JPWO2012029433A1 - Treatment material and treatment method - Google Patents

Abstract

Provided are a treatment material and a treatment method for effectively treating organic compounds, viruses, microorganisms and the like. The treatment material according to the present invention is a treatment material for decomposing an organic compound that can be hydrolyzed or a treatment material for inactivating one or more selected from the group consisting of viruses, microorganisms, proteins, and peptides. And having a single crystal fiber of cellulose.

Description

  The present invention relates to a treatment material and a treatment method, and more particularly, to treatment of organic compounds, viruses, microorganisms and the like using single crystal fibers of cellulose.

  Traditionally, cellulose has been used in a variety of applications. For example, Patent Document 1 describes that a hollow fiber made of cellulose acetate is used as a filtration membrane provided in a water treatment device.

JP 2006-167604 A

  However, conventionally, cellulose-based materials such as cellulose acetate that have been used for filtration membranes are obtained by chemically treating cellulose derived from nature, and are materials that themselves have a water treatment function. There wasn't.

  The present invention has been made in view of the above problems, and an object thereof is to provide a treatment material and a treatment method for effectively treating organic compounds, viruses, microorganisms and the like.

  The processing material which concerns on one Embodiment of this invention for solving the said subject is a processing material for decomposing | disassembling the organic compound which can be hydrolyzed, Comprising: It has the single crystal fiber of a cellulose, It is characterized by the above-mentioned. ADVANTAGE OF THE INVENTION According to this invention, the processing material which decomposes | disassembles an organic compound effectively can be provided.

  Moreover, the said organic compound is good also as being 1 or more types selected from the group which consists of an ester compound, an amide compound, and a urethane compound. The organic compound may be one or more selected from the group consisting of peptides, proteins, lipids, carbohydrates, DNA, and RNA. The organic compound may be an organic compound present on the surface of a virus or a microorganism.

  A treatment material according to an embodiment of the present invention for solving the above problems is a treatment material for inactivating one or more selected from the group consisting of viruses, microorganisms, proteins and peptides, It has a single crystal fiber. ADVANTAGE OF THE INVENTION According to this invention, the processing material which inactivates 1 or more types selected from the group which consists of a virus, microorganisms, protein, and a peptide effectively can be provided.

  Moreover, the said processing material is good also as being a molded object which has the said single crystal fiber of the cellulose on the surface. The treatment material may be an aqueous solution containing the single crystal fiber of cellulose.

  The processing method according to an embodiment of the present invention for solving the above-described problem decomposes the organic compound by bringing a processing target that is an organic compound that can be hydrolyzed into contact with a single crystal fiber of cellulose. It is characterized by that. ADVANTAGE OF THE INVENTION According to this invention, the processing method which decomposes | disassembles an organic compound effectively can be provided.

  Moreover, the said organic compound is good also as being 1 or more types selected from the group which consists of an ester compound, an amide compound, and a urethane compound. In the treatment method, the organic compound may be one or more selected from the group consisting of peptides, proteins, lipids, carbohydrates, DNA, and RNA. In the treatment method, the organic compound may be an organic compound present on the surface of a virus or a microorganism.

  The processing method according to an embodiment of the present invention for solving the above-described problem includes: a treatment target selected from the group consisting of a virus, a microorganism, a protein, and a peptide; and a single crystal fiber of cellulose. The treatment target is inactivated by contact with the substrate. ADVANTAGE OF THE INVENTION According to this invention, the processing method which inactivates 1 or more types selected from the group which consists of a virus, microorganisms, protein, and a peptide effectively can be provided.

  Moreover, in the said processing method, as the said process target and the molded object which has the said cellulose single crystal fiber on the surface are made to contact, the said process target and the said cellulose single crystal fiber are made to contact. Also good. In the treatment method, the treatment target and the cellulose single crystal fiber may be brought into contact with each other by mixing the treatment target and an aqueous solution containing the cellulose single crystal fiber.

  ADVANTAGE OF THE INVENTION According to this invention, the processing material and processing method which process effectively processing objects, such as an organic compound, a virus, and a microorganism, can be provided.

It is explanatory drawing which shows an example of the result of having evaluated decomposition | disassembly of a carboxylic acid ester compound in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated decomposition | disassembly of the phosphate ester compound in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated decomposition | disassembly of an amide compound in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows the other example of the result of having evaluated decomposition | disassembly of a carboxylic acid ester compound in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated the inactivation of protein in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated degradation of protein in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated inactivation of the virus in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated degradation of the protein of the virus in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows the other example of the result of having evaluated decomposition | disassembly of the protein of a virus in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows the further another example of the result of having evaluated degradation of the virus protein in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated the inactivation of microorganisms in the Example which concerns on one Embodiment of this invention. It is explanatory drawing which shows the other example of the result of having evaluated inactivation of microorganisms in the Example which concerns on one Embodiment of this invention.

  Hereinafter, an embodiment of the present invention will be described. Note that the present invention is not limited to this embodiment.

  As a result of intensive studies, the inventors of the present invention can efficiently decompose an organic compound by bringing an aqueous solution containing an organic compound that can be hydrolyzed into contact with a single crystal fiber of cellulose. In addition, the present inventors have uniquely found that a treatment target such as a virus, a microorganism, a protein, and other organic compounds and a single crystal fiber of cellulose are brought into contact with each other to efficiently inactivate the treatment target.

  The present invention has been made based on such uniquely obtained knowledge. That is, the present invention uses cellulose single crystal fibers as a functional material that effectively promotes the degradation of organic compounds and / or effectively inactivates treatment targets such as viruses, microorganisms, proteins, and peptides. To do.

  The mechanism by which the single crystal fiber of cellulose according to the present invention develops the activity (hydrolysis activity) for promoting the hydrolysis of organic compounds and / or the mechanism to inactivate treatment targets such as viruses, microorganisms, proteins, and peptides are still not available Although not fully elucidated, for example, in the crystal fiber of the cellulose, it is considered that the crystal plane showing the hydrolysis activity and / or the inactivation function is exposed. The crystal plane showing the hydrolytic activity and / or inactivation function is exposed by performing a process of dispersing individual single crystal fibers in the process for producing single crystal fibers of cellulose according to the present invention.

  Conventionally, a filtration membrane such as a hollow fiber manufactured from a cellulose-based material such as cellulose acetate has been used for the purpose of water treatment or the like. However, such a conventional cellulosic material is obtained by chemically processing naturally-derived cellulose to improve its processability, and its crystallinity is remarkably reduced. The material did not contain single crystal fiber. Conventionally, filter paper has been produced from pulp containing cellulose fibers having a certain degree of crystallinity, but in the production process, the cellulose fibers have not been sufficiently loosened. For this reason, the cellulose fiber which comprises the conventional filter paper was not what the crystal plane which shows a hydrolysis activity and / or an inactivation function exposed.

  In the treatment method according to the present embodiment (hereinafter referred to as “the present method”), for example, a treatment target that is an organic compound that can be hydrolyzed and a single crystal fiber of cellulose are brought into contact (for example, the organic compound is treated with The organic compound is decomposed by bringing the aqueous solution contained therein into contact with the single crystal fiber of the cellulose). That is, in this case, the present method is a method for decomposing an organic compound (for example, an organic compound contained in an aqueous solution) using cellulose single crystal fibers.

  In addition, in this method, for example, one or more treatment targets selected from the group consisting of viruses, microorganisms, proteins and peptides are brought into contact with cellulose single crystal fibers (for example, the viruses, microorganisms, proteins And an aqueous solution containing at least one selected from the group consisting of peptides and a single crystal fiber of the cellulose are brought into contact) to inactivate the treatment target. That is, in this case, the method uses one or more types selected from the group consisting of viruses, microorganisms, proteins and peptides (for example, viruses, microorganisms, proteins and 1 or more selected from the group consisting of peptides).

  The single crystal fiber of cellulose is, for example, selected from the group consisting of a single crystal fiber and / or a virus, a microorganism, a protein, and a peptide that decomposes the organic compound by bringing it into contact with a treatment target that is an organic compound that can be hydrolyzed. If it is the single crystal fiber which inactivates the said process target by making it contact with the process target which is 1 or more types, it will not be restricted in particular.

  That is, the cellulose single crystal fiber is selected from the group consisting of, for example, cellulose I type single crystal fiber, cellulose II type single crystal fiber, cellulose III type single crystal fiber, and cellulose IV type single crystal fiber. More than a seed.

The cellulose type I is, for example, cellulose I _α type or cellulose I _β type. Cellulose type III is, for example, cellulose III type _I or cellulose III type _II . The cellulose type IV is, for example, cellulose IV type _I or cellulose IV type _II .

The cellulose may be, for example, cellulose type I or crystalline cellulose derived from the cellulose type I. In this case, the cellulose is at least one selected from the group consisting of cellulose I _α- type, cellulose I _β- type, cellulose III _I- type and cellulose IV _I- type, preferably cellulose I _α- type, cellulose I _β- type and One or more types selected from the group consisting of cellulose III type _I.

The cellulose single crystal fibers may be, for example, naturally-derived cellulose single crystal fibers (that is, cellulose single crystal fibers obtained from natural raw materials). In this case, the naturally-derived cellulose is, for example, one or more selected from the group consisting of naturally-occurring cellulose I _α type, cellulose I _β type, cellulose III _I type, and cellulose IV _I type, preferably naturally derived. 1 type or more selected from the group consisting of cellulose I _α type, cellulose I _β type and cellulose III _I type.

  Note that naturally-derived cellulose single crystal fibers may contain trace components (for example, metals) other than cellulose derived from the raw materials and / or purification treatment. Therefore, it is speculated that this trace component may contribute to the decomposition of the organic compound by the single crystal fiber of cellulose.

  The crystallinity of the single crystal fiber of cellulose is not particularly limited as long as the single crystal fiber is formed, and is, for example, 60% or more, and preferably 80% or more. The diameter of the single crystal fiber of cellulose is not particularly limited, and is, for example, 3 to 100 nm, preferably 10 to 30 nm. The length of the single crystal fiber of cellulose is not particularly limited, and is, for example, 1 to 100 μm, preferably 10 to 30 μm.

  In this method, the organic compound to be decomposed is not particularly limited as long as it is an organic compound that can be hydrolyzed. That is, the organic compound is at least one selected from the group consisting of an ester compound, an amide compound, and a urethane compound, for example.

  The ester compound is not particularly limited as long as it is an organic compound having an ester bond. That is, the ester compound includes, for example, a carboxylic acid ester compound (a compound having a chemical structure in which a carboxylic acid and an alcohol are dehydrated and condensed), a phosphoric acid ester compound (a compound having a chemical structure in which a phosphoric acid and an alcohol are dehydrated and condensed), Thioester compounds (compounds having a chemical structure in which carboxylic acid and thiol are dehydrated and condensed), borate ester compounds (compounds having a chemical structure in which boric acid and alcohol are dehydrated and condensed), sulfate compounds (sulfuric acid and alcohol are dehydrated) Compound having a condensed chemical structure), nitrate ester compound (compound having a chemical structure in which nitric acid and alcohol are dehydrated and condensed), carbonate compound (compound having a chemical structure in which carbonic acid and alcohol are dehydrated and condensed), and sulfonate ester Compound (sulfonic acid and alcohol are dehydrated and condensed It was a compound having the chemical structure) one or more selected from the group consisting of.

  Carboxylic acid ester compounds include, for example, formic acid ester compounds, acetic acid ester compounds, propionic acid ester compounds, butyric acid ester compounds, valeric acid ester compounds, caproic acid ester compounds, enanthic acid ester compounds, caprylic acid ester compounds, pelargonic acid ester compounds, Capric acid ester compound, Lauric acid ester compound, Myristic acid ester compound, Palmitic acid ester compound, Margaric acid ester compound, Stearic acid ester compound, Oleic acid ester compound, Linoleic acid ester compound, Linolenic acid ester compound, Arachidonic acid ester compound, One or more selected from the group consisting of docosahexaenoic acid ester compounds and eicosapentaenoic acid ester compounds.

  The phosphate ester compound is, for example, one or more selected from the group consisting of a phosphate monoester compound, a phosphate diester compound, and a phosphate triester compound. More specifically, the phosphate ester compound is, for example, one or more selected from the group consisting of phosphorylated peptides, phosphorylated proteins, DNA, RNA, and phospholipids.

  The amide compound is not particularly limited as long as it is an organic compound having an amide bond. The amide bond may be, for example, a peptide bond. That is, the amide compound is at least one selected from the group consisting of peptides, proteins, lipids and carbohydrates, for example.

  The urethane compound is not particularly limited as long as it is an organic compound having a urethane bond. The urethane compound is, for example, at least one selected from the group consisting of carbamic acid, ethyl carbamate, N-methylcarbamic acid-2- (1-methylpropyl) phenyl, and polyurethane.

  The molecular weight of the organic compound is not particularly limited as long as the organic compound is decomposed by contact with the single crystal fiber of cellulose, and is, for example, 100 to 100,000 Da, or 200 to 2000 Da. Further, the organic compound is not limited to those present alone in the aqueous solution. That is, the organic compound may be an organic compound present on the surface of a virus or a microorganism, for example.

  Moreover, the processing target inactivated in this method will not be restricted especially if it is 1 or more types selected from the group which consists of a virus, microorganisms, protein, and a peptide.

  The virus to be inactivated is, for example, a DNA virus and / or an RNA virus. The virus is a virus that can infect animal cells, plant cells and / or microorganisms, for example. Moreover, a virus is a virus (for example, a virus that can be pathogenic due to infection) that may be life-threatening or impaired in health when taken into a living body (for example, a human body).

  The microorganisms to be inactivated include, for example, bacteria (for example, one or more selected from the group consisting of Escherichia coli, Staphylococcus, Pseudomonas aeruginosa, soil fungi and streptococci), algae, fungi (for example, mold, yeast, and One or more selected from the group consisting of mushrooms) and one or more selected from the group consisting of protozoa. In addition, microorganisms, for example, against microorganisms (eg, animal cells, plant cells and / or other microorganisms) that can be life-threatening or impaired in health when taken into a living body (eg, the human body). Microorganisms that can show pathogenicity (pathogenic microorganisms).

  The protein to be inactivated is, for example, a protein having an activity that can act on another substance. That is, protein is 1 or more types selected from the group which consists of an enzyme, a toxin, and an antigen, for example. Proteins can also be life-threatening or health-impaired proteins (eg, verotoxin, cholera toxin, botulinum toxin, tetanus toxin, diphtheria toxin, Staphylococcal enterotoxin, Clostridium perfringens alpha toxin, staphylococcal alpha toxin, Shiga Shigella toxin, Vibrio parahaemolyticus, pertussis toxin, dendrotoxin, and phospholipase A2). The molecular weight of the protein is not particularly limited, and is, for example, 10 kDa or more and 1000 kDa or less.

  The peptide to be inactivated is, for example, a peptide having an activity that can act on another substance. That is, the peptide is, for example, one or more selected from the group consisting of teitoustoxin, melittin, apamin, MCD-peptide, cyanotoxin, amanitin, conotoxin, ochratoxin A, α-bungarotoxin, erabutoxin, and cobrotoxin It is. Moreover, a peptide is a peptide (for example, toxin peptide) which, when taken in a living body (for example, human body), may threaten its life or impair health. The molecular weight of the peptide is not particularly limited, and is, for example, 0.2 kDa or more and less than 10 kDa.

  The aqueous solution to be treated in the present method (hereinafter referred to as “treatment solution”) contains an organic compound that can be hydrolyzed and / or one or more selected from the group consisting of viruses, microorganisms, proteins, and peptides. If it is aqueous solution to be used, it will not be restricted in particular. That is, the solution to be treated is, for example, clean water (for example, tap water), well water, rain water, river water, sea water, lake water, pond water, sewage, or waste water.

  Further, the solution to be treated may be an aqueous solution already purified by a method other than the present method. That is, the solution to be treated is, for example, an aqueous solution (for example, so-called purified water) purified in advance by a predetermined method, and an organic compound to be decomposed and / or a virus to be inactivated despite the purification. It may be an aqueous solution in which one or more selected from the group consisting of microorganisms, proteins and peptides remain.

  The solvent contained in the treatment target solution may be only water, but is not limited thereto, and the organic compound is decomposed in the treatment target solution and / or a group consisting of viruses, microorganisms, proteins and peptides. Water and other solvents (for example, organic solvents such as alcohol) may be contained as long as at least one selected from the ranges is inactivated. Further, the solution to be treated may be, for example, an aqueous solution containing only the organic compound to be decomposed as an organic compound that can be hydrolyzed. Moreover, the solution to be treated may be, for example, an aqueous solution containing only one or more selected from the group consisting of viruses, microorganisms, proteins and peptides to be inactivated.

  In addition, the gas to be treated in the present method (hereinafter referred to as “treatment gas”) is at least one selected from the group consisting of an organic compound that can be hydrolyzed and / or a virus, a microorganism, a protein, and a peptide. If it is a gas containing, it will not be restricted especially. That is, the processing target gas is, for example, air containing the above-described processing target.

  In this method, the mode in which the single crystal fiber of cellulose and the treatment target (for example, one or more selected from the group consisting of an organic compound to be hydrolyzed and / or a virus, a microorganism, a protein, and a peptide) are contacted, Treatment of the treatment target in the solution to be treated and / or the gas to be treated by contact with the single crystal fiber of the cellulose (for example, selected from the group consisting of decomposition of the organic compound and / or virus, microorganism, protein and peptide) It is not particularly limited as long as one or more inactivations are promoted. That is, in this method, for example, the processing material according to the present embodiment (hereinafter referred to as “the present processing material”) is used.

  This treatment material is, for example, a treatment material (organic compound decomposition material) for decomposing an organic compound that can be hydrolyzed, and has single crystal fibers of cellulose. Moreover, this processing material is a processing material (inactivation material) for inactivating 1 or more types selected from the group which consists of a virus, microorganisms, protein, and a peptide, for example, Comprising: It has the single crystal fiber of a cellulose .

  That is, this treatment material has a single crystal fiber of cellulose as a component having a function of decomposing an organic compound and / or a function of inactivating one or more selected from the group consisting of viruses, microorganisms, proteins and peptides. Therefore, the present treatment material is not particularly limited as long as it has single crystal fibers of cellulose so as to be in contact with the treatment object in the treatment object solution and / or the treatment object gas.

  The treatment material is, for example, a molded body having cellulose single crystal fibers on the surface thereof. That is, the present treatment material is a structure having a predetermined shape having single crystal fibers of cellulose on the surface that can be in contact with the treatment object in the treatment object solution and / or the treatment object gas.

  The treatment material is, for example, a filtering material. The filter medium is, for example, a filter. The filter is, for example, a molded body in which a large number of holes through which the solution to be processed and / or the gas to be processed pass are formed. Specifically, the filter is, for example, a membrane filter or a hollow fiber. The filter may be a mask, for example. Moreover, a filter medium is a granular material, for example. In this case, the filter medium is, for example, a large number of granular bodies assembled so that the solution to be processed passes through the gap.

  The treatment material, which is a molded body, is composed of a cellulose single crystal fiber, or a mixed material containing a cellulose single crystal fiber and another material. That is, the treatment material is, for example, a mixed material containing cellulose single crystal fiber and a material suitable for forming a molded body (for example, a polymer material such as pulp, regenerated cellulose, cellulose derivative, vinyl polymer). And the mixed material is manufactured into a predetermined shape (for example, a film shape, a hollow fiber shape, or a granular shape). In this case, at least a part of the cellulose single crystal fibers contained in the mixed material is exposed on the surface of the treatment material, and the treatment target treatment (for example, decomposition of organic compounds and / or viruses, microorganisms, proteins and peptides). One or more inactivation selected from the group consisting of:

  The treatment material is produced, for example, by molding a material suitable for forming a molded body into a predetermined shape and chemically bonding cellulose single crystal fibers to the surface of the obtained molded body. In this case, at least a part of the surface of the treatment material is coated with single crystal fibers of cellulose, and the single crystal fibers of cellulose are treated (for example, decomposition of organic compounds and / or viruses, microorganisms, proteins and peptides). One or more inactivation selected from the group consisting of:

  And when this processing material is a molded object, in this method, the processing target solution and / or processing target gas and the processing target gas are brought into contact with the processing target solution and / or processing target gas. The processing target contained is processed. That is, for example, when the treatment material is a filtering material, the treatment object solution and / or the treatment object gas is filtered with the treatment material. At this time, the single crystal fiber of cellulose present on the surface of the treatment material and the treatment target in the treatment target solution and / or the treatment target gas filtered by the treatment material come into contact with each other. (For example, decomposition of organic compounds and / or inactivation of one or more selected from the group consisting of viruses, microorganisms, proteins and peptides) proceed efficiently.

  The conditions for filtering the solution to be treated and / or the gas to be treated with the treatment material may be appropriately adjusted so that the treatment subject in the solution to be treated is efficiently processed. That is, for example, filtration of the solution to be treated and / or the gas to be treated with the treatment material may be repeated. In addition, for example, the treatment target solution that is filtered with the treatment material so that the single crystal fiber of cellulose included in the treatment material and the treatment target in the treatment target solution and / or the treatment target gas are in sufficient contact with each other. Alternatively, the flow rate of the processing target gas may be adjusted.

  The treatment material is an aqueous solution containing, for example, cellulose single crystal fibers. That is, the treatment material is an aqueous solution in which single crystal fibers of cellulose are dispersed. In this case, the concentration of the single crystal fiber of cellulose contained in the treatment material is not particularly limited as long as the treatment target is promoted when the treatment material is mixed with the treatment target solution. It may be 0.001 to 5 wt%, may be 0.01 to 5 wt%, may be 0.05 to 1 wt%, and may be 0.1 to 1 wt% Good.

  And when this processing material is aqueous solution, in this method, the processing target contained in the said processing target solution is processed by mixing a processing target solution and this processing material. That is, for example, the treatment material and the solution to be treated are mixed, and the obtained mixed solution is held at a predetermined temperature for a predetermined time. At this time, in the mixed solution, the single crystal fiber of cellulose and the treatment target come into contact with each other, and the treatment target treatment (for example, decomposition of organic compound and / or virus, microorganism, protein and peptide is selected from the group consisting of One or more inactivations proceed efficiently.

  In this method, the conditions (for example, temperature and time) for bringing the treatment target into contact with the single crystal fiber of cellulose are within a range in which the treatment of the treatment target is promoted as compared with the case where the single crystal fiber of cellulose is not used. It is not particularly limited if there is.

  That is, for example, the treatment target solution and the single crystal fiber of cellulose are brought into contact with each other until the content of the treatment target in the treatment target solution is equal to or lower than a predetermined threshold value. For example, when the solution to be treated contains a virus and / or microorganism, the solution to be treated and the single crystal fiber of cellulose until the pathogenicity of the virus and / or microorganism is sufficiently reduced or eliminated. Contact. In addition, the temperature which makes a process target and the single crystal fiber of a cellulose contact is 0-100 degreeC, for example, and 10-40 degreeC may be sufficient.

  In this method, an aqueous solution (hereinafter referred to as “treatment”) in which the content of organic compounds and / or one or more activities selected from the group consisting of viruses, microorganisms, proteins and peptides are reduced by the use of single crystal fibers of cellulose. "After-solution") is recovered. That is, when using this processing material which is a molded object, the post-processing solution is separated from this processing material and collected.

  Specifically, when the treatment material is a filter material, a filtrate obtained by filtering the solution to be treated with the treatment material is recovered as a post-treatment solution. Moreover, when using this processing material which is aqueous solution, the aqueous solution from which the single crystal fiber of the cellulose was removed is collect | recovered as a solution after a process. The removal of the single crystal fiber of cellulose can be performed, for example, by filtration with a filter having a hole having a diameter that does not allow the single crystal fiber to pass through, or by centrifugation.

  Thus, according to this method and / or this processing material, the process target contained in a process target solution can be processed effectively. For example, a hydrolyzable organic compound and / or one or more treatment targets selected from the group consisting of viruses, microorganisms, proteins and peptides contained in the treatment target solution are taken into a living body (for example, a human body). Organic compounds that are present on the surface of a virus and / or pathogenic microorganism and are involved in the expression of infection or pathogenicity that may be life threatening or health-damaging , Organic compounds, viruses, and pathogenic microorganisms that are accumulative in the living body), the treatment target is decomposed and / or inactivated by the method and / or the treatment material, and the treatment target is used. An aqueous solution in which toxicity, infectivity, pathogenicity, and accumulation are sufficiently reduced or eliminated can be efficiently produced. Therefore, according to this method and / or this treatment material, for example, safe and harmless drinking water and medical water can be efficiently produced.

  Next, specific examples according to the present embodiment will be described.

[Preparation of Cellulose I _α Type and Cellulose I _β Type]
Known technical literature (Vanderhart, DL;. Atalla, RH Macromolecules 1984, 17, 1465-1472) according to, Cladophora (Cladophora) derived from cellulose I _alpha type single crystal fibers, and ascidian (Halocynyhia) derived cellulose I _beta type A single crystal fiber was prepared.

First, a sample made of shiogusa or maboya was immersed in a 1M NaOH aqueous solution at room temperature overnight or more, and then thoroughly washed with tap water. Next, the washed sample was immersed in a 0.3% NaClO ₂ solution adjusted to pH 4.5 with 0.1 M acetate buffer at 80 ° C. for 2 hours, and then sufficiently washed with tap water. The above operation was repeated twice or three times until the sample color became white.

  Further, the sample was pulverized to a sufficiently fine fragment using a homogenizer (Physcotron, manufactured by Microtech), and then centrifuged at 10,000 rpm for 5 minutes to collect the precipitate.

  The recovered precipitate was suspended in an appropriate amount (for example, 300 mL when the recovered amount after homogenization was 100 g) of 4M HCl. Next, the resulting solution was hydrolyzed by stirring (500 rpm) at 80 ° C. for 8 hours, and then the same amount of distilled water as that solution was added to stop the reaction. The solution after the reaction was centrifuged at 10,000 rpm for 5 minutes, and the supernatant was removed. Distilled water was added to the precipitate and redispersed. This operation was repeated 3 to 5 times until the supernatant was completely neutralized.

Distilled water was added to the precipitate and centrifuged at 2000 rpm for 5 minutes. After confirming that the supernatant was cloudy and the solid was floating, the supernatant was collected. The supernatant was suction filtered using a glass filter washed with sulfuric acid, and the filtrate was recovered. Thus, an aqueous solution containing cellulose I _α- type single crystal fibers derived from rhododendron (hereinafter referred to as “cellulose I _α aqueous solution”) and an aqueous solution containing cellulose I _β- type single crystal fibers derived from maboya (hereinafter referred to as “cellulose”). I _β aqueous solution ”). These aqueous solutions were stored at 4 ° C.

Incidentally, the cellulose I _beta-type concentration of the cellulose I _alpha type concentration and cellulose I _beta in the aqueous solution in the cellulose I _alpha aqueous solution, collect the aqueous solution of 1mL sample tube 5 mL, dried at 105 ° C. 24 hours, It was determined by measuring the weight of the solid content after drying.

[Preparation of Cellulose III Type _I ]
According to a known technical document (Wada, M .; Nishiyama, Y .; Langan, P. Macromolecules 2006, 39, 2947-2952.), Cellulose III type _I single crystal fiber derived from Shiogusa was prepared.

First, a sample made of Shiogusa was soaked in a 1 M NaOH aqueous solution at room temperature overnight or more, and then thoroughly washed with tap water. Next, the washed sample was immersed in a 0.3% NaClO ₂ solution adjusted to pH 4.5 with 0.1 M acetate buffer at 80 ° C. for 2 hours, and then sufficiently washed with tap water. The above operation was repeated twice or three times until the sample color became white.

  Further, the sample was pulverized to a sufficiently fine fragment using a homogenizer (Physcotron, manufactured by Microtech), and then centrifuged at 10,000 rpm for 5 minutes to collect the precipitate. The collected precipitate was frozen with liquid nitrogen and freeze-dried overnight until it became damp. Then, the precipitate was further dried at 105 ° C. for 3 hours or more.

  About 1 g of the sample thus obtained was put in a pressure vessel and cooled to -13 ° C., and ammonia gas was filled in the pressure vessel over 30 minutes. This pressure vessel is heated to 140 ° C. in an oil bath to produce a supercritical fluid (critical point: 132.4 ° C., 11.28 MPa), and the pressure in the pressure vessel is maintained at 15-18 MPa for 1 hour of reaction. I let you. After removing the ammonia, the obtained solid was set in a filtration device, washed with 300 to 500 mL of methanol until the ammonia odor disappeared, and further washed with about 200 mL of acetone. The washed solid was transferred to a beaker and left overnight until the acetone evaporated.

  The obtained solid was suspended in an appropriate amount (for example, 300 mL when the amount recovered after homogenization was 100 g) of 4M HCl. Next, the obtained solution was hydrolyzed by stirring (500 rpm) at 80 ° C. for 8 hours, and then the same amount of distilled water as that solution was added to stop the reaction. The container after the reaction was centrifuged at 10,000 rpm for 5 minutes, and the supernatant was removed. Distilled water was added to the precipitate for redispersion, and centrifugation was again performed at 10,000 rpm for 5 minutes to remove the supernatant. This operation was repeated 3 to 5 times until the supernatant was completely neutralized.

Distilled water was added to the precipitate and centrifuged at 2000 rpm for 5 minutes. It was confirmed that the supernatant was cloudy and the solid was floating, and the supernatant was collected. The supernatant was suction filtered using a glass filter washed with sulfuric acid, and the filtrate was recovered. In this way, an aqueous solution containing cellulose III type _I single crystal fibers derived from rhododendron (hereinafter referred to as “cellulose III _I aqueous solution”) was obtained. This aqueous cellulose III _I solution was stored at 4 ° C. Cellulose III type _I single crystal fibers can also be prepared by the method described in JP-A-2008-161125.

The concentration of cellulose III type _{I in the} aqueous solution of cellulose III _I was determined by collecting 1 mL of the aqueous solution in a 5 mL sample tube, drying it at 105 ° C. for 24 hours, and measuring the weight of the solid content after drying.

[Acceleration of decomposition of carboxylic acid ester compound by cellulose I _α type]
Cellulose _Iα type single crystal fiber was used as the single crystal fiber of cellulose. That is, a predetermined amount of the cellulose I _α aqueous solution prepared in Example 1 was collected in a 1.7 mL tube and centrifuged to precipitate cellulose I _α type. Then, 1 mL of 0.1 M HEPES buffer solution (pH 7.4) was added so that the concentration of cellulose I _α type was 0.5 wt%, and cellulose I _α type was dispersed in the buffer solution. Thus, a cellulose I _α aqueous solution containing cellulose I _α type at a concentration of 0.5 wt% was prepared.

  On the other hand, 4-nitrophenylacetic acid, which is an acetic ester, was used as a carboxylic acid ester compound that is a substrate to be decomposed. That is, 4-nitrophenylacetic acid was dissolved in dimethyl sulfoxide to prepare a substrate solution containing 10 mM 4-nitrophenylacetic acid.

A cellulose / substrate solution was then prepared. That is, 1 mL of cellulose I _α aqueous solution and 10 μL of substrate solution were mixed in a 1.7 mL tube, and 10 mM HEPES buffer (pH 7.4) containing 0.5 wt% cellulose I _α type and 100 μM substrate. A cellulose / substrate solution consisting of

In addition, a cellulose / substrate solution incubation was performed. That is, the cellulose / substrate solution was incubated at 30 ° C. for a predetermined time while rotating with a rotator so that cellulose I _α type in the tube was uniformly dispersed. Incubation times were 1.5 hours, 3 hours, 4.5 hours, 6 hours and 7.5 hours.

The supernatant containing the degraded substrate was then recovered from the cellulose / substrate solution. That is, after incubation, the cellulose / substrate solution was centrifuged at 15000 rpm for 1 minute at 30 ° C. using a centrifuge. 200 μL of the supernatant of the cellulose / substrate solution (hereinafter referred to as “supernatant sample”) was collected and transferred to a new 1.7 mL tube. On the other hand, a precipitate containing cellulose I _α type obtained by centrifuging the cellulose / substrate solution after incubation was collected.

  Next, 400 μL of 10 mM HEPES buffer (pH 7.4) was added to the collected supernatant sample for dilution, and the absorption intensity at 400 nm was measured using an ultraviolet-visible spectrophotometer. Each supernatant sample was measured three times independently.

For comparison, a control solution consisting of 100 mM HEPES buffer (pH 7.4) containing 100 μM substrate but not cellulose I _α type was prepared. This control solution was also incubated in the same manner as the cellulose / substrate solution described above, and the absorption intensity at 400 nm was measured with an ultraviolet-visible spectrophotometer.

  FIG. 1 shows the measurement results. In FIG. 1, the horizontal axis represents the incubation time (hour), and the vertical axis represents the absorption intensity (−) at 400 nm. In FIG. 1, black circles indicate the measurement results of the supernatant sample collected from the cellulose / substrate solution after incubation, and white circles indicate the measurement results of the control solution after incubation.

  As shown in FIG. 1, it was confirmed that the decomposition of 4-nitrophenylacetic acid progressed at a significantly higher rate in the cellulose / substrate solution than in the control solution.

Next, based on the measurement result of the supernatant sample of the cellulose / substrate solution, ln ([A] / [A _max ]) (where [A] and [A _max ] represents the concentration of the product at each reaction time and the concentration of the product after completion of the reaction, respectively) and approximated by a straight line, and the reaction rate constant k ₁ = 0.20 h ⁻¹ from the slope of the approximate line. Got. On the other hand, based on the measurement result of the control solution, in the same manner to obtain a reaction rate constant _k spont = _0.027h ^-1. That is, the reaction rate constant calculated for the decomposition reaction of 4-nitrophenylacetic acid brought into contact with cellulose I _α- type single crystal fiber is the reaction rate constant calculated for the spontaneous decomposition reaction of 4-nitrophenylacetic acid. It was more than 7 times.

Further, the cellulose I _α form recovered after the reaction was washed with a HEPES buffer, and a cellulose / substrate solution was prepared and incubated in the same manner as described above, and the absorption intensity at 400 nm was measured. As a result, a reaction rate constant almost equivalent to the above case was obtained. That is, by reusing cellulose _Iα type single crystal fiber, the carboxylic acid ester compound could be repeatedly decomposed many times.

  In a preliminary experiment, 4-nitrophenyl galactoside, which is an ether compound, was not decomposed by the single crystal fiber of cellulose. Therefore, it was speculated that the decomposition of the organic compound by the single crystal fiber of cellulose is hydrolysis and a nucleophilic reaction by the hydroxyl group (OH group) of the cellulose.

[Acceleration of decomposition of phosphate ester compound and amide compound by cellulose I _α type and cellulose I _β type]
Cellulose I _α- type single crystal fibers and cellulose I _β- type single crystal fibers were used as the single crystal fibers of cellulose. That is, in the same manner as in Example 2 above, a cellulose I _α aqueous solution containing cellulose I _α type at a concentration of 0.5 wt% and a cellulose I _β aqueous solution containing cellulose I _β type at a concentration of 0.5 wt% were obtained. Prepared.

  On the other hand, 4-nitrophenyl phosphoric acid which is a phosphoric acid monoester compound was used as a phosphoric acid ester compound which is a substrate to be decomposed. Moreover, 4-nitroacetanilide was used as an amide compound which is a substrate to be decomposed. That is, in the same manner as in Example 2 described above, a substrate solution containing 10 mM 4-nitrophenyl phosphate and a substrate solution containing 10 mM 4-nitroacetanilide were prepared.

In addition, a cellulose / substrate solution was prepared and incubated. That is, in the same manner as in Example 2 above, a cellulose / substrate solution comprising a 10 mM HEPES buffer (pH 7.4) containing 0.5 wt% cellulose I _α form and 100 μM 4-nitrophenyl phosphate. And a cellulose / substrate solution consisting of 10 mM HEPES buffer (pH 7.4) containing 0.5 wt% cellulose I _β- form and 100 μM 4-nitroacetanilide. Incubated for days.

  Then, in the same manner as in Example 2 above, 400 μL of 10 mM HEPES buffer solution (pH 7.4) was added to the supernatant sample collected from the cellulose / substrate solution after incubation and diluted, and an ultraviolet-visible spectrophotometer was used. Used to measure the absorption intensity at 400 nm. Further, a control solution was prepared in the same manner as in Example 2 above, and incubation and absorption intensity were measured.

FIG. 2 shows the absorption intensity (−) at 400 nm when 4-nitrophenyl phosphate is used as the substrate. FIG. 3 shows the absorption intensity (−) at 400 nm when 4-nitroacetanilide is used as the substrate. In FIG. 2 and FIG. 3, “cellulose I _α ” indicates the results when a cellulose / substrate solution containing cellulose I _α type is used, and “cellulose I _β ” indicates cellulose / _α containing cellulose I _β type. The result when using the substrate solution is shown, and the “control” shows the result when using the control solution.

  As shown in FIGS. 2 and 3, in the cellulose / substrate solution, the degradation of the phosphoric monoester compound and amide compound activated with the nitrophenyl group proceeded at a significantly higher rate than the control solution. It was confirmed.

  In addition, according to the reaction rate constant obtained in the same manner as in Example 2 above, the decomposition reaction rate is the largest when the substrate is a carboxylic acid ester compound, and then the largest is the case of a phosphoric acid monoester compound, The smallest was the case of the amide compound.

[Acceleration of decomposition of carboxylic acid ester compound by cellulose I _β type and cellulose III _I type]
As the single crystal fibers of cellulose, cellulose I _β type single crystal fibers and cellulose III type _I single crystal fibers were used. That is, a predetermined amount of the cellulose I _β aqueous solution or cellulose III _I aqueous solution prepared in Example 1 was collected in a 1.7 mL tube and centrifuged to precipitate cellulose I _β type or cellulose III _I type. Then, 300 μL of 0.1 M HEPES buffer solution (pH 7.4) is added so that the concentration of cellulose I _β type or cellulose III _I type is 0.5 wt%, and cellulose I _β type or cellulose III _I type is added to the buffer. Dispersed in the liquid. Thus, to prepare a cellulose III _I aqueous solution containing a cellulose III _I type conductivity in a concentration of cellulose I _beta-type solution and 0.5 wt% containing cellulose I _beta type conductivity in a concentration of 0.5 wt%.

  On the other hand, 4-nitrophenylacetic acid, which is an acetic ester, was used as a carboxylic acid ester compound that is a substrate to be decomposed. That is, 4-nitrophenylacetic acid was dissolved in dimethyl sulfoxide to prepare a substrate solution containing 10 mM 4-nitrophenylacetic acid.

A cellulose / substrate solution was then prepared. That is, 300 μL of cellulose I _β- type aqueous solution or cellulose III _I aqueous solution and 3 μL of substrate solution were mixed in a 1.7 mL tube, and 10 mM HEPES containing 0.5 wt% cellulose I _β- type and 100 μM substrate. A cellulose / substrate solution consisting of a buffer (pH 7.4) and a 10 mM HEPES buffer (pH 7.4) containing 0.5 wt% cellulose III type _I and 100 μM substrate was prepared. .

In addition, a cellulose / substrate solution incubation was performed. That is, the cellulose / substrate solution was incubated at 30 ° C. for 7.5 hours while rotating with a rotator so that cellulose I _β type and cellulose III _I type were uniformly dispersed in the tube.

  Then, in the same manner as in Example 2 described above, a supernatant sample was collected from the cellulose / substrate solution after incubation, and the absorption intensity at 400 nm was measured.

FIG. 4 shows the measurement results. In FIG. 4, “cellulose I _β ” indicates the result when a cellulose / substrate solution containing cellulose I _β type is used, and “cellulose III _I ” indicates a cellulose / substrate solution containing cellulose III _I type. The results when used are shown, and “control” shows the results when using a control solution.

  As shown in FIG. 4, it was confirmed that the decomposition of the carboxylic acid ester compound proceeded at a significantly higher rate in the cellulose / substrate solution than in the control solution.

[Inactivation of protein by cellulose I _β type]
Cellulose _Iβ type single crystal fiber was used as the single crystal fiber of cellulose. That is, in the same manner as in the above example, 10 mM HEPES buffer (pH 7.4) was added so that the concentration of cellulose I _β type was 1 wt%, and cellulose I _β type was dispersed in the buffer solution. Thus, a cellulose I _β aqueous solution containing cellulose I _β type at a concentration of 1 wt% was prepared.

  On the other hand, β-galactosidase which is an enzyme was used as a protein to be treated. That is, β-galactosidase was dissolved in HEPES buffer to prepare a solution containing 4 nM β-galactosidase.

  Further, p-nitrophenol-β-D-galactopyranoside (hereinafter referred to as “PNPG”), which is a substrate of β-galactosidase, was replaced with 86 mM sodium phosphate, 116 mM 2-mercaptoethanol, and 1 mM magnesium chloride. A PNPG solution was prepared by dissolving in a phosphate buffer solution (pH 7.3).

A cellulose / β-galactosidase solution was then prepared. That is, 100 μL of cellulose I _β aqueous solution and 100 μL of β-galactosidase solution are mixed, and cellulose / β- consisting of 10 mM HEPES buffer containing 0.5 wt% cellulose I _β type and 2 nM β-galactosidase. A 200 μL galactosidase solution was prepared.

  Furthermore, incubation of the cellulose / β-galactosidase solution was performed. That is, the cellulose / β-galactosidase solution was allowed to stand and incubated at 30 ° C. for 2 hours.

  Then, the supernatant containing the treated β-galactosidase and / or β-galactosidase degradation product was recovered from the cellulose / β-galactosidase solution. That is, after incubation, the cellulose / β-galactosidase solution was centrifuged at 15000 rpm for 1 minute at 30 ° C. using a centrifuge. 50 μL of the supernatant of the cellulose / β-galactosidase solution (hereinafter referred to as “supernatant sample”) was collected and transferred to a 96-well microplate.

  Next, 50 μL of the PNPG solution is added to the collected 50 μL supernatant sample, and the absorption intensity at 405 nm (absorption wavelength specific for the enzymatic degradation reaction product of PNPG by β-galactosidase) is measured over time using a plate reader. Measured.

For comparison, a control solution consisting of 10 mM HEPES buffer (pH 7.4) containing 2 nM β-galactosidase but not cellulose I _β type was prepared. Then, this control solution was also incubated in the same manner as the above-mentioned cellulose / β-galactosidase solution, the supernatant was collected, the PNPG solution was added to the supernatant, and the absorption intensity at 405 nm was increased with time using a plate reader. Measured.

  FIG. 5 shows the measurement results. In FIG. 5, the horizontal axis represents the enzyme reaction time (that is, the time elapsed since the PNPG solution was added to the supernatant after incubation) (minutes), and the vertical axis represents the absorption intensity (−) at 405 nm. In FIG. 5, the black circles indicate the enzyme reaction times 0, 2.5, 5, 7.5, 10, 20, 30, 40, 90 of the supernatant sample collected from the cellulose / β-galactosidase solution after incubation. The measurement results at 120 minutes are shown, and white circles indicate the measurement results at the enzyme reaction times 0, 2.5, 5, 7.5, 10, 20, 30, 40 and 60 minutes of the control solution after incubation.

As shown in FIG. 5, in the supernatant of the control solution not containing cellulose I _β type (open circles), the enzymatic degradation reaction of PNPG by β-galactosidase progressed, and the amount of degradation products increased over time. On the other hand, in the supernatant (black circle) collected after the incubation of the cellulose / β-galactosidase solution, the enzyme degradation reaction product of PNPG was not substantially detected. This result was considered to be because β-galactosidase was inactivated by contact with cellulose I _β type.

[Protein degradation by cellulose I _β type]
Further, in the same manner as described above, three types of cellulose / β-galactosidase solutions comprising 10 mM HEPES buffer containing 0.5 wt% cellulose type I _β and 1 nM, 5 nM or 20 nM β-galactosidase. Was prepared, each cellulose / β-galactosidase solution was incubated, and a supernatant sample was collected.

  Next, 1 mL of the supernatant sample was concentrated to 100 μL using a centrifugal evaporator (hereinafter referred to as “concentrated supernatant sample”). Furthermore, 10 μL of the concentrated supernatant sample, 8 μL of 1M dithiothreitol, and 80 μL of loading buffer (200 mM Tris buffer (pH 6) containing 10% sodium dodecyl sulfate, 0.05% bromophenol blue, 50% glycerol) 8)) and the resulting mixture was incubated at 105 ° C. for 5 minutes.

  The mixed solution after the incubation was subjected to electrophoresis at 150 mV for 60 minutes using 4% acrylamide gel. Β-galactosidase was identified using the molecular weight marker migrated at the same time as an index.

For comparison, does not contain a cellulose I _beta type, 1 nM, 3 types consisting of HEPES buffer 10mM containing β- galactosidase 5nM or 20nM of the control solution was prepared, the supernatant of each control solution Similarly, electrophoresis was carried out.

  FIG. 6 shows a photograph of the gel after electrophoresis. FIG. 6 shows a case of using a cellulose / β-galactosidase solution containing a molecular weight marker (“marker” in FIG. 6), 1 nM, 5 nM or 20 nM β-galactosidase (“1” in “cellulose treatment” in FIG. 6). , “5” and “20”), and when using a control solution containing 1 nM, 5 nM or 20 nM β-galactosidase (“untreated” “1”, “5” and “20” in FIG. 6) The results are shown.

β-galactosidase shows a band around 116 kDa. In this regard, as shown in FIG. 6, the band of β-galactosidase when _β -galactosidase was treated with cellulose I _β type (“cellulose treatment” in FIG. 6) at all concentrations of β-galactosidase was β -Galactosidase was significantly thinner than that when it was not treated with cellulose I _β form ("no treatment" in Figure 6).

Furthermore, as shown in FIG. 6, when 1 nM β-galactosidase was treated with cellulose I _β type (“1” of “cellulose treatment” in FIG. 6), the treatment with cellulose I _β type was not performed ( A new broad band of 29 kDa or less that was not found in “1” of “No treatment” in FIG. 6 was detected. When 5 nM or 20 nM β-galactosidase is treated with cellulose I _β type (“5” and “20” in “cellulose treatment” in FIG. 6), the treatment with cellulose I _β type is not performed (FIG. 6). A new broad band of 44.3 kDa or less that was not found in “5” and “20” of “No treatment” in 6) was detected.

That is, it was confirmed that by bringing _β -galactosidase into contact with cellulose I _β type, the amount of the β-galactosidase is remarkably reduced and a substance having a smaller molecular weight is generated. This result was considered to indicate that β-galactosidase was decomposed by contact with cellulose I _β type, and a decomposed product having a smaller molecular weight was produced.

[Inactivation of virus by cellulose type I _β ]
Cellulose _Iβ type single crystal fiber was used as the single crystal fiber of cellulose. That is, in the same manner as in the above example, 10 mM HEPES buffer solution (pH 7.4) was added so that the concentration of cellulose I _β type was 0.1 wt%, and cellulose I _β type was dispersed in the buffer solution. It was. Thus, a cellulose I _β aqueous solution containing cellulose I _β type at a concentration of 0.1 wt% was prepared.

On the other hand, a filamentous virus (hereinafter referred to as “phage”) was used as a virus to be treated. That is, the phage was dispersed in 10 mM HEPES buffer (pH 7.4) so as to be 2.0 × 10 ¹⁰ pfu to prepare a phage solution.

A cellulose / phage solution was then prepared. That is, a cellulose / phage solution composed of 10 mM HEPES buffer containing 0.05 wt% cellulose and 1.0 × 10 ¹⁰ pfu phage was mixed with 150 μL cellulose I _β aqueous solution and 150 μL phage solution. Prepared.

  In addition, incubation of the cellulose / phage solution was performed. That is, the cellulose / phage solution was allowed to stand and incubated at 20 ° C. for 10 minutes, 2 hours, or 24 hours.

  The supernatant containing the treated phage was then recovered from the cellulose / phage solution. That is, after incubation, the cellulose / phage solution was centrifuged at 15000 rpm for 1 minute at 20 ° C. using a centrifuge. 200 μL of the supernatant of the cellulose / phage solution (hereinafter referred to as “supernatant sample”) was collected and serially diluted with 50 mM Tris buffer (pH 7.5) containing 150 mM NaCl and 0.1% gelatin. .

  Next, 10 μL of the serially diluted phage solution and 200 μL of E. coli ER2738 culture solution cultured until the absorption intensity at 660 nm reached 0.5 were mixed and incubated for 5 minutes. Mixed with the prepared LB agar medium.

  Furthermore, the total amount of the resulting mixture contains isopropyl β-thiogalactopyranoside (IPTG) and 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal). The mixture was solidified on an agar medium and cultured at 37 ° C. overnight. Thereafter, the remaining phages contained in the supernatant sample were quantified by counting the number of plaques formed by phages formed on the agar medium. For each supernatant sample, the number of plaques was independently measured three times.

  FIG. 7 shows the measurement results. In FIG. 7, the horizontal axis represents the incubation time (hour) of the cellulose / phage solution, and the vertical axis represents the residual phage amount (pfu).

As shown in FIG. 7, it was confirmed that the number of plaque-forming phages contained in the supernatant of the cellulose / phage solution decreased as the incubation time of the cellulose / phage solution increased. This result was considered to be because the phage was inactivated by contact with cellulose type I _β .

[Degradation of viral protein by cellulose type I _β ]
Moreover, the supernatant sample of the cellulose / phage solution incubated for 10 minutes or 120 minutes at 20 degreeC was collect | recovered similarly to the above-mentioned case. Next, 10 μL of the supernatant sample, 8 μL of 1M dithiothreitol, and 80 μL of loading buffer (10 mM sodium dodecyl sulfate, 0.05% bromophenol blue, 200 mM Tris buffer containing 50% glycerol (pH 6. 8)) and the resulting mixture was incubated at 105 ° C. for 5 minutes.

  Then, the mixed solution after the incubation was subjected to electrophoresis at 200 mV for 30 minutes using a Bio-Rad mini-PROTEAN TGX gel AnykD. Using the molecular weight markers migrated at the same time, major outer shell proteins and non-major outer shell proteins directly related to infection were identified and quantified.

  FIG. 8 shows a photograph of the gel after electrophoresis. FIG. 8 shows a molecular weight marker (“marker” in FIG. 8), no incubation (“0” in FIG. 8), 10 minutes incubation (“10” in FIG. 8), 120 minutes The results of the above incubation (“120” in FIG. 8) are shown.

As shown in FIG. 8, when the phage is treated with cellulose I _β type for 10 minutes or 120 minutes (“10” or “120” in FIG. 8), the major outer protein band is treated with cellulose I _β type. Compared with the case of not performing ("0" of FIG. 8), it became remarkably thin.

Furthermore, as shown in FIG. 8, when the phage was treated with cellulose I _β type for 10 minutes or 120 minutes (“10” or “120” in FIG. 8), the treatment with cellulose I _β type was not performed ( A new band (“fragment” in FIG. 8) having a molecular weight of 6500 Da or less that was not found in “0” in FIG. 8 was detected.

That is, it was confirmed that by bringing the phage into contact with cellulose type I _β, the amount of the main outer shell protein of the phage was significantly reduced and a substance having a smaller molecular weight was produced. This result was considered to indicate that the main outer shell protein of the phage was degraded by contact with cellulose type I _β, and a degradation product having a smaller molecular weight was produced.

FIG. 9 shows the result of quantifying the amount of protein remaining after incubation of cellulose I _β type and phage for 10 minutes or 120 minutes. In FIG. 9, the horizontal axis indicates the incubation time (minutes), and the vertical axis indicates the ratio (%) of the remaining protein. In FIG. 9, white circles indicate the residual ratio of the major outer protein of the phage, and black circles indicate the residual ratio of the non-major outer protein of the phage.

As shown in FIG. 9, it was confirmed that as the incubation time increased, the proportion of residual protein decreased for both the major and non-major outer phage proteins. That is, it was confirmed that the amount of the major outer shell protein and the non-major outer shell protein of the phage decreased with time by bringing the phage into contact with cellulose type I _β .

Also, FIG. 10 shows that when the phage was not incubated with cellulose I _β type (“0” in FIG. 8), there was no molecular weight of 6500 or less newly appeared after 10 minutes or 120 minutes of incubation. 9 shows the result of quantification of the fragmented protein bands (“10” and “fragment” of “120” in FIG. 8). In FIG. 10, the horizontal axis indicates the incubation time (minutes), and the vertical axis indicates the amount of fragmented protein (au).

As shown in FIG. 10, the amount of protein fragments that were not detected at all before incubation (incubation time “0”) increased over time during the incubation time. This result was considered to indicate that the outer shell protein of the phage was degraded and fragmented by incubation with cellulose type I _β .

[Inactivation of microorganisms by cellulose I _β type]
Cellulose _Iβ type single crystal fiber was used as the single crystal fiber of cellulose. That is, cellulose type I _β was dispersed in ultrapure water so that its concentration was 0.5 wt%. Thus, a cellulose I _β aqueous solution containing cellulose I _β type at a concentration of 0.5 wt% was prepared.

  On the other hand, three types of E. coli (ER2738 strain, XL1-Blue strain and Rosetta strain) were used as microorganisms to be treated. That is, each strain was cultured in LB medium until the absorption intensity at 660 nm reached 0.5 to prepare an E. coli solution. Thereafter, the E. coli solution was centrifuged at 3500 rpm at 4 ° C. to precipitate E. coli, and the medium was removed. Then, 10 mM phosphate buffer solution (pH 7.4) containing 150 mM NaCl is added so that the absorption intensity at 660 nm is 0.45 or 0.0045, and the precipitated E. coli is dispersed again, so that the concentration of E. coli is different. Different types of E. coli solutions were prepared.

A cellulose / E. Coli solution was then prepared. That was centrifuged cellulose I _beta aqueous 300 [mu] L, the supernatant was removed. Then, 300 μL of E. coli solution (one of two types of E. coli solutions) is mixed, cellulose I _β type is dispersed again, and consists of a 10 mM phosphate buffer containing 0.5 wt% cellulose and E. coli. A cellulose / E. Coli solution was prepared.

  Furthermore, incubation of the cellulose / E. Coli solution was performed. That is, the cellulose / E. Coli solution was allowed to stand and incubated at 20 ° C. for 7 days.

  Then, the cellulose / E. Coli solution was serially diluted with a phosphate buffer, layered on an agar medium, solidified, and cultured at 37 ° C. overnight. Thereafter, the remaining E. coli contained in the cellulose / E. Coli solution was quantified by counting the number of E. coli colonies formed on the agar medium. For each sample, the number of colonies was independently measured three times.

For comparison, a control solution consisting of a 10 mM phosphate buffer containing E. coli but not containing cellulose type I _β was prepared. This control solution was also incubated and cultured on an agar medium in the same manner as the cellulose / E. Coli solution described above, and the number of colonies was counted to quantify the remaining E. coli.

  11 and 12 show the measurement results. FIG. 11 shows the results when using an E. coli solution in which E. coli is dispersed at a concentration at which the absorption intensity at 660 nm is 0.45, and FIG. The result when using the Escherichia coli solution in which is dispersed is shown. 11 and 12, the horizontal axis indicates the type of E. coli strain, and the vertical axis indicates the amount of residual E. coli (cfu). In FIGS. 11 and 12, the open bars show the results for E. coli without incubation, the hatched bars show the results for E. coli after 7 days incubation in the control solution, and the black bars Shows the results for E. coli after 7 days incubation in cellulose / E. Coli solution.

As shown in FIGS. 11 and 12, for any of the three strains, as compared with the case not contacted with the cellulose I _beta type (bar hatched in FIGS. 11 and 12), and the cellulose I _beta type It was confirmed that the colony forming ability of Escherichia coli when contacted (black bars in FIGS. 11 and 12) was significantly reduced. This result was thought to be due to the inactivation of E. coli by contact with cellulose type I _β .

Further, as shown in FIGS. 11 and 12, when the E. coli having a concentration at which the absorption intensity at 660 nm is 0.0045 is brought into contact with cellulose I _β type (FIG. 12), the residual E. coli amount is 0. The concentration was significantly reduced compared to that obtained when E. coli at a concentration of .45 was contacted with cellulose _Iβ type (FIG. 11).

Here, if Escherichia coli was inactivated by simply adsorbing to cellulose _Iβ type, the higher the concentration of Escherichia coli, the more efficient the adsorption. As a result, E. coli was more effectively inactivated. Should be activated. However, as described above, when the concentration of E. coli was low (FIG. 12), E. coli was inactivated more effectively than when the concentration of E. coli was high (FIG. 11). Therefore, it was considered that the inactivation of Escherichia coli by contact with cellulose type I _β was not caused by a simple adsorption phenomenon but by a decomposition reaction as in the case of the above-described proteins and viruses.

Claims (14)

A treatment material for decomposing an organic compound that can be hydrolyzed,
A treatment material comprising a single crystal fiber of cellulose. The treatment material according to claim 1, wherein the organic compound is at least one selected from the group consisting of an ester compound, an amide compound, and a urethane compound. The treatment material according to claim 1 or 2, wherein the organic compound is at least one selected from the group consisting of peptides, proteins, lipids, carbohydrates, DNA, and RNA. The treatment material according to any one of claims 1 to 3, wherein the organic compound is an organic compound present on a surface of a virus or a microorganism. A treatment material for inactivating one or more selected from the group consisting of viruses, microorganisms, proteins and peptides,
A treatment material comprising a single crystal fiber of cellulose. The treated material according to any one of claims 1 to 5, wherein the treated material has a single crystal fiber of cellulose on its surface. The treatment material according to claim 1, wherein the treatment material is an aqueous solution containing single crystal fibers of cellulose. A treatment method comprising decomposing the organic compound by bringing a treatment target, which is an organic compound that can be hydrolyzed, into contact with a single crystal fiber of cellulose. The processing method according to claim 8, wherein the organic compound is at least one selected from the group consisting of an ester compound, an amide compound, and a urethane compound. The processing method according to claim 8 or 9, wherein the organic compound is at least one selected from the group consisting of peptides, proteins, lipids, carbohydrates, DNA, and RNA. The processing method according to any one of claims 8 to 10, wherein the organic compound is an organic compound present on a surface of a virus or a microorganism. A treatment method comprising inactivating a treatment target by bringing the treatment target selected from the group consisting of a virus, a microorganism, a protein, and a peptide into contact with a single crystal fiber of cellulose. . 13. The treatment object and the cellulose single crystal fiber are brought into contact with each other by bringing the treatment object into contact with a molded body having the cellulose single crystal fiber on the surface thereof. The processing method described in any one of. The said process target and the aqueous solution containing the said single crystal fiber of the cellulose are mixed, The said process target and the single crystal fiber of the cellulose are made to contact. The one of the Claims 8 thru | or 12 characterized by the above-mentioned. The processing method described in 1.

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