KR102092141B1 - Self-decontamination material - Google Patents

Abstract

The present application relates to a self-detoxifying substance using a support and an enzyme.

Description

Self-detoxifying substance {SELF-DECONTAMINATION MATERIAL}

This application relates to self-decontamination materials, in particular self-decontamination materials using supports and enzymes.

The possibility of exposure to toxic agents, including chemical agents used as war gases and toxic substances for industrial use, poses a danger not only to soldiers but also to civilians. Chemical agents are still in possession of various countries, and some terrorist groups can easily make or obtain them, and until recently, they have been used in the Middle East and other regions of Syria to report damages, including civilians.

Since chemical agents are generally highly likely to be sprayed in the form of fine aerosol fumes, they can be deposited on the surface of various military equipment and weapons, including military uniforms, as well as inhalation by breathing. In order to minimize the risk of contact when the surfaces of various military equipment and weapons are contaminated with chemical agents, the toxic chemical agents should be quickly removed.

In particular, it usually penetrates into the inhalation or skin, such as sarin (also called sarin or GB), soman (also called soman or GD), and VX, and acts on the nervous system of people and animals, causing sudden paralysis and eventually dying within a short time There is a need to develop technologies that can quickly remove organophosphorus compounds, which are neuroactive agents that lead to distilled mustard (HD), a persistent blister agent.

In addition to the above chemical agents as well as the need to decontaminate industrial chemicals, for example, pesticides that can cause paralysis of the nervous system, that is, organic agents, parathion (parathion), paraoxon (paraoxon) and actin as pesticides It has become very important to effectively detoxify precision equipment and surface contamination by various toxic substances that are not limited to toxic substances including malathion and organophosphorous agents used as neuroactive agents.

Currently, the most commonly used neuroactive detoxifying agent, including the U.S. military, is the DS2 solution.The composition of DS2 is caustic soda (NaOH) 2% by weight, 28% ethylene glycol monomethyl ether, And diethylenetriamine (diethylenetriamine) is composed of 70%.

DS2 is an effective detoxifying agent for the organophosphorous agent, which is a neuroactive agent, but it is quite toxic, flammable, corrosive and toxic by-product by itself, as well as detoxification in the case of the main component of diethylenetriamine. Known as a substance, there is a possibility of causing health hazards in use or production. In addition, when decontamination using DS2, the decontamination process is completed only after decontamination using water, so the decontamination process is completed, which increases the burden of logistics by storing or transporting a lot of water in the decontamination area.

Another decontamination agent in operation in the US military is the XE555 resin, which is used to quickly decontaminate surfaces contaminated with chemical agents. Although XE555 is effective in removing chemical agents, there is a disadvantage that it does not have the ability to sufficiently neutralize the absorbed toxic chemical agents, and thus there is a problem that steam of toxic substances can escape from the XE555 resin after detoxification.

Republic of Korea Patent Publication No. 10-2014-0140144, which is the background technology of the present application, relates to a dehalogenation reaction activity for an organic halogen compound and a variant of the haloalkan dehalogenase with improved thermal stability and its use. The publication discloses a configuration of improving a variant of DhaA having excellent haolalakne dehalogenase (HDL) activity by expressing the variant on the cell surface.

The present application is to solve the problems of the above-described prior art, it is an object to provide a self-detoxifying material using a support and an enzyme.

However, the technical problems to be achieved by the embodiments of the present application are not limited to the technical problems as described above, and other technical problems may exist.

As a technical means for achieving the above technical problem, the first aspect of the present application, the support; And a hydrolytic enzyme immobilized through a physical bond to the support, wherein the support adsorbs a chemical agent, and the hydrolytic enzyme hydrolyzes the chemical agent to provide an auto-detoxifying material.

According to one embodiment of the present application, the support is made of cyclodextrin, calixarenes, corannulene, glass silica beads, activated carbon, graphene, and combinations thereof. It may be to include materials selected from the group, but is not limited thereto.

According to one embodiment of the present application, the support may be a poly beta cyclodextrin (poly-β-cyclodextrin), but is not limited thereto.

According to the exemplary embodiment of the present application, the poly beta cyclodextrin may be synthesized by a combination of beta cyclodextrin (β-cyclodextrin) and hexamethylene diisocyanate, but is not limited thereto.

According to one embodiment of the present application, the support may be one having a porous nano-structure, but is not limited thereto.

According to the exemplary embodiment of the present application, the support may be to perform a chaperone function, but is not limited thereto.

According to one embodiment of the present application, the hydrolase is an organophosphorous hydrolase (OPH), an organophosphorous acid anhydrolase (OPAA), a haloalkane dehalogenase (HD) ) And combinations thereof, but may be an enzyme selected from the group.

According to one embodiment of the present application, the chemical agent may include, but is not limited to, an agent selected from the group consisting of neuroactive agents, blistering agents, blood agents, choking agents, and combinations thereof.

According to one embodiment of the present application, the neuroactive agent comprises a material selected from the group consisting of methyl paraoxon (MPO), soman, sarin, tabun, and combinations thereof. It may be, but is not limited thereto.

According to the exemplary embodiment of the present application, the self-detoxifying material may be one that can remove and reuse by-products hydrolyzed by an organic solvent, but is not limited thereto.

According to the exemplary embodiment of the present application, the organic solvent may include a material selected from the group consisting of methanol, acetone, ethanol, isopropanol, acetonitrile, and combinations thereof, but is not limited thereto.

The above-described problem solving means are merely exemplary and should not be construed as limiting the present application. In addition to the exemplary embodiments described above, additional embodiments may exist in the drawings and detailed description of the invention.

According to the above-mentioned problem solving means of the present application, the self-detoxifying material according to the present application has long-lasting activity of hydrolytic enzymes by adsorbing a wide range of hydrophilic or hydrophobic chemical agents and toxic industrial chemicals that support hydrophobic interaction. It is possible to increase the adsorption and decontamination performance by maintaining the time.

Furthermore, by controlling the reversible binding process of the support, for example, poly beta cyclodextrin and the hydrolase, the hydrolyzed enzyme is not denatured in the process of physically adsorbing to the poly beta cyclodextrin and prevents degeneration. As a tool to make use of the hydrophilic interior space. Since most of the poisonous substances, especially biochemical poisonous substances, are hydrophilic or extremely hydrophobic, it is easy to induce reversible adsorption and desorption when desired, thereby enhancing the detoxification function.

In addition, even when the hydrolytic enzyme has been used for a long time, the three-dimensional structure of the hydrolytic enzyme is deformed and thus loses its function as a hydrolytic enzyme, the support performs a chaperone function at an appropriate pH condition and refolds it again. Since it returns to the catalytic ability of the initial hydrolase, the hydrolase can be recycled.

The self-decontamination material according to the present application can be utilized in various fields such as military operations, protection facilities and equipment applications, civil environment protection and purification due to its ability to adsorb and decompose chemical agents and pesticides. In particular, it is a system that appropriately combines environmentally friendly substances differently from the existing toxic detoxifying substances, and it is possible to maintain an environmentally friendly system by recycling self-detoxifying substances.

1 is an image schematically showing a self-decontamination material according to an embodiment of the present application.
2 is a schematic diagram for synthesizing poly beta cyclodextrin according to one embodiment of the present application.
3 is a scanning electron microscope (SEM) photograph of poly beta cyclodextrin according to an embodiment of the present application.
4 is a schematic diagram of a hydrolysis enzyme of an auto-detoxicant according to an embodiment of the present application to hydrolyze a chemical agent.
Figure 5 is a schematic diagram of the support of the self-decontamination material according to an embodiment of the present application (sorption) hydrolyzed by-products.
Figure 6 (a) is a decomposition evaluation result graph of the hydrolytic enzyme of the self-detoxifying material according to an embodiment of the present application, Figure 6 (b) is a decomposition evaluation result graph of the hydrolysis enzyme according to a comparative example of the present application to be.

Hereinafter, embodiments of the present application will be described in detail with reference to the accompanying drawings so that those skilled in the art to which the present application pertains may easily practice.

 However, the present application may be implemented in various different forms and is not limited to the embodiments described herein. In addition, in order to clearly describe the present application in the drawings, parts irrelevant to the description are omitted, and like reference numerals are assigned to similar parts throughout the specification.

Throughout this specification, when a part is "connected" to another part, it includes not only "directly connected" but also "electrically connected" with another element in between. do.

Throughout the present specification, when a member is positioned on another member “on”, “on the top”, “top”, “bottom”, “bottom”, and “bottom”, it means that a member is on another member. This includes cases where there is another member between the two members as well as when in contact.

Throughout the present specification, when a part “includes” a certain component, it means that the component may further include other components, not to exclude other components, unless specifically stated to the contrary.

As used herein, the terms “about”, “substantially”, and the like are used in or near the numerical values when manufacturing and substance tolerances unique to the stated meanings are presented, to aid understanding of the present application The exact or absolute value of the hazard is used to prevent unscrupulous use of the disclosed disclosure by unconscientious intruders. In addition, throughout the present specification, "step of" or "step of" does not mean "step for".

Throughout the present specification, the term “combination of these” included in the expression of the marki form means one or more mixtures or combinations selected from the group consisting of the components described in the expression of the marki form, the component. It means to include one or more selected from the group consisting of.

 Throughout this specification, the description of “A and / or B” means “A or B, or A and B”.

Throughout the present specification, “self-decontamination” refers to decontamination made of the applied material itself without using a special decontamination agent by coating a material surface that can be decontaminated by itself on the material surface.

Hereinafter, with reference to embodiments and examples and drawings for the self-detoxifying material of the present application will be described in detail. However, the present application is not limited to these embodiments and examples and drawings.

The first aspect of the present application includes a support; And a hydrolytic enzyme immobilized through a physical bond to the support, wherein the support adsorbs a chemical agent, and the hydrolytic enzyme hydrolyzes the chemical agent to provide an auto-detoxifying material.

1 is an image showing a self-decontamination material according to an embodiment of the present application.

According to one embodiment of the present application, the support is cyclodextrin (cyclodextrin; CD), calixarenes, coranulene, glass silica beads, activated carbon, graphene, and combinations thereof It may be to include a material selected from the group consisting of, but is not limited thereto.

Cyclodextrin is a cyclic oligosaccharide having a toric shape, and 6, 7, and 8 glucopyranoside units (α-D-glucopyranose units) are bound to α-cyclodextrin (α-CD) ), β-cyclodextrin (β-CD), and γ-cyclodextrin (γ-CD). The size of the hydrophobic internal cavity of the cyclodextrin is 0.57 nm for α-CD, 0.78 nm for β-CD, and 0.95 nm for γ-CD. Purification of wastewater because the hydrophobic internal cavity of the cyclodextrin can form an inclusion complex by performing host-guest interaction through various hydrophobic compounds and hydrophobic interactions, It is applied to various fields such as food and pharmaceutical industries.

According to one embodiment of the present application, the support may be a poly beta cyclodextrin (poly-β-cyclodextrin), but is not limited thereto.

The poly beta cyclodextrin was synthesized by polymerizing cyclodextrin using a spacer. In the synthesis, the length of the spacer molecule (for example, hexyl, octyl, etc.) and the reaction equivalent weight are controlled differently to perform a crosslinking reaction, and the poly beta cyclodextrin prepared accordingly has its own self-adsorption function. It can be used as a maximizing support. By maximizing the hydrophobic interaction of the poly beta cyclodextrin, the adsorption power to the chemical agent can be dramatically improved compared to the conventional cyclodextrin.

Furthermore, by physically controlling the reversible binding process between the poly beta cyclodextrin and the hydrolase, the hydrolyzed enzyme is not denatured in the physical adsorption process with the poly beta cyclodextrin, and is a very small tool as a tool for preventing denaturation. Take advantage of the castle's interior space.

Moreover, most neuroactive agents are organic compounds with high vapor pressure and high hydrophobicity. Even if the reactivity of the hydrolytic enzyme is high at room temperature, if the neuroactive agent is not held on the interface for a period of time, there is a problem that the hydrolytic enzyme cannot react with the neuroactive agent. The present inventors recognize this problem and self-detox to provide time to allow the reaction of the hydrolytic enzyme and the neuroactive agent to occur by holding the neuroactive agent for a period of time using a support for imparting hydrophobic interaction to solve the problem. The material was developed.

According to one embodiment of the present application, the poly beta cyclodextrin may be synthesized by a combination of beta cyclodextrin (β-cyclodextrin) and hexamethylene diisocyanate (HDI), but is not limited thereto.

2 is a schematic diagram for synthesizing poly beta cyclodextrin according to one embodiment of the present application.

Referring to FIG. 2, it can be confirmed that the poly beta cyclodextrin is synthesized by a polymerization reaction between the beta cyclodextrin and the hexamethylene diisocyanate.

 According to one embodiment of the present application, the support may be one having a porous nano-structure, but is not limited thereto.

3 is a scanning electron microscope (SEM) photograph of poly beta cyclodextrin according to an embodiment of the present application.

Referring to Figure 3, it can be seen that the poly beta cyclodextrin as the support has a porous nanostructure. Dimethylformamide (DMF) or dimethylacetamide (DMAC), a solvent used in the synthesis reaction of poly beta cyclodextrin as the support, can form a porous nanostructure by acting as a kind of porogen during synthesis. . The bucket structure inside the beta cyclodextrin also contributes to the formation of the porous nanostructure of the poly beta cyclodextrin, but can form a porous nanostructure of different size depending on the bonding method with the spacer. Specifically, the support may have a porous nano-structure to increase the hydrolytic enzyme that physically binds the support. Due to this, the hydrolytic enzyme can hydrolyze the chemical agent more effectively.

According to the exemplary embodiment of the present application, the support may be to perform a chaperone function, but is not limited thereto.

Chaperone refers to a protein that assists in the folding and unfolding of other proteins, or the incorporation and disassembly of a large protein in which several proteins are combined. Specifically, even when the hydrolytic enzyme has been used for a long time to deform the three-dimensional structure of the hydrolytic enzyme and lose its function as a hydrolytic enzyme, the support performs a chaperone function at an appropriate pH condition and refolds again. By returning to the catalytic ability of the initial hydrolase, the hydrolase can be recycled.

According to one embodiment of the present application, the hydrolase is an organophosphorous hydrolase (OPH), an organophosphorous acid anhydrolase (OPAA), a haloalkane dehalogenase (HD) ) And combinations thereof, but may be an enzyme selected from the group. Preferably, the hydrolase may be an organophosphate hydrolase.

Organic phosphatase is derived from Pseudomonas diminuta or Flavobacterium species, a single dimer organophosphoresterase that can degrade a wide range of toxic organophosphate compounds. (homodimeric organophosphotriesterase). The organophosphate hydrolase can hydrolyze various phosphate-ester bonds including P-O, P-F, P-CN, and P-S bonds. The organic phosphatase has an advantage of high turnover rate.

According to one embodiment of the present application, the chemical agent may include, but is not limited to, an agent selected from the group consisting of neuroactive agents, blistering agents, blood agents, choking agents, and combinations thereof. Preferably, the chemical agent may include a neuroactive agent.

Chemical agents are compounds that are used to kill, disable, or inflict severe damage on the basis of their chemical properties, and mainly refer to toxic chemicals. Chemical agents are classified according to their physical state, purpose of use, and physiological action. Among them, they are classified into neuroactive agents, suffocating agents, blood agents, and blister agents.

When a neuroactive agent is absorbed into the body through the respiratory system, digestive system, and skin, it inhibits the action of cholinesterase enzymes in the body and accumulates acetylcholine, causing symptoms such as pupil contraction, dyspnea, and muscle spasms. To protect against neuroactive agents, gas masks and protective clothing should be worn. If it is contaminated with a neuroactive agent, it can be treated by injecting a neuroactive antidote.

According to one embodiment of the present application, the neuroactive agent comprises a material selected from the group consisting of methyl paraoxon (MPO), soman, sarin, tabun, and combinations thereof. It may be, but is not limited thereto. Preferably, the neuroactive agent may include methyl paraoxone.

4 is a schematic diagram of a hydrolysis enzyme of a self-detoxifying substance according to an embodiment of the present application to hydrolyze a chemical agent, and FIG. 5 is a sorption of a by-product of which the support of the auto-detoxicating substance according to an embodiment of the present application is hydrolyzed sorption).

4 and 5, the chemical agent, for example, methyl paraoxone, is hydrolyzed by the hydrolase, for example, an organophosphate hydrolase. The hydrolyzed by-products, for example p-nitrophenol, are sorbed by the support, for example poly beta cyclodextrin and bound to the interior space of the poly beta cyclodextrin.

Specifically, the poly-beta cyclodextrin can constrain the hydrolyzed by-product p-nitrophenol to the internal space, so that the self-detoxifying material may not only simply detoxify, but may also serve to protect from highly toxic by-products.

According to the exemplary embodiment of the present application, the self-detoxifying material may be one that can remove and reuse by-products hydrolyzed by an organic solvent, but is not limited thereto.

According to the exemplary embodiment of the present application, the organic solvent may include a material selected from the group consisting of methanol, acetone, ethanol, isopropanol, acetonitrile, and combinations thereof, but is not limited thereto. Preferably, the organic solvent may include methanol.

The self-detoxifying material adsorbed by the by-products can no longer adsorb and decompose the chemical agent by the by-products. In order to solve this, by-products adsorbed to the self-detoxifying material may be removed using the organic solvent, for example, methanol. The self-decontamination material from which the by-products have been removed can restore the performance of the initial self-decontamination material to adsorb and decompose chemical agents. According to this, since the self-decontamination material is recyclable, it has a positive effect in cost and environmental aspects.

The present invention will be described in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present application.

[Example] Preparation of self-decontamination material

First, 5 ml of DMF was added to an aqueous solution in which 1 g of beta cyclodextrin and 1.274 ml of hexamethylene diisocyanate solution were mixed, and then polymerized at 70 ° C. to synthesize poly beta cyclodextrin (PCD). Subsequently, 1 g of an organic phosphatase (OPH) and 10 ml of a 1 mM phosphorus CHES buffer solution were mixed, and then filtered to remove dust. After adding 2 g of poly beta cyclodextrin to the aqueous solution and filtering, a self-detoxifying substance in which poly beta cyclodextrin and organic phosphatase was combined was obtained. The self-decontamination material was freeze dried.

[Comparative example]

In order to confirm how the poly beta cyclodextrin, which is the most important feature of the present application, has an effect on the detoxification properties of the self-detoxifying substance, a hydrolytic enzyme of a comparative example without binding to the support was prepared. The hydrolase of the comparative example means organophosphate hydrolase (OPH).

[Experimental Example]

The properties of the self-decontamination material prepared in the above example were confirmed, and the results are shown as (a) and (b) of FIG. 6.

Figure 6 (a) is a decomposition evaluation result graph of a hydrolytic enzyme of a self-detoxifying material according to an embodiment of the present application, Figure 6 (b) is a decomposition evaluation result graph of a hydrolysis enzyme according to a comparative example of the present application to be. Figure 6 (a) is an experiment in which 20 ml of a methyl paraoxone, which is 200 mg and 50 μM of the self-detoxifying material according to the above example, is reacted, and FIG. 6 (b) is 7.5 mg, 3.3 mg, 1.65 mg, 0.825 mg Phosphorus is an experiment in which the hydrolase according to the comparative example and 50 ml of methyl paraoxone, respectively, are reacted with each.

Referring to (a) and (b) of FIG. 6, it can be confirmed that the organophosphate hydrolase decomposes methyl paraoxone within 20 minutes. In addition, the initial rate of hydrolysis of the self-detoxifying material according to the embodiment is higher than the initial rate of hydrolysis of the hydrolase according to the comparative example when considering the amount of p-nitrophenol and methyl paraoxone initially adsorbed to poly beta cyclodextrin. You can see it is fast.

The above description of the present application is for illustrative purposes, and those skilled in the art to which the present application pertains will understand that it is possible to easily modify to other specific forms without changing the technical spirit or essential features of the present application. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present application is indicated by the claims below, rather than the detailed description, and it should be interpreted that all changes or modifications derived from the meaning and scope of the claims and equivalent concepts thereof are included in the scope of the present application.

Claims (11)

In self-detoxifying substances,
A support comprising a poly beta cyclodextrin (poly-β-cyclodextrin); And
A hydrolytic enzyme immobilized through a physical bond to the support;
Including,
The support adsorbs the chemical agent, the hydrolytic enzyme hydrolyzes the chemical agent,
The chemical agent includes an agent selected from the group consisting of a neuroactive agent, a blister agent, a blood agent, a choking agent, and combinations thereof,
The hydrolase contains an organophosphorous hydrolase (OPH),
By-products formed by the hydrolysis and sorbed in the self-detoxication material can be removed and reused by an organic solvent,
The by-product of the hydrolyzed chemical agent is bound to the interior space of the poly beta cyclodextrin,
Self-detoxifying substance.
delete delete According to claim 1,
The poly beta cyclodextrin is synthesized by the combination of beta cyclodextrin (β-cyclodextrin) and hexamethylene diisocyanate (hexamethylene diisocyanate), self-detoxifying material.
According to claim 1,
The support is to have a porous nano-structure, self-decontamination material.
According to claim 1,
The support is to perform a chaperone (chaperone) function, self-decontamination material.
delete delete According to claim 1,
The neuroactive agent is a methyl paraoxon (methyl paraoxon; MPO), soman (soman), sarin (sarin), tabun (tabun) and includes a substance selected from the group consisting of self-detoxifying substances.
delete According to claim 1,
The organic solvent comprises a material selected from the group consisting of methanol, acetone, ethanol, isopropanol, acetonitrile, and combinations thereof.

Images