KR101652263B1 - Adhesive Protein Comprising Antimicrobial Peptide and Antimicrobial Coating Composition Comprising the Same - Google Patents

Abstract

The present invention relates to an antibacterial adhesive protein in which an antibacterial peptide is fused at the end of a self-adhesive adhesive protein, an antibacterial coating composition comprising the antibacterial adhesive protein, and an antibacterial film coated with the antibacterial coating composition. The antimicrobial adhesive protein of the present invention and the antimicrobial coating composition containing the antimicrobial adhesive protein of the present invention can be coated on the surface of a liquid crystal protective film, a food packaging material, a medical instrument, etc. of a mobile phone to ensure safety against harmful microorganisms. By maintaining the antimicrobial activity constantly, it is possible to prevent the antibacterial activity from being reduced by the elution of the antimicrobial substance.

Description

[0001] The present invention relates to an adhesive protein comprising an antibacterial peptide and an antibacterial coating composition comprising the same. ≪ Desc / Clms Page number 1 >

The present invention relates to an adhesive protein comprising an antibacterial peptide, an antibacterial coating composition comprising the adhesive protein, an antibacterial film coated with the antibacterial coating composition and a coating method. More specifically, the present invention relates to an adhesive protein and an antibacterial coating composition comprising the same, wherein the antibacterial peptide is fused between the C-terminal or N-terminal or hybrid adhesive protein of the adhesive protein.

A variety of daily necessities such as electronic products used in everyday life are easily exposed to harmful bacteria or viruses due to dust, fingerprints, saliva, and grease. Furthermore, due to the high interest in environmental hygiene and the upgrading of lifestyle, production of antibacterial properties of petrochemical materials used for industrial products such as food packaging materials, storage containers, toothbrushes, chopping boards, And demand is increasing.

Many antimicrobial compositions have been commercialized. However, since most of the antimicrobial compositions are mixed with liquid or polymeric substances (see Patent Document 1), the antimicrobial component is gradually removed during use, and the persistence of the antimicrobial activity is raised. In addition, conventional antimicrobial techniques have mainly been using silver nanoparticles or inorganic particles, and there has been controversy about human hazards due to dislocation of fine particles, and thus there is an increasing demand for environmentally friendly antibacterial technology. Organic antimicrobial substances such as quaternary ammonium salts, triazine, and benzimidazole, and natural antimicrobial substances such as essential oil and extract of horseradish have been added to the polymer substance simply by adding an antimicrobial substance to the polymer substance, (Patent Document 2 and Patent Document 3), technical limitations such as the self-toxicity of the organic antimicrobial substance, the elution of the antimicrobial substance and the reduction of the antibacterial activity thereof have been pointed out (Non-Patent Document 1) When injected or extruded, the antimicrobial material may cause pyrolysis and yellowing. Furthermore, when the antimicrobial film is packaged together with the food, a serious side effect such as causing tolerance due to elution of the antimicrobial substance mentioned above may occur.

Methods for immobilizing the antimicrobial agent or increasing the antimicrobial persistence by developing a substance that prevents the chemical structure of the substance from killing the bacteria or preventing the bacteria from adhering to the surface have been studied (Patent Document 4, Non-Patent Document 2 And non-patent document 3). In addition, antimicrobial techniques including two or more antimicrobial agents have been developed to give a broad antibacterial power (Patent Document 5).

Accordingly, the present invention provides a water-soluble coating composition composed of a pure protein and having an adhesive strength, and having an antibacterial effect on various microorganisms. The antimicrobial composition according to the present invention has a strong antibacterial activity, It has excellent biodegradability and excellent environment-friendliness without using an organic solvent.

Korean Patent Publication No. 2003-0066424 Korean Patent Publication No. 2009-0024467 Korean Patent Publication No. 2008-0110578 Korean Patent Publication No. 2008-0039460 U.S. Published Patent No. 8,062,691

 K. Glinel et al., Acta Biomaterialia 8, 1670 (2012)  Mo S, Krunica, et al., J. Nat. Prod. 2009, 72 (11): 2043-5  Bin Wang, et al., J. Appl. Polym. Sci. 130 (5), p3489-3497, 2013

Disclosure of the Invention The present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide an antibacterial coating composition comprising the adhesive protein in which the antibacterial peptide is fused to the C- The purpose is to provide.

The present invention also relates to a method for chemically modifying tyrosine residues of an adhesive protein with 3,4-dihydroxyphenylalanine (DOPA), which can be coated with an antimicrobial surface by a simple method, The present invention also provides an environmentally friendly coating composition capable of sustaining a coating for a long period of time.

It is another object of the present invention to provide a photocurable antimicrobial coating composition which can be applied to various materials such as a liquid crystal film through mixing of a photocurable urethane acrylate and an antimicrobial peptide fusion adhesive protein.

Certain advantages and novel features of the present invention will become more apparent from the following detailed description and preferred embodiments with reference to the accompanying drawings.

The present invention provides an antibacterial adhesive protein to which an antibacterial peptide is genetically added to an adhesive protein capable of self-adhesion in a humid environment or a dry environment, and an antibacterial coating composition comprising the same.

Further, the present invention provides an antibacterial adhesive protein having an excellent antibacterial function by adding at least one antimicrobial peptide to the C-terminal or N-terminal of the adhesive protein.

According to one embodiment of the present invention, an antimicrobial coating composition capable of being simply coated on various surfaces can be provided by replacing the tyrosine portion of the antibacterial mussel adhesive protein to which the antibacterial peptide is added with DOPA.

The present invention also provides a UV light curable coating composition comprising a water soluble urethane acrylate and an adhesive protein to which an antimicrobial peptide has been added.

Hereinafter, the present invention will be described in detail.

The present invention provides an antimicrobial coating composition comprising an antimicrobial adhesive protein alone or a mixture of the antimicrobial adhesive protein and a photocurable urethane acrylate-based adhesive resin. The antimicrobial coating composition provided in the present invention is composed of an antibacterial adhesive protein functionalized with an antimicrobial peptide present naturally or artificially, and the basic structure of the antibacterial adhesive protein is composed of X-P-Y. X and Y are antimicrobial peptides, and P is an adhesive protein. Any suitable antimicrobial adhesive protein may be used in the coating compositions of the present invention.

The adhesive protein applied to the antimicrobial adhesive protein of the present invention includes all proteins capable of self-adhesion without limitation. Self-adhesive proteins include any protein that inherently has adhesive properties or is adhered via chemical modification. Examples of commercially available self-adhesive proteins include, but are not limited to MAPTrix ™, a mussel-derived recombinant adhesive protein marketed by Colodis Biosciences, Inc., North Ogsta, South Carolina. An example of an adhesive protein to which adhesiveness has been imparted through chemical modification is a protein modified with an acrylate. Examples include, but are not limited to, acrylated collagen or mussel adhesive proteins. One embodiment of the present invention provides an antimicrobial coating composition comprising an acrylated mussel adhesive protein.

MAPTrix (TM) provided in one embodiment of the present invention is a recombinantly functionalized mussel adhesive protein. In the present invention, the mussel adhesive protein may be used as such or may be a foot protein (FP-5) (SEQ ID NO: 10, 11, 12 or 13) (SEQ ID NO: 1), FP-3 (SEQ ID NO: 1), FP-3 described in SEQ ID NO: 2 (SEQ ID NO: 4), FP-4 (SEQ ID NO: 9), and fragments of the respective proteins can be used as the fusion protein fused with at least one second peptide. Preferably, the first peptide is FP-5 comprising the amino acid sequence of SEQ ID NO: 10, 11, 12 or 13 and the second peptide is FP-1 comprising the amino acid sequence of SEQ ID NO: 1, 2 or 3 to be. According to one embodiment of the present invention, the mussel adhesive protein preferably has an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 19, but is not limited thereto. In the present invention, the antimicrobial peptide is necessary to add an antibacterial function to the adhesive protein, preferably the mussel adhesive protein.

The antimicrobial peptide may be added at the C-terminal, N-terminal, or both, or between hybrid mussel adhesive proteins of a mussel adhesive protein by gene recombination technology. For example, an antimicrobial peptide may be added between FP-1 and FP-5 in the case of the fusion protein FP-151 having a structure in which one FP-5 is bound between two FP-1. In addition, different antimicrobial peptides can be added between both ends or the fusion protein. For example, the adhesive proteins comprising the antimicrobial peptides of the present invention can be used in combination with the alpha-helical 23 amino acid peptides Magainin or Dermaseptin isolated from the skin of the African frog Genopus laevis But are not limited to, antimicrobial peptides such as human defensin, cathelicidin LL-37, and histatin. The antimicrobial peptides of the present invention may be used alone or in combination.

The antimicrobial peptide fused to the adhesive protein in the present invention may be any natural peptide or artificially synthesized peptide without limitation. According to one embodiment of the present invention, the antimicrobial peptide exhibits an antimicrobial effect through a mechanism that disrupts the cell membrane of the microorganism or permeates the cell membrane to inhibit the metabolic function. According to another embodiment of the present invention, any antimicrobial peptide exhibiting an antimicrobial effect through a mechanism of destroying the cell membrane of a microorganism can be used in the present invention. Preferably, the antimicrobial peptide to be fused to the adhesive protein may be selected from among antimicrobial peptides effective against gram-positive bacteria as well as gram-negative bacteria. KLWKKWAKKWLKLWKA (SEQ ID NO: 20), FALALKALKKL (SEQ ID NO: 21), ILRWPWWPWRRK (SEQ ID NO: 22), AKRHHGYKRKFH (SEQ ID NO: 23), KWKLFKKIGAVLKVL (SEQ ID NO: 24), LVKLVAGIKKLKWK (SEQ ID NO: 25), IWSILAPLGTTLVKLVAGIGQQKRK 26), GIGAVLKVLTTGLPALISWI (SEQ ID NO: 27), SWLSKTAKKGAVLKVL (SEQ ID NO: 28), KKLFKKILKYL (SEQ ID NO: 29), GLKKLISWIKRAAQQG (SEQ ID NO: 30) and GWLKKIGKKERVGQHTRDATIQGLGIAQQAANVAATAR (SEQ ID NO: 31).

According to one embodiment of the present invention, there is provided a coating composition based on an antibacterial adhesive protein wherein different antibacterial peptides are added to the C-terminus and N-terminus of an adhesive protein in order to provide an antibacterial adhesive protein having a broad antibacterial effect. The present invention provides a broad range of antimicrobial coating compositions, preferably wherein the antimicrobial coating composition comprises a peptide having a peptide FALALKALKKL (SEQ ID NO: 21) at the C-terminus that is effective against Gram-negative bacteria and having an effect on gram- AKRHHGYKRKFH (SEQ ID NO: 23).

In the present invention, the antimicrobial peptide fusion mussel adhesive protein is converted to DOPA and further to DOPA quinone through chemical modification of the tyrosine residues in the constituent amino acids. The modified DOPA and DOPA quinone can play a very important role in adhesion to the surface have. According to one embodiment of the present invention, a chemical modification of the recombinant mussel adhesive protein can be carried out using a mushroom-derived tyrosinase enzyme capable of mediating such chemical modification. The chemically modified antimicrobial mussel adhesive protein can be attached to a variety of materials such as metals, plastics, and glass.

The concentration of the coating composition based on the antimicrobial adhesive protein can be adjusted according to the added antimicrobial peptide. Preferably, the concentration of antimicrobial adhesive protein in the antimicrobial coating composition is from 0.01 to 1% (wt / wt), more preferably from 0.01 to 0.3% (wt / wt).

The water-soluble acrylate-based antimicrobial coating composition mixed with the antimicrobial adhesive protein of the present invention is a UV light-curable antimicrobial coating agent. At this time, the antimicrobial adhesive protein and the acrylate can be mixed first. As the photoinitiator, benzoyl peroxide and Irgacure 184 may be used, but the present invention is not limited thereto. The weight ratio of the acrylate to the antibacterial adhesive protein is mixed at a ratio of 1: 0.01, preferably 1: 0.001.

According to one embodiment of the present invention, the acrylate used in the present invention may be any water soluble acrylate without any limitation. The antimicrobial coating composition provided in the present invention is composed of a water-soluble acrylate and an antibacterial adhesive protein selected from at least one of urethane acrylate, polyester acrylate and epoxy acrylate.

The present invention also provides an antibacterial functional film comprising a UV photocurable antimicrobial coating composition comprising an antibacterial adhesive protein.

The film used to produce the antimicrobial film coated with the antimicrobial composition may be a polyethylene or polypropylene film commonly used for food packaging, mobile phones, liquid crystal protection of smart phones, and the like, but is not limited thereto. The amount of the UV photocurable antimicrobial composition to be applied can be appropriately selected in consideration of the amount of the antibacterial adhesive protein, the kind of the coating object, and the like.

According to one embodiment of the present invention, an antimicrobial coating film can be prepared by applying a urethane acrylate containing an antibacterial adhesive protein to a polystyrene film, followed by photocuring. According to another embodiment of the present invention, the antimicrobial activity of the antimicrobial coating film can be confirmed by examining the antimicrobial activity of Gram-negative bacteria, for example, E. coli, on the uncoated surface and the coated surface.

The photocurable antimicrobial coating composition of the present invention, in which the antimicrobial adhesive protein alone or in combination with the above-mentioned antimicrobial adhesive protein and acrylate is coated on the surface of a liquid crystal protective film, a food packaging material, a medical instrument or the like of a mobile phone, There is an effect that can be secured. In particular, the antimicrobial adhesive protein of the present invention maintains antimicrobial activity on the basis of the strong adhesive force of the mussel adhesive protein, thereby preventing the antimicrobial activity from being reduced due to elution into the antimicrobial substance.

The above-described and other features and advantages of the present invention will be apparent upon consideration of the following embodiments and drawings provided for illustrative purposes.
1 is a diagram showing the basic structure of an antibacterial adhesive protein which is the core of an antibacterial coating composition.
2 is a photograph showing the antibacterial activity of an antibacterial adhesive protein fused with the antimicrobial peptide A against E. coli.
Fig. 3 is a photograph showing the antibacterial activity of the antibacterial adhesive protein fused with the antimicrobial peptide B against E. coli. Fig.
4 is a photograph showing the antibacterial activity of an antibacterial adhesive protein fused with the antimicrobial peptide C against E. coli.
5 is a photograph showing the antibacterial activity of an antibacterial adhesive protein fused with the antimicrobial peptide D against E. coli.
FIG. 6 is a photograph showing the antibacterial activity against E. coli on the surface after the mussel adhesive protein fused with the chemically modified antibacterial peptide A is coated on the polystyrene surface.
7 is a photograph showing the antimicrobial persistence of E. coli on the surface after the mussel adhesive protein fused with the chemically modified antimicrobial peptide A is coated on the polystyrene surface.
8 is a photograph showing antibacterial effect on an antimicrobial coating surface prepared by using peroxide as an antimicrobial coating composition composed of a water-soluble acrylate and an antibacterial adhesive protein.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Also, as disclosed in the present invention, all functionally equivalent products, compositions and methods are included within the scope of the present invention.

Example  1. Preparation of Expression Vector of Mussel Adhesion Protein Fused with Antibacterial Peptides

In order to prepare a mussel adhesive protein fused with an antimicrobial peptide, a genetic sequence to which a conventional antimicrobial peptide sequence was added at the C-terminal or N-terminal site of the mussel adhesive protein was designed and expressed in Novacell Technology (Korea) I commissioned the production of a vector. The constructed vector was transformed into E. coli BL21 (DE3), and the added sequence is shown in Table 1.

Fusion peptide Amino acid sequence Fusion site in mussel adhesive protein A KLWKKWAKKWLKLWKA (SEQ ID NO: 20) C-terminus B FALALKALKKL (SEQ ID NO: 21) N-terminal C ILRWPWWPWRRK (SEQ ID NO: 22) C-terminus D AKRHHGYKRKFH (SEQ ID NO: 23) C-terminus

Example  2. Preparation of Mussel Adhesion Protein Fused with Antibacterial Peptides

2.1. E. coli BL21 ( DE3 ) Culture

E. coli BL21 (DE3) was cultured in LB (5 g / L yeast extract, 10 g / L Tryptone and 10 g / L NaCl) medium. When the absorbance of the culture reached 0.6 at 600 ㎚, 1 mM to induce the expression of recombinant antimicrobial peptide fusion mussel adhesive protein. The E. coli BL21 (DE3) culture broth was centrifuged at 13,000 rpm for 4 to 10 minutes to obtain cell pellets, which were stored at -80 ° C.

2.2. Expression of Mussel Adhesion Protein Fused with Antibacterial Peptides

The cell pellet was diluted with 100 μg of SDS-PAGE buffer (0.5 M Tris-HCl, pH 6.8, 10% glycerol, 5% SDS, 5% β-mercaptoethanol, 0.25% bromophenol blue) and boiled at 100 ° C. for 5 minutes to denature . For SDS-PAGE, the sample was electrophoresed on a 15% SDS-polyacrylamide gel and protein bands were detected using Coomassie blue staining.

2.3. Purification of mussel adhesive proteins fused with antimicrobial peptides

The cell pellet obtained in Example 2.1 was stirred using a dissolution buffer (2.4 g / L sodium phosphate monobasic, 5.6 g / L sodium phosphate dibasic, 10 mM EDTA and 1% Triton X-100) To disrupt the cells. The lysate was centrifuged at 9,000 rpm for 20 minutes to obtain an insoluble protein aggregate containing mussel adhesive protein. The mussel adhesive protein fused with the antimicrobial peptide was extracted from the insoluble protein aggregate using 25% acetic acid and centrifuged at 9,000 rpm for 20 minutes to recover the supernatant containing the mussel adhesive protein. The recovered supernatant was raised to pH 12.8 with 10 N NaOH, and the supernatant was recovered by centrifugation under the same conditions. The supernatant was neutralized to pH 6-7 with acetic acid and then centrifuged under the same conditions to obtain a precipitate of the antibacterial peptide fusion mussel adhesive protein. The obtained precipitate was dissolved in an appropriate amount of purified water and then lyophilized to obtain an antifungal peptide fusion mussel adhesive protein freeze-dried product having a purity of 90% or more.

Example  3. Antibacterial activity of mussel adhesive protein with antimicrobial peptide fused

First, mussel adhesive proteins A, B, C and D, in which antimicrobial peptides were fused, were prepared by concentration. The concentration for the antimicrobial activity test was adjusted to 10-0.01 ㎎ / ㎖ by using PBS (phosphate buffered saline) buffer solution. Escherichia coli, which is a Gram-negative bacterium, was used as a test strain for the antimicrobial activity. Escherichia coli was cultured in LB medium at 37 DEG C and 150 rpm for an absorbance of 1.0. The E. coli culture broth was diluted with PBS to 10 ⁴ CFU / ml at an absorbance of 1.0, mixed with a pre-prepared antimicrobial peptide fusion protein and a sterilized tube at a ratio of 9: 1, incubated at 37 ° C for 1 hour Respectively. One hour later, 100 쨉 l of E. coli was collected from each tube, plated on an agar medium, and cultured for 24 hours under the same conditions.

As a result, as shown in Fig. 1 to Fig. 4, the antimicrobial peptide fusion mussel protein of the present invention showed an antibacterial effect of 99.99% as compared with the control group.

As shown in FIG. 3, the concentration effect of the antimicrobial adhesive was confirmed. As a result, it was confirmed that the effect of the antimicrobial adhesive was excellent even at a concentration of 1 mg / ml which is lower than the antibiotic concentration.

Example  4. Tyrosinase enzyme treatment for self-adhesion of antimicrobial protein

Each of the lyophilized antimicrobial peptide fusion mussel proteins was dissolved at a concentration of 1 mg / ml in 0.1 M sodium acetate buffer containing 25 mM ascorbic acid and 20 mM sodium borate, The oxygen in the solution was saturated. After that, tyrosinase was added at a concentration of 40 to 1 占 퐂, preferably 5 to 1 占 퐂 / 1 mg of the antibacterial adhesive protein, followed by stirring for 1 hour while injecting oxygen. After 1 hour, 5% acetic acid was added to terminate the chemical modification reaction. The reaction-completed mussel protein solution was lyophilized and obtained as a powder. Through the above process, the tyrosine residue of the antibacterial adhesive protein was modified to DOPA.

The antimicrobial activity of the enzyme - treated antimicrobial adhesive protein was examined in order to examine the effect of the self - adhesive force, surface coating test and enzyme treatment on the antimicrobial activity. First, an antimicrobial peptide fusion mussel protein A was prepared for each concentration. The concentration for the antimicrobial activity test was adjusted to 10-0.01 mg / ml using PBS buffer solution. The solution was spread on a plate made of polystyrene for cell culture as a whole, and the remaining solution was recovered. After drying for 24 hours under Laminar flow, the surface was washed three times with sterile PBS and dried again for 24 hours. When the surface was completely dried, the pre-cultured Escherichia coli test solution was diluted with PBS to 10 ⁴ CFU / ml, and then 1 ml each was dispensed onto the surface-coated plate. After incubation for 1 hour at 37 ° C in a thermo-hygrostat, 100 μl of each plate coliform solution was aliquotted onto an agar medium and incubated for 24 hours under the same conditions.

As a result, the antimicrobial peptide fusion mussel protein showed an antimicrobial effect of 99.99% as compared with the control group when the 0.1% protein solution was coated on the surface (FIG. 5). In order to test the antimicrobial persistence, the plate in which the E. coli test solution was dispensed was washed with the same procedure after one week, and the same test was repeated to confirm that the antibacterial activity was maintained (FIG. 6).

Example  5. Containing antibacterial adhesive proteins UV  Light-curing urethane Acrylate  Preparation of composition

1 g of urethane acrylate was dissolved in 0.5 to 1 ml, preferably 0.5 ml of ethanol, and then mixed with 0.5 ml of an aqueous ethanol solution in which 5 mg of antibacterial mussel protein A was dissolved. 0.2 ml of an acetone / ethanol solution in which 30 mg of benzyl peroxide was dissolved was added to the antibacterial mussel protein-acrylate mixture, and then exposed to two UV bulbs in a UV room (wavelength: 285 nm) heated to 90 ° C and subjected to a curing reaction for 10 minutes An antimicrobial film was prepared. The antibacterial activity of the prepared antimicrobial film was tested for E. coli according to the same method and procedure as in Example 3 above.

As a result, as compared with the uncoated control group (A) as shown in Fig. 8, E. coli growth on the photocurable acrylate-coated film surface (B, C) containing the antibacterial peptide fusion mussel adhesive protein was not observed I did.

<110> Kollodis Bioscience Co. Ltd. <120> Adhesive Protein Comprising Antimicrobial Peptide and          Antimicrobial Coating Composition Comprising the Same <130> 2014-dpa-0823 <160> 31 <170> Kopatentin 2.0 <210> 1 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> peptide peptide of the tandem repeat decapeptide derived from foot          protein 1 (FP-1, Mytilus edulis) <400> 1 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys   1 5 10 <210> 2 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> 2 times repeated sequence derived from foot protein 1 (FP-1,          Mytilus edulis) <400> 2 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro   1 5 10 15 Pro Thr Tyr Lys              20 <210> 3 <211> 60 <212> PRT <213> Artificial Sequence <220> <223> 6 times repeated sequence derived from foot protein 1 (FP-1,          Mytilus edulis) <400> 3 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro   1 5 10 15 Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys              20 25 30 Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr          35 40 45 Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys      50 55 60 <210> 4 <211> 39 <212> PRT <213> Artificial Sequence <220> Partial sequence of foot protein type 2 (FP-2, Mytilus          californianus) <400> 4 Glu Val His Ala Cys Lys Pro Asn Pro Cys Lys Asn Asn Gly Arg Cys   1 5 10 15 Tyr Pro Asp Gly Lys Thr Gly Tyr Lys Cys Lys Cys Val Gly Gly Tyr              20 25 30 Ser Gly Pro Thr Cys Ala Cys          35 <210> 5 <211> 52 <212> PRT <213> Artificial Sequence <220> Foot protein type 3 (FP-3, Mytilus edulis) <400> 5 Ala Asp Tyr Tyr Gly Pro Lys Tyr Gly Pro Pro Arg Arg Tyr Gly Gly   1 5 10 15 Gly Asn Tyr Asn Arg Tyr Gly Gly Ser Arg Arg Tyr Gly Gly Tyr Lys              20 25 30 Gly Trp Asn Asn Gly Trp Lys Arg Gly Arg Trp Gly Arg Lys Tyr Tyr          35 40 45 Glu Phe Glu Phe      50 <210> 6 <211> 46 <212> PRT <213> Artificial Sequence <220> Foot protein type 3 (FP-3, Mytilus galloprovincialis: mgfp-3A) <400> 6 Ala Asp Tyr Tyr Gly Pro Lys Tyr Gly Pro Pro Arg Arg Tyr Gly Gly   1 5 10 15 Gly Asn Tyr Asn Arg Tyr Gly Arg Arg Tyr Gly Gly Tyr Lys Gly Trp              20 25 30 Asn Asn Gly Trp Lys Arg Gly Arg Trp Gly Arg Lys Tyr Tyr          35 40 45 <210> 7 <211> 50 <212> PRT <213> Artificial Sequence <220> Foot protein type 3 (FP-3, Mytilus edulis: mefp-3F) <400> 7 Ala Asp Tyr Tyr Gly Pro Asn Tyr Gly Pro Pro Arg Arg Tyr Gly Gly   1 5 10 15 Gly Asn Tyr Asn Arg Tyr Asn Gly Tyr Gly Gly Gly Arg Arg Tyr Gly              20 25 30 Gly Tyr Lys Gly Trp Asn Asn Gly Trp Asn Arg Gly Arg Arg Gly Lys          35 40 45 Tyr Trp      50 <210> 8 <211> 44 <212> PRT <213> Artificial Sequence <220> Foot protein type 3 (FP-3, Mytilus californianus) <400> 8 Gly Ala Tyr Lys Gly Pro Asn Tyr Asn Tyr Pro Trp Arg Tyr Gly Gly   1 5 10 15 Lys Tyr Asn Gly Tyr Lys Gly Tyr Pro Arg Gly Tyr Gly Trp Asn Lys              20 25 30 Gly Trp Asn Lys Gly Arg Trp Gly Arg Lys Tyr Tyr          35 40 <210> 9 <211> 60 <212> PRT <213> Artificial Sequence <220> Partial sequence from foot protein type 4 (Mytilus californianus) <400> 9 Gly His Val His Arg His Arg Val Leu His Lys His Val His Asn His   1 5 10 15 Arg Val Leu His Lys His Leu His Lys His Gln Val Leu His Gly His              20 25 30 Val His Arg His Gln Val Leu His Lys His Val His Asn His Arg Val          35 40 45 Leu His Lys His Leu His Lys His Gln Val Leu His      50 55 60 <210> 10 <211> 75 <212> PRT <213> Artificial Sequence <220> Foot protein type 5 (FP-5, Mytilus edulis) <400> 10 Ser Ser Glu Glu Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Ala Tyr His   1 5 10 15 Tyr His Ser Gly Gly Ser Tyr His Gly Ser Gly Tyr His Gly Gly Tyr              20 25 30 Lys Gly Lys Tyr Tyr Gly Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr Lys          35 40 45 Asn Ser Gly Lys Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg      50 55 60 Lys Gly Tyr Lys Lys Tyr Tyr Gly Gly Ser Ser  65 70 75 <210> 11 <211> 76 <212> PRT <213> Artificial Sequence <220> &Lt; 223 > Foot protein 5 (FP-5, Mytilus edulis) <400> 11 Ser Ser Glu Glu Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Thr Tyr His   1 5 10 15 Tyr His Ser Gly Gly Ser Tyr His Gly Ser Gly Tyr His Gly Gly Tyr              20 25 30 Lys Gly Lys Tyr Tyr Gly Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr Lys          35 40 45 Asn Ser Gly Lys Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg      50 55 60 Lys Gly Tyr Lys Lys Tyr Tyr Gly Gly Gly Ser Ser  65 70 75 <210> 12 <211> 71 <212> PRT <213> Artificial Sequence <220> &Lt; 223 > Foot protein 5 (FP-5, Mytilus coruscus) <400> 12 Tyr Asp Asp Tyr Ser Asp Gly Tyr Tyr Pro Gly Ser Ala Tyr Asn Tyr   1 5 10 15 Pro Ser Gly Ser His Trp His Gly His Gly Tyr Lys Gly Lys Tyr Tyr              20 25 30 Gly Lys Gly Lys Lys Tyr Tyr Tyr Lys Phe Lys Arg Thr Gly Lys Tyr          35 40 45 Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg Lys Gly Tyr Lys Lys      50 55 60 His Tyr Gly Gly Ser Ser Ser  65 70 <210> 13 <211> 76 <212> PRT <213> Artificial Sequence <220> <223> mussel adhesive protein protein type 5 from (Mytilus          galloprovincialis) <400> 13 Ser Ser Glu Glu Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Thr Tyr His   1 5 10 15 Tyr His Ser Gly Gly Ser Tyr His Gly Ser Gly Tyr His Gly Gly Tyr              20 25 30 Lys Gly Lys Tyr Tyr Gly Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr Lys          35 40 45 Asn Ser Gly Lys Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg      50 55 60 Lys Gly Tyr Lys Lys Tyr Tyr Gly Gly Gly Ser Ser  65 70 75 <210> 14 <211> 99 <212> PRT <213> Artificial Sequence <220> <223> mussel adhesive protein foot protein type 6 <400> 14 Gly Gly Asn Tyr Arg Gly Tyr Cys Ser Asn Lys Gly Cys Arg Ser   1 5 10 15 Gly Tyr Ile Phe Tyr Asp Asn Arg Gly Phe Cys Lys Tyr Gly Ser Ser              20 25 30 Ser Tyr Lys Tyr Asp Cys Gly Asn Tyr Ala Gly Cys Cys Leu Pro Arg          35 40 45 Asn Pro Tyr Gly Arg Val Lys Tyr Tyr Cys Thr Lys Lys Tyr Ser Cys      50 55 60 Pro Asp Phe Tyr Tyr Tyr Asn Asn Lys Gly Tyr Tyr Tyr Tyr Asn  65 70 75 80 Asp Lys Asp Tyr Phe Asn Cys Gly Ser Tyr Asn Gly Cys Cys Leu Arg                  85 90 95 Ser Gly Tyr             <210> 15 <211> 194 <212> PRT <213> Artificial Sequence <220> <223> hybrid mussel adhesive protein (FP-151, MEFP-5 based) <400> 15 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro   1 5 10 15 Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys              20 25 30 Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr          35 40 45 Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ser Ser Glu Glu      50 55 60 Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Ala Tyr His Tyr His Ser Gly  65 70 75 80 Gly Ser Tyr His Gly Ser Gly Tyr His Gly Gly Tyr Lys Gly Lys Tyr                  85 90 95 Tyr Gly Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr Lys Asn Ser Gly Lys             100 105 110 Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg Lys Gly Tyr Lys         115 120 125 Tyr Tyr Gly Gly Ser Ser Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys     130 135 140 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro 145 150 155 160 Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys                 165 170 175 Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr             180 185 190 Tyr Lys         <210> 16 <211> 196 <212> PRT <213> Artificial Sequence <220> <223> hybrid mussel adhesive protein (FP-151, MGFP-5 based) <400> 16 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro   1 5 10 15 Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys              20 25 30 Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr          35 40 45 Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ser Ser Glu Glu      50 55 60 Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Thr Tyr His Tyr His Ser Gly  65 70 75 80 Gly Ser Tyr His Gly Ser Gly Tyr His Gly Gly Tyr Lys Gly Lys Tyr                  85 90 95 Tyr Gly Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr Lys Asn Ser Gly Lys             100 105 110 Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg Lys Gly Tyr Lys         115 120 125 Lys Tyr Tyr Gly Gly Gly Ser Ser Ala Lys Pro Ser Tyr Pro Pro Thr     130 135 140 Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser 145 150 155 160 Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys                 165 170 175 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro             180 185 190 Pro Thr Tyr Lys         195 <210> 17 <211> 192 <212> PRT <213> Artificial Sequence <220> <223> hybrid mussel adhesive protein (FP-151, MCFP-5 based) <400> 17 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro   1 5 10 15 Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys              20 25 30 Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr          35 40 45 Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Tyr Asp Gly Tyr      50 55 60 Ser Asp Gly Tyr Tyr Pro Gly Ser Ser Ala Tyr Asn Tyr Ser Ser Ser Ser Ser  65 70 75 80 His Gly Tyr His Gly His Gly Tyr Lys Gly Lys Tyr Tyr Gly Lys Gly                  85 90 95 Lys Lys Tyr Tyr Tyr Lys Tyr Lys Arg Thr Gly Lys Tyr Lys Tyr Leu             100 105 110 Lys Lys Ala Arg Lys Tyr His Arg Lys Gly Tyr Lys Lys Tyr Tyr Gly         115 120 125 Gly Gly Ser Ser Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys     130 135 140 Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr 145 150 155 160 Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser                 165 170 175 Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys             180 185 190 <210> 18 <211> 177 <212> PRT <213> Artificial Sequence <220> <223> hybrid mussel adhesive protein (FP-131) <400> 18 Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro   1 5 10 15 Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys              20 25 30 Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr          35 40 45 Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Gly Cys Arg Ala      50 55 60 Asp Tyr Tyr Gly Pro Lys Tyr Gly Pro Pro Arg Arg Tyr Gly Gly Gly  65 70 75 80 Asn Tyr Asn Arg Tyr Gly Gly Ser Arg Arg Tyr Gly Gly Tyr Lys Gly                  85 90 95 Trp Asn Asn Gly Trp Lys Arg Gly Arg Trp Gly Arg Lys Tyr Tyr Glu             100 105 110 Phe Glu Phe Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro         115 120 125 Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr     130 135 140 Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr 145 150 155 160 Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Lys                 165 170 175 Leu     <210> 19 <211> 180 <212> PRT <213> Artificial Sequence <220> <223> hybrid mussel adhesive protein (FP-251) <400> 19 Met Glu Val His Ala Cys Lys Pro Asn Pro Cys Lys Asn Asn Gly Arg   1 5 10 15 Cys Tyr Pro Asp Gly Lys Thr Gly Tyr Lys Cys Lys Cys Val Gly Gly              20 25 30 Tyr Ser Gly Pro Thr Cys Ala Cys Ser Ser Glu Glu Tyr Lys Gly Gly          35 40 45 Tyr Tyr Pro Gly Asn Ser Asn His Tyr His Ser Gly Gly Ser Tyr His      50 55 60 Gly Ser Gly Tyr His Gly Gly Tyr Lys Gly Lys Tyr Tyr Gly Lys Ala  65 70 75 80 Lys Lys Tyr Tyr Tyr Lys Tyr Lys Tyr Leu                  85 90 95 Lys Lys Ala Arg Lys Tyr His Arg Lys Gly Tyr Lys Lys Tyr Tyr Gly             100 105 110 Gly Ser Ser Glu Phe Glu Phe Ala Lys Pro Ser Tyr Pro Pro Thr Tyr         115 120 125 Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr     130 135 140 Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala 145 150 155 160 Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro                 165 170 175 Thr Tyr Lys Lys             180 <210> 20 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 20 Lys Leu Trp Lys Lys Trp Ala Lys Lys Trp Leu Lys Leu Trp Lys Ala   1 5 10 15 <210> 21 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 21 Phe Ala Leu Ala Leu Lys Ala Leu Lys Lys Leu   1 5 10 <210> 22 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 22 Ile Leu Arg Trp Pro Trp Trp Pro Trp Arg Arg Lys   1 5 10 <210> 23 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 23 Ala Lys Arg His His Gly Tyr Lys Arg Lys Phe His   1 5 10 <210> 24 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 24 Lys Trp Lys Leu Phe Lys Lys Ile Gly Ala Val Leu Lys Val Leu   1 5 10 15 <210> 25 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 25 Leu Val Lys Leu Val Ala Gly Ile Lys Lys Phe Leu Lys Trp Lys   1 5 10 15 <210> 26 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 26 Ile Trp Ser Ile Leu Ala Pro Leu Gly Thr Thr Leu Val Lys Leu Val   1 5 10 15 Ala Gly Ile Gly Gln Gln Lys Arg Lys              20 25 <210> 27 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 27 Gly Ile Gly Ala Val Leu Lys Val Leu Thr Thr Gly Leu Pro Ala Leu   1 5 10 15 Ile Ser Trp Ile              20 <210> 28 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 28 Ser Trp Leu Ser Lys Thr Ala Lys Lys Gly Ala Val Leu Lys Val Leu   1 5 10 15 <210> 29 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 29 Lys Lys Leu Phe Lys Lys Ile Leu Lys Tyr Leu   1 5 10 <210> 30 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 30 Gly Leu Lys Lys Leu Ile Ser Trp Ile Lys Arg Ala Gln Gln Gly   1 5 10 15 <210> 31 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Anti-microbacterial peptide <400> 31 Gly Trp Leu Lys Lys Ile Gly Lys Lys Ile Glu Arg Val Gly Gln His   1 5 10 15 Thr Arg Asp Ala Thr Ile Gln Gly Leu Gly Ile Ala Gln Gln Ala Ala              20 25 30 Asn Val Ala Ala Thr Ala Arg          35

Claims (13)

An amino acid sequence selected from the group consisting of SEQ ID NOS: 20 to 31 at the C-terminal, N-terminal, or C-terminal and N-terminal of the mussel adhesive protein having the amino acid sequence selected from the group consisting of SEQ ID NOS: An antimicrobial adhesive protein to which an antimicrobial peptide having a sequence is fused. delete delete delete delete delete delete An antimicrobial coating composition comprising the antibacterial adhesive protein of claim 1. The method of claim 8,
Wherein the concentration of the antimicrobial adhesive protein in the antimicrobial coating composition is 0.01 to 1% (wt / wt). The method of claim 8,
The antimicrobial coating composition further comprising a water-soluble acrylate. The method of claim 10,
Wherein said water soluble acrylate is selected from the group consisting of urethane acrylate, polyester acrylate and epoxy acrylate. An antimicrobial film coated with the antimicrobial coating composition of claim 8. The method of claim 12,
The antimicrobial film is used for food packaging or liquid crystal protection of mobile phones.

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