KR20150087542A - Novel antimicrobial peptide from Pinus densiflora Sieb. et Zucc. and use thereof - Google Patents

Abstract

The present invention relates to novel antimicrobial PN-5 peptides isolated from pine needles and a usage thereof. More specifically, the present invention confirms the novel peptides having an amino acid sequence described as SEQ ID NO: 1 shows a significant antifungal effect against pathogenic strains and food-poisoning pathogenic strains and does not have cytotoxicity. Accordingly, the novel antimicrobial peptides isolated from the pine needles can be used as antimicrobial and pharmaceutical compositions, cosmetic compositions, pesticides, dietary supplements, preservative compositions, and quasi.

Description

Novel antimicrobial peptide from Pinus densiflora Sieb. et Zucc. and use thereof}

The invention pine (Pinus densiflora Sieb. et < RTI ID = 0.0 > Zucc. < / RTI >

Functional antibiotics such as pesticides, food preservatives, cosmetics and medicines are used in a broad sense to prevent the function of pesticides in agricultural products, the deterioration, decay, discoloration and chemical change of foods and medicines, including fungicides and antioxidants In a narrow sense, it inhibits the growth of microorganisms such as bacteria, fungi and yeast, and inhibits the growth of microorganisms or sterilizes the microorganisms in pesticides, foods and medicines. Ideal conditions such as pesticide, food preservative, cosmetics and medicines should not be toxic and should be effective in trace amount.

Plants or animals produce naturally occurring peptide antibiotics (Bevins et al ., Ann . Rev. Biochem . , 59, 395-414, 1990). The peptide antibiotics are structurally divided into three groups: the first is a cysteine-rich β-sheet peptide, the second is an α-helical amphipathic molecule, the third is a cysteine- Is a proline-rich peptide (Mayasaki et al., Int . J. Antimicrob . Agents , 9, 269-280, 1998). These antimicrobial peptides are known to play an important role in the host defense and innate immune system (Boman, HG, Cell, 65 , 205, 1991;... Boman, HG, Annu Rev Microbiol, 13, 61, 1995). These antimicrobial peptides have a variety of structures depending on the amino acid sequence. The most abundant structure of these antimicrobial peptides is a structure having no cysteine residue and forming an amphipathic alpha helix. The antimicrobial peptides cecropin ) Have this structure.

The use of natural food preservatives is one of the biggest issues in the food industry as a new food preservation method that can meet the increased desire of consumers for their high quality and safety. In general, foodstuffs that are susceptible to corruption and deterioration in the natural state are very important because they can be contaminated by food-spoilage microorganisms at all stages of human consumption, such as harvesting, storage, and distribution. Conventional food preservation technologies have been used simply by methods such as heat treatment, salt treatment, acid treatment, drying, and chemical synthetic preservatives that focus on the extension of the storage period and sterilization of corrupt microorganisms. These food preservatives can be classified into four types. First, chemical preservatives have the advantage that they can be stored for years. However, there are disadvantages in that they can eat chemicals together when they are consumed, which is a safety problem. Pasteurization has the advantage of being able to store in a short time without destroying nutrients, but it has the disadvantage of not being able to kill all bacteria. Irradiation kills all microorganisms and does not affect the taste and texture of the food, but it has the disadvantage that it can denature the chemical part of the food. Finally, dehydration has the disadvantage of changing the taste and texture of the food, although it can be stored for a long time. Therefore, new alternatives to these are needed.

The pine leaves used in the present invention include moisture (65-70%), fiber (9-14%), crude protein, crude fat, phosphorus, iron and the like as general components, and vitamins having anticancer effect and antioxidant effect A and vitamin C, and contains benzoic acid with antimicrobial activity. It has also been reported that turpentine, which is known to be the main component of pine needle, has a large amount of unsaturated fatty acid, which prevents the accumulation of cholesterol and prevents arteriosclerosis. Although pine needle extract has antimicrobial activity in the previous studies, research on small size proteins and peptides that have antimicrobial activity rather than substance using pine needle is still lacking.

As a result of efforts to develop natural antibiotics, the present inventors have found that a novel PN-5 peptide can be obtained by extracting, isolating and purifying from pine needles, and the peptide has remarkable antibacterial effect against pathogenic microorganisms and remarkable antifungal effect And confirming that there is no cytotoxicity through measurement of hemolytic activity in human red blood cells, the present invention has been completed by showing that the peptide can be effectively used as an effective ingredient of a natural antibiotic.

An object of the present invention is pine (Pinus densiflora Sieb. et Zucc.), a process for producing the antibiotic peptide PN-5, and an antibiotic composition containing the antibiotic peptide PN-5 as an active ingredient.

In order to accomplish the object of the present invention, the present invention provides an antibiotic peptide comprising the amino acid sequence of SEQ ID NO: 1.

In addition,

1) pulverizing the pine needle and extracting it with an extraction solvent;

2) centrifuging the extract of step 1) and recovering the supernatant fraction;

3) separating a fraction having a molecular weight of 10 KDa or less in the supernatant fraction of step 2);

4) eluting the separated fractions of step 3) at a concentration gradient of acetonitrile B of from 5 to 65% in the mobile phase in a chromatography packed with a _C18 column. A method for producing an antibiotic peptide is provided.

The present invention also provides an antibiotic pharmaceutical composition comprising an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

In addition, the present invention provides a composition for nasal preparation comprising an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The present invention also provides an antibiotic cosmetic composition comprising an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

In addition, the present invention provides an external preparation for antibiosis comprising an antimicrobial peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The present invention also provides an antibiotic health food containing an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The present invention also provides an antibiotic pesticide containing an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

In addition, the present invention provides an antibiotic drug product containing an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The novel PN-5 peptide isolated from the pine needle of the present invention exhibits a remarkable antifungal effect against pathogenic microorganisms and a remarkable antifungal effect against food poisoning strains, does not destroy human red blood cells and is safe in cytotoxicity, And can be useful as a pharmaceutical composition, a cosmetic composition, an agricultural chemical, a health food, a composition for a nasal cavity, and a medicine preservative.

1 is a pine (Pinus densiflora Sieb. et Zucc.), a process for isolating and purifying the antibiotic-functional peptide, PN-5.
Figure 2 shows a peptide obtained by reverse phase high performance liquid chromatography (reverse-phase HPLC) on a sample separated at 40% acetonitrile.
FIG. 3 is a graph showing the results of Tricine SDS-PAGE electrophoresis analysis on the total extract and protein composition extracted from pine needle leaf.
FIG. 4 is a graph showing the results of Tricine SDS-PAGE electrophoresis analysis on the composition of samples 1 to 10 separated by reverse phase high performance liquid chromatography.
FIG. 5 is a graph showing the antimicrobial activity against the pathogenic strain of the total extract extracted from pine needle leaf.
FIG. 6 is a graph showing the antimicrobial activity of a total extract extracted from pine needle against a food poisoning strain. FIG.
FIG. 7 is a graph showing the antimicrobial activity against a pathogenic strain of a sample obtained by dividing the total extract extracted from pine needle by size.
FIG. 8 is a graph showing the antimicrobial activity of a sample obtained by dividing the total extract extracted from pine needle by size, against the food poisoning strain.
FIG. 9 shows antimicrobial activity against a pathogenic strain of a sample obtained by dividing the pine needle extract by size into 40% acetonitrile through a solid phase extraction column. FIG.
FIG. 10 shows antifungal activity against a food poisoning strain of a sample obtained by dividing the pine leaf extract by size into 40% acetonitrile through a solid phase extraction column. FIG.
FIG. 11 is a graph showing antimicrobial activity against pathogenic microorganisms of samples 1 to 10 isolated by reverse phase high performance liquid chromatography. FIG.
FIG. 12A is a graph showing the degree of cytotoxicity against mouse macrophages Raw 264.7 of samples 4 to 10 separated using reverse phase high performance liquid chromatography. FIG.
FIG. 12A is a graph showing the degree of cytotoxicity against human skin cells HaCaT of samples No. 4 to No. 10 isolated by reverse phase high performance liquid chromatography. FIG.
FIG. 13 is a graph showing the degree of cell destruction (Hemolytic) against human red blood cells of samples 4 to 10 separated by reverse phase high performance liquid chromatography.
14 is a graph showing the molecular weights of 5, 7, 8, and 10 samples having antimicrobial activity and low toxicity among the samples separated by reverse phase high performance liquid chromatography.

Hereinafter, the present invention will be described in detail.

The present invention provides an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1.

The peptide is called Pinus densiflora Sieb. et Zucc.), but is not limited thereto. The pine leaves may be cultivated or sold without limitation.

The peptide preferably has a molecular weight of 1907.0 Da, but is not limited thereto.

The peptide may be an Escherichia coli ), Staphylococcus ( Staphylococcus aureus) aureus , Staphylococcus epidermis , and Pseudomonas aeruginosa , but the present invention is not limited thereto.

The peptides of Salmonella typhimurium (Salmonella typhimurium ) and Listeria monocytogenes ( Listeria monocytogenes ), but the present invention is not limited thereto.

In a specific embodiment of the present invention, the inventors of the present invention have found that, in order to extract an antifungal peptide, a protein extract is obtained from a pine needle, and the extract is centrifuged to recover the supernatant fraction according to size and hydrophilic hydrophobic properties, C ₁₈ column reverse phase HPLC was used to obtain the peptide.

Further, the present inventors confirmed the presence of about 1-3.5 kDa peptide in the active fraction of the present invention by confirming the molecular weight by the Tricine SDS-PAGE method using the extract, the peptide and the standard protein obtained by the above production method ( 2 and 3) .

Further, the present inventors have found that Escherichia coli, which is a pathogenic microorganism of the extract obtained by the above- coli , Staphylococcus aureus , Staphylococcus ( Staphylococcus aureus ), Staphylococcus epidermis) and Pseudomonas her rugi Labor (Pseudomonas aeruginosa , and food poisoning strains Salmonella typhimurium and Listeria monocytogenes . As a result, it was confirmed that the pine leaf extract had a strong antimicrobial activity against pathogenic strains and food poisoning strains (see FIGS. 5 and 6).

In addition, the present inventors confirmed the antimicrobial activity against pathogenic strains and food poisoning strains of> 30 kDa, 10-30 kDa, and <10 kDa samples obtained by isolating the extracts obtained by the above-mentioned method, It was confirmed that it has a strong antimicrobial activity against food poisoning strains, and particularly excellent antimicrobial activity of <10 KDa samples (see FIGS. 7 and 8).

In order to identify small-sized peptides having antimicrobial activity in the fraction samples obtained by the above-described method, 40% sample of acetonitrile was separated using a solid phase column at <10 KDa sample, and a pathogenic strain and food poisoning strain , It was confirmed that 40% sample of acetonitrile has a strong antimicrobial activity against pathogenic strains and food poisoning strains (see FIGS. 9 and 10).

In addition, the present inventors confirmed the antimicrobial activity against pathogenic strains of 10 fraction samples separated from the pine leaf extract, and as a result, it was confirmed that samples 4-10 fraction had antimicrobial activity against pathogenic strains (see FIG. 11).

In addition, the present inventors confirmed cytotoxicity to animal cells and cytotoxicity through erythrocyte hemolytic activity of each of the active fractions 4-10, and as a result, 4-10 fraction samples showed low cytotoxicity against animal cells , And it was confirmed that there was no degree of destruction against red blood cells (see Figs. 12A and 12B and Fig. 13).

In addition, the present inventors analyzed the molecular weight of Samples 5, 7, 8, and 10, which have excellent antimicrobial activity and low cytotoxicity among the 4-10 fraction samples. As a result, the molecular weight was 1907.0 Da and the N-amino terminal sequence This peptide was identified as XXXXFKFLARTGKFL (SEQ ID NO: 1), which was named "PN-5" (see FIG. 14).

Therefore, the novel PN-5 peptide isolated from the pine needle of the present invention exhibits remarkable antibacterial effect against pathogenic microorganisms and remarkable antifungal effect against food poisoning strains, does not destroy human red blood cells, and is safe in cytotoxicity, Antibiotic pharmaceutical compositions, cosmetic compositions, pesticides, health foods, compositions for oral administration, and pharmaceutical preservatives.

In addition,

1) pulverizing the pine needle and extracting it with an extraction solvent;

2) centrifuging the extract of step 1) and recovering the supernatant fraction;

3) separating a fraction having a molecular weight of 10 KDa or less in the supernatant fraction of step 2);

4) eluting the separated fractions of step 3) at a concentration gradient of acetonitrile B of from 5 to 65% in the mobile phase in a chromatography packed with a _C18 column. A method for producing an antibiotic peptide is provided.

In the above preparation method, the extraction solvent of step 1) is preferably a buffer solution consisting of 10 mM HEPES and 10 mM NaCl, but is not limited thereto.

In the above production method, the separation of step 2) is preferably performed using an ultrafiltration filter, but not limited thereto.

As a method for the separation, it is preferable to use a separation method customary in the art such as ultrafiltration and chromatography. Specifically, anion exchange chromatography, size exclusion chromatography using chromatographic methods including size exclusion chromatography, reaverse-phase chromatography, high performance liquid chromatography (HPLC) or reverse phase high performance liquid chromatography And more particularly, C ₁₈ reverse phase HPLC method is most preferably used, but not limited thereto.

The peptide is preferably derived from pine needle, but is not limited thereto. The pine leaves may be cultivated or sold without limitation.

The peptide preferably has a molecular weight of 1907.0 Da, but is not limited thereto.

The peptide preferably has an antibacterial activity against E. coli, Staphylococcus aureus, Staphylococcus epidermis and Pseudomonas aeruginosa, but is not limited thereto.

The peptide preferably has antifungal activity against Salmonella typhimurium and Listeria monocytogenes, but is not limited thereto.

The novel PN-5 peptide isolated from pine needle by the production method of the present invention exhibits remarkable antibacterial effect against pathogenic microorganisms and remarkable antifungal effect against food poisoning strains, does not destroy human red blood cells, and is safe in cytotoxicity , Pharmaceutical compositions for antibiotics, cosmetic compositions, agricultural chemicals, health foods, compositions for oral administration, and pharmaceutical preservatives.

The present invention also provides an antibiotic pharmaceutical composition comprising an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The peptide is preferably derived from pine needle, but is not limited thereto. The pine leaves may be cultivated or sold without limitation.

The peptide preferably has a molecular weight of 1907.0 Da, but is not limited thereto.

The peptide preferably has an antibacterial activity against E. coli, Staphylococcus aureus, Staphylococcus epidermis and Pseudomonas aeruginosa, but is not limited thereto.

The peptide preferably has antifungal activity against Salmonella typhimurium and Listeria monocytogenes, but is not limited thereto.

The novel PN-5 peptide isolated from the pine needle of the present invention exhibits a remarkable antifungal effect against pathogenic microorganisms and a remarkable antifungal effect against food poisoning strains, does not destroy human red blood cells and is safe in cytotoxicity, May be useful as pharmaceutical compositions.

The peptide of the present invention can be administered parenterally at the time of clinical administration and can be used in the form of a general pharmaceutical preparation. Parenteral administration may mean administration through an administration route other than oral, such as rectal, intravenous, peritoneal, muscular, arterial, transdermal, nasal, inhalation, ocular and subcutaneous.

That is, the antibiotic peptide of the present invention can be administered in various parenteral formulations. In the case of formulation, the antibiotic peptide is prepared using a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous agents, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the suspending agent. As a suppository base, witepsol, macrogol, tween 61, cacao paper, Liu ling, glycerogelatin and the like can be used.

 In addition, the antibiotic peptide of the present invention can be used in combination with various carriers permitted as medicines such as physiological saline or an organic solvent, and can be used in combination with carbohydrates such as glucose, sucrose or dextran, Antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers may be used as pharmaceuticals.

The effective dose of the antibiotic peptide of the present invention is 0.01 to 100 mg / kg, preferably 0.1 to 10 mg / kg, and can be administered once to three times a day.

The total effective amount of the novel peptide of the present invention in the pharmaceutical composition of the present invention may be administered to the patient in a single dose by bolus injection or by infusion for a relatively short period of time, Multiple doses may be administered by a fractionated treatment protocol administered over a prolonged period of time. Since the concentration of the effective dose of the patient is determined in consideration of various factors such as the route of administration and the number of treatments as well as the age and health condition of the patient, in view of this point, Lt; RTI ID = 0.0 &gt; of &lt; / RTI &gt; novel peptide as a pharmaceutical composition.

The present invention also provides an antibiotic cosmetic composition comprising an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The peptide is preferably derived from pine needle, but is not limited thereto. The pine leaves may be cultivated or sold without limitation.

The peptide preferably has a molecular weight of 1907.0 Da, but is not limited thereto.

The peptide preferably has an antibacterial activity against E. coli, Staphylococcus aureus, Staphylococcus epidermis and Pseudomonas aeruginosa, but is not limited thereto.

The peptide preferably has antifungal activity against Salmonella typhimurium and Listeria monocytogenes, but is not limited thereto.

The novel PN-5 peptide isolated from the pine needle of the present invention exhibits a remarkable antifungal effect against pathogenic microorganisms and a remarkable antifungal effect against food poisoning strains, does not destroy human red blood cells and is safe in cytotoxicity, It can be usefully used as a cosmetic composition.

The cosmetic composition of the present invention includes components commonly used in cosmetic compositions in addition to the antibiotic peptides described above, and includes customary adjuvants such as sulfating agents, stabilizers, solubilizing agents, vitamins, pigments and fragrances, and carriers.

In the cosmetic composition of the present invention, the peptide of the present invention may be added to the cosmetic composition usually contained in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion (skin), a nutritional lotion (milk lotion), a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder .

When the formulation of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tragacantha, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tragacantha, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyltaurate, sarcosinates, fatty acid amides Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The present invention also provides an antibiotic health food containing an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The peptide is preferably derived from pine needle, but is not limited thereto. The pine leaves may be cultivated or sold without limitation.

The peptide preferably has a molecular weight of 1907.0 Da, but is not limited thereto.

The peptide preferably has an antibacterial activity against E. coli, Staphylococcus aureus, Staphylococcus epidermis and Pseudomonas aeruginosa, but is not limited thereto.

The peptide preferably has antifungal activity against Salmonella typhimurium and Listeria monocytogenes, but is not limited thereto.

The novel PN-5 peptide isolated from the pine needle of the present invention exhibits a remarkable antifungal effect against pathogenic microorganisms and a remarkable antifungal effect against food poisoning strains, does not destroy human red blood cells and is safe in cytotoxicity, It can be useful as a health food.

There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include various foods such as confectionery, bread, and noodles, drinks such as water, soft drinks and fruit drinks, gum, tea, vitamin complex, seasoning, Health functional foods in a conventional sense.

The antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 of the present invention may be added to the food as it is or may be used in combination with other food or food ingredients, and may be suitably used according to conventional methods. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (for prevention or improvement). Generally, the amount of the compound in the health food may be from 0.01 to 15% by weight, preferably from 0.1 to 5% by weight of the total food, and 0.01 to 5.0 g, preferably 0.01 to 5% by weight, 1.0 g. However, in the case of long-term intake for the purpose of health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

The health functional beverage composition of the present invention is not particularly limited to the other ingredients other than the peptides as essential ingredients in the indicated ratios and may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings (taumakin, stevia extract, etc.) and synthetic flavorings (saccharin, aspartan, etc.) can be advantageously used as flavorings other than those described above. The ratio of the natural carbohydrate is generally about 0.1 to 2.0 g, preferably about 0.1 to 1.0 g per 100 of the composition of the present invention.

The health functional food of the present invention can be prepared by adding the antifungal peptide consisting of the amino acid sequence of the present invention described above to the food manufacturing process of the base material or by adding the antifungal peptide comprising the amino acid sequence of the present invention And adding the antifungal peptide to the composition. At this time, a taste and odor corrector may be added as needed.

In addition to the above, the antifungal peptide consisting of the amino acid sequence of the present invention can be used as a nutritional supplement, a vitamin, a mineral (electrolyte), a flavor such as a synthetic flavor and a natural flavor, a colorant and an aggravating agent (cheese, A salt thereof, alginic acid and its salt, an organic acid, a protective colloid concentrating agent, a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in a carbonated drink, and the like. In addition, the antifungal peptide consisting of the amino acid sequence of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of these additives is not so important, it is general that the antifungal peptide composed of the amino acid sequence of the present invention is selected in a range of about 20 parts by weight per 100 parts by weight.

In addition, the present invention provides a composition for nasal preparation comprising an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The present invention also provides an antibiotic pesticide containing an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

In addition, the present invention provides an antibiotic drug product containing an antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

The peptide is preferably derived from pine needle, but is not limited thereto. The pine leaves may be cultivated or sold without limitation.

The peptide preferably has a molecular weight of 1907.0 Da, but is not limited thereto.

The peptide preferably has an antibacterial activity against E. coli, Staphylococcus aureus, Staphylococcus epidermis and Pseudomonas aeruginosa, but is not limited thereto.

The peptide preferably has antifungal activity against Salmonella typhimurium and Listeria monocytogenes, but is not limited thereto.

The novel PN-5 peptide isolated from the pine needle of the present invention exhibits remarkable antibacterial effect against pathogenic microorganisms and remarkable antifungal effect against food poisoning strains, does not destroy human red blood cells, and is safe in cytotoxicity, Compositions, antibiotics external preparations, antibiotic pesticides and antibiotics.

The preservative composition includes a cosmetic preservative or a pharmaceutical preservative. Preservatives, cosmetic preservatives, and pharmaceutical preservatives of foods mentioned above are additives used to prevent deterioration, decay, discoloration and chemical change of medicines. These include antiseptics and antioxidants and inhibit the growth of microorganisms such as bacteria, fungi and yeast, And functional antibiotics such as inhibiting the growth of microorganisms or sterilizing microorganisms in pharmaceuticals. Ideal conditions for such preservative compositions should be non-toxic and effective in trace amounts.

When the composition of the present invention is used as a quasi-drug additive, the extract or fraction may be added as it is or may be used together with other quasi-drugs or quasi-drugs, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.

The quasi-drug composition of the present invention may be a disinfectant cleaner, a shower foam, a gagrin, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment agent, a patch or a filter filler.

Hereinafter, the present invention will be described in detail with reference to Examples and Production Examples.

However, the following examples and preparations are merely illustrative of the present invention, and the content of the present invention is not limited by the following examples and production examples.

< Example  1> Protein Extraction from Pine Needle

Pine Needle (Pinus densiflora Sieb. et Zucc.) was pulverized into powder using a mortar and mortar in the presence of liquid nitrogen. The powder was added to protein extraction buffer solution (10 mM HEPES, 10 mM NaCl) and mixed well Proteins were extracted using a centrifuge. At this time, the protein extraction method of the present invention was modified by a known method [Ana Segura et al., Mol Plant Microbe Interact., 12 (1): 16-23, 1999].

< Example  2> Of peptide  refine

The extract obtained from the above Example 1 was centrifuged at 6,000 x g for 30 minutes and then the supernatant was taken and the spectra / Por 7 (MWCO: 1,000) membrane (Spectrum Laboratories, Inc.) And dialyzed with distilled water at 4 캜 for purification. The purified sample was then separated into samples of sizes> 30 kDa, 10-30 kDa and <10 kDa using Centricon with 30 kDa and 10 kDa membranes. Of the three samples, the <10 kDa sample was again separated into acetonitrile 10%, 40%, and 100% samples using hydrophilic and hydrophobic properties through a solid phase extraction column. The above acetonitrile 40% sample Purification was carried out using Shim-pack VP-ODS, C18 column (4.6 x 250 mm; 4.6 0.3 particles; 12.0 1.0 nm pore; Shimadzu) reverse phase HPLC. At this time, elution was carried out with a linear gradient (5 to 65%, 2% / min, 30 min) of buffer solution (acetonitrile B containing 0.1% TFA (FIG. 2) And the antimicrobial activity and cell activity were measured.

< Example  3> Confirm molecular weight of protein

<3-1> Identification of Protein in Pine Needle Extract

Tricine SDS-PAGE was performed to identify proteins in the pine leaf extract obtained from Example 1 above.

Specifically, Tricine SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) was performed according to the Schagger and von Jagow method using a slab gel obtained by the method described in Example 1 above . After electrophoresis, the gel was stained with silver staining to identify the band of the protein. At this time, the molecular weight standard protein is 26.6 kDa triosephosphate isomerase, 17 kDa of muscle pigment, Myoglobin 17 kDa, which is a protein isolated from rabbit muscle, and α-Lactalbumin ) 14.2 kDa, aprotinin 6.5 kDa extracted from bovine lung, insulin chain B (insulin chain B) 3.5 kDa, one of the insulins, and Bradykinin 1 kDa, the main component of the poison.

As a result, as shown in Fig. 3, it was confirmed that the peptides derived from the pine needle extract were present at about 1-3.5 KDa in molecular weight (Fig. 3).

<3-2> Identification of purified protein isolated from pine needle extract

Tricine SDS-PAGE was performed to identify the protein in the sample separated and purified from the above <Example 2>.

Specifically, using the reverse phase high performance liquid chromatography as the last step of separation and purification as in the above <Example 2>, 10 samples from the separation step and the composition of the standard protein were compared with those in Example <3- 1>, silver staining was performed after Tricine SDS-PAGE electrophoresis.

As a result, as shown in Fig. 4, the peptides derived from the pine needle extract of the present invention exist at about 1-3.5 kDa in terms of molecular weight by Tricine SDS-PAGE when the protein low molecular markers are compared with each other (Fig. 4).

< Example  4> Confirmation of antifungal activity

<4-1> Confirmation of antibacterial activity of pine needle extract

In order to measure the antibacterial activity of the pine leaf extract obtained from <Example 1>, the following experiment was conducted.

Specifically, Escherichia coli, an anti-pathogenic microorganism, coli ), Staphylococcus ( Staphylococcus aureus) aureus , Staphylococcus epidermis) and Pseudomonas Ke rugi labor (Pseudomonas aeruginosa) (using the FIG. 5), and the Middle East type strain of Salmonella typhimurium (Salmonella typhimurium), and Listeria monocytogenes to Ness (Listeria monocytogenes (Fig. 6) were used. The number of the microorganisms was diluted so as to be 2 × 10 ⁵ cells per well on a 96-well microtiter plate, and 100 μl of each medium containing the bacteria was dispensed on a plate. Only the bacteria were dispensed into the negative control, One plate was inoculated with the bacterium and the pine needle extract obtained by the method described in Example 1, and cultured in a 36 ° C incubator for 4 hours. Thereafter, to confirm the antifungal activity directly by the eye, the cells were plated on agar medium and then cultured in a 36 ° C incubator for 24 hours.

As a result, as shown in FIG. 5 and FIG. 6, the pine needle extract had a strong antimicrobial activity against pathogenic strains in a concentration-dependent manner (FIG. 5), and also against Salmonella typhimurium and Listeria monocytogenes It was confirmed to have a strong antimicrobial activity (Fig. 6).

<4-2> Isolation from pine needle extract Of peptide  Identify antimicrobial activity

In order to measure the antimicrobial activity of a sample separated and purified by size in the pine leaf extract according to the method described in <Example 2>, the following experiment was conducted.

Specifically, Escherichia coli and Staphylococcus aureus, which are pathogenic microorganisms, were dispensed in the same manner as in the above Example <4-1>, and the strains were classified into 30 KDa, 10-30 KDa, &Lt; 10 KDa sample (Fig. 7).

In addition, Salmonella typhimurium and L. monocytogenes, which are food poisoning strains, were dispensed and diluted with <10 KDa samples separated and purified by the method described in <Example 2> as described in Example <4-1> Respectively.

As a result, as shown in FIG. 7, it was confirmed that three samples separated by size showed strong antimicrobial activity against pathogenic bacteria, and exhibited remarkable antimicrobial activity particularly in the <10 kDa sample (FIG. 7).

In addition, as shown in FIG. 8, it was confirmed that a sample of < 10 kDa had an excellent antimicrobial activity against a food poisoning strain (FIG. 8).

<4-3> Isolation and purification from pine needle extract Of peptide  Identify antimicrobial activity

In order to measure the antimicrobial activity of a 40% sample of acetonitrile isolated from the <10 KDa sample obtained from <Example 2>, the following experiment was conducted.

Specifically, as in Example <4-1>, Escherichia coli and Staphylococcus aureus, which are pathogenic strains, were mixed and mixed with a 40% sample of acetonitrile separated by the method described in <Example 2> (Fig. 9).

Salmonella typhimurium and Listeria monocytogenes, which are food poisoning strains, were also dispensed and mixed with a 40% sample of acetonitrile separated by the method described in <Example 2> as in Example <4-1> (Fig. 10).

In addition, Escherichia coli and Lokocus aureus, which are pathogenic strains, were dispensed in the same manner as in Example <4-1>, and purified by reversed phase HPLC on a 40% acetonitrile sample by the method described in Example 2 And fractionated with fractions 1 to 10 (FIG. 11).

As a result, as shown in FIG. 9 and FIG. 10, it was confirmed that 40% sample of acetonitrile isolated from pine needle extract showed strong antimicrobial activity against pathogenic strain and food poisoning strain (FIGS. 9 and 10).

As a result, as shown in FIG. 11, it was confirmed that up to 4 to 10 of the 10 fraction samples obtained by purifying the 40% sample of acetonitrile using reverse phase high performance liquid chromatography had antibacterial activity (FIG. 11) .

< Example  5> The pine needle extract Of peptide  Cytotoxicity check

MTT assay was performed to confirm cytotoxicity of the peptides derived from the pine needle to animal cells.

Specifically, mouse macrophage Raw 264.7 cells (FIG. 12A) and human skin cells HaCat cells (FIG. 12B) were divided into 2 × 10 ⁴ cells in a 96-well microtiter plate and cultured in a 37 ° C. CO ₂ incubator And cultured for 24 hours. Subsequently, samples 4 to 10 having antimicrobial activity among the samples purified using the reversed-phase HPLC obtained by the method described in Example 2 above were placed in a 96-well microtiter plate ), Placed in a 37 ° C CO ₂ incubator, and reacted for 24 hours. Then, 10 μl of 5 mg / ml MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) was added to each well and the mixture was placed in a 37 ° C CO ₂ incubator for 4 hours Lt; / RTI &gt; Then, the supernatant was removed and 50 쨉 l of DMSO was added to each well. After reacting for 30 minutes, the absorbance at 570 nm was measured to confirm the degree of toxicity in the cells.

As a result, as shown in FIGS. 12A and 12B, it was confirmed that fraction samples 4 to 10 for rat macrophages and human skin cells had low cytotoxicity (FIGS. 12A and 12B).

< Example  6> Pine needle extract Of peptide  Identification of erythrocyte hemolytic activity

In order to confirm the cytotoxicity through human erythrocyte hemolytic activity against PN-5 peptide derived from pine needle, the following experiment was performed.

Specifically, human erythrocytes were washed three times with PBS buffer solution (phosphate-buffered saline: 35 mM phosphate buffer / 0.15 M NaCl, pH 7.0), and diluted with PBS buffer solution to 8.0% 100 μl of the red blood cell solution was loaded on a 96-well microtiter plate, and 100 μl of the peptide solution isolated and purified by the method described in Example 2 was added to each well, followed by incubation at 37 ° C for 1 hour After incubation, the absorbance at 414 nm was measured. Also, as a control, the same procedure was carried out for melittin, a known peptide. At this time, the value obtained when 0.1% Triton X-100 was treated was calculated as 100% hemolysis, and the hemolysis rate of the peptide, that is, hemolysis was calculated by the following formula (1). In the following, A is the absorbance at 141 nm in the peptide solution, B is the absorbance at 141 nm in 0.1% Triton X-100, and C is the absorbance at 141 nm in the PBS solution.

As a result, as shown in Fig. 13, in the case of melittin, a known peptide, the hemolysis rate of the cells was 30% even at a peptide concentration of 0.31 μM and 100% of the red blood cells were destroyed under the condition of the peptide concentration of 5 μM. On the other hand, in the case of the peptides derived from pine needle, it was confirmed that the red blood cells were not destroyed at the same concentration condition, so that the peptides isolated from the pine needle extract had no cytotoxicity (FIG. 13).

< Example  7> Determination of amino acid sequence and molecular weight

From the results of Examples 4 to 6, fractions 4 to 10 having antimicrobial activity derived from pine needle and having no cytotoxicity were dissolved in 6N-HCl and hydrolyzed at 110 ° C for 22 hours After drying, the amino acid sequence was confirmed by an amino acid analyzer and the molecular weight was confirmed by MALDI-MS (Kratos Analytical Ins).

As a result, as shown in Fig. 14, it was confirmed that the amino acid sequence appeared only in the No. 5 fraction sample, the N-amino terminal sequence was XXXXFKFLARTGKFL (SEQ ID NO: 1), the molecular weight was 1907.0 Da and this was named "PN-5" (Fig. 14).

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

< Manufacturing example  1> Preparation of pharmaceutical preparations

<1-1> Sanje  Produce

20 mg of the peptide of the present invention

20 mg of lactose

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

<1-2> Preparation of tablets

10 mg of the peptide of the present invention

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

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

&Lt; 1-3 > Preparation of capsules

10 mg of the peptide of the present invention

3 mg of crystalline cellulose

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> Liquid  Produce

20 mg of the peptide of the present invention

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

<1-5> Preparation of Injection

10 [mu] g / ml of the peptide of the present invention

Diluted hydrochloric acid BP until pH 7.6

Sodium chloride up to 1 ml for injection

The peptide of the present invention was dissolved in an appropriate volume of injectable sodium chloride BP. The pH of the resulting solution was adjusted to pH 7.6 using diluted hydrochloric acid BP, and the volume was adjusted using main sodium chloride BP and mixed thoroughly. The solution was filled in a 5 ml type I ampoule made of transparent glass, sealed in an upper lattice of air by dissolving the glass, sterilized by autoclaving at 120 DEG C for 15 minutes or longer, and an injection solution was prepared.

< Manufacturing example  2> Manufacture 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 8.0 glycerin 5.0 Squalane 10.0 Polyoxyethylene sorbitan monooleate 2.0 Nursery oil 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

<2-3> Essence

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

ingredient Content (% by weight) The peptide of the present invention 0.1 to 30% Sitosterol 1.7 The polyglyceryl 2-oleate 1.5 Ceramide 0.7 Setares-4 1.2 cholesterol 1.5 Dicetyl phosphate 0.4 Concentrated glycerin 5.0 Carboxyvinyl polymer 0.2 Xanthan gum 0.2 antiseptic a very small amount Spices a very small amount Purified water To 100

<2-4> Facial wash ( Cleansing Foam )

To prepare an antimicrobial cleanser (cleansing foam) containing the peptide of the present invention, it may be formulated as described in [Table 4] below and may 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

<2-5> 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 5] below.

Compounding 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

<2-6> Massage Cream

In order to prepare an antimicrobial massage cream containing the peptide of the present invention, it is prepared according to a conventional method in the field of cosmetics as described in the following [Table 6].

Compounding ingredient Content (% by weight) The peptide of the present invention 0.1 to 30% Propylene glycol 6.0 glycerin 4.0 Triethanolamine 0.5 Wax 2.0 Tocophenylacetate 0.1 Polysorbate 60 3.0 Sorbitan sesquioleate 2.5 Cetearyl alcohol 2.0 Liquid paraffin 30.0 Xanthan gum 0.5 Incense, preservative a very small amount Purified water To 100

<2-7> Pack

To prepare an antimicrobial pack containing the peptide of the present invention, the preparation is carried out according to the conventional production method in the field of cosmetics as shown in Table 7 below.

Compounding ingredient Content (% by weight) The peptide of the present invention 0.1 to 30% Propylene glycol 2.0 glycerin 4.0 Polyvinyl alcohol 10.0 ethanol 7.0 Paiji-40 Hydrogenated Castor Oil 0.8 Triethanolamine 0.3 Incense, preservative a very small amount Purified water To 100

< Manufacturing example  3> Manufacturing of food

Foods containing the peptides of the present invention were prepared as follows.

<3-1> Production of flour food

0.5 to 5.0 parts by weight of the peptide of the present invention was added to wheat flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture to prepare a health food for antifungal drugs.

<3-2> Dairy products ( dairy products )

5-10 parts by weight of the peptide of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<3-3> Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The peptide of the present invention was concentrated under reduced pressure in a vacuum concentrator, dried by spraying and dried in a hot-air drier, and pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder.

The grains, seeds, and dry powder of the peptides of the present invention prepared above were blended in the following proportions.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

The peptide (3 parts by weight) of the present invention,

(0.5 parts by weight), and

Rhubarb (0.5 parts by weight).

<3-4> Production of health supplement foods

100 mg of the peptide of the present invention

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

0.13 mg vitamin B1

0.15 mg of vitamin B2

0.5 mg vitamin B6

0.2 [mu] g vitamin B12

10 mg vitamin C

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

24.8 mg of magnesium chloride

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

< Manufacturing example  4> Health drink  Produce

100 mg of the peptide of the present invention

Citric acid 100 mg

100 mg of oligosaccharide

2 mg of plum concentrate

100 mg taurine

Purified water was added to 500 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the resulting solution was filtered to obtain a sterilized 1 liter container, sealed sterilized, It is used in the production of the health beverage composition of the invention.

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. And can be used in the manufacture of an ointment which can be applied thinly to human body according to its effect, which is included in the purpose and scope of the present invention as defined in the appended claims.

<110> Industry-Academic Cooperation Foundation Chosun University <120> Novel antimicrobial peptide from Pinus densiflora Sieb. et Zucc.          and use thereof <130> 14p-01-33 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 15 <212> PRT <213> Pinus densiflora Sieb. et Zucc. <400> 1 Xaa Xaa Xaa Xaa Phe Lys Phe Leu Ala Arg Thr Gly Lys Phe Leu   1 5 10 15

Claims (15)

An antibiotic peptide consisting of the amino acid sequence of SEQ ID NO: 1.
The method of claim 1, wherein the peptide is pine (Pinus densiflora Sieb. et Zucc. &lt; / RTI &gt;
The antibiotic peptide according to claim 1, wherein the peptide has a molecular weight of 1907.0 Da.
The peptide of claim 1, wherein the peptide is selected from the group consisting of Escherichia coli ), Staphylococcus ( Staphylococcus aureus) aureus , Staphylococcus epidermis , and Pseudomonas aeruginosa , which have antimicrobial activity.
The method of claim 1, wherein the peptide is Salmonella typhimurium (Salmonella typhimurium), and Listeria monocytogenes to Ness (Listeria monocytogenes ). &lt; RTI ID = 0.0 &gt; Antibody &lt; / RTI &gt;
1) pulverizing the pine needle and extracting it with an extraction solvent;
2) centrifuging the extract of step 1) and recovering the supernatant fraction;
3) separating a fraction having a molecular weight of 10 KDa or less in the supernatant fraction of step 2);
4) eluting the separated fractions of step 3) at a concentration gradient of acetonitrile B of from 5 to 65% in the mobile phase in a chromatography packed with a _C18 column. A method for producing antibiotic peptides.
An antibiotic pharmaceutical composition comprising the antibiotic peptide of claim 1 as an active ingredient.
8. The method of claim 7, wherein the peptide is selected from the group consisting of Escherichia coli ), Staphylococcus ( Staphylococcus aureus) aureus , Staphylococcus epidermis) and Pseudomonas Ke rugi labor (Pseudomonas aeruginosa) A pharmaceutical composition for antibiotic, characterized in that it has an antimicrobial activity against.
The method of claim 7, wherein the peptide is Salmonella typhimurium (Salmonella typhimurium), and Listeria monocytogenes to Ness (Listeria monocytogenes of the present invention have an antifungal activity.
11. An antidandruff composition comprising the antimicrobial peptide of claim 1 as an active ingredient.
An antibiotic cosmetic composition comprising the antibiotic peptide of claim 1 as an active ingredient.
An external preparation for antibiotics comprising the antibiotic peptide of claim 1 as an active ingredient.
An antibiotic health food containing the antimicrobial peptide of claim 1 as an active ingredient.
An antibiotic pesticide containing the antibiotic peptide of claim 1 as an active ingredient.
An article of antibiotics containing the antimicrobial peptide of claim 1 as an active ingredient.

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