98 g of ethanol (Wako Special Grade), and the temperature was raised slowly. The mixture was heated at 60 ° C. for 4 hours and taken out. The removed pellets were changed to be flexible and hard to break like rubber. When this was further dried at 60 ° C., pellets having a compressive strength of about 1.4 MPa were obtained.

With respect to the pellets after polymerization and curing, when this was heated to 600 ° C. by thermal analysis, the organic polymer was decomposed around 400 ° C., and the weight loss due to decomposition was 50%. The content of is considered to be about 50%. Further, according to the nitrogen adsorption isotherm, the pore volume in the compact 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 to the amount of MCM particles containing 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 put into 0.7 mL of dimethyl sulfoxide (Wako Special Grade), dissolved by heating and stirring, and mesoporous titanium oxide having a particle size of 10 to 100 nm and an average pore size of 4.2 nm. (Alfa Aesar No. 43250) (Raw material particles B) 0.20 g was mixed to prepare a slurry. This slurry was dripped into a beaker containing room temperature water and solidified, and dried at 60 ° C. to form mesoporous TiO ₂ —EVOH pellets.

  According to the 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 particle content in the pellet is 65%. %. Further, when the pore volume in the molded body pretreated at 100 ° C. and calculated by the nitrogen adsorption measurement result is converted into the amount of mesoporous titanium oxide particles contained, the mesoporous volume of mesoporous titanium oxide in the molded body is 64%. It is thought that the degree is maintained.

(Example 5)
Example 2 is the same as 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 size of 5.7 nm is used as the raw material particles. Thus, mesoporous alumina-PES pellets were formed. According to the 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 size of 8 nm was used as the raw material particles. According to the thermal analysis, the content of particles in the pellet was about 55%.

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

(Example 8)
As an organic polymer, 34.5 g of ethylene-vinyl alcohol copolymer resin having an ethylene content of 48 mol% used in Example 1 was mixed with 181.71 g of dimethyl sulfoxide used in Example 1, and overnight at 48 ° C. and 200 rpm. Stir to dissolve. While the mixture was heated and stirred, 68.01 g of the synthetic high silica zeolite (raw material particles E) used in 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 to the compact was 67% by weight. After stirring overnight, using a double-mouth nozzle with a slurry discharge port diameter of 6 mmφ and a core liquid discharge port diameter of 2.5 mmφ, the resulting slurry was placed in a cylinder pump and placed in a 34 ° C. water bath at 40.0 ml / min. While injecting, 25 ° C. water was flowed at 70.0 ml / min as a core solution to form a hollow fiber.

  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 compact is considered to be 64% by weight. According to mercury porosimetry, the porosity after drying at 60 ° C. overnight is 40%, and after heating at 150 ° C. for 2 hours to crystallize EVOH, the porosity is 37%, and the pore size distribution curve According to this, 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 60 ° C. drying treatment was 89% or more of the powder sample in terms of the amount of zeolite particles.

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 particle F) having a particle size of 10 μm, a crystal particle size of 2 μm to 4 μm, and a pore size of 0.58 nm was added. Thus, 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 charged amount of zeolite particles to the compact was 63% by weight. After stirring overnight, the resulting slurry was put into a cylinder pump at 22.3 ml / min using a double-mouth nozzle in a dry-wet spinning device, and injected into a 25 ° C. water bath as a core solution. Water at 1 ° C. was flowed at 11.7 ml / min to form a hollow fiber.

  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 compact is considered to be 64% by weight. After drying at 60 ° C. overnight and further heating at 150 ° C. for 2 hours, the porosity was 40% as measured with a mercury porosimeter, and according to the pore size distribution curve, the porous body had a macropore of 0.43 μm. Further, according to the nitrogen adsorption measurement, the total pore volume in the compact after drying at 60 ° C. is 100% or more of the powder sample in terms of the amount of zeolite particles, of which the low pressure part (relative pressure P / Po << The nitrogen adsorption amount of 0.12) was 64% or more of the powder sample.

(Example 10)
The organic polymer was used in the same manner as in Example 9, except that polysulfone (Udel PSF p-1700, Solvay Advanced Polymers Co., Ltd.) was used as the organic polymer, and dimethylformamide of Example 3 was used as a good solvent for the slurry. A slurry for molding was prepared by adding the raw material particles F while stirring the previously dissolved solvent, and using the same nozzle as in Example 9, spinning of the hollow fiber was performed under the same conditions.

  The produced hollow fiber membrane had an outer diameter of 1.8 mm and an inner diameter of 1.0 mm. The content of zeolite particles in the compact by thermal analysis was 59% by weight, and it is considered that there was a loss of fine particles of 4-5% with respect to the charging ratio during spinning. It was dried at 60 ° C. overnight and measured with a mercury porosimeter. The porosity was 66%. According to the pore size distribution curve, it was a multi-element porous body having macropores of 0.22 μm and 1.43 μm. Further, according to the nitrogen adsorption measurement, the total pore volume in the compact after drying at 60 ° C. is 85% of the powder sample in terms of the amount of zeolite particles, of which the low pressure part (relative pressure P / Po <0 .12) was 77% or more of the powder sample.

(Example 11)
In a state where 203.50 g of dimethylformamide used in Example 3 was heated and stirred, 20.24 g of the methyl methacrylate resin used in Example 3 was gradually mixed as an organic polymer and stirred overnight at 330 rpm at 50 ° C. And dissolved. While the mixture was heated and stirred, 40.22 g of the synthetic high silica zeolite (raw material particles F) used in Example 9 was added, and stirring was further continued for 4 to 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 charged amount of the zeolite particles to the compact was 67% by weight. After stirring for 5 hours, using the double-neck nozzle used in Example 1, the obtained slurry was put into a cylinder pump at 20.6 ml / min and injected into a 30 ° C. hot water bath, and the core liquid was 25. A film was formed into a hollow fiber shape by flowing water 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 the thermal analysis, the weight loss when heated to 600 ° C. is 39% by weight, and the content of zeolite particles in the compact is considered to be 61% by weight. According to mercury porosimetry, the porosity after drying overnight at 60 ° C. was 55%. According to the pore diameter distribution curve, it was a multi-element porous body having macropores of 2.5 μm diameter and 6 μm diameter. According to the nitrogen adsorption measurement, the total pore volume in the compact after drying at 60 ° C. was 82% of the powder sample in terms of the amount of zeolite particles, of which the low pressure part (relative pressure P / Po <0.12) due to the zeolite pores. ) Was 64% or more of the powder sample.

(Example 12)
In order to improve the strength and flexibility of the hollow fiber obtained in 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 Example 3. The hollow fiber after drying overnight at room temperature was put into the container, and it heated at 40-50 degreeC for 2 hours, and repolymerized. After further washing and drying at room temperature, heat treatment was performed again in ethanol under reduced pressure for 2 hours. According to mercury porosimetry, the porosity was 40% due to the polymerization treatment, which was reduced from that before the treatment, and macropores were generated 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 compact after vacuum drying at 150 ° C. for 24 hours was 97% of the powder sample in terms of fine particle content, of which the low pressure part (relative pressure P / Po) due to the micropores. The nitrogen adsorption amount of <0.12) was 77% or more of the powder sample, and the pore retention rate also increased compared to the hollow fiber before treatment. In addition, 0.3087 g of this hollow fiber 60 ° C. dried product 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.

(Example 13)
In a state where 110.95 g of a normal methylpyrrolidone solution (Vilomax HR-16NN, Toyobo Co., Ltd.) containing 14% by weight of a polyamideimide resin as an organic polymer was heated and stirred at 45 ° C. and 335 rpm, Examples 9-12 31.19 g of the synthetic high silica zeolite (raw material particles F) used in 1 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 to the compact was 67% by weight. After stirring for 5 hours, using the double-neck nozzle used in Example 1, the obtained slurry was put into a cylinder pump and injected into a 30 ° C. hot water bath at 21.4 ml / min. Water at 10.degree. C. was flowed at 10.4 ml / min to form a hollow fiber film.

  The produced hollow fiber membrane had an outer diameter of 2.6 mm, an inner diameter of 1.0 mm, and a bending strength of about 4-5 MPa. According to thermal analysis, the content of zeolite particles in the compact was around 65% by weight. According to mercury porosimetry, the porosity after drying overnight at 60 ° C. was 50%. According to the pore diameter distribution curve, it was a multi-element porous body having mesopores having a diameter of 20 nm or less and macropores having a diameter of about 0.2 μm. According to the nitrogen adsorption measurement, the total pore volume in the compact 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 nitrogen adsorption amount of <0.12) was 24% or less of the powder sample and the pore volume of the mesopores was large, but the micropores were considerably crushed.

(Example 14)
For the polyamideimide hollow fiber obtained in Example 13, in order to improve the micropore function, in the same manner as in Example 12, the hollow fiber after drying at room temperature was subjected to heat treatment in ethanol under reduced pressure at 40-50 ° C. For 2 hours. According to the thermal analysis, the zeolite content was not changed to 65% by weight. According to mercury porosimetry, the porosity increased to 59% compared to before the treatment, and according to the pore size distribution curve, macropores were generated with a diameter of 0.43 μm instead of 0.2 μm. According to the nitrogen adsorption measurement, the total pore volume in the compact after vacuum drying at 150 ° C. for 24 hours was 0.17 cm ³ / g, 1.4 times that of the raw material particles, of which the low-pressure part (relative The amount of nitrogen adsorbed at a pressure P / Po <0.12) was 84% or more of the powder sample, and the pore retention rate was significantly increased compared to the hollow fiber before treatment. Further, when the strength of this hollow fiber was measured, the bending strength was 4-6 MPa, and no decrease in strength was observed compared to before the treatment. FIG. 5 shows a comparison of nitrogen adsorption isotherms at room temperature for the raw material particles F of Example 13 and Example 14 (modified product of Example 13). In addition, it pre-processes on 150 degreeC 24 hours pressure reduction conditions, and is conversion by content of particle | grains F.

(Example 15)
In Example 2, 20.10 g of synthetic high silica zeolite (SiO ₂ / Al ₂ O ₃ = 52) (raw material particles G) having a particle size of 0.5-1 μm, a crystal particle size of 50 nm, and a pore size of 0.58 nm was used as the raw material particles. The mixture was previously mixed with 50.10 g of dimethyl sulfoxide used, and 9.98 g of the resin used in Example 2 was added as an organic polymer while stirring with heating at 200 rpm at 47 ° C. 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 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 to the compact was 67% by weight. Using the double-neck nozzle used in Example 1, spinning of the hollow fiber was attempted while flowing the slurry at 19 to 22.5 mL / min under the condition of an idle running distance of 5 cm and a hot water bath of 27 ° C. When flowing at -2 mL / min, the injection of the slurry was interrupted and became intermittent, so that hollow fiber molding could not be performed, and a pellet-shaped molded body was obtained instead.

  When the pellet-shaped molded body after washing and drying was measured with a mercury porosimeter, the porosity was about 69%. The particle content estimated from the thermal analysis was 67%, which was as charged. According to the nitrogen adsorption measurement, the total pore volume in the compact is 56% of the powder sample in terms of 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 polyethersulfone 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. A pellet was prepared in the same manner as in Example 2 except that the molding slurry was prepared. At this time, the fine particle concentration in the slurry was 16% by weight, and the charged ratio of the fine particles to the compact 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 substances was approximately 20%, and the content of fine particles was 80%, which was higher than the charging ratio. According to mercury porosimetry, the porosity of the molded body was 69%, and according to the pore size distribution curve, it was a multi-element porous body having mesopores in a region around 5 nm in addition to a plurality of macropores. Furthermore, according to the nitrogen adsorption measurement, the total pore volume in the compact was about 50% of the powder sample in terms of the fine particle content.

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

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

(Comparative Example 2)
1.15 g of polysulfone used in Example 10 as an organic polymer is placed in 20.36 g of dimethylformamide used in Examples 3, 10 and 11 at the same time as 1.15 g of raw material particles A and stirred to form a molding slurry. Except for the preparation, pellet molding was attempted in the same manner as in Example 2, but could not be molded.

(Comparative Example 3)
Except for using an ethylene-vinyl alcohol copolymer resin (Soarnol D2908, Nippon Synthetic Chemical Industry Co., Ltd.) having an ethylene content of 29 mol% as an organic polymer, an attempt was made to produce pellets in the same manner as in Example 7. The prepared slurry spreads 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 a solvent for the slurry, a mixed solvent of 95.5 g of dimethylformamide and 54.83 g of dimethylacetamide (Wako Special Grade) used in Examples 3, 10 and 11 was used, and 60.12 g of raw material particles F were previously dispersed therein. A slurry for forming a porous body was prepared in the same manner as in Example 10 except that polysulfone as an organic polymer was added while heating and stirring, but the polysulfone was not mixed in the slurry, and a 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 dimethylformamide used in 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 was not obtained.

(Comparative Example 6)
As described in Example 15, the molding slurry was prepared by the method of mixing the raw material particles G in advance, 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 Examples 3 and 11 as an organic polymer was dissolved in 160.69 g of dimethyl sulfoxide by heating and stirring at 45 ° C., and then stirred with natural mordenite powder from Kamiaiko (New Tohoku) 40.0 g of Chemical Industry Co., Ltd., raw material particles H) was added while sieving with a 30 mesh SUS wire mesh to prepare a molding slurry. At this time, the concentration of fine particles in the slurry was 18% by weight, and the charged amount of zeolite particles in the compact was 71% by weight. After stirring overnight, using the double-mouth nozzle used in Example 1 in a dry / wet spinning device, the resulting slurry was put into a cylinder pump and injected into a 40 ° C. hot water bath at 22.8 ml / min. Then, 25 ° C. water was flowed at 10.1 ml / min as a core solution to form a hollow fiber.

  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 was put in ethanol (Wako special grade 99.5%) 1L 791.82 g) and dissolved in the same manner as in Example 3. And heated at 40-60 ° C. for 2 hours for repolymerization. The hollow fiber after the treatment was washed and dried, and thermal analysis showed that the weight loss due to the decomposition of the organic matter was 40% by weight, and the content of zeolite particles in the molded body was 60% by weight. It is thought that the loss at the time of adding the raw material to was large. According to mercury porosimetry, the porosity was 46%. According to the nitrogen adsorption measurement, the total pore volume in the compact after vacuum drying at 150 ° C. for 24 hours is 24% of the powder sample, particularly the low pressure part (relative pressure P / Po) due to micropores in terms of fine particle content. The nitrogen adsorption amount of <0.12) was 5% or less of the powder sample, and it was the same even when the hollow fiber before the polymerization treatment was measured.

  Table 1 shows the physical properties of the functional fine particles used in Examples and Comparative Examples.

  In Tables 2-4, the physical property of the (porous) molded object obtained by the Example or the comparative example is shown.

Example 11 Cobalt (CoCl ₂₎ adsorption performance of ethanol after treatment of the molded body of Comparative Example 7 and Comparative Example 7, at room temperature for 24 hours, by a batch test, a comparison was made with其's raw material particles. The results are shown in Table 5. Each of the adsorbents was added by adding 0.3 g to 30 mL of cobalt aqueous solution 50 ppm. The raw materials F and H have different Si / Al ratios in the zeolite. In the zeolite particles, the substitution of Si in the structure with Al causes ion exchange sites, so the Co adsorption capacity of the raw material H is considerably larger than that of F. In Example 11, the amount of cobalt adsorbed on the molded body by the PMMA resin is higher than that of the raw material F, but the amount of adsorption of the PMMA molded body of Comparative Example 7 is considerably lower than that of the raw material H. When Comparative Example 7 was heat-treated in ethanol under reduced pressure as in Examples 12 and 14, the ability to adsorb cobalt corresponding to the raw material particles was recognized as shown in Table 5.

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

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

(Example 19)
Polyethersulfone as an organic polymer (Veradel PES 3000P Solvay Advanced Polymer Co., Ltd., 0.54 g of weight average molecular weight (Mw) = 57,000) was completely added to 5.50 g of dimethyl sulfoxide (Wako special grade) at 50 ° C. Dissolved and mixed with 0.55 g of synthetic mesoporous silica (raw material particles I) used in Example 18 to prepare a slurry. At this time, the concentration of particles in the slurry was 8.3% by weight, the amount of polyethersulfone added was 8.2% by weight, and the amount of MCM particles charged to the compact was 50% by weight. This slurry was dropped into a beaker containing hot water of about 32 ° C., solidified for 2 hours or more, dried at 60 ° C., stirred and washed at room temperature for 25 hours or more until no turbidity was observed, and “MCM-41-PES pellets” Was molded.

When 0.0288 g of the pellet after stirring and washing until the cloudiness disappeared was stirred in 3 mL of water at room temperature for 72 hours or more and the dry weight was measured, almost no change in weight was observed. When this was heated to 600 ° C. by thermal analysis, the organic polymer was 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. (It is considered that a reversible endothermic peak and weight loss not found in the raw material particles and the organic polymer were observed around 120 ° C., and that a new adsorption site derived from molding was generated.) When measured with a mercury porosimeter, the porosity was 78%. According to the pore size distribution curve, it was a multi-element porous body having 4 nm to 10 nm diameter nanopores derived from MCM-41 and 0.1 μm to 1 μm macropores. Further, according to the nitrogen adsorption isotherm, the pore volume of the compact 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. It is considered that almost all the mesopore volume of MCM-41 in the molded body is retained in terms of the amount of MCM-41 particles contained in the compact.

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

(Example 21)
“FSM-22-6 nm-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 size of 6 nm was used as the raw material particles. According to the thermal analysis, the content of particles in the pellet was about 52%.

(Example 22)
“FSM-22-8 nm-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 size of 8 nm was used as the raw material particles. According to the thermal analysis, the content of particles in the pellet was about 55%.

(Example 23)
“SBA-15-4 nm-PES pellets” were molded in the same manner as in Example 19 except that SBA-15 type synthetic mesoporous silica (raw material particles M) having a pore size of 4 nm was used as the raw material particles. According to the thermal analysis, the content of particles in the pellet was about 50%. Moreover, the external appearance image of the composite_body | complex of the SBA-15 type synthetic mesoporous silica (raw material particle | grains M) used in Example 23, the obtained molded object, and the said molded object and an enzyme (lipase) is shown in FIG. (A) is a powder (10 mg) of typical mesoporous silica (SBA-15 type, raw material particles M), (b) is a molded body (20 mg), and (c) is the molded body and It is a complex with an enzyme (lipase). (A) and (b) are white and (c) is brown.

(Example 24)
“SBA-15-6 nm-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 size of 6 nm was used as the raw material particles. According to the thermal analysis, the content of particles in the pellet was about 50%.

(Example 25)
“SBA-15-8 nm-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 size of 8 nm was used as the raw material particles. According to the thermal analysis, the content of particles in the pellet was about 50%.

(Example 26)
“SBA-16-5 nm-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 size of 5 nm was used as the raw material particles. According to the 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 put in 5.55 mL of dimethyl sulfoxide (Wako Special Grade), dissolved by heating and stirring, and an MSU-X type synthetic mesoporous having an average pore diameter of 5.8 nm. 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 dripped into a beaker containing room temperature water to be solidified 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)
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, the same as in Example 19. Thus, “mesoporous Al ₂ O ₃ -PES pellets” were formed. According to the 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), dissolved by heating and stirring, and mesoporous titanium oxide having a particle size of 10 to 100 nm and an average pore size of 4.2 nm. (Alfa Aesar No. 43250) (Raw material particle R) 0.20 g was mixed to prepare a slurry. This slurry was dripped into a beaker containing room temperature water to be solidified 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%. Moreover, the specific surface area in the molded body which was pretreated at 100 ° C. and calculated from the vapor adsorption measurement result 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, when converted in terms of the amount of mesoporous titanium oxide particles contained, it is considered that almost all mesoporous volumes of mesoporous titanium oxide in the compact were retained.

(Comparative Example 8)
A “PES pellet”, which is a pellet of polyethersulfone only, was prepared in the same manner as in Example 19 except that the raw material particles were not added and double the amount of polyethersulfone, which is an organic polymer, was added. did. According to mercury porosimetry, the pellet was a porous body with a porosity 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 fine particles used in the production of a porous molded body of mesoporous fine particles.

  Table 7 shows a porous molded body of mesoporous fine particles obtained in Examples 18 to 29, and a molded body manufactured using only polyether sulfone, which is an organic polymer, without using raw material particles in Example 19. The physical properties of are shown.

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

(1-1) Production Method of AzoR-MCM-41-EVOH Hollow Fiber Composite The MCM-41-EVOH hollow obtained in Example 18 was used as a porous molded body of mesoporous fine particles for the enzyme immobilization support. Yarn (average pore size: 2.66 nm) was used. For comparison, MCM-41 type synthetic mesoporous silica (raw material particles I) was used as the mesoporous silica fine particles before molding. As the azo reductase, Escherichia coli-derived AzoR (dimer, amino acid residue number: 201, molecular weight (monomer): about 23 kD) was used.

  First, an AzoR gene amplified from Escherichia coli chromosomal DNA was inserted into a pET100 / D-TOPO vector to prepare a circular plasmid DNA for AzoR expression. Next, the circular plasmid DNA was introduced into recombinant E. coli, and AzoR was produced using an E. coli protein expression system.

  When immobilizing the enzyme, according to the procedure described in FIG. 8, 1 mL of a buffer solution (25 mM Tris-HCl (pH 7.5)) containing AzoR (0.387 mg) was cut in advance to a length of 5 mm, MCM-41-EVOH hollow fiber (15 or 30 mg) weighed into a microtube and equilibrated with the same buffer was combined by gently mixing for 20 hours at room temperature using a rotator, The AzoR-MCM-41-EVOH hollow fiber composite was obtained by performing the centrifugation operation and the washing operation using the same buffer twice. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used without being washed, 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 the enzyme, a washing step is performed together with the following centrifugation step.

Specifically, centrifugation (4 ° C., 18,000 G, 10 minutes) was performed on the dispersion of the AzoR-MCM-41-EVOH hollow fiber composite (not washed), and all the supernatant was collected. . Subsequently, 1 mL of a washing buffer (using the buffer selected at the time of immobilization) was added, and the mixture was stirred for about 5 seconds at room temperature using a Vortex Mixer to resuspend the complex of AzoR and the mesoporous silica compact. After that, centrifugation (4 ° C., 18,000 G, 10 minutes) was performed, and a washing operation for collecting all the supernatant was performed. Again, the washing operation was repeated using 1 mL of the washing buffer, and finally, an AzoR-MCM-41-EVOH hollow fiber composite (washed) was obtained. Hereinafter, it is referred to as “AzoR-MCM-41-EVOH hollow fiber”.
For comparison, a composite was obtained by the same procedure using MCM-41 type synthetic mesoporous silica (raw material particles I) (10 mg). Hereinafter, it is referred to as “AzoR-MCM-41-powder”.

  The amount of enzyme immobilized on the molded body 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 collected liquid of the centrifugation step and the washing step. Was evaluated by subtraction and division. FIG. 9 shows the amount of AzoR immobilized on MCM-41-powder (typical mesoporous silica (MCM-41 type, raw material particle I), or MCM-41-EVOH hollow fiber of Example 18). Show.

  From FIG. 9, it was recognized that the amount of AzoR immobilized on the MCM-41-EVOH hollow fiber and the rate of immobilization tend to increase in proportion to the amount of the molded body. Considering that the content of MCM-41 particles in the molded body was 66% by weight from Example 18, MCM-41 particles contained in 15 mg and 30 mg of MCM-41-EVOH hollow fiber were 10 mg and 20 mg, respectively. It seems to be equivalent. Here, when MCM-41-powder (10 mg) and MCM-41-EVOH hollow fiber (15 mg) were compared, the amount of MCM-41 particles was equal but the hollow fiber molded body was more than the powder. However, it was found that the enzyme immobilization amount and the immobilization rate were reduced, but the molded body retained the enzyme immobilization ability corresponding to 82.5% of the powder.

(1-2) Evaluation of methyl red degradation activity The enzyme activities of AzoR-MCM-41-EVOH hollow fiber and AzoR-MCM-41-powder obtained in (1-1) were used as azo dyes as reaction substrates. Evaluation was based on the decomposition rate (decoloration rate) of (methyl red).
Specifically, as shown in FIG. 8, the following enzyme reaction is caused for each complex, and the change in absorbance of various reaction substrates (methyl red and coenzyme (NADH)) is examined to determine the enzyme. Activity was evaluated.

The enzyme reaction was performed on the reaction substrate (methyl red, NADH, etc.) for the AzoR-MCM-41-EVOH hollow fiber and the AzoR-MCM-41-powder obtained in (1-1). 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 volume: 1 mL”. For comparison, free AzoR that does not use an immobilized support was also reacted by mixing with the same reaction substrate.
The reaction conditions were that the temperature was maintained at 37 ° C. for 30 minutes with gentle 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 collected solution was measured at 340 nm or 430 nm using a microplate reader (SpectraMax M2e; manufactured by Molecular Devices). Thus, the consumption rate of NADH 30 minutes after the reaction in the methyl red decomposition reaction or the decomposition rate (decolorization rate) of methyl red was evaluated. FIG. 10 and Table 8 show the change in absorbance of NADH and methyl red in the reductive decomposition of methyl red with AzoR-MCM-41-EVOH hollow fiber 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 degradation of azo dye (methyl red) by AzoR-mesoporous silica complex (powder or molded product). FIG. 10 (c) shows the degradation rate of methyl red.

  From FIGS. 10 (a) and (b), NADH and methyl were found in any of free AzoR, AzoR-MCM-41-powder, or AzoR-MCM-41-EVOH hollow fiber without using an immobilized support. There was a tendency for the absorbance of red to decrease significantly. This indicates that also in the immobilized enzyme, the degradation reaction of methyl red accompanying consumption of NADH proceeds efficiently.

  Interestingly, focusing on the change in absorbance after the enzyme reaction in FIG. 10 (a), a greater decrease in absorbance was shown for the immobilized enzyme compared to the free enzyme. From this, it is presumed that NADH was consumed more efficiently or that the immobilized enzyme and NADH formed a complex.

  Further, the absorbances of NADH and methyl red in the absence of enzyme showed comparable values when using either MCM-41-powder or MCM-41-EVOH hollow fiber as compared to the reaction solution itself. Therefore, the reaction substrate itself, that is, NADH and methyl red molecules are hardly adsorbed on the mesopores of the MCM-41-powder and the mesopores of the MCM-41-EVOH hollow fiber and the organic polymer part. It was suggested.

  Also, from FIG. 10 (c), the degradation rate of methyl red by the enzyme immobilized on the MCM-41-powder and MCM-41-EVOH hollow fiber was equivalent to that of the unimmobilized free enzyme, It was found that the original enzyme activity was retained even in the immobilized state.

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

(1-1) Production Method of Composite of AzoR and Various Porous Molded Body Eleven types of moldings obtained in Examples 18 to 28 were used as a porous molded body of mesoporous fine particles on an AzoR immobilization support. Used the body. Moreover, the “PES pellet” obtained in Comparative Example 8 was used as a molded body that did not contain raw material particles for comparison. Furthermore, nine types of synthetic mesoporous silica (raw material particles I to Q) were used as 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) into each well of a 96-well deep bottom type plate in advance. 11 types of molded bodies that had been equilibrated with the liquid and “PES pellets” (corresponding to 20 mg) were mixed using a stirrer (TAITEC, Deep Well Maximizer, BioShaker M • BR-022UP) (25 And a composite of AzoR and various porous molded bodies was obtained by performing the centrifugation operation and the washing operation using the same buffer twice. . The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used without washing, but in this example, in order to evaluate the binding stability of the enzyme to the molded body. In addition, the washing process is performed together with the following centrifugation process.

  Specifically, centrifugation (25 ° C., 4,700 rpm, 10 minutes) was performed on the dispersion of the complex of AzoR and various porous molded bodies (unwashed), and the entire supernatant was collected. . 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 body was stirred for 1 hour at 25 ° C. using the stirring device. Then, centrifugation (25 ° C., 4,700 rpm, 10 minutes) was performed, and a washing operation for collecting all the supernatant was performed. Again, the washing operation was repeated using 1 ml of the washing buffer solution, and finally, composites (washed) of AzoR and various porous molded bodies were obtained. Hereinafter, it is referred to as “AzoR-mesoporous fine particle compact”.

  For comparison, nine types of synthetic mesoporous silica (raw material particles I to Q) (equivalent to 10 mg) were used to prepare a microtube, a rotator, and a microtube centrifuge (25 ° C., 20,000 G, 10 The composite was obtained by the same procedure as that of the molded body except that the minute) was used. Hereinafter, it is referred to as “AzoR-mesoporous fine particle powder”.

  The amount of enzyme immobilized on various mesoporous fine-particle compacts and powders is determined based on the amount of enzyme before immobilization (0.328 mg in 1 mL), and the free enzyme contained in the collected liquid in the centrifugation step and washing step. Calculated by subtracting the amount. FIG. 11 shows the amount of AzoR immobilized on a mesoporous fine particle molded body, a molded body not containing raw material particles, or mesoporous fine particle powder. (A) is the amount of AzoR immobilized on various molded products of mesoporous fine particles. (B) is the amount of AzoR immobilized on various powders of mesoporous fine particles.

11 (a) and 11 (b), AzoR, which is generally equivalent to the immobilized support other than mesoporous Al ₂ O ₃ -EVOH pellets, SBA-16-5nm-powder, or mesoporous Al ₂ O ₃ powder. The amount of immobilization of was shown. From Table 7, considering that the content of mesoporous fine particles in 11 types of molded products obtained in Examples 18 to 28 is 52% by weight in average, the mesoporous fine particles contained in 20 mg of the molded product correspond to 10 mg. The immobilization amount of AzoR shown in FIG. 11 (a) is equivalent to the immobilization amount of AzoR with respect to the mesoporous fine particle powder (10mg) in FIG. 11 (b). This suggests that the enzyme immobilization ability is equivalent to that of powder. Here, SBA-16-5nm- powder, or a mesoporous _Al 2 _{O 3} powder, containing these powders, SBA-16-5nm-PES pellets, mesoporous _Al 2 _O 3 EVOH pellets, or mesoporous Al _In comparison with ₂ O ₃ -PES pellets, among the above three types of molded products, polyether salmon was particularly used as the organic polymer, although almost no enzyme was immobilized on the above two types of powders. In the case of SBA-16-5 nm-PES pellets and mesoporous Al ₂ O ₃ -PES pellets molded using Hong (PES), the same enzyme immobilization as in the case of using PES pellets containing no raw material particles The amount of conversion was shown. From this, it was found that the enzyme (AzoR) adsorbs nonspecifically on the PES surface portion of the molded body, and when a molded body using polyethylene polyvinyl alcohol copolymer resin (EVOH) as the organic polymer was used. Suggests the possibility of specific immobilization in the mesopores contained in the compact. Therefore, from the results of Example 30 and FIG. 11, it was revealed that a molded body using EVOH is suitable for immobilizing AzoR, such as MCM-41-EVOH hollow fiber.

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

Various AzoR-mesoporous fine particle powders are transferred from a microtube to each well of a 96-well deep bottom type plate that already contains AzoR-mesoporous fine particle compacts and AzoR-PES pellets (without raw material particles) before the enzyme reaction. Moved. The enzyme reaction contains methyl red against the AzoR-mesoporous particulate compact, AzoR-PES pellets (without raw material particles), and AzoR-mesoporous particulate powder in each well of the 96 well deep bottom type plate. First, Tris buffer solution (0.966 mL) containing only NADH and FMN was added, and the mixture was stirred (30 ° C., 1,200 rpm, 10 minutes) using the above stirring device, and then the methyl red solution (0. 034 mL) was added (first reaction).
At that time, the reaction composition of the reaction substrate was adjusted to “25 mM Tris-HCl (pH 7.5), 0.05 mM methyl red, 0.2 mM NADH, 1 μM FMN, reaction volume: 1 mL”. In the second and subsequent reactions, the reaction was started by adding a buffer solution, a substrate, and 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 that does not use an immobilized support was also reacted by mixing with the same reaction substrate.
The reaction conditions were that the temperature was maintained at 30 ° C. for 10 minutes while mixing (1,200 rpm) using the stirring device. Subsequently, after 10 minutes, all the supernatant was recovered by centrifugation in a heated state (30 ° C., 4,700 rpm, 5 minutes), and the absorbance at 430 nm of the recovered solution was measured using the microplate reader. As a result, the decomposition rate (decolorization rate) of methyl red was evaluated. FIG. 12 shows the decolorization rate of methyl red when repeatedly using various AzoR complexes (5 reactions). The degradation rate (decoloration rate) of methyl red by the AzoR-mesoporous fine particle composite (a: molded product, b: powder) and the durability (5 reactions) in repeated use of the immobilized enzyme were shown.

  FIG. 13 (a) shows the molded product of Example 23, and FIG. 13 (b) shows the composite of the molded product after the methyl red decomposition reaction and AzoR (AzoR-SBA-15-4nm-PES pellets). The photograph of the external appearance of the composite_body | complex (AzoR-PES pellet (without raw material particle | grains)) of the said molded object and AzoR after completion | finish of a methyl red decomposition | disassembly reaction is shown to c) for the molded object of the comparative example 8, respectively.

  12 (a) and 12 (b) show a decolorization rate of about 90% of methyl red in the same manner as FIG. 10 (c) except for SBA-15-6nm-powder, and high durability in repeated use. Was recognized.

  On the other hand, FIG. 14 shows the results of evaluating the enzyme activity of free AzoR (enzyme amounts: 0, 1, 10, 100 μg). FIG. 14 shows the effect of unfixed free AzoR concentration on the degradation rate (decolorization rate) of methyl red. From FIG. 14, it was found that the optimum amount of enzyme that enables a decolorization 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 enzyme is immobilized on various AzoR complexes, a part of the immobilized enzyme, that is, 1 to 10 μg of enzyme is involved in the reaction. It is inferred that From this, it was suggested that when using a high concentration enzyme, the activity of the free enzyme is remarkably reduced, but the original enzyme activity can be stably expressed in the immobilized state.

  Interestingly, when the appearance of the AzoR-mesoporous fine particle compact and the AzoR-PES pellet (without raw material particles) after the reaction in FIG. 12A was confirmed, the mesoporous fine particle compact exhibited a pink color. The PES pellets (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 body, a complex of AzoR-NADH-methyl red is formed in the mesopores in the molded body. On the other hand, in AzoR-PES pellets (without raw material particles), it is considered from FIG. 11 that a considerable amount of AzoR is adsorbed nonspecifically on the PES surface portion of the molded body. It is assumed that no complex is formed. That is, it is considered that the decomposition reaction of highly active methyl red by the AzoR-PES pellet (without raw material particles) shown in FIG. 12A is due to the contribution of a small amount of free AzoR released from the molded body. Therefore, it was suggested that AzoR immobilized on mesopores in the molded product greatly contributed to the enzyme reaction by the AzoR-mesoporous fine particle molded product.

(Example 32: Production of complex of GDH and various porous molded bodies and enzyme reaction)
In this example, the mesoporous fine-particle porous molded body obtained in Examples 18 to 28 and the molding produced in Comparative Example 8 using only polyether sulfone, which is an organic polymer, without using 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) Production Method of Composite of GDH and Various Porous Molded Body Eleven types of moldings obtained in Examples 18 to 28 are used as a porous molded body of mesoporous fine particles on the GDH immobilization support. Used the body. Moreover, the “PES pellet” obtained in Comparative Example 8 was used as a molded body that did not contain raw material particles for comparison. Furthermore, nine types of synthetic mesoporous silica (raw material particles I to Q) were used as 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) into each well of a 96-well deep-bottom type plate in advance. 11 types of molded bodies that had been equilibrated with the liquid and “PES pellets” (equivalent to 20 mg) were mixed by mixing (25 ° C., 1,200 rpm, 24 hours) using the above stirring device. The composite of GDH and various porous molded bodies was obtained by performing the centrifugation operation and the washing operation using the same buffer twice. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used without washing, but in this example, in order to evaluate the binding stability of the enzyme to the molded body. In addition, the washing process is performed together with the following centrifugation process.

  Specifically, centrifugation (25 ° C., 4,700 rpm, 10 minutes) was performed on the dispersion of the complex of GDH and various porous molded bodies (unwashed), and the entire supernatant was collected. . Subsequently, 1 mL of a washing buffer solution (using the buffer solution selected at the time of immobilization) was added, and the complex of GDH and mesoporous silica molded body was stirred at 25 ° C. for 1 hour using the stirring device. Then, centrifugation (25 ° C., 4,700 rpm, 10 minutes) was performed, and a washing operation for collecting all the supernatant was performed. Again, the washing operation was repeated using 1 mL of the washing buffer solution, and finally, a complex (washed) of GDH and various porous molded bodies was obtained. Hereinafter, it is referred to as “GDH-mesoporous fine particle compact”.

  For comparison, nine types of synthetic mesoporous silica (raw material particles I to Q) (equivalent to 10 mg) were used to prepare a microtube, a rotator, and a microtube centrifuge (25 ° C., 20,000 G, 10 The composite was obtained by the same procedure as that of the molded body except that the minute) was used. Hereinafter, it is referred to as “GDH-mesoporous fine particle powder”.

  The amount of enzyme immobilized on various mesoporous fine-particle compacts and powders is determined based on the amount of enzyme before immobilization (0.363 mg in 1 mL), and the free enzyme contained in the collected liquid in the centrifugation step and washing step. Calculated by subtracting the amount. FIG. 15 shows the amount of GDH immobilized on a mesoporous fine particle molded body, a molded body not containing raw material particles, or mesoporous fine particle powder. FIG. 15 (a) shows the amount of glucose dehydrogenase (GDH) immobilized on a molded body of various mesoporous fine particles. (B) has shown the fixed amount of glucose dehydrogenase (GDH) with respect to the powder of various mesoporous microparticles | fine-particles.

15 (a) and 15 (b), as a whole, it is compared with a molded body and powder of SBA-16-5 nm, a molded body and powder of mesoporous Al ₂ O ₃ , or a PES pellet containing no raw material particles. When a molded body and powder of synthetic mesoporous silica of MCM-41, FSM-22, or SBA-15 type were used, a larger amount of GDH was shown. When the molded product of MCM-41, FSM-22, or SBA-15 type synthetic mesoporous silica was compared with the powder, the amount immobilized on the fine particle powder was slightly larger, but FSM-22 or SBA- Focusing on 15 types of synthetic mesoporous silica, it was recognized that the amount of GDH immobilized was increased in proportion to the pore diameter as a whole for both the compact and the powder. This is thought to be because the amount of immobilization depended on the pore size because GDH having a larger molecular size was not saturated and adsorbed as compared with saturated adsorption of AzoR in Example 31. That is, it suggests that the mesoporous fine particles contained in the compact retain the enzyme immobilization ability equivalent to that of the powder. Here, when considering the amount of GDH immobilized on PBA pellets containing no SBA-16-5 nm molded body and powder, mesoporous Al ₂ O ₃ molded body and powder, or raw material particles, EVOH is used as an organic polymer. It is inferred that more GDH was adsorbed nonspecifically to the PES surface portion of the molded body using PES than to the molded body using PES. Nevertheless, the MCM-41, FSM-22, or SBA-15 type synthetic mesoporous silica molded body has an immobilization amount more than twice that of the case where PES pellets (without raw material particles) are used. From this, it was found that a considerable amount of GDH was fixed to the mesopores of the molded body.

(1-2) Evaluation of NADH production activity 11 types of GDH-mesoporous fine particles, GDH-PES pellets (without raw material particles) obtained in (1-1), and 9 types of GDH-mesoporous fine particles Each enzyme activity was evaluated by the molar concentration of reduced NADH produced from a coenzyme (oxidized NAD ⁺ ) as a reaction substrate. At this time, durability (reaction 5 times) in repeated use of the composite was also evaluated.
Specifically, each of the complexes was subjected to the following enzyme reaction, and the change in absorbance of NADH was examined to determine the generated NADH concentration.

Various GDH-mesoporous fine particle powders are transferred from a microtube to each well of a 96-well deep bottom type plate that already contains GDH-mesoporous fine particle compacts and GDH-PES pellets (without raw material particles) before the enzyme reaction. Moved. The enzyme reaction does not contain glucose with respect to the GDH-mesoporous fine particle compact, GDH-PES pellet (without raw material particles), and GDH-mesoporous fine particle powder in each well of the 96-well deep bottom type plate. , Tris buffer solution (0.967 mL) containing only NAD ⁺ was added, and the mixture was stirred (30 ° C., 1,200 rpm, 10 minutes) using the above stirring device, and then the glucose solution (0.033 mL) was added. Started by addition (first reaction).

At that time, the reaction composition of the reaction substrate was adjusted to “25 mM Tris-HCl (pH 7.5), 0.5 mM NAD ⁺ , 10 mM glucose, reaction volume: 1 mL”. In the second and subsequent reactions, the reaction was started by adding a buffer solution, a substrate, and 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 using an immobilized support was also reacted by mixing with the same reaction substrate.

  The reaction conditions were that the temperature was maintained at 30 ° C. for 10 minutes while mixing (1,200 rpm) using the stirring device. Subsequently, after 10 minutes, all the supernatant was recovered by centrifugation (30 ° C., 4,700 rpm, 5 minutes) in a heated state, and the absorbance at 340 nm of the recovered solution was measured using the microplate reader. The NADH production efficiency (NADH concentration) was evaluated. FIG. 16 shows the generated NADH concentration when various GDH complexes are repeatedly used (5 reactions). FIG. 16 (a) shows the NADH concentration produced by the molded product of the GDH-mesoporous fine particle composite and the durability (5 reactions) in repeated use of the immobilized enzyme. (B) shows the NADH concentration produced by the powder of the GDH-mesoporous fine particle complex and the durability (5 reactions) in repeated use of the immobilized enzyme.

16 (a) and 16 (b), the overall shape of MCM-41 type synthetic mesoporous silica and GDH immobilized on the powder remained low, but FSM-22 or SBA- 15 types of synthetic mesoporous silica compacts and GDH immobilized on powder showed a tendency for enzyme activity to increase in proportion to the pore diameter. Further, from FIG. 16 (a), comparing the activities of the GDH-mesoporous fine particle molded product and the GDH-PES pellet (without raw material particles), most of the GDH-mesoporous fine particle molded products are from GDH-PES pellets (without raw material particles). Also clearly expressed high activity, suggesting the high stability of the enzyme immobilized in the mesopores of the compact. In GDH-PES pellets (without raw material particles), it is considered from FIG. 15 that a considerable amount of GDH is adsorbed nonspecifically on the PES surface portion of the molded body, but from FIG. It was found that the activity of GDH was very low. From the above, it was suggested that GDH immobilized in the mesopores in the compact greatly contributed to the highly active NADH production reaction by the GDH-mesoporous fine particle compact.
Further, when comparing the molded body with the powder, in the case of GDH immobilized on the molded body, although the activity of the first reaction was low, a tendency was found that the activity gradually increased with the number of repeated use. This is because, in the initial stage of the reaction, diffusion of the reaction substrate in the macropores and mesopores in the molded body is rate limiting, and the contact frequency between the enzyme immobilized on the mesopores and the reaction substrate is low. It is assumed that there is.

  On the other hand, FIG. 17 shows the results of evaluating the enzyme activity of free GDH (enzyme amounts: 0, 1, 10, 100 μg). FIG. 17 shows the effect of unfixed free GDH concentration on the generated 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 generated NADH concentration by various GDH complexes is about 0 to 0.4 mM, and from FIG. 15, considering that about 100 to 300 μg of enzyme is immobilized on the GDH complex, It is inferred that a part of the immobilized enzyme, that is, an enzyme corresponding to 1 to 10 μg is involved in the reaction.

(Example 33: Production of complex of lipase and various porous molded bodies and enzyme reaction)
In this example, the mesoporous fine-particle porous molded body obtained in Examples 18 to 28 and the molding produced in Comparative Example 8 using only polyether sulfone, which is an organic polymer, without using 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) Manufacturing method of complex of lipase and various porous molded bodies Eleven types of moldings obtained in Examples 18 to 28 as a porous molded body of mesoporous fine particles were used as the lipase immobilization support. Used the body. Moreover, the “PES pellet” obtained in Comparative Example 8 was used as a molded body that did not contain raw material particles for comparison. Furthermore, nine types of synthetic mesoporous silica (raw material particles I to Q) were used as 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) in advance in each well of a 96-well deep-bottom plate, and use the same buffer. 11 types of molded bodies that had been equilibrated with the liquid and “PES pellets” (equivalent to 20 mg) were mixed by mixing (25 ° C., 1,200 rpm, 24 hours) using the above stirring device. The composite of lipase and various porous molded bodies was obtained by performing centrifugation operation and washing operation using the same buffer twice. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used without washing, but in this example, in order to evaluate the binding stability of the enzyme to the molded body. In addition, the washing process is performed together with the following centrifugation process.

  Specifically, centrifugation (25 ° C., 4,700 rpm, 10 minutes) was performed on the dispersion of the complex of lipase and various porous molded bodies (unwashed), and the entire 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 lipase and mesoporous silica molded body was stirred at 25 ° C. for 1 hour using the above stirring device. Then, centrifugation (25 ° C., 4,700 rpm, 10 minutes) was performed, and a washing operation for collecting all the supernatant was performed. Again, the washing operation was repeated using 1 ml of the washing buffer solution, and finally a complex (washed) of lipase and various porous molded bodies was obtained. Hereinafter, it is referred to as “lipase-mesoporous fine particle molded product”.

  For comparison, nine types of synthetic mesoporous silica (raw material particles I to Q) (equivalent to 10 mg) were used to prepare a microtube, a rotator, and a microtube centrifuge (25 ° C., 20,000 G, 10 The composite was obtained by the same procedure as that of the molded body except that the minute) was used. Hereinafter, it is referred to as “lipase-mesoporous fine particle powder”.

  The amount of enzyme immobilized on various mesoporous fine-particle compacts and powders is determined based on the amount of enzyme before immobilization (0.456 mg in 1 mL), and the free enzyme contained in the collected liquid in the centrifugation step and washing step. Calculated by subtracting the amount. FIG. 18 shows the amount of lipase immobilized on a mesoporous fine particle molded body, a molded body not containing raw material particles, or mesoporous fine particle powder. FIG. 18 (a) shows the amount of lipase immobilized on a molded body of various mesoporous fine particles. (B) shows the amount of lipase immobilized on various mesoporous fine particle powders.

From FIGS. 18 (a) and (b), a synthetic mesoporous of MCM-41, FSM-22, SBA-15 type, SBA-16 type, or mesoporous aluminum oxide type excluding mesoporous Al ₂ O ₃ -EVOH pellets. Comparing the molded body of the silica and the powder, the amount of lipase immobilized on the molded body and the powder is the same. If attention is paid to the synthetic mesoporous silica of the FSM-22 or SBA-15 type, both the molded body and the powder Overall, a tendency for the amount of immobilized lipase to increase in proportion to the pore diameter was observed. This is similar to the tendency of GDH immobilization in Example 32, and since the lipase is not saturatedly adsorbed to the mesopores, the immobilization amount is considered to depend on the pore diameter. That is, it suggests that the mesoporous fine particles contained in the compact retain the enzyme immobilization ability equivalent to that of the powder.

Further, from FIG. 18 (a), only the mesoporous Al ₂ O ₃ -EVOH pellets had a low lipase immobilization amount, but other MCM-41, FSM-22, SBA-15 type, SBA When a molded body of synthetic mesoporous fine particles of -16 type or mesoporous aluminum oxide type is used, the amount of lipase immobilized is about 2-3 times that of PES pellets that do not contain raw material particles. It was. From this, it was found that a considerable amount of lipase was immobilized on the mesopores of the molded body.

(1-2) Evaluation of hydrolyzing activity of triglyceride 11 types of lipase-mesoporous fine particles obtained in (1-1), lipase-PES pellets (without raw material particles), and 9 types of lipase-mesoporous fine particles The enzyme activity of each powder was evaluated by the molar concentration of a fluorescent substance (pyrene) bonded to a fatty acid produced by hydrolysis of fluorescent triglyceride as a reaction substrate. At this time, durability (reaction 5 times) in repeated use of the composite was also evaluated.
Specifically, the following enzyme reaction was caused for each of the complexes, and the concentration of free pyrene was determined by examining the fluorescence intensity of pyrene.

  Various lipase-mesoporous fine particle powders are transferred from a microtube to each well of a 96-well deep bottom type plate that already contains a lipase-mesoporous fine particle compact and a lipase-PES pellet (without raw material particles) before the enzyme reaction. Moved. The enzyme reaction was carried out using a phosphate buffer solution for the lipase-mesoporous fine particle compact, lipase-PES pellet (without raw material particles), and lipase-mesoporous fine particle powder in each well of the 96-well deep bottom type plate. (0.9 mL) was added, and the mixture was stirred (30 ° C., 1,200 rpm, 10 minutes) using the above stirring apparatus, and then started by adding a fluorescent triglyceride solution (0.1 mL) (Reaction 1 Second time).

  At that time, the reaction composition of the reaction substrate was adjusted to “150 mM PBS (pH 8.2), 1 μM fluorescent triglyceride, reaction volume: 1 mL”. In the second and subsequent reactions, the reaction was started by adding a buffer solution, a substrate and 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 not using an immobilized support was also reacted by mixing with a similar reaction substrate.

  The reaction conditions were that the temperature was maintained at 30 ° C. for 10 minutes while mixing (1,200 rpm) using the stirring device. Subsequently, after 10 minutes, all the supernatant was collected by centrifugation in a heated state (30 ° C., 4,700 rpm, 5 minutes), and the fluorescence intensity of the collected solution was measured using the microplate reader ( (Measurement wavelength: excitation 342 nm / fluorescence 400 nm), the production efficiency (pyrene concentration) of fatty acid was evaluated. In FIG. 19, the free pyrene density | concentration at the time of the repeated use (reaction 5 times) of various lipase complexes is shown. (A) of FIG. 19 shows the free pyrene concentration in the triglyceride degradation by the molded product of the lipase-mesoporous fine particle composite and the durability (reaction 5 times) in repeated use of the immobilized enzyme. (B) of FIG. 19 shows the free pyrene concentration in triglyceride degradation by the lipase-mesoporous fine particle powder and durability in repeated use of the immobilized enzyme (5 reactions).

As shown in FIGS. 19 (a) and (b), the overall concentration of free pyrene was found to be lower when the lipase-mesoporous fine particle compact was used as compared with the lipase-mesoporous fine particle powder. No broad increase / decrease in pyrene concentration was observed 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 in five repeated reactions. Further, from FIG. 19 (a), the activity of the lipase-PES pellets (without the raw material particles) gradually decreased with the number of repeated use, and almost no activity was expressed at the fifth reaction. In lipase-PES pellets (without raw material particles), it is considered from FIG. 18 that a considerable amount of lipase is adsorbed nonspecifically on the PES surface portion of the molded product, but the stability of the lipase in the adsorbed state is extremely low. It has been found. On the other hand, most of the lipase-mesoporous fine particle compacts exhibited sustained enzyme activity in the reaction 1 to 5 times, which means that the stability of the enzyme immobilized in the mesopores of the compacts is high. It suggests. From the above, it was suggested that the lipase immobilized in the mesopores in the molded product greatly contributed to the hydrolysis reaction of the highly active triglyceride by the lipase-mesoporous fine particle molded product.
FIG. 20 shows the results of evaluating the enzyme activity of free lipase (enzyme amounts: 0, 1, 10, 100 μg). FIG. 20 shows the effect of the unfixed free lipase concentration on the concentration of the fluorescent substance (pyrene) released by triglyceride decomposition. FIG. 20 shows that the pyrene concentration tends to increase in proportion to the enzyme concentration. The amount of enzyme that gives the concentration of pyrene produced (20 to 40 nM) by the lipase-mesoporous fine particle molded article shown in FIG. 19A is considered to be in the range of 50 to 100 μg from FIG. From FIG. 18, considering that about 100 to 250 μg of enzyme is immobilized on the lipase complex, it is inferred that about half of the immobilized enzyme is involved in the reaction. The

(Example 34: Production of complex of protease and various porous molded bodies and enzyme reaction)
In this example, porous molded bodies of mesoporous fine particles obtained in Examples 22 and 25, and protease N (PN: derived from genus Streptomyces), Nagase Chem for activated carbon pellets (derived from rice husks, Dexerials Corporation) for comparison Tex Co., Ltd.) and enzyme activity were evaluated.

(1-1) Production Method of Complex of Protease and Various Porous Molded Body Two types of moldings obtained in Examples 22 and 25 were used as a porous molded body of mesoporous fine particles on the protease support. Used the body. Moreover, “activated carbon pellets” having a larger mesopore volume than conventional activated carbon were used as porous bodies having different surface affinity for comparison. Furthermore, two types of synthetic mesoporous silica (raw material particles L and O) and “activated carbon powder” were used as mesoporous silica fine particles before molding for comparison.

  When immobilizing the enzyme, 1 mL of a buffer solution (100 mM Tris-HCl (pH 7)) containing protease (0.271 mg), two types of molded bodies previously measured in a microtube, and “activated carbon pellet” (Equivalent to 20 mg) is mixed using a rotator (4 ° C., 15 hours), and then subjected to centrifugation and washing using the same buffer twice. A composite with a green compact was obtained. The complex of the enzyme of the present invention and the porous molded body of mesoporous fine particles can be used without washing, but in this example, in order to evaluate the binding stability of the enzyme to the molded body. In addition, the washing process is performed together with the following centrifugation process.

  Specifically, centrifugation (4 ° C., 20,000 G, 10 minutes) is performed on a dispersion (unwashed) of a complex of the protease and various porous molded bodies (or activated carbon pellets), All supernatant was collected. Subsequently, 1 mL of a washing buffer (using the buffer selected at the time of immobilization) is added, and a complex of protease and mesoporous silica compact (or activated carbon pellet) is used at room temperature for about 5 seconds using a Vortex Mixer. After stirring, centrifugation (4 ° C., 20,000 G, 10 minutes) was performed, and a washing operation for collecting all the supernatant was performed. Again, the washing operation was repeated using 1 ml of the washing buffer solution, and finally a complex (washed) of 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”.

  For comparison, two types of synthetic mesoporous silica (raw material particles L and O) and “activated carbon powder” (corresponding to 10 mg) were used to obtain a composite by the same procedure as that for the molded body. Hereinafter, it is referred to as “protease-mesoporous fine particle powder” or “protease-activated carbon powder”.

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

  21 (a) and 21 (b), FSM-22 (raw material particle L), SBA-15 (raw material particle O) type synthetic mesoporous silica and activated carbon are compared. The tendency for the amount of immobilized protease to increase was observed in the order of “22> SBA-15”. Comparing the compact with the powder, overall, when the compact was used, the amount of protease immobilized was low, but the protease immobilization behavior depending on the immobilization support used powder. This suggests that the mesoporous microparticles contained in the compact retain approximately the same enzyme immobilization ability as the powder.

(1-2) Evaluation of hydrolysis activity of gelatin Protease complex obtained in (1-1) (two types of protease-mesoporous fine particles, protease-activated carbon pellets, two types of protease-mesoporous fine particles, and , Protease-activated carbon powder) was evaluated for enzyme activity. The evaluation was performed based on the relative activity based on the fluorescence intensity of the unfixed free enzyme with the fluorescence intensity increasing with hydrolysis of the fluorescent gelatin as the reaction substrate as an index. At this time, durability (reaction 10 times) in repeated use of the composite was also evaluated.
Specifically, the following enzyme reaction was caused for each of the complexes to determine the degradation activity of gelatin.

  The enzyme reaction is performed on the protease complex (protease-mesoporous fine particle compact, protease-activated carbon pellet, protease-mesoporous particulate powder, and protease-activated carbon powder) in each microtube obtained in (1-1). The reaction was started by adding Tris buffer (0.4 mL) containing fluorescent gelatin, sodium chloride, and calcium chloride (first reaction).

  At that time, the reaction composition of the reaction substrate was adjusted to “45 mM Tris-HCl (pH 7.6), 0.1 mg / mL fluorescent gelatin, 135 mM sodium chloride, 4.5 mM calcium chloride”. In the second and subsequent reactions, the reaction was started by adding the 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. For comparison, a free protease not using an immobilized support was also reacted by mixing with a similar reaction substrate.

  The reaction conditions were that the temperature was maintained at 37 ° C. for 30 minutes while mixing using a rotator. Subsequently, all the supernatant is collected by centrifugation (4 ° C., 20,000 G, 5 minutes), and the fluorescence intensity of the collected solution is measured using the microplate reader (measurement wavelength: excitation 495 nm / fluorescence 520 nm). Thus, the degradation efficiency of gelatin was evaluated. FIG. 22 shows the relative activity based on the degradation efficiency of gelatin by free protease during repeated use of various protease complexes (10 reactions). (A) of FIG. 22 shows the degradation activity of gelatin by the molded product of the protease-mesoporous fine particle composite and the durability (reaction 10 times) in repeated use of the immobilized enzyme. FIG. 22 (b) shows the gelatin degradation activity by the protease-mesoporous fine particle powder and the durability (10 reactions) in repeated use of the immobilized enzyme.

22 (a) and 22 (b), it was observed that the gelatin degradation activity tends to be lower when the protease-mesoporous fine particle compact is used as a whole, as compared with the protease-mesoporous fine particle powder. Both the molded body and the powder showed a significantly superior enzyme activity expression rate and repeated durability when compared with activated carbon (pellet and powder).
Specifically, the degradation activity of gelatin in the 10th reaction was 3.9% when protease-activated carbon powder was used, but about 100% when protease-mesoporous fine particle powder was used. (FSM type: 100%, SBA type: 98.3%), which showed extremely high repetition durability (FIG. 22B).
On the other hand, when protease-activated carbon pellets were used, gelatin degradation activity in the 10th reaction was 41.1%, which was higher than that of activated carbon powder. Was found to have higher activity (FSM type: 92.4%, SBA type: 84.6%) (FIG. 22 (a)).

From the results of FIG. 21 and FIG. 22, activated carbon (pellet and powder) having high surface hydrophobicity has an enzyme activity higher than that of mesoporous silica (molded body and powder), although the protease immobilization rate is high. The extremely low value indicated the superiority of the mesoporous silica powder and the molded product in this reaction system.
Comparing the FSM type and SBA type mesoporous silica, it was observed that the gelatin and the powder as a whole had a tendency to increase the degradation activity of gelatin in the order of “FSM-22> SBA-15”. This is considered to be a result depending on the amount of protease immobilized (FSM-22> SBA-15) shown in FIG.
Further, from the results shown in FIG. 21, in view of the fact that the amount of protease immobilized was lower (70 to 80% in the case of powder) when the molded body was used than the powder, It is presumed that the mesoporous fine particles contained in the body retain the enzyme activity expression ability equivalent to that of the powder.

  As described in detail above, the porous molded body of micro or mesoporous fine particles according to the present invention is a porous material that is easy to handle by mixing a porous fine particle powder having adsorbability with an organic polymer resin in a phase separation method. It can be obtained by molding into In addition, according to the present invention, a microfiltration membrane composed of a porous molded body of the above micro to mesoporous fine particles can be provided. The porous molded body of the present invention can be suitably used as a treatment agent such as industrial waste water.

Claims (23)

A functional fine particle having a pore diameter of 0.3 nm to 50 nm having micro to mesopores, mainly composed of silicate, aluminosilicate, alumina or titania and having a particle diameter of 0.01 μm to 200 μm, and an organic polymer A porous molded body comprising:
Macropores with a pore diameter of 50 nm to 100 μm,
A porous body having a porosity of 34% or more;
The content of the functional fine particles in the molded body is 46% by mass or more,
At least part of the surface of the functional fine particles is exposed from the molded body,
The micro volume or mesopore volume of the functional fine particles is maintained at 40% or more of the raw material functional fine particles,
A porous molded body of micro or mesoporous fine particles characterized by the above.   The porous molded body of micro to mesoporous fine particles according to claim 1, wherein a weight change before and after stirring of the porous molded body when continuously stirred in water for 24 hours or more is 1% or less.   The porous molded body of micro to mesoporous fine particles according to claim 1 or 2, characterized in that the shape of the porous molded body is granular, hollow particle or hollow fiber.   The porous molded body of micro to mesoporous fine particles according to any one of claims 1 to 3, wherein the functional fine particles and / or the organic polymer is a mixture of plural kinds.   The function of the raw material functional fine particles of the functional fine particles is an adsorption performance, and the characteristic expression of the function is maintained after the formation of the porous molded body. A porous molded body of micro or mesoporous fine particles as described in the item.   The porous molded body of micro to mesoporous fine particles according to any one of claims 1 to 5, wherein particles having a size of several microns can be filtered under pressure from the dispersion.   The porous molding of micro or mesoporous fine particles according to any one of claims 1 to 6, wherein the organic polymer is polyethersulfone, polysulfone, EVOH resin, PMMA resin, or polyamideimide resin. body. A step of obtaining a raw material slurry by dispersing functional fine particles mainly composed of silicate, aluminosilicate, alumina or titania having micro to mesopores in an organic solvent in which the organic polymer is previously dissolved;
The method for producing a porous molded body of micro to mesoporous fine particles according to any one of claims 1 to 7, comprising a step of molding the raw material slurry by injecting the raw material slurry into a non-solvent.   The method for producing a porous molded body of micro to mesoporous fine particles according to claim 8, wherein the organic polymer has a hydrophilic group.   The micro or mesoporous microporous material according to claim 8 or 9, further comprising a step of treating the organic polymer with a polymerization accelerator or a curing accelerator after molding, wherein the organic polymer is a PMMA resin or a polyamideimide resin. A method for producing a molded product.   The porous molded body of the micro or mesoporous fine particles is further subjected 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 micro to mesoporous fine particles as described in 1.   The functional fine particles are mesoporous fine particles having mesopores having a pore diameter of 2 nm to 50 nm and a particle diameter of 0.01 μm to 200 μm, and the function of the raw material functional fine particles is one type or two or more types. The porous molded body according to any one of claims 1 to 7, comprising an enzyme and / or protein immobilizing ability, wherein the content of the functional fine particles in the molded body is 46% by mass or more. An enzyme carrier.   A complex of the enzyme-supporting carrier according to claim 12 and an enzyme.   14. The complex according to claim 13, wherein the enzyme is an oxidoreductase, hydrolase, transferase, eliminase, isomerase, and / or synthase.   The complex according to claim 13 or 14, wherein the enzyme reaction of the enzyme is an oxidation-reduction reaction, hydrolysis reaction, transfer reaction, elimination reaction, isomerization reaction, and / or synthesis reaction. The composite according to any one of claims 13 to 15, wherein each of one or more enzymes and / or proteins involved in the enzyme reaction of the enzyme is immobilized in a mesopore having a pore diameter of 2 nm to 50 nm. A method of manufacturing 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 adjusted to pH 3-11.   Furthermore, the manufacturing method of Claim 16 including the washing | cleaning process of wash | cleaning the porous molded object of the mesoporous fine particle after completion | finish of immobilization several times with the buffer solution adjusted to pH 3-11. The complex according to any one of claims 13 to 15, wherein each of one or more enzymes and / or proteins involved in the enzyme reaction is immobilized on a porous molded body of mesoporous fine particles. An enzymatic reaction method,
An immobilization step of immobilizing the enzyme and / or protein on a porous molded body of mesoporous fine particles in a buffer adjusted to pH 3-11;
A reaction substrate is added to a buffer solution having a pH of 3 to 11 containing a complex of a porous compact of mesoporous fine particles obtained in the immobilization step and an enzyme, or mesoporous fine particles obtained in the immobilization step An enzyme reaction method comprising an enzyme reaction step in which a complex of a porous molded body and an enzyme is added to a buffer solution having a pH of 3 to 11 containing a reaction substrate to perform an enzyme reaction involving the enzyme and / or protein. The complex according to any one of claims 13 to 15, wherein each of one or more enzymes and / or proteins involved in the enzyme reaction is immobilized on a porous molded body of mesoporous fine particles. An enzymatic reaction method,
An immobilization step of immobilizing the enzyme and / or protein on a porous molded body of mesoporous fine particles in a buffer adjusted to pH 3-11;
A washing step of washing a complex of a porous molded body of mesoporous fine particles obtained in the immobilization step and an enzyme with a buffer solution of pH 3-11;
An enzyme reaction step of performing an enzyme reaction involving the enzyme and / or protein in a reaction solution containing a reaction substrate, a complex of a porous molded body of mesoporous fine particles obtained after the washing step and the enzyme obtained in the washing step; Enzymatic reaction method comprising.   20. The enzymatic reaction method according to claim 18 or 19, wherein the enzymatic reaction is a method for producing a functional useful substance from a reaction substrate.   The enzyme reaction method according to claim 18 or 19, wherein the enzyme reaction is a method of decomposing an environmental pollutant serving as a reaction substrate present in the environment.   The enzyme reaction method according to claim 18 or 19, wherein the enzyme reaction method is a method for detecting or quantifying a reaction substrate present or possibly present in a test sample.   A kit, sensor or apparatus for enzyme reaction involving the enzyme and / or protein, comprising the complex according to any one of claims 13 to 15.