JPH048290A - Microorganism adsorbent - Google Patents

Abstract

PURPOSE:To inexpensively improve adsorption ability, by attaching a specific polymer to the surface of a plate substrate. CONSTITUTION:4-Vinylpyridine is mixed with a monovinyl monomer in a ratio of (10-90): (90-10), polymerized in >=300 degree of polymerization to give a vinyl- based polymer (A) shown by the formula (R1 is benzyl, 4-16C alkyl or pentafluorophenylmethyl; R2 is H or 1-3C alkyl; X is halogen atom; Y is H, 1-2C alkyl, benzyl, ether, carboxyl, carboxylic ester or aryl). Then the component A is dissolved in an alcohol to give a solution (B). The solution B is sprayed on a plate substrate (C) such as asbestos plate and attached to the substrate in such a way that thickness of the component A attached to the surface of the component C is 0.001-1mum and pick-up is 0.001-100wt.% to produce a microorganism adsorbent.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to a microbial adsorbent. More specifically, the present invention relates to a microbial adsorbent used as an adsorbent for microorganisms and a carrier for holding microbial cells in a bioreactor or a biosensor.

(Prior art) Conventionally, as this type of microbial adsorbent,
There is an insoluble polymer compound made of cross-linked polyvinylpyridium halide described in Japanese Patent No. 1641.

This compound is an excellent microbial adsorbent that can capture microorganisms in a living state with high efficiency and does not pollute the environment because it is insoluble in water.

However, the above-mentioned cross-linked polyvinylpyridinium halide is insoluble not only in water but also in ordinary organic solvents, so it can only be handled as a solid and is difficult to process.
There was rWIM, which had a small surface area per unit weight and poor adsorption efficiency.

(Problems to be Solved by the Invention) The present invention has been made in order to eliminate the drawbacks of the above-mentioned prior art, and is insoluble in water and can be adsorbed without reducing the activity of microorganisms. The object of the present invention is to provide a microbial adsorbent having a large surface area and high microbial adsorption efficiency.

(Means and effects for solving the problems) In order to solve the above problems, the present invention provides the following: "General formula: R2 alkyl group or pentafluorophenylmethyl group, R2 is a hydrogen atom or a C5-C3 alkyl group, X is Halogen atom, Y
is a hydrogen atom, a vinyl copolymer represented by a CI-C3 alkyl group, benzyl group, ether group, carboxyl group, carboxylic acid ester group, or aryl group) is attached to the surface of the plate-like base material. The gist of the project was ``a microbial adsorbent featuring the following characteristics.''

That is, although the vinyl copolymer used in the microbial adsorbent of the present invention has the same excellent microbial adsorption ability as cross-linked polyvinylpyridinium pallide, it is insoluble or sparingly soluble in water. Since it is soluble in organic solvents and can be made into a solution, processing such as impregnation or coating on other substrates, which was not possible with cross-linked polyvinylpyridinium halide, is possible.

The microorganism adsorbent of the present invention utilizes the properties of this copolymer to attach vinyl copolymer units having excellent microbial adsorption properties to the surface of a plate-like base material. It greatly increases the surface area per weight and enhances the ability to adsorb microorganisms.

In the present invention, a vinyl copolymer represented by the following general formula is used, which is obtained by copolymerizing 4-vinylpyridine and a monovinyl monomer and then reacting with a halide.

General formula "R" However, in the formula, R1 is a benzyl group, a C4-CI8 alkyl group or a pentafluorophenylmethyl group, R2 is a hydrogen atom or a 01-C3 alkyl group, X is a halogen atom, Y is a hydrogen atom, 01 ~C3 alkyl group, benzyl group, ether group, carboxyl group, carboxylic acid ester group, or aryl group.The vinyl copolymer is a random copolymer or a block copolymer.

Monovinyl monomers used for copolymerization include monoolefins such as ethylene, propylene, and butene, styrene, vinyl acetate, acrylic acid, acrylic esters, methacrylic acid, methacrylic esters, aliphatic vinyl esters, acrylonitrile, and derivatives thereof. However, the present invention is not limited to this, and various types can be used alone or in combination.

However, if a monovinyl monomer having a highly hydrophilic functional group is used, the resulting copolymer may become water-soluble depending on the degree of polymerization, which is not desirable.

The ratio of this 4-vinylpyridine to monovinyl monomer, that is, the ratio n:m, varies depending on the type of monovinyl monomer used and the degree of polymerization, but is approximately 10
:90 to 90:10 is desirable. If the proportion of 4-vinylpyridine is less than this range, sufficient microbial adsorption ability will not be obtained, and if it is greater than this, the resulting copolymer will be highly water-soluble. A particularly desirable ratio of n:m is in the range of 10:90 to TO:30.

Further, the degree of polymerization of the vinyl copolymer is at least 30
It is desirable that the degree of polymerization is 0 or more, and if the degree of polymerization is lower than this, the resulting copolymer will have high water solubility.

A copolymer of a monovinyl monomer and 4-vinylpyridine is produced by quaternizing pyridine by reacting with a halide such as an alkyl halide, a benzyl halide, or a pentafluorophenylmethyl halide, and is expressed by the following formula. Forms a functional group.

Formula: Since it is positively charged and the cell surface of microorganisms is generally negatively charged, it is presumed that electrostatic interaction is one important factor.

The vinyl copolymer thus obtained is substantially insoluble or sparingly soluble in water, but soluble in organic solvents. Taking advantage of this property, the vinyl polymer is dissolved in an organic solvent to form a solution. This organic solvent is
Alcohols, esters, phenols, ethers, etc. can be used, but from the viewpoint of ease of handling, it is preferable to use alcohols.

Next, a solution in which the vinyl copolymer is dissolved is applied to the plate-shaped substrate by means such as impregnation or spray coating.

Thereafter, the vinyl copolymer is adhered to the plate-like base material through a drying process.

The amount of the vinyl copolymer adhered to the plate-like base material is not particularly limited, but the adhesive thickness of the vinyl copolymer is 0.001
Since it is desirable that the thickness is in the range of -1 μm, it is preferable that the adhesion range is approximately 0.001 to 100% by weight on the base material. If it is less than 0.001% by weight, a sufficient adsorption effect for microorganisms will not be obtained, and if it is more than 100% by weight, the adsorption effect will not increase even though the amount of vinyl polymer used increases, and it will become uneconomical.

Various materials such as an asbestos board, a wooden board, a plastic board, a concrete board, a wire mesh, a corrugated board, a perforated board, etc. can be used as the plate-like base material of the present invention. Further, the size, shape, and specific gravity of this plate-like base material can be freely selected depending on the purpose and usage conditions. As a result, the surface area of the film formed on the surface of the plate-like base material by the adhesion of the vinyl copolymer can be changed as appropriate depending on the usage conditions and applications. For example, when carrying out water treatment by bringing sewage into contact with the microbial adsorbent, if the shape of the plate-like base material is a porous plate, the surface area of the film formed on the surface of the base material due to the adhesion of the vinyl copolymer will be growing. As a result, the area of contact with the treated water becomes larger, and the ability to adsorb microorganisms is dramatically improved. It is also possible to laminate a plurality of thin plate-shaped substrates, bond them together, and then form a coating of vinyl copolymer on the surface of the bonded plate-shaped substrates. . In this case, the surface area of the film due to the adhesion of the vinyl copolymer can be dramatically increased, and the microbial adsorption ability of the microbial adsorbent can also be dramatically improved.

Moreover, by selectively using materials such as metal, concrete, and plastic for the plate-like base material, the strength of the microbial adsorbent can be increased and the weight can be reduced. Note that the plate-like base material may be made of a composite of many kinds of materials, such as a wire mesh with a plastic coating on the surface.

The microorganisms to be adsorbed by the microorganism adsorbent of the present invention include bacteria, fungi, algae, viruses, etc.
When the microbial adsorbent of the present invention is used as a water treatment material or an air treatment material, the water or air is purified by adsorbing and removing these microorganisms contained in the water or air to be treated. In addition, when used in bioreactors, etc., bacteria with specific functions are adsorbed in a live or highly active state, and the enzymes contained in these bacteria are activated to produce reaction products. obtain or sort substances.

The experiments conducted by the present inventors have revealed that the vinyl copolymer of the present invention requires moisture in order to fully exhibit its characteristic microbial adsorption ability.

Therefore, when the microbial adsorbent of the present invention is used as a filter material for an air filter, it is necessary to use alkaline earth metal salts such as calcium chloride, magnesium chloride, lithium chloride, potassium metasilicate, etc. together with vinyl copolymers.
Plates of deliquescent substances such as titanium sulfate, water-soluble polymers such as polyvinyl alcohol, polyacrylates, and vinyl pyrrolidone, and moisturizing agents such as silica gel, zeolite, collagen, niphosphorus pentoxide, magnesium oxide, calcium oxide, and ethylene glycol. It is attached to the surface of the base material. As a result, the microbial adsorbent exhibits excellent microbial adsorption ability as in the case of a water-based adsorbent.

In addition, when the microbial adsorbent of the present invention is used only for the removal of microorganisms, if a bactericide is attached to the surface of the plate-like base material together with the vinyl copolymer of the present invention, the microbial adsorption ability and , it is possible to obtain a microbial adsorbent that also has sterilizing performance. Examples of such disinfectants include antibiotic polymyxins, positive surfactants such as quaternary ammonium salts, amphoteric surfactants such as alkylaminoethylglycine, biguanides such as chlorhexidine and polyhexamethylene biguanidine, and undecylenic acid. Higher fatty acids, metals and metal ions, phenols, etc. can be used.

(Example) Hereinafter, the present invention will be explained in detail according to examples.

Example 1 After copolymerizing 4-vinylpyridine and styrene at a ratio of l:2.5 mol, a vinylpyridinium-based copolymer was obtained by quaternizing with benzyl halide in an equimolar amount to 4-vinylpyridine. Ta.

Next, 10cwxlOOc* of 325 mesh wire mesh was immersed in the ethanol solution of the vinylpyridinium copolymer and dried at 80°C to obtain a microbial adsorbent with a copolymerization rate of 0.15% on the wire mesh. I got it.

Put the spirally wound microbial adsorbent into a beaker, and add E. coli to the beaker with 0.85% sterile saline at a concentration of 1.7 x 10'll/-1! ! 1000w+I of the cloudy liquid was added and stirred with a magnetic stirrer. The number of viable bacteria remaining in the solution was measured at regular intervals using the agar plate pour method, and the eradication rate was determined and shown in Table 1.

Comparative Example 1 Example! Instead of using the vinylpyridinium copolymer used in step 1, place only a spirally wound 325 mesh wire mesh 10cm x 100cs+ into a beaker, and add 0.85% of E. H bacteria [1.0% of saline solution] to this beaker. 7xlO
10,100 O of a suspension at a concentration of 1/1 was added and stirred with a magnetic stirrer. The number of viable bacteria remaining in the solution was measured at regular intervals using the agar plate pour method, and the eradication rate was determined and shown in Table 1.

(The following is a blank space) The adsorbent in Table 1 exhibited excellent microbial adsorption ability and a high sterilization rate compared to Comparative Example 1, in which no vinylpyridinium copolymer was attached.

Example 2 The spirally wound microbial adsorbent of Example 1 was placed in a beaker, and 0.8% of Staphylococcus aureus was added to the beaker.
A solution of 10100 (1) suspended in 5% sterile saline at a concentration of 1.2xlo' cells/1 was added and stirred with a magnetic stirrer. It was measured using the dilution method, and the eradication rate was determined and shown in Table 2.

(Margin below) As is clear from Table 1, Microorganisms of Example 1 Table 2 The microbial adsorbent of the present invention also showed good adsorption of bacteria.

As is clear from Table 2, Example 4 4-vinylpyridine and styrene were copolymerized at a ratio of 1:2.5 mol using different types of bacteria, and then 4-vinylpyridine and styrene were copolymerized in an equimolar amount. A vinylpyridinium copolymer was obtained by quaternization treatment with a halide.

On the other hand, 10 pieces of acrylic disks with a roughened surface and a diameter of 12 cm+ were placed so that the acrylic disks were parallel to each other and there was a gap of 5.
A multilayer rotating body was produced by fixing the rotating body to a shaft passing through the center so that the rotational speed was 100 ms.

Next, this multilayer rotating body was immersed in an ethanol solution of the vinylpyridinium copolymer, and then dried at 80°C, and the adhesion rate of the copolymer to the acrylic plate was 1.5!
% microbial adsorbent was obtained.

The microorganism adsorbent using the above multilayer rotating body was used to absorb Escherichia coli at 0.
The E. coli suspension was immersed in 1200 ml of a suspension of 85% sterile saline at a concentration of 1.6 x 1O'/1, and the E. coli suspension was stirred by rotating around the shaft. The number of viable bacteria remaining in the solution was measured at regular intervals using the agar plate pour method, and the eradication rate was determined and shown in Table 3.

Comparative Example 2 The vinylpyridinium copolymer used in Example 4 was not used, but only the multilayer rotating body used in Example 4 was used, and E. coli was incubated at 0.
The E. coli suspension was immersed in 1,200 ml of a suspension of 1.6 x 10 6 cells/ml in sterile saline solution, and the E. coli suspension was stirred by rotating around the shaft. The number of remaining viable bacteria was measured at regular intervals using the agar plate pouring method, and the eradication rate was determined and shown in Table 3.

(Leaving space below) Table 3 (Effects of the Invention) As mentioned above, the microbial adsorbent of the present invention is a copolymer that is insoluble in water and soluble in organic solvents and has excellent microbial adsorption ability. This is attached to a plate-like base material.

Therefore, by appropriately selecting the size and shape of the plate-shaped substrate, it is possible to increase the surface area of the film formed on the surface of the plate-shaped substrate by the adhesion of the vinyl copolymer, and the microbial adsorbent can dramatically increase the microbial adsorption capacity of Furthermore, it is easy to manufacture, and costs can be reduced accordingly. Furthermore, since the vinyl copolymer used in the present invention is insoluble in water, it will not dissolve in water and pollute the environment. Furthermore, the strength of the microbial adsorbent of the present invention can be increased and the weight of the microbial adsorbent can be reduced by selectively using the material of the plate-like base material.

Furthermore, the microbial adsorbent of the present invention can be easily adapted to the intended use and usage conditions, since the shape, size, and specific gravity of the thick microbial adsorbent of the plate-like base material can be freely designed.

Furthermore, when the microorganism adsorbent of the present invention is used as a biosensor or bioreactor, microorganisms can be immobilized in a living and highly active state with high efficiency, so that the excellent biocatalytic function can be fully utilized. be able to.

Claims (1)

[Claims] There are general formulas, ▲mathematical formulas, chemical formulas, tables, etc.▼ (However, in the formula, R_1 is a benzyl group, C_4 to C_1_
6 alkyl group or pentafluorophenylmethyl group, R_2 is a hydrogen atom or C_1 to C_3 alkyl group, X is a halogen atom, Y is a hydrogen atom, C_1 to C_3 alkyl group, benzyl group, ether group, carboxyl group, carbon A microbial adsorbent characterized in that a vinyl copolymer represented by an acid ester group or an aryl group is attached to the surface of a plate-like base material.