KR20180078853A - Antimicrobial Peptide Derived From The Mytilus coruscus And Its Use - Google Patents

Abstract

The present invention relates to antimicrobial peptide derived from Mytilus coruscus, and an antimicrobial pharmaceutical composition containing the same. The antimicrobial peptide according to the present invention is derived from the gills of Mytilus coruscus, and has low hemolytic activity and low toxicity to humans, and excellent antimicrobial activity.

Description

Antimicrobial peptides derived from mussels and uses thereof <br> <br> Patents Register Register Login 5,657,858 Kind Code:

The invention mussels (Mytilus coruscus , and antimicrobial pharmaceutical compositions containing the antimicrobial peptides, a composition for preventing or treating food poisoning, a cosmetic composition, a feed additive or a sanitary article.

Most vertebrate and invertebrate animals, including humans, have an immune system to defend themselves against the ingress of external agents such as bacteria, viruses, fungi, etc. (Zasloff, M. Antimicrobial peptides of multicellular organisms. Nature 2002; 415: 389 -95). These immune systems can be divided into innate immunity, which plays a primary defense regardless of the intruder, and host defense, which reacts specifically to the intruder. In vertebrates, innate immunity and adaptive immunity are well developed, but invertebrate animals contain only innate immunity (Tincu, JA, Taylor, SW Antimicrobial peptides from marine invertebrates, Antimicrob. Agents Chemother. 2004; 48: 3645-3654) . Especially, congenital immunity induces early defense and adaptive immunity against external invasive substances, and therefore, interest in its function and role is increasing. Some of the immune-related substances responsible for innate immunity include lectin, prophenoloxidase activating factor, cytokines, enzymes, and antimicrobial peptide / protein. They are directly or indirectly responsible for the primary elimination of non-magnetic substances in the body from the outside. In particular, the antimicrobial peptide (AMP) among various innate immune factors has the property that it is difficult for the pathogens to acquire tolerance for self defense and has the characteristics of nonspecific and rapid reactivity. Therefore, Crit Rev Biotechnol. 2012; 32 (1): 323-322. [CrossRef], [Web of Science ®] 143-71).

To date, AMP studies on plant and terrestrial organisms have been actively under way, but studies from marine invertebrates have been limited. In fact, most of the marine invertebrates are largely dependent on the innate immune system as a defense against external intrusion, so the role of AMP is more important. Unlike terrestrial environments, marine life in the marine environment is always exposed to pathogenic infections such as bacteria, viruses, parasites, and fungi, and it is likely to include an efficient defense mechanism against the infectious agent to overcome this environmental condition Seo, JK, Crawford, JM, Stone, KL, Noga, EJ Purification of a novel arthropod defensin from the American oyster, Crassos-trea virginica Biochem. Biophys. Res. Com-mun 2005; 338: 1998-2004).

In the meantime, peptidic antibiotics have been isolated and tested in a variety of organisms to find antibiotics that can replace conventional antibiotics. However, there are very few cases where it is commercialized because the safety of the peptide antibiotics when injected into the body and the safety of the human body are raised.

To solve these difficulties, it is necessary to find a structure that can efficiently kill microorganisms without harming the human body by comparing and analyzing the amino acid sequence and structure of peptides having antimicrobial activity. In order to do this, information on the amino acid sequence and structure of various antimicrobial peptides is required.

Recently, it has been reported that antimicrobial active peptides play an important role as an effective defense mechanism against external infectious agents in the gills or mucous membranes of oysters, abalones, and the military. These reports are important for research on the search and purification of antimicrobial active substances that function as innate immunity as gill and / or mantle, which play a primary defense role against the external environment, play an important role in the immune mechanism of the shellfish. However, efforts to develop a new antimicrobial peptide using this as a main target are insufficient.

As a prior art document related to the present invention, Korean Patent No. 10-1374267 (Patent Document 1) is disclosed, and Patent Document 1 relates to an antibiotic peptide having a novel amino acid sequence, and more particularly, The present invention relates to a peptide having excellent antimicrobial activity against E. coli or Staphylococcus aureus. Patent Document 1 discloses only the content of antimicrobial peptides isolated from abalone, but does not disclose or suggest any antimicrobial peptides derived from peptides purified from mussels.

Patent Document 1. Korean Patent No. 10-1374267

The invention mussels (Mytilus coruscus , which has low hemolytic activity and is excellent in antimicrobial activity.

The present invention also provides a pharmaceutical composition for antimicrobial use comprising the above-mentioned antimicrobial peptide as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating food poisoning comprising the above-mentioned antimicrobial peptide as an active ingredient.

The present invention also provides a cosmetic composition comprising the above-mentioned antimicrobial peptide as an active ingredient.

The present invention also provides a feed additive comprising the antimicrobial peptide as an active ingredient.

The present invention also provides a sanitary article comprising the antibacterial peptide as an active ingredient.

The invention mussels (Mytilus The present invention relates to an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus .

The antimicrobial peptide may be selected from the group consisting of Bacillus subtilis subtilis ) and E. coli . The antimicrobial activity of the present invention is not particularly limited.

In another aspect, the present invention mussels (Mytilus The present invention relates to an antimicrobial pharmaceutical composition containing an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an effective ingredient.

In another aspect, the present invention mussels (Mytilus The present invention relates to a pharmaceutical composition for preventing or treating food poisoning using an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an effective ingredient.

In another aspect, the present invention mussels (Mytilus The present invention relates to an antimicrobial cosmetic composition containing an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an effective ingredient.

The cosmetic composition may be in the form of a solution, powder, emulsion, lotion, spray, ointment, aerosol, cream or foam.

In another aspect, the present invention mussels (Mytilus The present invention relates to an antimicrobial food additive containing an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an effective ingredient.

In another aspect, the present invention mussels (Mytilus The present invention relates to a feed additive containing an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an active ingredient.

In another aspect, the present invention mussels (Mytilus The present invention relates to a hygiene product containing an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an active ingredient.

The present invention relates to an antimicrobial peptide which is derived from the gills of mussels and has a low hemolytic activity and is low in toxicity to human body and excellent in antimicrobial activity and which is useful as a pharmaceutical composition for antimicrobial use, Can be utilized.

FIG. 1 is a photograph showing the composition distribution of peptides using the extract of Example 1 in Experimental Example 1 and the results of performing Acid Urea-PAGE and bug-blotting in order to confirm their antibacterial activity.
Fig. 2 is a graph showing reversed-phase HPLC results for purifying an antimicrobial peptide from the extract of Example 1. Fig.
3 is a graph showing the results of cation exchange HPLC for purifying an antimicrobial peptide from the extract of Example 1. Fig.
4 is a graph showing the final purification results of the antimicrobial peptide using the reversed phase HPLC from the extract of Example 1. Fig.
Fig. 5 shows the result of confirming the molecular weight of the antibacterial peptide of Example 2 by MALDI-TOF method in Experimental Example 2. Fig.
6 shows the amino acid sequence (SEQ ID NO: 1) of the antimicrobial peptide of the present invention which was finally purified.
Fig. 7 shows the result of prediction of the secondary structure of the antimicrobial peptide of Example 2 in Experimental Example 3. Fig.
Fig. 8 shows the results of measurement of the binding strength of the peptide of Example 2 with DNA using the EMSA method. Fig.
Fig. 9 shows the result of confirming the antimicrobial activity of the antimicrobial peptide of Example 2. Fig.
Fig. 10 shows the results of analysis of hemolytic activity of the antimicrobial peptide of Example 2. Fig.

The antimicrobial peptides of the present invention are useful as mussel 1 &lt; / RTI &gt; derived from E. coli .

The antimicrobial peptide of SEQ ID NO: 1 is an antimicrobial peptide which does not show hemolytic activity and thus has no cytotoxicity problem. Bacillus subtilis subtilis ) and Escherichia coli ( E. coli ).

A pharmaceutical composition for the antimicrobial of the present invention, mussels (Mytilus The antimicrobial peptide having the amino acid sequence of SEQ ID NO: 1 derived from coruscus is included as an active ingredient.

The method of administration of the pharmaceutical composition is not particularly limited, but it is preferably injected into an artery or vein, subcutaneously, rectally, nasally, and any other parenteral route, It is preferable to be injected intravenously, orally, or directly into muscle cells.

In addition, the dosage level selected from the composition will depend on the activity of the compound, the route of administration, the severity of the condition being treated, and the condition and previous history of the patient being treated. It is within the knowledge of the art, however, to gradually increase the dose until the desired effect is achieved, starting at a dose of the compound at a level lower than that required to achieve the desired therapeutic effect, , Body shape, and body weight. The composition may be further processed before being formulated into a pharmaceutically acceptable pharmaceutical formulation and may be preferably ground or ground into smaller particles. Also, the composition will vary depending on the condition and the patient being treated, but it can be determined non-ingeniously.

In addition, the antimicrobial peptide can prevent or treat diseases such as food poisoning, which is a typical disease caused by Bacillus subtilis and E. coli , which exhibit antibacterial activity, As a pharmaceutical composition for preventing or treating the above diseases.

The antimicrobial peptide may be used as an active ingredient of a cosmetic composition for antimicrobial use. At this time, the cosmetic composition may be in the form of a solution, powder, emulsion, lotion, spray, ointment, aerosol, cream or foam.

The antimicrobial peptide may be used as an active ingredient of an antimicrobial food additive or a feed additive.

The antimicrobial peptide may also be used as an active ingredient of a sanitary article such as a wet tissue, a hand sanitizer, an oral cleanser, an oral disinfectant, and a toothpaste additive.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Example

&Lt; Example 1: Tissue Extraction of Mussel >

The mussel used in the experiment was purchased from the Haenmangdong Marine Products Research Center, Gunsan City, Jeonbuk Province, and was immediately taken to the laboratory for tissue harvesting and extraction. Each extract was stored at -70 ℃ until use.

Tryptic soy broth (TSB) and agarose type I (Low EEO Agar) for the determination of antimicrobial activity were obtained from Merck (Darmstadt, Germany) and Sigma (St. Louis, MO, USA) Were purchased and used. The HPLC water and acetonitrile (CH ₃ CN) used in the purification of the compound were purchased from Tedia (Ohio, USA), and all reagents except the reagents were used for the Exp.

After extracting the gill tissue of the mussel, the extraction process using weak acid was carried out immediately. The collected tissues were added to 1% acetic acid (v / v) corresponding to a tissue volume of 4 times, heated, boiled for 5 minutes, and completely cooled while being stored in ice. The cooled tissue was completely disrupted using a homogenizer and then centrifuged. The supernatant was taken and stored at -70 ° C until used for antibacterial activity searches and peptide purification.

Experimental Example 1: Confirmation of Distribution of Peptide Substances [

Acid Urea-PAGE and bug-blot were performed to determine the distribution of the peptide material in the extract of Example 1.

In order to perform AU-PAGE, 20 μl of each extract was mixed with 2 × sample buffer to gel containing acetic acid and urea, and then introduced into gel, followed by electrophoresis at 150 V for 50 minutes 1 μg of histone H1 (~ 21 kDa), lysozyme (~ 11 kDa, 2 μg), aprotinin (~6.5 kDa, 3 μg) and synthetic peptide piscidin 1 (~2.5 kDa, 1 μg) were used. After electrophoresis, gel was stained with coomassie brilliant blue (CBB) R-250 dye and the results were confirmed.

In order to perform the bug-blotting, the gel was rinsed without electrophoresis and then blotted on an LB plate containing E. coli D31, and the gel was firstly cultured for 3 hours at 37 ° C. And the result of clearing zone and stained gel after second incubation at 37 ℃ for 16-18 hours.

The results of the above-mentioned AU-PAGE and bug-blot are shown together in FIG.

As a result of the above-mentioned AU-PAGE and bug-blot, the mussel gill extract according to the present invention showed strong antimicrobial activity in the low molecular weight substance and the high molecular weight substance part. These results indicate that there are various sizes of proteinaceous antimicrobial substances with positive net-charge in mussel gill extract.

&Lt; Example 2: Isolation and purification of antimicrobial peptide >

The mussel gill extract of Example 1 was separated and purified by HPLC (YL9100 HPLC system, Younglin Instrument, Korea). For cation exchange HPLC, 100 mM Tris-HCl (pH 7.5) was added at 0 ° C to 1.0 M NaCl with 10 mM phosphate buffer (pH 6.0) and PB containing 1.0 M NaCl (pH 6.0) using TSKgel SP-5PW And the separation process was performed at 220 nm at a flow rate of 1 mL / min. In addition, 95% A solvent (0.1% TFA in HPLC grade water ) and 5% B solvent _{(0.1% TFA in HPLC CH 3} CN) was CapCell-Pak C18 column (4.6 mm × 250 mm equilibrated in order to carry out a reverse phase HPLC, Shiseido, Japan) and then eluted with a gradient of solvent B (5-65%) for 60 minutes. At this time, the flow rate was maintained at 1.0 mL / min, and the separated fractions were measured for antibacterial activity against B. subtilis KCTC1021 by URDA method. Active peaks were purified to a single peak after the final step with the same conditions.

The purified extract was introduced into a CapCell-Pak C ₁₈ column (4.6 mm x 250 mm, Shiseido, Japan), and the separated fractions were assayed for antibacterial activity against B. subtilis by the URDA method. Fig. 2 is a graph showing the results of reversed phase HPLC for purifying antimicrobial peptides from mussel gill extract. Fig. In FIG. 2, the fractions eluted at 14 minutes showed antibacterial activity, and the peaks showing antibacterial activity against B. subtilis KSTC1021 were indicated by arrows.

The active fractions were collected and introduced into a cation exchange column, TSK-gel SP-5PW column (7.5 mm x 75 mm, Tosoh, Japan), and the separation procedure was performed. 3 is a graph showing the results of cation exchange HPLC for purifying an antimicrobial peptide from mussel gill extract. In FIG. 3, peaks showing antibacterial activity in B. subtilis KSTC1021 are indicated by arrows. As a result of the above experiment, the antimicrobial activity was confirmed in the fraction eluted at a concentration of 0.83M NaCl.

The active fractions obtained from the cation exchange column having the antibacterial activity confirmed were collected and purified to a pure CapCell-Pak C ₁₈ column (4.6 mm x 250 mm) in order to finally purify the antimicrobial peptide. 4 is a graph showing the final purification results of antimicrobial peptides using reverse phase HPLC from mussel gill extract. In FIG. 4, peaks showing antibacterial activity against B. sutilis KCTC1021 are indicated by arrows.

Experimental Example 2: Determination of molecular weight and sequence of peptide

The molecular weight of the antimicrobial peptide of Example 2 was measured using a MALDI-TOF / TOF 5800 system (AB SCIEX). The matrix used was α-Cyano-4-hydroxycinnamic acid (CHCA) dissolved at a concentration of 5 mg / mL (0.1% TFA / 50% acetonitrile). In the measurement of the molecular weight, the peptide was dissolved in 10 μL of a 0.1% TFA solvent, and the mixture was mixed with the matrix at a ratio of 1: 1 (v / v), and the molecular weight was measured using a MS Reflector (Positive) operation mode. 2 was used.

As a result of the molecular weight measurement, it was confirmed that the molecular weight of the purified peptide was 5540.4 Da. Fig. 5 shows the result of measuring the molecular weight of the peptide purified from mussel gills. At this time, the molecular weight of the peptide was measured in a linear mode using an UltrafleXtreme MALDI-TOF mass spectrometer equipped with a pulsed smart beam II.

To determine the amino acid sequence of the antimicrobial peptide purified from the mussel gill extract, the Procise 491 HT protein sequencer (Applied Biosystems, USA) in Korea Basic Science Research Institute was used and analyzed by Edman decomposition method. As a result, it was found that the peptide was composed of 56 amino acids.

Figure 6 shows the amino acid sequence (SEQ ID NO: 1) of the final purified antimicrobial peptide.

<Experimental Example 3: Secondary structure of peptide>

The secondary structure of the antimicrobial peptide of Example 2 was predicted using the Garnier method of EMBOSS GUI ( http://imed.med.ucm.es/EMBOSS/ ). As a result, the purified peptide was predicted to take a predominantly α-helix structure, including a partially turn structure (see FIG. 7).

Experimental Example 4: Confirmation of homology of peptide

Homology confirmation of the purified peptides was performed using Genome Net's BLASTP 2.2.10 and TBLASTN 2.2.10. As a result, the final purified peptides did not show high homology with known antimicrobial peptides / proteins to date. These results indicate that the peptide of the present invention purified from mussel gill is a new antimicrobial peptide that has not been reported so far.

Experimental Example 5: DNA-Binding Assay

After obtaining extracts from mussels gill and mantle, DNA-binding assay was performed to confirm the DNA-binding ability of each of the purified antimicrobial peptides from each extract.

The DNA-binding assay of each peptide was confirmed to inhibit DNA migration in agarose-gel electrophoresis after incubation of DNA and peptides. To this end, 1 Kb DNA marker, λ-Hind III-digested DNA (50 ng) (Roche, Basel, Switzerland) and each peptide (1 ug) were mixed and reacted at 37 ° C. for 60 minutes. Then 0.5 μg / mL ethidium bromide ) On a 1.0% agarose gel.

The binding strength of the peptide purified from the peptide (Gill) and the mussel extract of the mussel of Example 2 to the DNA was measured using an electrophoretic mobility shift assay (EMSA) method (see FIG. 8). Specifically, the DNA-binding ability of each peptide was determined by mixing 1 kb DNA ladder with 1 μg piscidin or 5 μl peptide solution, reacting at 37 ° C for 60 minutes, and then introducing into 1% agarose gel for electrophoresis It is.

As a result of the measurement, the peptide purified from the peptide (Gill) and the Mantle extract of Example 2 slightly inhibited the electrical transfer of DNA, and piscidin 1, an antimicrobial peptide used as a positive control, showed a strong DNA-binding ability . These results indicate that the peptide of Example 2 contains a substance with DNA-binding ability at a somewhat lower level than piscidin 1.

<Experimental Example 6: Confirmation of antimicrobial activity of peptide>

To confirm the antimicrobial activity of the antimicrobial peptide of Example 2, URDA (ultrasensitive radial diffusion assay) was used for E. coli D31 and B. subtilis KCTC1021.

The strain used for URDA was first cultured for 18 hours at each medium and incubation temperature, and the bacterial concentration was adjusted to 84% T (1 x 10) using a colorimeter (Product No. 52-1210, BioMerieux, Inc., USA) ⁸ CFU / mL). Then, add 0.5 mL of the diluted bacterial solution to each well of the underlay gel containing 9.5 mL of 0.03% TSB, 1% Type agarose and 10 mM phosphate buffer (pH 6.5), mix well and pour flat onto the plate Hardened. After punching a 2.5 mm diameter well using a punch on a hard plate, 5 μL of peptide solution (1000 ug / mL in 0.01% HAc) was introduced. All of the samples were confirmed to be free of solvent by using 5 μL of 0.01% acetic acid. When each sample (peptide solution) permeates into the culture medium, the culture is firstly incubated for 3 hours, then overlay gel containing 10 mL of 6% TSB, 1% Type agarose and 10 mM phosphate buffer (pH 6.5) Followed by secondary culture at the same temperature for 18 hours. The size of the clearing zone around the well was measured the next day to confirm the antimicrobial activity.

The results of confirming the antimicrobial activity of the peptide solution are shown in FIG.

As shown in FIG. 9, the antimicrobial peptide of Example 2 purified from mussel gill extract showed strong antimicrobial activity against the two strains used for the measurement. These results also indicate that the peptide of Example 2 exhibits antimicrobial activity against both Gram- (+) and Gram- (-) bacteria.

Experimental Example 7: Characterization of hemolysis of peptide

In order to confirm the presence or absence of toxic substances contained in the antimicrobial peptide of Example 2, the hemolytic activity of the peptide against human erythrocytes (blood type: type B) was measured (see FIG. 10).

Specifically, the same amount of phosphate buffered saline (PBS, 50 mM sodium phos-phate, 150 mM NaCl, pH 7.4) was added to human blood, and the mixture was centrifuged at 8000 x g for 1 minute at 4 ° C to remove the supernatant. After this washing procedure was repeated 3 times, the obtained red blood cells were added with PBS to make 3% hematocrit. To measure hemolytic activity, 90 μL of 3% hematocrit was added to the e-tube and 10 μL of each concentration of the peptide solution was added. Each e-tube was reacted at 37 ° C for 1 hour and centrifuged at 13,000 x g for 10 minutes at 4 ° C. 70 μL of each supernatant was transferred to a 96-well microtiter plate and the degree of hemoglobin leaching was measured at 542 nm. Triton X-100 (0.1%) was used as an experimental control for 100% hemolysis of red blood cells, and hemolytic activity was calculated using the following equation.

% Absolute = 542 nm in buffer) / (Abs 542 nm in 0.1% Triton X-100 - Abs 542 nm in buffer)] x 100

Piscidin 1 was used as a comparative substance and 0.1% Triton-X 100 was used as a standard material for 100% hemolysis. As a result, piscidin 1, which is an antimicrobial peptide used as a control, exhibited strong hemolytic activity of 70% or more even at a low concentration (50 μg / mL), whereas it was purified from the antimicrobial peptide and mussel coat extract of Example 2 purified from mussel gill extract The peptide showed little hemolytic activity. These results indicate that the peptide contains few substances exhibiting cytotoxicity.

The preparation examples for the peptides of the present invention are illustrated below.

&Lt; Preparation Example 1 > Preparation of pharmaceutical preparations

<1-1> Preparation of powder

The peptide of the present invention 20 mg

Lactose 20 mg

After mixing the above components, the mixture was packed in an airtight container to prepare a powder.

<1-2> Preparation of tablets

The peptide of the present invention 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

&Lt; 1-3 > Preparation of capsules

The peptide of the present invention 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

<1-4> Production of liquid agent

The peptide of the present invention 20 mg

Isomer 10g

Mannitol 5g

Purified water Suitable amount

Each component was added to purified water in accordance with the conventional liquid preparation method and dissolved. After the lemon flavor was added in an appropriate amount, the above components were mixed, and purified water was added thereto to adjust the whole volume to 100 ml. The solution was filled in a brown bottle and sterilized, .

<1-5> Preparation of Injection

The peptide of the present invention 10 mu g / ml

Dilute hydrochloric acid BP until pH 7.6

Injection sodium chloride BP Up to 1 ml

The peptide of the present invention was dissolved in an appropriate volume of injectable sodium chloride BP, and the pH of the resulting solution was adjusted to 7.6 with dilute hydrochloric acid BP. The volume was adjusted by using sodium chloride solution for injection and mixed thoroughly. The solution was filled in 5 ml Type I ampoule made of transparent glass, sealed in the upper lattice of the air by dissolving the glass, and autoclaved at 120 ° C for 15 minutes or longer to prepare an injection solution.

&Lt; Preparation Example 2 > Preparation of cosmetics

<2-1> Flexible longevity (skin)

In order to prepare an antibacterial softening water containing the peptide of the present invention, it may be formulated as described in the following [Table 1] and can be produced according to a conventional manufacturing method in the field of cosmetics.

ingredient Content (% by weight) The peptide of the present invention 0.1 to 30% 1,3-butylene glycol 3.0 glycerin 5.0 Polyoxyethylene (60) Hardened castor oil 0.2 ethanol 8.0 Citric acid 0.02 Sodium citrate 0.06 antiseptic a very small amount Spices a very small amount Purified water To 100

<2-2> Nourishing lotion (lotion)

To prepare an antimicrobial nutrition lotion containing the peptide of the present invention, it may be formulated as described in [Table 2] below and may be manufactured according to a conventional manufacturing method in the field of cosmetics.

ingredient Content (% by weight) The peptide of the present invention 0.1 to 30% 1,3-butylene glycol 3.0 glycerin 5.0 Squalane 10.0 Polyoxyethylene sorbitan monooleate 2.0 Nursery oil 0.1 to 30% Polyoxyethylene (60) Hardened castor oil 0.2 ethanol 8.0 Citric acid 0.02 Sodium citrate 0.06 antiseptic a very small amount Spices a very small amount Purified water To 100

<2-3> Cleanser (cleansing foam)

To prepare an antimicrobial cleanser (cleansing foam) containing the peptide of the present invention, it may be formulated as described in [Table 3] below and can be produced according to a conventional production method in the field of cosmetics.

ingredient Content (% by weight) The peptide of the present invention 0.1 to 30% Sodium N-acylglutamate 20.0 glycerin 10.0 PEG-400 15.0 Propylene glycol 10.0 POE (15) oleyl alcohol ether 3.0 Laurin derivative 2.0 Methyl paraben 0.2 EDTA-4Na 0.03 Spices 0.2 Purified water To 100

<2-4> Nourishing cream

In order to prepare an antimicrobial nutrition cream containing the peptide of the present invention, the preparation is carried out according to a conventional method in the field of cosmetics as described in [Table 4].

ingredient Content (% by weight) The peptide of the present invention 0.1 to 30% vaseline 7.0 Liquid paraffin 10.0 Wax 2.0 Polysorbate 60 2.5 Sorbitan sesquioleate 1.5 Squalane 3.0 Propylene glycol 6.0 glycerin 4.0 Triethanolamine 0.5 Xanthan gum 0.5 Tocophenylacetate 0.1 Incense, preservative a very small amount Purified water To 100

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. It is natural.

<110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION, KUNSAN NATIONAL UNIVERSITY <120> Antimicrobial Peptide Derived From The Mytilus coruscus And Its          Use <130> HPC-6798 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 56 <212> PRT <213> Mytilus coruscus <400> 1 Ala Ala Thr Lys Gly Arg Gly Arg Pro Lys Lys Ala Ala Ala Ala Ala   1 5 10 15 Gly Asp Ser Ala Ala Lys Ala Lys Ala Pro Lys Lys Ser Pro Lys Lys              20 25 30 Gly Ala Lys Lys Ala Val Lys Arg Ala Gly Ala Lys Lys Thr Ala Lys          35 40 45 Lys Ala Gly Lys Gys Lys Lys Lys      50 55

Claims (9)

Mytilus 1. An antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus .
The method according to claim 1,
The antimicrobial peptide may be selected from the group consisting of Bacillus subtilis wherein the antimicrobial peptide has an antimicrobial activity against at least one selected from the group consisting of subtilis and E. coli .
Mytilus 1. An antimicrobial pharmaceutical composition comprising an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an active ingredient.
Mytilus The antimicrobial peptide having the amino acid sequence of SEQ ID NO: 1 derived from coruscus is an effective ingredient for preventing or treating food poisoning.
Mytilus 1. An antimicrobial cosmetic composition comprising an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an active ingredient.
6. The method of claim 5,
Wherein the composition is in the form of a solution, powder, emulsion, lotion, spray, ointment, aerosol, cream or foam.
Mytilus 1. An antimicrobial food additive comprising an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an active ingredient.
Mytilus The present invention relates to a feed additive comprising an antimicrobial peptide having an amino acid sequence of SEQ ID NO: 1 derived from coruscus as an active ingredient.
Mytilus 1. A sanitary article comprising as an active ingredient an antimicrobial peptide having the amino acid sequence of SEQ ID NO: 1 derived from coruscus .

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