KR101350016B1 - Antifouling composites and paints and manufacturing method thereof - Google Patents

Abstract

The antifouling composition comprising the fiber matrix complex of the antifouling enzyme-three-dimensional network structure of the present invention and the antifouling coating composition comprising the same are applied to the existing antifouling enzyme-substrate complex which was limited in industrial application due to the activity of the antifouling enzyme being too low. Compared with this, it is possible to obtain improved antifouling performance, thereby increasing the industrial applicability, and solving economic problems by using an inexpensive support.

Description

Antifouling paint composition and its manufacturing method {Antifouling composites and paints and manufacturing method

The present invention relates to a paint composition for antifouling, and more particularly, a much larger amount of antifouling enzyme is immobilized on a fiber of a three-dimensional network structure, and the antifouling enzyme activity is stable for a long time, as compared with a conventional antifouling composition including an antifouling enzyme. It is to provide an antifouling composition comprising a fiber matrix complex having a sustainable antifouling enzyme-three-dimensional network structure, and an antifouling coating composition comprising the same.

The antifouling paint composition is intended to prevent or control the attachment and growth of marine organisms causing surface contamination of the seawater-deposited coating film. The hull bottom (aquatic bottom) and aquatic structures are attached to the surface of various aquatic organisms such as shellfish, oysters, mussels, shellfish such as mussels and shellfish, vegetation such as seaweeds, and aquatic bacteria. As a result, the appearance becomes poor or the function is lost. In particular, when these aquatic organisms attach to and propagate on the bottom, the surface roughness of the entire ship's outer shell is increased, resulting in a decrease in ship speed, thereby increasing fuel consumption and operating costs. Removing aquatic creatures from the bottom requires a lot of effort and long working hours. In addition, when mucus (mud matter) or slime is attached to bacteria that attach to and propagate to underwater structures, bacteria are spoiled, or large organisms that are tacky adhere to and propagate in underwater structures (for example, steel structures). In the case of damaging the anti-corrosion coating of the underwater structure, there is a problem in reducing the strength or function of the underwater structure to significantly shorten the life.

In order to prevent such a problem, it is general practice to coat an antifouling coating composition containing copper (tributyltin methacrylate, methyl methacrylate, etc.) and copper oxide (Cu ₂ O) as an antifouling paint excellent in antifouling property on the bottom part to be.

The copolymer constituting the antifouling paint composition is hydrolyzed in seawater and is converted into bistributyltin oxide (Bu ₃ Sn-OSnBu ₃ (Bu is butyl group)) or tributyltin halide (Bu ₃ SnX (X is halogen atom )), Thereby exhibiting an antifouling effect. Since the copolymer is a hydrolyzable self-polishing paint, the hydrolyzed copolymer itself is also converted into water-soluble and dissolved in seawater, so that the resin remains on the surface of the bottom coating film and remains active Keep the surface.

However, the abovementioned organic tin compounds are very toxic and highly likely to cause marine pollution, malformed fish or malformed clams, and destruction of aquatic ecosystems through the food chain. Due to these problems, it has been required to develop a tin-free antifouling paint which can replace the conventional organotin-based antifouling paints.

Recently, in order to replace tin, various materials such as zinc, copper, sylil, silicone, copper dioxide, and materials having good wear properties have been used. Conventional paints use a binder partially gelled by mixing a metal ester compound such as metal acetate with an acid group-containing polymer. Such a binder has poor film-forming ability and storage stability to cause cracking, poor abrasiveness, and difficulty in application as a high viscosity. In addition, conventional acrylic polymers such as zinc acrylate mixed with a metal ester compound have a problem in that they are not well controlled in abrasion and are insecure in long-term abrasion.

In addition, these substances are also reported to adversely affect the ecosystem because they contain heavy metals, etc. and these environmental problems are increasing the limit of their use.

Thus, Korean Patent Application No. 2001-7010883 discloses a composition for ship antifouling including an antifouling enzyme or microorganisms producing the same in epoxy or glass fiber. In addition, Korean Patent Application No. 2002-7012611 discloses an antifouling composition comprising rosin and antifouling enzyme, and rosin has a function of immobilizing antifouling enzyme and prevents antifouling enzymes from being released to the outside. However, all of these prior arts have a problem that the amount of antifouling enzymes fixed to the support is extremely low, and thus the antifouling effect is low as well as the antifouling enzyme is easily leaked to the outside as time passes, and the antifouling effect rapidly drops.

The present invention has been made to overcome the above-mentioned problems, the problem to be solved of the present invention, compared to the antifouling composition comprising a conventional antifouling enzyme contains a significantly larger amount of antifouling enzymes and antifouling enzymes over time The present invention provides an antifouling composition and an antifouling coating composition comprising the same, in which activity is maintained without being almost leaked to the outside.

In order to solve the above problems, the present invention, the cross-linked antifouling enzyme aggregate and the surface of the three-dimensional network fibers carried in the pores of the porous matrix including the three-dimensional network fibers and the diameter larger than the inlet size of the pores It provides an antifouling paint composition comprising; fiber matrix complex of the enzyme three-dimensional network structure comprising a shell comprising surrounding cross-linked antifouling enzymes.

According to a preferred embodiment of the present invention, the three-dimensional network fibers may be microfibers or nanofibers.

According to another preferred embodiment of the present invention, the three-dimensional network fiber is made of polyvinyl alcohol, polyacrylonitrile, nylon, polyester, polyurethane, polyvinyl chloride, polystyrene, cellulose, chitosan, polylactic acid, Polylactic-co-glycolic acid, polyglycolic acid polycaprolactone, collagen, polypyrrole, polyaniline and poly (styrene-co-maleic anhydride).

According to another preferred embodiment of the present invention, the shell may be a multilayer.

According to another preferred embodiment of the present invention, the antifouling enzyme is acylase, lactonase, protease, peroxidase, aminopeptidase, phosphatase, transaminase, serine-endopeptidase, cysteine-endopeptidase and It may be any one or more selected from the group consisting of metalloendopeptidase.

According to another preferred embodiment of the present invention, the antifouling paint composition may comprise 0.001 to 50% by weight of the composite.

According to another preferred embodiment of the present invention, (1) adding an antifouling enzyme to the porous matrix comprising three-dimensional network fibers; (2) adding a precipitation agent to the matrix to prevent leakage of antifouling enzymes added into the pores of the matrix; (3) preparing an antifouling enzyme-fiber matrix complex by adding a crosslinking agent to form an enzyme aggregate to form crosslinking between the antifouling enzymes; And (4) provides a method for producing an antifouling paint composition comprising the step of adding the complex to the paint.

According to another preferred embodiment of the present invention, the precipitation agent is methanol, ethanol, 1-propanol, 2-propanol, butyl alcohol, acetone, polyethylene glycol, ammonium sulfate, sodium chloride, sodium sulfate, sodium phosphate, potassium chloride , Potassium sulfate, potassium phosphate and aqueous solutions thereof may be used alone or in combination.

According to another preferred embodiment of the present invention, the crosslinking agent is selected from the group consisting of a diisocyanate, a dianhydride, a diepoxide, a dialdehyde, a diimide, 1-ethyl-3-dimethylaminopropylcarbodiimide, a glutardialdehyde , Bis (imido ester), bis (succinimidyl ester), and diacid chloride.

According to another preferred embodiment of the present invention, it may further comprise the step of removing the precipitation agent and crosslinking agent between the step (3) and (4).

The terms used in the present invention will be described.

The term antifouling enzyme refers to an enzyme contained in an antifouling composition that prevents contaminants from adhering to and growing on the surface of a substance through biological antifouling action. For example, surface attachment and growth of biological contaminants is driven by a unique quorum sensing system, while quorum sensing materials are compounds of the N-acyl homoserine lactone (AHL) family, This compound acts to hydrolyze and quorum quenching the signal. Meanwhile, the antifouling enzyme disclosed in Korean Patent Application No. 2001-7010883 and Korean Patent Application No. 2002-7012611 can be included in the present invention.

The term 'substantially free of covalent bonds between fibers and enzymes' refers to functional groups capable of covalently bonding with enzymes naturally formed on the surface of the fibers (e.g., they remain on the surface of the fibers without reacting during the polymerization and spinning of the fibers). Except for the formation of covalent bonds between amino groups, etc.) and enzymes, it does not form functional groups on the surface of the fiber through modification of the fiber surface, etc. It means that no covalent bond is formed.

The antifouling coating composition of the present invention can obtain improved antifouling performance as compared with the conventional antifouling enzyme-substrate complex which was restricted by industrial application due to the activity of the antifouling enzyme too low, thereby increasing the industrial applicability, and supporting the support having low cost. Economic problems can also be solved.

In addition, if it is mixed with various polymer materials widely used as paint materials and developed and used as paint materials, it can be effectively applied to various industrial fields (sea port industry, fluid transportation industry, water treatment industry, etc.) where antifouling materials are used. Therefore, it will be possible to use as a material to replace the existing chemical antifouling substances that are limited use due to environmental pollution.

1 is a schematic diagram showing a process for producing polyaniline nanofibers.
2 is a schematic diagram showing a step of immobilizing antifouling enzymes on a fiber matrix.
FIG. 3 is a SEM photograph of polyaniline nanofibers (PANFs) prepared in Preparation Example 1. FIG.
Figure 4 is a perspective view and a cross-sectional view of the polyaniline nanofiber matrix composite of the antifouling enzyme three-dimensional network structure according to the present invention.
5 is a polymer nanofiber complex (EA-AC / PANF) adsorbed by antifouling enzymes using UV / Vis-spectrophotometer (EA-AC / PANF), a polymer nanofiber composite (EAC-AC / PANF) cross-linked after adsorption of antifouling enzymes After the adsorption and precipitation, the initial activity of the crosslinked polymer nanofibers (EAPC-AC / PANF) composite is measured.
Figure 6 is a graph of the stability results confirmed by measuring the initial activity of antifouling enzyme-polymer nanofibers.
Figure 7 is a paper tape sample (A) untreated, paper tape sample (B) coated only latex paint, and antifouling enzyme-polymer nanofiber composite and latex paint mixture to confirm the antifouling effect of the antifouling coating composition It is the photograph which prepared the paper tape sample (C) which apply | coated the antifouling coating composition using this.
FIG. 8 is an antifouling using a paper tape sample (A) which is not treated at all, a paper tape sample (B) coated only with a latex paint, and an antifouling enzyme-polymer nanofiber composite and a latex paint mixture under the condition of contamination. It is an image when the surface of the paper tape sample (C) which apply | coated the coating composition was observed with the optical microscope.

Hereinafter, the present invention will be described in more detail.

As described above, the antifouling compositions containing antifouling enzymes exhibited antifouling effects, but due to their excessively low activity, antifouling performance was notably low when applied to actual industrial applications. There was a problem falling.

Therefore, in the present invention, the crosslinked antifouling enzyme aggregate and the crosslinked antifouling enzymes surrounding the surface of the 3D network fiber are supported in the pores of the porous matrix including the 3D network fibers and the diameter is larger than the inlet size of the pores. An antifouling coating composition comprising an antifouling enzyme-containing three-dimensional network structure including a shell containing the shell; Through this, the antifouling coating composition of the present invention can obtain improved antifouling performance as compared with the conventional antifouling enzyme-substrate complex which was restricted by industrial application due to the activity of the antifouling enzyme so low, as well as increasing the industrial applicability, manufacturing cost Inexpensive supports also solve economic problems.

First, the fiber matrix complex of the antifouling enzyme three-dimensional network structure included in the coating composition of the present invention will be described. First, the complex of the present invention comprises the steps of: (1) adding an antifouling enzyme to a porous matrix comprising three-dimensional network fibers; (2) adding a precipitation agent to the matrix to prevent leakage of antifouling enzymes added into the pores of the matrix; And (3) forming an enzyme aggregate by adding a crosslinking agent to form crosslinks between the antifouling enzymes, thereby preparing an antifouling enzyme-fiber matrix complex.

First, as step (1), the antifouling enzyme is adsorbed and / or covalently bonded to the porous matrix including the three-dimensional network fibers. If the surface of the three-dimensional network fiber contains a functional group capable of covalently binding to the antifouling enzyme, it is adsorbed if a covalent bond occurs between the antifouling enzyme and the fiber.

Fibers used in the present invention can be used without limitation as long as the three-dimensional network is formed during the spinning can form voids between the fibers and fibers, polyvinyl alcohol, polyacrylonitrile, nylon, polyester, Polyurethane, polyvinyl chloride, polystyrene, cellulose, chitosan, polylactic acid, polylactic-co-glycolic acid, polyglycolic acid polycaprolactone, collagen, polypyrrole, polyaniline and poly (styrene-co-maleic anhydride) It may be any one or more selected from the group consisting of, more preferably in consideration of cost and efficiency it is advantageous to use polyaniline fibers, but is not limited thereto.

The kind of spinning for producing the fiber can also utilize a conventional polymerization and / or spinning process if it has a three-dimensional network structure between the fiber and the fiber, and both electrospinning and melt spinning can be used. The diameter of the fibers can also be applied to both nanofibers and microfibers, but it may be more advantageous to use nanofibers in view of the size of the enzyme aggregates described below and the size of pores formed between the fibers. However, carbon nanotubes do not fall within the scope of the present invention because they cannot form a three-dimensional network even if they are made of fibers.

The porous matrix according to one embodiment of the present invention may include a part or all of the three-dimensional network fibers, and the porosity refers to a space (gap) between the fibers and the fibers forming the three-dimensional network.

The fibers of the present invention form a network of three-dimensional networks through which a fiber matrix structure can be made, and the matrix structure can be an amorphous structure in which the fibers are intricately intertwined.

The antifouling enzyme included in the present invention can be used without limitation as long as it can be used in a paint composition for antifouling, preferably, acylase, lactonase, protease, peroxidase, aminopeptidase, phosphatase, transamina An azede, serine-endopeptidase, cysteine-endopeptidase and metalloendopeptidase can be used alone or in combination.

On the other hand, the fiber matrix of the present invention can be applied to a fiber that substantially includes or does not contain any functional groups (eg, amino groups, carboxyl groups) that can be covalently bonded to the antifouling enzymes on the surface thereof. It may be covalently bonded to one another or adsorbed between the surface of the fiber or the void (space) formed between the fiber and the fiber. Therefore, if the antifouling enzyme is simply adsorbed on the surface of the matrix and the external impact such as water washing is applied, most of the adsorbed antifouling enzymes are separated from the matrix, and as a result, the fixation rate of the enzyme is significantly reduced.

In step (2), the matrix is subjected to enzyme precipitation in order to prevent the outflow of adsorbed and / or covalently bound enzymes in the pores of the matrix. The adsorbed enzyme is so small that it is hardly visible to the naked eye. When the precipitating agent is added to the matrix to precipitate the adsorbed enzyme, the adsorbed enzymes are aggregated with each other and become large in size. Eventually, the adsorbed enzyme is precipitated between pores (spaces) . This allows the formation of crosslinked antifouling enzyme aggregates having a diameter larger than the inlet size of the inter-fiber pores, and if the precipitation agent is not added, the aggregate size becomes smaller than the pore inlet size, which facilitates the subsequent integration. It breaks out between the interfiber voids.

The precipitating agent that can be used at this time can be used without limitation as long as it can precipitate the enzyme with little effect on the activity of the enzyme. Preferably, the precipitating agent is selected from the group consisting of methanol, ethanol, 1-propanol, 2-propanol, But are not limited to, acetone, polyethylene glycol, ammonium sulfate, sodium chloride, sodium sulfate, sodium phosphate, potassium chloride, potassium sulfate, potassium phosphate and aqueous solutions thereof.

Next, in step (3), a crosslinking agent is added to form crosslinks between the antifouling enzymes to form an enzyme aggregate. Specifically, since the size of the antifouling enzyme precipitated is relatively small compared to the size of the pores (space) formed between the fiber and the fiber, the antifouling enzyme precipitated by external impact such as water washing may flow out of the pore. However, when crosslinking is formed between the precipitated antifouling enzyme and the antifouling enzyme by adding a crosslinking agent, the crosslinked antifouling enzymes form an aggregate, and the antifouling enzyme aggregate thus formed almost fills the inside of the pores. As a result, the size of the antifouling enzyme aggregate formed is larger than that of the inlet of the pores, so that the cross-linked enzyme aggregate does not easily flow out of the pores even by an external impact such as washing with water. As a result, since the antifouling enzyme assembly is located in the pores over time, even if a direct binding relationship such as covalent bond between the fiber and the antifouling enzyme is not formed, the antifouling enzyme assembly may be provided in the fiber matrix for a long time. In addition, since the complex formed between the antifouling enzyme-3D network fiber matrix has a much higher amount of fixed enzyme than the conventional complex, the performance of the conventional complex is improved even when it is used in a biosensor or a biofuel cell. It can be significantly improved compared to.

On the other hand, the effect of adding the crosslinking agent after the treatment with the precipitating agent is remarkably enhanced as compared with the case where only the crosslinking agent is added. When the cross-linking agent is added after adsorption of the antifouling enzyme, the concentration of the antifouling enzymes becomes equal to the surrounding concentration even if the inside of the pores formed between the fibers and the fibers is not filled or the filling of the housework. As antifouling enzymes at the same concentration crosslink, they do not form a larger mass than pores in the pores in the nanofibers, and the crosslinked antifouling enzymes are likely to leak out during washing. Covalent bonds, except for covalently bound antifouling enzymes, do not form a large mass and are more likely to be washed later.

However, in the present invention, enzymes are forced to more closely fill pores in nanofibers through precipitation after adsorption of antifouling enzymes, and the enzymes filled in the cooperatives form a large mass through crosslinking. As a result, losses in the washing process are expected to be less than EAC.

Furthermore, in the case of polyaniline nanofibers having few functional groups capable of covalently binding to antifouling enzymes on the surface of the fiber, antifouling enzymes are very hard to be immobilized on the surface of the fiber. However, in the present invention, since the antifouling enzymes deposited on the surface of the fiber are crosslinked to form a shell surrounding the surface of the fiber, even if a covalent bond is not substantially formed between the fiber and the antifouling enzyme like a hot dog, Large amounts of antifouling enzymes can form and form a shell to be immobilized.

The crosslinking agent that can be used in the present invention can be used without limitation as long as it can form crosslinking between enzymes without inhibiting the activity of antifouling enzymes, but is preferably diisocyanate, dianhydride, diepoxide, dialdehyde At least one selected from the group consisting of diimide, 1-ethyl-3-dimethyl aminopropylcarbodiimide, glutar dialdehyde, bis (imido ester), bis (succinimidyl ester) and diacid chloride Compounds may be used, and more preferably, glutar dialdehyde may be used, but is not limited thereto. It will be apparent to those skilled in the art to use without limitation crosslinking agents known in the art.

On the other hand, after the step (3), preferably the step of removing the added cross-linking agent and precipitation agent by washing the antifouling enzyme three-dimensional network fiber matrix complex may be further performed. In the case of the matrix complex prepared by the conventional manufacturing method during the washing step, the antifouling enzyme immobilized between the pores is substantially leaked out of the pores, but the antifouling enzyme-3D network fiber matrix complex prepared according to the present invention is inside the pores. Since the size of the antifouling enzyme aggregate formed in the pores is larger than the inlet of the pores, the antifouling enzyme aggregates can be fixed inside the pores without being leaked to the outside despite the washing process.

The fiber matrix composite of the antifouling enzyme-three-dimensional network structure of the present invention prepared through this is supported in the pores of the porous matrix including the three-dimensional network fibers, and the crosslinked antifouling enzyme aggregate having a diameter larger than the inlet size of the pores; And a shell comprising crosslinked antifouling enzymes surrounding the surface of the three-dimensional network fiber.

Figure 4 is a perspective view and a cross-sectional view of the polyaniline nanofiber matrix composite of the antifouling enzyme three-dimensional network structure according to an embodiment of the present invention. In this case, a crosslinked enzyme aggregate having a diameter larger than the inlet size of the pores is carried inside the pores of the fiber matrix of the three-dimensional network structure. In addition, a shell containing an antifouling enzyme aggregate surrounds the surface of the three-dimensional network fiber. 4 is a cross-sectional view of the shell of the shell including the enzyme aggregate covering the surface of the three-dimensional network fiber as if the covalent bond is not substantially formed between the fiber and the enzyme aggregate forming the shell. Shells containing enzyme aggregates, such as hot dogs, surround the surface of three-dimensional network fibers.

As a result, the antifouling enzyme-three-dimensional network fiber matrix complex of the present invention has a sufficiently large size of the antifouling enzyme complex formed inside the pore so as not to flow out from the pores formed between the fiber and the fiber, compared to the conventional antifouling enzyme complex. Remarkably large amounts of antifouling enzymes can be immobilized on the matrix to form complexes. In other words, the size of the antifouling enzyme complex formed inside the pore is larger than the size of the inlet of the pore, which is a passage that flows out from the pores formed between the fibers and the outside, so that the antifouling enzyme complex may be formed even when there is an external stimulus such as water washing. It can be provided in the interior of the complex, which allows the complex to be maintained for a long time without forming a direct binding relationship between the antifouling enzyme and the matrix.

Furthermore, in the case of polyaniline nanofibers having few functional groups capable of covalently binding to antifouling enzymes on the surface of the fiber, antifouling enzymes are very hard to be immobilized on the surface of the fiber. However, in the present invention, since the antifouling enzymes deposited on the surface of the fiber are crosslinked to form a shell surrounding the surface of the fiber, even if a covalent bond is not substantially formed between the fiber and the antifouling enzyme like a hot dog, Large amounts of antifouling enzymes can form and immobilize shells, which can form multilayers.

Next, the coating composition including the antifouling enzyme-three-dimensional network fiber matrix complex described above will be described. The paint that can be used in the present invention can be used without limitation so long as it is a kind that can be used conventionally. For example, synthetic resin paints such as keshu-based, oil-based enameled natural fibers, nitrocellulose, alkyd resins, vinyl resins, polyester resins, epoxy resins, emulsion-based (eg vinyl acetate groups), water-soluble baking resin-based paints, plastics Stysol paint, powder paint and latex paint can be used without limitation. The composite of the present invention may be included in 0.001 to 50% by weight relative to 100% by weight of the coating composition. In addition, it will be apparent to those skilled in the art that the components of the coating composition, other than the antifouling enzyme-three-dimensional network fiber matrix complex of the present invention, may use materials used in conventional antifouling compositions and antifouling coating compositions.

In general, when untreated natural antifouling enzymes are mixed with a coating material, the loss of enzymatic activity rapidly progresses, and the antifouling performance is notably low, and the stability is low, and thus the antifouling performance is lost in a short time. However, by immobilizing antifouling enzymes in the three-dimensional fiber matrix complex, it was confirmed through application experiments that a large amount of antifouling enzymes showed high antifouling activity and high antifouling activity even if mixed with paint material. In addition, the synthesized antifouling enzyme composition was evenly dispersed in an aqueous solution and did not sink to the bottom of the coating composition, but was evenly dispersed in a thinly coated coating composition and an external contact site, thereby exhibiting an antifouling effect. As a result, the antifouling enzyme exposed to the outside does not easily decrease its activity through immobilization, and shows a high activity for a long time, so that the effect of the antifouling coating composition can be maintained for a long time.

The paint composition of the present invention will be applicable to the surface of any object that can be applied paint. For example, in the sea port business, it can show antifouling function by applying to bottom, deck, propeller, etc.In the business of transporting fluids, the operation cost is increased due to the impediment of fluid flow through the application of pipes and connection parts of various materials. Can be prevented. In addition, in the water treatment industry (water quality management, seawater desalination, etc.), the efficiency of water quality management can be increased by applying to various reactor inner surfaces such as water tanks, reactors, aerators for aeration, and impellers for mixing. Using the proven paint material, it can be used for artificial organs inserted into the human body and medical equipments inserted and inserted into the human body, so that it can function without deterioration of performance for a long time.

Preparation Example Preparation of Polyaniline Nanofiber Matrix of 3D Network Structure

In the same manner as in Figure 1 was prepared a polyaniline nanofiber matrix of a three-dimensional network structure.

Specifically, the electrically conductive nanofibers polyaniline nanofibers for antifouling enzyme fixation were prepared through oxidative polymerization using ammonium persulfate as an initiator. The oxidation polymerization used a rapidly mixing reaction method that controls the amount of ammonium persulfate to prevent the polyaniline overgrowth. Ammonium persulfate was prepared by dissolving 0.1 M in 1 M HCl solution. Add 1.5 ml of aniline to 8.5 ml of 1 M HCl and mix well. 10 ml of the prepared ammonium persulfate solution was added to a 10 ml solution containing aniline and HCl and mixed well. The well mixed solution was stirred at 200 rpm at room temperature for 24 hours. After the polymerization process, the solution was centrifuged, the supernatant was discarded, distilled water was put in, and the mixture was well stirred and washed several times before being stored at 4 ° C. The polyaniline nanofibers prepared were synthesized by controlling the amount of ammonium persulfate to prevent overgrowth. Nanofibers are intricately connected like corals to form a three-dimensional network structure, and depending on the concentration of aniline, the nanofibers have many pores in the inner part as well as pores in the outer part of the nanofibers. The pores of the nanofibers (spaces formed between fibers) play a large role in antifouling enzyme immobilization. The Brunauer-Emmett-Teller equation (BET) surface area of the polyaniline nanofibers was 58.4 m ² / g and the average pore diameter was 12.26 nm. Total pore volume was 0.179 cm ^{3 /} g (p / p0 = 0.990).

<Comparative Example 1> Preparation of antifouling enzyme-polymer nanofiber complex according to antifouling enzyme fixation method (EA) using antifouling enzyme adsorption method

The polyaniline polymer nanofibers prepared in the preparation example and antifouling enzymes (acylase, acylase, AC) were mixed to synthesize polymer nanofiber composites having enzymatic adsorption. Specifically, 2 mg of polyaniline polymer nanofibers prepared in the above preparation example and 1 ml of antifouling enzyme were mixed in a glass vial of 4 ml, wherein the antifouling enzyme was prepared at a concentration of 10 mg / ml in 100 mM phosphate buffer solution. Used. The prepared antifouling enzyme-polymer nanofiber mixture was stirred at 150 rpm using a stirrer for 2 hours at room temperature. Through this process, antifouling enzymes adsorb to the pores of polymer nanofibers and the surface of nanofibers. To remove unattached antifouling enzymes and capping the remaining unreacted antifouling enzymes on the polymer nanofibers, remove the supernatant after centrifugation and stir at 200 rpm using a stirrer for 30 minutes using 100 mM Tris buffer. I let you. After the completion of the capping reaction, it was washed with 100 mM phosphate buffer to remove the remaining Tris buffer.

<Comparative Example 2> Preparation of antifouling enzyme-polymer nanofiber complexes according to the antizyme adsorption and crosslinking (EAC) method using crosslinking method after adsorption of antifouling enzyme

The polyaniline polymer nanofibers of the preparation example and antifouling enzymes (acylase, acylase, AC) were mixed to synthesize the polymer nanofiber composites in which antifouling enzymes were adsorbed and crosslinked after adsorption. Specifically, 2 mg of polyaniline polymer nanofibers prepared in the above preparation example and 1 ml of antifouling enzyme were mixed in a glass vial of 4 ml, wherein the antifouling enzyme was prepared at a concentration of 10 mg / ml in 100 mM phosphate buffer solution. Used. The prepared antifouling enzyme-polymer nanofiber mixture was stirred at 150 rpm using a stirrer for 2 hours at room temperature. Through this process, antifouling enzymes adsorb to the pores and surfaces of polymer nanofibers. Then, in order to induce crosslinking through covalent bonds between adsorbed antifouling enzymes, glutaaldehyde, a crosslinking agent, was added to 0.5% of the total volume. Stir at 50 rpm using a rocker for 17 hours at 4 ° C for sufficient crosslinking. To remove unattached antifouling enzymes and capping the remaining unreacted antifouling enzymes on the polymer nanofibers, remove the supernatant after centrifugation and stir at 200 rpm using a stirrer for 30 minutes using 100 mM Tris buffer. I let you. After the completion of the capping reaction, it was washed with 100 mM phosphate buffer to remove the remaining Tris buffer.

<Example 1> Preparation of antifouling enzyme-polymer nanofiber composite according to antifouling enzyme fixing method (Enzyme adsorption, precipitation and crosslinking, EAPC) using crosslinking method after adsorption and precipitation of antifouling enzyme

The polyaniline polymer nanofibers of the Preparation Example were mixed with antifouling enzymes (acylase, acylase, AC) to synthesize polymer nanofiber composites in which antifouling enzymes were adsorbed, precipitated, and crosslinked. Specifically, 2 mg of polyaniline polymer nanofibers prepared in the above preparation example and 1 ml of antifouling enzyme were mixed in a glass vial of 4 ml, wherein the antifouling enzyme was prepared at a concentration of 10 mg / ml in 100 mM phosphate buffer solution. Used. The prepared antifouling enzyme-polymer nanofiber mixture was stirred at 150 rpm using a stirrer for 2 hours at room temperature. Through this process, antifouling enzymes adsorb to the pores of the polymer nanofibers. Then, ammonium sulfate solution, which is a precipitation agent, is added so that the concentration of ammonium sulfate in the solution is 45% (v / v), and the ammonium sulfate solution is dissolved in distilled water so that the composition of the ammonium sulfate solution is 60% (w / v). do. The solution mixed with the precipitation agent was stirred at 150 rpm at room temperature using a stirrer for 30 minutes. Then, in order to induce crosslinking through covalent bonds between the antifouling enzymes deposited after adsorption, a crosslinking agent, glutaaldehyde, was added to 0.5% of the total volume. For sufficient crosslinking, the mixture was stirred at 50 rpm using a rocker for 17 hours at 4 ° C. To remove unattached antifouling enzymes and capping the remaining unreacted antifouling enzymes on the polymer nanofibers, remove the supernatant after centrifugation and stir at 200 rpm using a stirrer for 30 minutes using 100 mM Tris buffer. I let you. After the completion of the capping reaction, it was washed with 100 mM phosphate buffer to remove the remaining Tris buffer. The antifouling enzyme prepared through this process is adsorbed and precipitated, and then the cross-linked polymer nanofiber composite (EAPC-AC / PANF) can be stored until use without storage loss if stored at 4 ° C. until use.

Example 2 Antifouling enzyme activity and stability measurement of antifouling enzyme-polymer nanofiber complex.

Antifouling enzyme-adsorbed polymer nanofiber composite (EA-AC / PANF) prepared in Example 1 and Comparative Examples 1 and 2, cross-linked polymer nanofiber composite (EAC-AC / PANF) after antifouling enzyme was adsorbed The antifouling enzyme activity of the crosslinked polymer nanofiber (EAPC-AC / PANF) complex after the adsorption and precipitation of the antifouling enzyme was measured using a UV / Vis-visspectrophotometer (Shimadzu UV-2450).

Specifically, the antifouling enzyme-polymer nanofiber complex stored in 4 was prepared by diluting to 0.05 mg / ml based on the polymer nanofiber weight using 100 mM phosphate buffer. In addition, 10 mM N-acetyl-L-methionine is prepared as a reaction solution, and the solution composition is used by dissolving 1.9125 mg of N-acetyl-L-methionine in 100 mM phosphate buffer. do. The activity of the antifouling enzyme was measured using the two solutions thus prepared. Absorbance at 238 nm using a UV / Vis-spectrophotometer, mixed with 250 ul antifouling enzyme-polymer nanofiber complex and 750 ul 10 mM N-acetyl-L-methionine solution to measure antifouling enzyme activity Was measured.

5 is a polymer nanofiber composite (EA-AC / PANF, Comparative Example 1) adsorbed antifouling enzyme using a UV / Vis-spectrophotometer, cross-linked polymer nanofiber composite (EAC-AC / PANF after adsorptive enzyme adsorption) , Comparative Example 2), the antifouling enzyme was measured by the adsorption, precipitation of the cross-linked polymer nanofibers (EAPC-AC / PANF, Example 1) measured the initial activity (initial rate) of the complex, Figure 6 is a graph It is shown. The activities of EA, EAC and EAPC are 51.56, 51.87 and 185.19 mM / min, respectively. EAPC activity was found to be 2.7 times greater than EA or EAC. This is due to the antifouling enzymes precipitated using the precipitation method, the larger amount of antifouling enzymes are immobilized and the higher activity is obtained by using a crosslinking agent.

5 is a table showing the experimental results of FIG. 6, wherein the results of measuring the activity of EA-AC / PANF, EAC-AC / PANF, and EAPC-AC / PANF samples are summarized in the table of FIG. 5, and of FIG. 6. The graph on the left shows this, and the graph on the right shows the relative value of the activity when the value of activity on the first day of confirmation of activity is 100.

As can be seen from FIG. 6, the antifouling enzyme was adsorbed and precipitated, and the crosslinked polymer nanofibers (EAPC-AC / PANF) complex was confirmed to have high stability as well as higher activity than other materials. The antifouling enzyme-polymer nanofiber complex was measured at 200 rpm using a stirrer in a 100 mM phosphate buffer at room temperature, and the activity was measured. The activity half-life of EA and EAC was less than 3 days and 4 days, respectively, but EAPC The activity has been stabilized by more than 90% of activity up to 150 days.

These results are expected to show sustained antifouling performance when antifouling enzymes are used after the adsorption and precipitation of crosslinked polymer nanofibers (EAPC-AC / PANF) complexes modified and modified with antifouling materials.

Example 4 Antifouling enzyme activity and stability of antifouling coating composition using antifouling enzyme-polymer nanofiber complex and latex paint mixture.

Preparation of the antifouling coating composition using the antifouling lake-polymer nanofiber (EAPC-AC / PANF) composite of the Example 1 and latex (latex) and the contamination is induced to confirm the antifouling effect to form a biofilm (Biofilm) Confirmation experiment was carried out under the condition that the effect was observed through a microscope.

Specifically, a paper tape capable of applying paint was prepared to make the surface rough with sandpaper so that the paint could be applied well. In order to confirm the antifouling effect of the antifouling coating composition, a paper tape sample (A) which was not treated, a paper tape sample (B) coated only with a latex paint, and an antifouling enzyme-polymer nanofiber composite (Example 1) and latex A paper tape sample (C) coated with an antifouling coating composition using a paint mixture was prepared and used for the experiment (FIG. 7). The composition of the antifouling coating composition using the antifouling enzyme-polymer nanofiber composite and the latex paint mixture was 0.5 mg of the antifouling enzyme-polymer nanofiber composite solution based on the weight of the polymer nanofiber, and the latex of acrylamide resin. 2 ml of paint was prepared by mixing

Each sample described above was stored in a solution containing 40 ml of synthetic wastewater and 1 ml of microbial (Pseudomonas Aureginosa) in a condition in which biofilms causing contamination and biofouling were well formed, followed by stirring at 200 rpm at room temperature. The reaction was carried out under the following conditions. The surface of the sample thus stored was observed through an optical microscope and the image magnified 100 times can be confirmed through FIG. 8.

As shown in FIG. 8, the results of the experiment using the sample prepared in FIG. 7, in the case of the paper tape sample A which was not treated, small spherical spot-shaped aggregates began to be observed after 1 day. After 3 days, lightly observed aggregates were observed in several places, and after 5 days, the aggregates gathered around were connected. This phenomenon appeared to be worse at 7 days, the formed biofilm was confirmed to continue to grow over time.

In the case of the paper tape sample (B) coated only with latex paint, the same image as when the experiment was first started was confirmed until day 3, and after 5 days, small spherical spot-like aggregates began to be observed. Lightly observed aggregates began to be concentrated in several places. These results showed the antifouling effect to some extent by applying only latex paint on the surface of the material, but after one week, it was observed that the aggregates started to be observed, which could not prevent the biofouling phenomenon continuously.

In the case of the paper tape sample (C) coated with the antifouling coating composition using the antifouling enzyme-polymer nanofiber composite and the latex paint mixture, the surface was not contaminated for 4 weeks while maintaining the surface as it was originally observed. Could. This prevents the formation of biofilm on the surface by the anti-biofouling effect of the antifouling enzyme-polymer nanofiber complex included in the antifouling coating composition, which is excellent in all fields where an actual antifouling coating composition can be applied. It can be confirmed that the antifouling action.

Since the present invention has excellent antifouling effect, it can be usefully used in the field of antifouling paint composition.

Claims (10)

Shell containing crosslinked antifouling enzymes contained within the interfiber pores of a matrix comprising three-dimensional network fibers and having a diameter larger than the inlet size of the pores and the crosslinked antifouling enzymes surrounding the surface of the three-dimensional network fibers. An antifouling paint composition comprising; fiber matrix complex of the enzyme three-dimensional network structure comprising a. The antifouling coating composition according to claim 1, wherein the three-dimensional network fibers are microfibers or nanofibers. 3. The method of claim 2,
The three-dimensional network fibers are polyvinyl alcohol, polyacrylonitrile, nylon, polyester, polyurethane, polyvinyl chloride, polystyrene, cellulose, chitosan, polylactic acid, polylactic-co-glycolic acid, polyglycolic acid Polycaprolactone, collagen, polypyrrole, polyaniline and poly (styrene-co-maleic anhydride) at least one selected from the group consisting of antifouling paint composition. The method of claim 1,
The shell is an antifouling paint composition, characterized in that it comprises a multilayer enzyme. The method of claim 1,
The antifouling enzyme is any selected from the group consisting of acylase, lactonase, protease, peroxidase, aminopeptidase, phosphatase, transaminase, serine-endopeptidase, cysteine-endopeptidase and metallo-endopeptidase. Antifouling paint composition, characterized in that one or more. The method of claim 1,
Antifouling coating composition comprising 0.001 to 50% by weight of the complex. (1) adding antifouling enzymes to a matrix comprising three-dimensional network fibers;
(2) adding a precipitation agent to the matrix to prevent leakage of antifouling enzymes added into the pores of the matrix; And
(3) preparing an antifouling enzyme-fiber matrix complex by adding a crosslinking agent to form an enzyme aggregate to form crosslinking between the antifouling enzymes; And
(4) A method for producing an antifouling paint composition comprising the step of adding the composite to the paint. The method of claim 7, wherein
The precipitation agent is methanol, ethanol, 1-propanol, 2-propanol, butyl alcohol, acetone, polyethylene glycol, ammonium sulfate, sodium chloride, sodium sulfate, sodium phosphate, potassium chloride, potassium sulfate, potassium phosphate and aqueous solutions thereof alone Or a method for producing a paint composition for antifouling, characterized in that mixed. The method of claim 7, wherein
The crosslinking agent is diisocyanate, dianhydride, diepoxide, dialdehyde, diimide, 1-ethyl-3-dimethyl aminopropylcarbodiimide, glutar dialdehyde, bis (imido ester), bis (succinimi A method for producing an antifouling coating composition, comprising any one or more compounds selected from the group consisting of dill esters) and diacid chlorides. The method of claim 7, wherein
Method of producing a paint composition for antifouling, characterized in that further comprising the step of removing the precipitation agent and crosslinking agent between the step (3) and (4).

Images