JPH0430272B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to a carrier suitable for immobilizing cells or microorganisms and a method for producing the same, and more specifically, it relates to a carrier suitable for immobilizing cells or microorganisms, and more specifically, to a carrier suitable for immobilizing cells or microorganisms, and more specifically, to a carrier suitable for immobilizing cells or microorganisms, and more specifically, to The present invention relates to a water-insoluble carrier used for immobilization and a method for producing the same. (Prior art) Conventionally, as a method for immobilizing cells or microorganisms, there is a method of physically adsorbing cells or microorganisms onto a water-insoluble carrier (hereinafter simply referred to as a carrier). Inorganic supports such as ceramics, metal oxides, or activated carbon have been used. (Problem to be solved by the invention) However, in the case of the above carrier,
It has been extremely difficult to control the microstructure, such as the pore diameter or regular arrangement of pores, to suit the wide range of cell and microbial colony sizes and growth forms. For this reason, it is not possible to obtain carriers with pores that are suitable for the size and growth form of cell and microorganism colonies, and carriers with micropores that are not suitable for the size and growth form of cell and microorganism colonies are used. , cells and microorganisms were more likely to fall off from the carrier. Furthermore, it has not been possible to control the higher-order structure of this carrier, including its shaping or processing. For example, it has been extremely difficult to glue or cut the pair into a predetermined shape, such as a three-dimensional structure, a disk shape, or a plate shape, to match the type of bioreactor. Furthermore, when these carriers are mixed into biochemical reaction products, especially when the products are foods or medicines, they are extremely harmful to the human body, with the exception of activated carbon. The object of this invention is to provide carriers for cells and microorganisms that can control the fine structure of the pore size and regular arrangement of pores, have pores that are suitable for the size and growth form of colonies of cells and microorganisms, and It is an object of the present invention to provide a carrier that can be prevented from falling off, has high strength, and can be easily bonded or cut into a predetermined shape. (Means and effects for solving the problem) In view of the above-mentioned problems, the creator has conducted extensive research and has found a new carrier and its manufacturing method that completely solves the above-mentioned problems. Successful. That is, this first invention consists of carbonized plant canal tissue that retains its form and is reinforced with carbonized organic matter impregnated into the walls forming the plant canal tissue. A carrier suitable for immobilizing cells or microorganisms. In addition, the second invention impregnates the walls forming the canal tissue of plants with organic matter,
This organic matter is treated to be infusible, and then heated to carbonize the plant canal tissue and the organic matter, thereby retaining and reinforcing the form of the plant canal tissue or microorganisms. This is a method for producing a carrier suitable for immobilization. The present invention is characterized in that the form of the plant's canal tissue is maintained as it is and is used as a carrier. The plant tube tissue constituting this carrier and the organic matter impregnated into the wall forming the tube tissue will be explained below. First, plant canal tissues, i.e., tissues with pores, preferably open pores, such as tracheids, ducts, or fiber cells, have pore diameters, distribution, arrangement, etc. that differ from cells after carbonization by heating. Select one suitable for immobilizing microorganisms. For example, in woody plants such as angiosperms, gymnosperms, and bamboos, the main pores are tracheids, and in conifers such as pine, cedar, and cypress, the main pores are tracheids. , occupies more than 90% of all tissues, and its size ranges from 1 to 70μ, depending on the type of plant. On the other hand, in broad-leaved trees, ducts and fiber cells are the main pores, and their sizes range from 20 to 500μ for ducts and 10 to 50μ for fiber cells, and vary depending on the type of plant. Therefore, the pore size of the carrier obtained by the present invention is within the range of 1 to 500μ, and within this range, it depends on the type of cells and microorganisms to be immobilized, and the size of the pore size is determined depending on the type of cells and microorganisms to be immobilized. When forming a colony, the optimal type of plant canal tissue can be selected in consideration of the size of the colony. The organic substances that are impregnated into the walls that form the plant's tube tissue include thermosetting resins such as phenolic resins, furan resins, epoxy resins, unsaturated polyester resins, urea resins, bismaleimide triazine resins, and polydivinylbenzene resins. vinyl chloride resin,
Thermoplastic resins such as vinyl fluoride resin, vinylidene chloride resin, acrylonitrile resin, petroleum, etc.
One or more selected from coal-based heavy oil, asphalt, coal tar, pitts, sugars, and their solvent-soluble components, with a carbonization rate of 3
% or more, preferably 10% or more. These organic substances are impregnated into the walls forming the plant canal tissue. Impregnation here means that organic matter coats the surface of the wall that forms the canal tissue or penetrates into the inside of the wall that forms the pores of the canal tissue, improving the carbonization rate of the canal tissue. This means that it is possible to improve the strength and durability after carbonization while maintaining the original pore shape of the tube structure. Next, a method for manufacturing a carrier according to the present invention will be explained. First, as described above, form the optimal plant canal tissue that is selected depending on the type of cells and microorganisms to be immobilized, and if they form a colony, the size of the colony. Thermosetting resins such as phenolic resins, furan resins, epoxy resins, unsaturated polyester resins, urea resins, bismaleimide triazine resins, polydivinylbenzene resins, vinyl chloride resins, vinyl fluoride resins, vinylidene chloride resins, etc. , thermoplastic resins such as acrylonitrile resin, petroleum/coal-based heavy oils, asphalt, coal tar, pitch,
One or more selected from sugars and their solvent-soluble components, with a carbonization rate of 3% or more,
Preferably 10% or more of the substance is impregnated. At this time, the impregnation rate of the organic matter is 1 for the canal tissue.
-100% by weight, and is impregnated in an amount necessary to increase the carbonization rate of the canal structure while maintaining the pore morphology of the canal structure. In addition, during impregnation, in order to increase permeability,
An appropriate solvent-soluble component may be used, or an organic substance diluted with a solvent may be used. Impregnation is preferably carried out at room temperature and reduced pressure so as not to damage the pore structure of the tube tissue, and excess organic matter adhering to the surface is removed by an appropriate method such as washing with a solvent or wiping. preferable. Next, after impregnation, an infusible treatment is performed. That is,
In order to prevent this organic material from redissolving in the solvent or softening and fluidizing, if the organic material is a thermosetting resin, it is heated to form a three-dimensional structure, and then the thermoplastic resin or petroleum - In the case of coal-based heavy oil, asphalt, coal tar, pitch, sugars, etc., add an appropriate hardening agent or oxidizing agent to make them infusible. Next, the impregnated plant tube tissue that has been rendered infusible as described above is heated and carbonized. Carbonization is performed at a heating temperature of 450 degrees or higher in a non-oxidizing atmosphere. The carbonization yield tends to increase as the heating rate decreases, but it is preferable to select the optimum heating rate depending on the type of organic substance impregnated. In this way, depending on the type of cells and microorganisms to be immobilized, and if they form a colony, the optimal plant canal tissue is selected in consideration of the size of the colony. The walls forming the tube tissue can be impregnated with organic matter, and after impregnation, the tissue can be treated to be infusible and then heated and carbonized to provide a suitable carrier for microorganisms and cells. Furthermore, this carrier has pores suitable for the size and growth form of colonies of cells and microorganisms, and can prevent cells and microorganisms from falling off from the carrier. In addition, this carrier can be easily adhered or cut into a predetermined shape to match the bioreactor in any state before impregnation, after impregnation, after infusibility, or after carbonization. For example, it can be processed into a thin plate and cells or microorganisms can be immobilized on both sides of the plate. In this case, it is necessary that the pores pass through. It can also be used in granular form as a packed fixed bed or a fluidized bed. Note that in many plants, the pores are oriented in one direction, so when bonding or cutting, it is desirable that the pores be oriented perpendicular to the immobilization surface. In addition, in the ductal tissues of plants such as broad-leaved trees, bamboo, and wisteria, liquid can be actively passed through the pores by utilizing the through pores derived from relatively large ducts, or it can be used in the form of granules and filled type. It may be used as a fixed bed or a fluidized bed. Note that since the support of the present invention is carbonaceous, oxygen-containing surface functional groups can be easily introduced through an oxidation reaction. Further, this oxygen-containing surface functional group can be substituted with another functional group, such as a methyl group, an amino group, a nitro group, etc., by an organic chemical method. Therefore, it is possible to select a functional group suitable for the cell or microorganism to be immobilized and replace it with an oxygen-containing surface functional group, thereby further improving the affinity with the cell or microorganism. In addition, since the carrier made of carbon material of the present invention has electrical conductivity, it can be used by generating heat by applying electricity or by forming an electric field, thereby further improving the affinity with cells or microorganisms. can. (Example) Next, the present invention will be described in more detail by showing an example. Example 1 Four types of broad-leaved trees, red wax and kapor, and coniferous trees, Japanese pine and Japanese cypress, were dried at 105°C for 10 hours, and commercially available resol resin (carbonization rate 33%) was mixed with methanol as an organic material. The mixture was diluted to a volume ratio of 7:3, and four types of plants were impregnated with this under reduced pressure. The impregnated product was heat treated at 170 degrees for 1 hour to make the resol resin infusible. That is, this resol resin was made into a three-dimensional network structure and cured. Each resol resin impregnation rate is lauan
7.4%, rice pine 13.5%, and rice cypress 26.0%. After the infusibility treatment, it was fired to 800 degrees Celsius in a non-oxidizing atmosphere at a heating rate of 50 degrees Celsius/hr to obtain a carrier which is a porous carbon material that retains the shape of the tube structure as it is. These exhibited regular microstructures with various pore sizes as shown in the scanning electron micrographs of FIGS. 1-8. In addition, compared to carbonized products without impregnating them with resol resin, (1) There is no loss of tissue even when ultrasonic cleaning is carried out in water. (2) Does not show any coloration even if rubbed strongly with paper, etc. It showed characteristics such as. Next, Table 1 below shows the bending strength of the carrier according to the present invention obtained in Example 1, compared with that of a non-impregnated comparative example.

[Table] Example 2 The invention product obtained from red lauan and rice cypress in Example 1 was applied to glassy carbon, stainless steel, mullite,
The immobilization state of each material, vinyl chloride and zeolite, and a mixture of methanogens, acetogens, and acidic bacteria was compared. Each sample is 30×
It was made into a plate shape of 10 x 2 mm. Collected from anaerobic digested sludge. These fungi were added to an initial concentration (as organic matter VS) of 200PPM.
In addition, a culture solution was prepared in which peptone and glycol were added as substrates at an initial concentration (in terms of organic carbon content) of 500 PPM. This culture solution was maintained at 37 degrees, and the various carriers described above were immersed in it, and the solution was passed through each carrier at a flow rate of 6.5/min. The immersion was carried out for 50 days, and the amount of bacteria immobilized on the various carriers was compared as the amount of organic carbon. The results are shown in Table 2.

[Table] In Table 2, the amount of immobilization on the 25th and 50th day is
The decrease on the 50th day is thought to be due to shedding. Furthermore, Table 2 shows that the material of the example shows a good immobilization amount compared to other materials, indicating that an optimal pore distribution exists. Separately, red lauan and rice cypress products were immersed in the culture solution for 8 days, and the immobilized state of bacterial cells was observed using a scanning electron microscope.
It is shown in FIG. Here, in FIGS. 9 and 10, it was observed that the colonies were developed and the bacterial bodies were immobilized. Moreover, in FIGS. 11 and 12, it was observed that the particles were immobilized inside the pores. (Effects of the Invention) As described in detail above, the first invention provides carbon fibers that are impregnated into the walls forming the canal tissue of plants and reinforced with carbonized organic matter, while retaining their form. It is a carrier suitable for immobilizing cells or microorganisms made of vascularized plant tissue. or,
The second invention is to impregnate the walls that form the plant's canal tissue with organic matter, treat this organic matter to make it infusible, and then heat it to carbonize the plant's canal tissue and the organic matter to change its form. This is a method for producing a carrier suitable for immobilizing cells or microorganisms, which is made of plant canal tissue and is characterized by being retained as is and reinforced. Therefore, by appropriately selecting the type of plant passage tissue to be used, it is possible to obtain a carrier having pores suitable for the size and growth form of colonies of various cells and microorganisms. In addition, since the entire carrier was made of carbon material,
It has a high affinity with cells and microorganisms, can prevent cells and microorganisms from falling off the carrier, and is harmless to the human body. On the other hand, since the carrier is made by reinforcing the tube tissue of a plant with carbon derived from an organic substance, its strength is high and the carrier itself can be prevented from falling off. Furthermore, since this material is reinforced with carbon derived from an organic substance, it is easy to bond or cut the carrier into various shapes.

[Brief explanation of drawings]

Figures 1 and 2 are micrographs showing the morphology of carbonized organisms that are carriers derived from red lauan obtained according to the present invention, and Figures 3 and 4 are photographs derived from Kapol obtained according to the present invention. FIGS. 5 and 6 are micrographs showing the morphology of carbonized organisms on the carrier derived from Japanese pine obtained according to the present invention, and FIGS. Figures 7 and 8 are micrographs showing the morphology of carbonized organisms of carriers derived from rice cypress obtained by this invention, and Figures 9 and 10 are micrographs of carriers derived from red lauan obtained by this invention. Figures 11 and 12 are micrographs showing the morphology of living organisms on which microorganisms have been immobilized after being carbonized on a carrier derived from rice cypress obtained according to the present invention. This is a micrograph showing the morphology of a transformed organism.

Claims (1)

[Scope of Claims] 1. Cells or cells made of carbonized plant canal tissue while retaining their form, which are impregnated into the walls forming the plant canal tissue and reinforced with carbonized organic matter. A carrier suitable for immobilizing microorganisms. 2 The organic substance is a thermosetting resin such as phenolic resin, furan resin, epoxy resin, unsaturated polyester resin, urea resin, bismaleimide triazine resin, polydivinylbenzene resin, vinyl chloride resin, vinyl fluoride resin, vinylidene chloride, etc. One or more types selected from resins, thermoplastic resins such as acrylonitrile resins, petroleum/coal heavy oils, asphalt, coal tar, pitches, sugars, and their solvent-soluble components,
2. A carrier suitable for immobilizing cells or microorganisms according to claim 1, which is a substance having a carbonization rate of 3% or more. 3. The carrier suitable for immobilizing cells or microorganisms according to claim 1, wherein the content of the organic matter is in the range of 1 to 100% based on the vascular tissue of the plant. 4. Impregnating the walls that form the plant's canal tissue with organic matter, treating the organic matter to make it infusible, and then heating it to carbonize the plant's canal tissue and the organic matter;
A method for producing a carrier suitable for immobilizing cells or microorganisms from plant tube tissue, which is characterized by retaining its shape and reinforcing it. 5 Claim 4, characterized in that the heating temperature is 450°C or higher and the heating is in a non-oxidizing atmosphere.
A method for producing a carrier suitable for immobilizing cells or microorganisms as described in 2.