JP2008238011A - Functional composite membrane formed by photoresist method, its manufacturing method and method of manufacturing useful substance by utilizing functional composite membrane - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a functional composite membrane formed by a photoresist method and two or more layers of phases laminated and having different functions, a method of manufacturing the composite membrane and a method of manufacturing a useful substance by using an enzyme, a protein, etc. and utilizing the composite membrane. <P>SOLUTION: The composite membrane is formed by a photoresist method comprising dissolving a substance containing glutamate decarboxylase in photo-sensitive polyvinyl alcohol, applying the resultant solution to one side of a cation exchange membrane and drying, and irradiating the resultant membrane with light containing the wavelengths hardening the photo-sensitive resin to polymerize. When the formed composite membrane, CM, is laid at the center of a diffusion dialysis vessel consisting of a chamber F on the Feed side and a chamber S on the Strip side and dialysis is carried out by causing a substrate solution performing an enzymatic reaction to flow on one composite surface to which a functional substance is fixed while causing an electrolyte to flow on the other composite surface, a target substance is produced on the composite surface to which the functional substance is fixed, and a part of the product is recovered in the electrolyte solution on the other side through the composite membrane, leading to simultaneous execution of reaction and separation. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a functional composite membrane using a photoresist method in which a plurality of phases having different functions are laminated, a method for producing the same, and a functional composite membrane using enzymes, proteins, cyclodextrins, etc. And a method for producing useful substances.

  When producing useful substances using enzymes, etc., not only enzymatic reactions but also many steps such as separation operation, concentration operation, purification operation, etc. are aimed at efficient production including cost reduction. It is important to introduce a highly efficient process capable of realizing a plurality of steps in one unit, and to reduce the number of processes and units to obtain as a product.

As one of such techniques, operation by a combined reactor that simultaneously performs reaction and separation can be an effective means.
In particular, when a membrane separation method with low energy consumption is employed in the solid-liquid separation operation, the membrane separation method is one of attractive separation operations because a stable operation can be performed without performing complicated operations.
However, when a substance having a large molecular weight such as a solid or colloidal substance is contained in the liquid, the scale adheres to the film surface and the performance of the film is deteriorated. Therefore, when performing the chemical reaction and the separation operation at the same time, it is necessary to perform the operation by immobilizing a useful substance on a gel-like or adherent carrier so that scale does not directly form on the membrane surface.

In general, in such a system, the immobilization carrier and the functional separation membrane often exist as separate units in one system.
Therefore, problems such as an increase in the size of the reaction tank or wear due to contact between carriers occur.
One method for solving this problem is to use a composite membrane in which a phase containing another functional substance is formed on the surface of the functional separation membrane. For example, an ion exchange membrane that is a kind of functional separation membrane is used. Various methods have been devised for combining technologies (see Patent Documents 1 to 4).
However, these methods use a method of chemically synthesizing or coating a phase containing the functional substance when the functional substance is combined with the base material film. In the case of producing a composite membrane containing various proteins, enzymes, etc., it is considered that the activity of useful substances is reduced during the complexation reaction, and sufficient functions are not always expressed.
JP 2005-068396 A JP 2004-139837 A Japanese Patent Application Laid-Open No. 2004-139836 Japanese Patent Laid-Open No. 10-312815

  In the present invention, there is provided a new creation method in which a phase containing a functional substance having thermal and chemical stress is combined with a film having other functionality without reducing its chemical and physical functions. An object of the present invention is to provide a method for producing a useful substance by expressing the function of the composite membrane.

The functional composite membrane of the present invention uses a polymer ion exchange membrane as a base material to be a functional composite membrane, and uses a substance having an azide-based photosensitive group as a photosensitive material for PVA as a photosensitive material. The amount of the photosensitive material is an amount to be solidified by irradiation with light, and the water-soluble material to be immobilized is mixed with the photosensitive material-containing PVA as a lyophilized powder to be mixed or increased in content as a solution, The amount of the immobilized substance such as an enzyme with respect to the photosensitive substance is 0.1-30 wt% after being dissolved or dispersed.
The immobilizing substance contains a single protein or a complex protein such as glutamic acid decarboxylase and amylase for producing a physiologically active substance.

The method for producing a functional composite film using the photoresist method of the present invention uses a material having photosensitivity and a polymer functional film, and a chemical and physical interaction such as an enzyme in the photosensitive material. After dissolving, dispersing, coating, impregnating, and drying on the functional polymer film, the photosensitive material is irradiated with a light source including the wavelength of the region where the photosensitive material undergoes chemical change, and then development and fixing are performed. A high-performance composite film having a plurality of functions in one film is manufactured by using a photoresist method to enhance mechanical bonding between different materials.
The method for producing a useful substance using the functional composite membrane of the present invention uses a composite membrane in which a heterogeneous phase containing an enzyme or the like is combined on a polymer functional membrane, and expresses the function of the enzyme or the like. Production and separation of substances are performed simultaneously.

In the method for producing a functional composite film using the photoresist method of the present invention, since the composite film is prepared using a photosensitive material, the functional material has a reduced functionality compared to a chemical synthesis method or the like. Therefore, composite membranes containing enzymes, proteins, cyclodextrins and the like can be easily produced. In addition, since a lithographic method, which is one of photoresist techniques, is used, a composite film having an arbitrary pattern can be designed by developing and fixing operations.
Further, in the method for producing a useful substance using the functional composite membrane of the present invention, a raw material liquid is supplied to the reaction phase of the obtained composite membrane, and a recovery liquid is passed through the other phase. It can be said that this is an industrially excellent production method because the function of the composite membrane can be expressed by itself and the substance produced in the reaction phase can be separated to the recovery side.
Furthermore, when a phase containing a substance that specifically acts on a specific cell or protein is combined, it is considered possible to separate allergens, viruses, etc. Applications can also be expected.

The inventors of the present invention have created a new creation in which a phase containing a functional substance having thermal and chemical stress is combined with a film having other functionality without deteriorating its chemical and physical functions. In order to provide a method, intensive studies were conducted.
As a result, a substance containing glutamic acid decarboxylase is dissolved in photosensitive polyvinyl alcohol, for example, water-soluble polyvinyl alcohol having an azide-based photosensitive group, and after coating and drying on one side of the cation exchange membrane, the photosensitive resin is cured. The inventors have conceived of a technique for producing a composite film by using a photoresist technique that is polymerized by irradiating a light source including a wavelength to achieve the present invention.
That is, the present invention uses a photosensitive material containing a functional substance to irradiate a light source including a wavelength region in which the photosensitive material undergoes a chemical change without undergoing a thermal polymerization reaction or a synthetic reaction. This is a technology that combines the existing functional membrane with the existing functional membrane, and the technology that allows the function of the composite membrane produced by the invented technology to be expressed and the production / separation operation of the substances simultaneously.
Note that the existing functional film is not limited to an organic film, and may be an inorganic film, cellulose, or the like.

  Since the composite membrane obtained in the present invention has different functionality depending on the composite phase, for example, a reaction solution is provided on each surface of the composite membrane in which a phase having reactivity and a phase having separation characteristics are combined. It is also possible to construct a system in which when a solution for recovering the reactant is flowed, a product is generated in the reaction phase and the product is simultaneously separated to the opposite surface.

In the method for producing a composite film according to the present invention, a functional substance is dissolved in a photosensitive material solution or dispersed as uniformly as possible in a colloidal state so that the composite film obtained after photocrosslinking is also functional. The substance is present uniformly. The content of the functional substance can be adjusted by adjusting the amount of the target substance to be dissolved.
A photosensitive resin solution containing a functional substance is applied and dried on the other film surface to be combined, but the number of times of application or the coating phase thickness can be controlled by spin coating. It is relatively easy to control the laminated structure of the chemical film.
When a negative photosensitive material is used on the coated and dried surface, photopolymerization is performed by irradiating a light source including a wavelength in a region where the material undergoes a chemical change, such as a light source including an ultraviolet region. Since the polymerization can be sufficiently performed even with a light source having a peak wavelength of 352 nm, even a substance that is easily damaged by ultraviolet rays can be immobilized while suppressing a decrease in its function as much as possible.

At this time, when a negative photosensitive material is used, when the irradiation light source is irradiated in an arbitrary shape by the lithographic technique, only the portion is polymerized and cured.
After that, when the developing operation is performed by washing with water or other developing solution, only the non-photosensitive part flows out, so that polymerization according to the pattern of the light source is possible, and a structure that is difficult to obtain by a chemical synthesis method Can be obtained. Note that when a positive type photosensitive material solution is used, only the portion irradiated with light dissolves when the development operation is performed. Therefore, this is opposite to the pattern control method when a negative type photosensitive material is used. It becomes the manufacturing method.
When the functional film that is combined by photopolymerization has water content, a part of the photosensitive material monomer is also impregnated in part of the functional film. Since the polymerization also proceeds in a part of the film, the adhesion is improved as compared with the case where the functional films are separately prepared and bonded.

When a composite material composited by the above-described method is used to produce a useful substance by expressing its function, the composite membrane CM thus produced is divided into 2 of the Feed side F and the Strip side S as shown in FIG. Placed in the center of a diffusion dialysis chamber consisting of a chamber, the substrate solution that performs the enzyme reaction flows on the composite surface on which the functional substance is immobilized, and the electrolyte solution is flowed on the other composite surface to perform the dialysis. The target product is generated on the composite surface, and a part of the product permeates the composite membrane and is collected in the opposite electrolyte solution, so that the reaction and the separation can be performed simultaneously. B is a thermostatic bath.
At this time, when performing an enzyme reaction, the solution may be adjusted to an optimum pH and temperature according to the type of enzyme.

Hereinafter, the method for producing a functional composite film using the photoresist method of the present invention will be described with reference to examples, but the present invention is not limited thereto.
(1) An ion exchange membrane having a thickness of about 100 to 200 μm is used as a functional polymer membrane serving as a base.
(2) A PVA aqueous solution (for example, water-soluble polyvinyl alcohol having an azide-based photosensitive group) having negative photosensitivity and containing a gel-like photosensitizer was used as the immobilizing agent.
(3) Although the weight ratio of the photosensitive material was 2 to 5 wt% with respect to the weight of the aqueous solution, it was not fixed at this concentration.
(4) The substance to be immobilized was treated by a freeze-drying method, then mixed and dissolved in a PVA aqueous solution having a photosensitive function, and uniformly dispersed. The specific operation in the examples was that a crude protein aqueous solution containing glutamate decarboxylase leached from rice bran was freeze-dried, and a predetermined amount was dissolved in PVA having a photosensitive function.
(5) The content ratio of the crude protein containing the enzyme and the like with respect to the aqueous solution (PVA aqueous solution having a photosensitivity function) was about 28%, but from the viewpoint of uniform dispersion, preferably from 0.1 wt% It is the range of the saturated solubility with respect to the aqueous solution (PVA aqueous solution having a photosensitive function).
(6) The aqueous solution (PVA aqueous solution having a photosensitivity function) containing the crude protein or the like was applied onto a cellulose membrane or an ion exchange membrane, impregnated, and then dried. In the examples, a cellulose membrane and a cation exchange membrane were used for coating and impregnation on one side, but the membrane to be combined is not limited to these two types.
(7) After drying, a light source having a peak wavelength of 352 nm was irradiated for 30 minutes in order to advance the chemical reaction. However, the light is not fixed to this peak wavelength as long as it includes a wavelength region in which a photochemical reaction occurs. Further, the energy output of the light source, the distance between the light source and the composite film, and the irradiation time are not fixed to specific values as long as the gel is sufficiently solidified.
(8) The thickness of the immobilization phase on the composite membrane obtained by the above method was about 300 μm, but in order to increase the immobilization amount per unit membrane area, the operation of coating, impregnation and photoreaction When the operation is repeated, the thickness of the film changes, so that the thickness is not fixed.
(9) FIG. 2 shows an image of a membrane in which a PVA phase containing glutamate decarboxylase is combined on a cation exchange membrane. 1 is an ion exchange group, 2 is a cation exchange membrane phase, 3 is an immobilization material, 4 is a bonded phase of a complexed membrane, and 5 is a phase containing a crude protein immobilized with a photosensitive material.

Hereinafter, although the Example demonstrates the manufacturing method of the useful substance using the functional composite film of this invention, this invention is not limited to this.
(1) Immobilization of a crude protein containing glutamate decarboxylase produced by the method of Example 1 to a PVA phase and a complex membrane obtained by complexing with a cellulose membrane express 1 mol / m of the function of glutamate decarboxylase. ³ was immersed in an aqueous solution of glutamic acid, and the reaction of the substance produced at a temperature of 40 degrees was followed.
(2) FIG. 3 is a graph showing the change with time of the amino acid concentration in the glutamic acid aqueous solution. ● is γ-aminobutyric acid (GABA), ○ is glutamic acid, Δ is aspartic acid, and □ is alanine.
(3) From FIG. 3, the concentration of glutamic acid in the immersion liquid rapidly decreases, but the concentration of γ-aminobutyric acid generated by the enzyme reaction correspondingly increases. From this result, it is understood that the function of the crude protein containing the enzyme immobilized on the water-soluble photosensitive material having PVA as a basic skeleton and complexed with the cellulose membrane can be expressed even when complexed.
(4) The glutamic acid aqueous solution used in this example is a substance supplied as a substrate for glutamic acid decarboxylase. When expressing the function of a protein such as another enzyme, a substance corresponding to the substrate specificity is used. It is desirable to supply Further, conditions such as temperature and pH are not particularly fixed as long as they do not inhibit the reaction.

(1) A dialysis composed of two chambers as shown in FIG. 1 is obtained by immobilizing a crude protein containing glutamic acid decarboxylase prepared by the method of Example 1 in a PVA phase and complexing it on one side of a cation exchange membrane. Arranged in the center of the tank.
(2) A crude protein-containing phase containing glutamic acid decarboxylase or the like was placed on the Feed side F shown in FIG. 1, and an ion exchange membrane phase was placed on the opposite Strip side S.
(3) The glutamic acid aqueous solution serving as a substrate was fed to the Feed side F, and the hydrochloric acid aqueous solution was flowed to the Strip side S, so that the enzyme reaction and the separation operation were performed simultaneously. The solution to be fed to the Feed side is not particularly fixed as long as it is a substrate solution that expresses functions such as enzymes. In addition, the solution that flows to the strip side is not fixed to hydrochloric acid as long as the function of the functional membrane can be expressed. In this embodiment, the operation mode is a batch circulation type operation. However, when a sufficient membrane area and a sufficient amount of solution are ensured, an operation by a continuous operation is also possible.
(4) The results are shown in a graph showing the change over time in the amino acid concentration in the Strip chamber of FIG. ● is γ-aminobutyric acid (GABA), ○ is glutamic acid, Δ is aspartic acid, and □ is alanine.
(5) From FIG. 4, although the membrane permeation of glutamic acid as a substrate was observed at the initial stage of dialysis, the produced γ-aminobutyric acid permeated the composite membrane and was separated and collected on the hydrochloric acid side.
Therefore, it was experimentally demonstrated that the function of the newly produced composite membrane was expressed, and that the product could be separated and collected on the opposite side just by flowing the substrate to one side of the composite membrane using the composite membrane. It was done.

  Applications in a wide range of fields such as food industry, pharmaceutical manufacturing, medical field, analytical chemistry, electronics industry including sensors, and wastewater treatment can be expected.

It is the schematic of the apparatus for expressing the function of a composite membrane and performing enzyme reaction and separation operation simultaneously. It is an image figure of the film | membrane which compounded the PVA phase containing an enzyme on the one side of a cation exchange membrane surface. It is a graph which shows a time-dependent change of the amino acid concentration in the solution at the time of immersing the membrane which combined the PVA phase containing glutamic acid decarboxylase with the cellulose in the glutamic acid aqueous solution. 1 shows the time-dependent change in amino acid concentration in the Strip chamber when a membrane in which a PVA phase containing glutamic acid decarboxylase is combined on one side of a cation exchange membrane is attached to the apparatus shown in FIG. FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Ion-exchange group 2 Cation-exchange membrane phase 3 Immobilization substance 4 Junction phase of complexed membrane 5 Phase containing crude protein immobilized with photosensitive substance

Claims (4)

  A polymeric ion-exchange membrane is used as the base material for the functional composite film, and a material having an azide-based photosensitive group, which is a photosensitive material, is used as the photosensitive material in PVA. The amount of the photosensitive material is irradiated with light. The water-soluble substance to be fixed is mixed, dissolved or dispersed in a photosensitive substance-containing PVA as a freeze-dried powder to increase the content or mixed as a solution. A functional composite membrane characterized in that the amount of an immobilized substance such as an enzyme is 0.1-30 wt%.   The functional substance according to claim 1, wherein the immobilized substance contains a single protein or a complex protein such as glutamic acid decarboxylase and amylase for producing a physiologically active substance. Composite membrane.   Using photosensitive materials and functional polymer membranes, highly functional materials that have chemical and physical interactions such as enzymes or cyclodextrins are dissolved and dispersed in the photosensitive materials. After coating, impregnating, and drying on the molecular functional film, mechanically between different materials using a photoresist method that develops and fixes by irradiating a light source with a wavelength in a region where the photosensitive material undergoes a chemical change A method for producing a functional composite film using a photoresist method, wherein a highly functional composite film having a plurality of functions is produced in a single film with enhanced bonding properties.   A functional composite membrane characterized by using a composite membrane in which different types of phases containing enzymes and the like are combined on a functional polymer functional membrane, and simultaneously generating and separating substances by expressing the functions of the enzymes and the like. A method for producing useful substances using

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