KR20190051472A - Oxyasin-5 peptide isolated from Oxya chinensis sinuosa and antimicrobial, antimycotic and antiallergic composition comprising it - Google Patents

Abstract

The present invention relates to an Oxyasin-5 peptide derived from Oxya chinensis and an antimicrobial, antimycotic or antiallergic composition containing the same. The Oxyasin-5 peptide exhibits excellent antimicrobial, antimycotic and antiallergic activities with little cytotoxicity, and thus can be provided as a composition for antibiotics, food preservatives, cosmetic preservatives, pharmaceutical preservatives and treatment of various allergic diseases. In the peptide technology related to Oxya chinensis, studies on the antimicrobial, antimycotic or antiallergic diseases with peptides having the same amino acid sequence as the present invention have not been conducted yet.

Description

The present invention relates to an oxyacyl-5 peptide derived from a rice-grasshopper, and an antibacterial, antifungal or antiallergic composition containing the same, and an antimicrobial, antifungal or antiallergic composition containing the same,

The present invention relates to an oxyacyl-5 peptide derived from a rice plant and a composition thereof, and more particularly to a novel oxyacyl-5 peptide derived from a rice plant and showing excellent antibacterial, antifungal or antiallergic activity, ≪ / RTI >

Infection with pathogenic microorganisms is one of the most common and fatal causes of human disease, unfortunately the antibiotic resistance of pathogenic microorganisms has been caused by the abuse of antibiotics. In fact, the rate at which pathogenic microorganisms are resistant to new antibiotics is much faster than the rate at which analogs of new antibiotics are developed. For example, pathogenic microbial species such as Enterococcus faecalis , Mycobacterium tuberculosis , and Pseudomonas aeruginosa , which can pose a life threat, I have developed resistance to all antibiotics. Tolerance to antibiotics is distinct from resistance to antibiotics, which was first discovered in the 1970s by Pneumococcus sp. And provided important clues to the mechanism of action of penicillin. Resistant species stop growing in the presence of the usual concentration of antibiotics but do not eventually die. This resistance occurs because antibiotics inhibit the cell wall synthetase when autolytic enzymes such as autolysin do not activate, which makes penicillin an endogenous hydrolytic enzyme. Activation kills bacteria and bacteria also inhibit their activity, resulting in the survival of antibiotics.

It is clinically very important that pathogenic microorganisms have resistance to antibiotics, because the inability to eradicate the resistant microorganisms is ineffective in antibiotic treatment in clinical infections. In addition, tolerance is considered to be a prerequisite for resistance to antibiotics, which is due to the survival of strains despite antibiotic therapy. These strains acquire new genetic elements that are resistant to antibiotics and continue to grow in the presence of antibiotics. Since virtually all pathogenic microorganisms showing resistance are known to have resistance, development of new antibiotics capable of killing pathogenic microorganisms resistant to such antibiotic resistance is urgent.

As described above, it is necessary to develop a new antibiotic to prevent damage caused by pathogenic microorganisms showing resistance to antibiotics, and to develop a new antibiotic that acts independently of autolysin activity. There is also a need to provide pharmaceutical compositions for effectively treating such new antibiotics with pathogenic microorganisms.

On the other hand, some of the substances that play an important role in maintaining the homeostasis of organisms are physiologically active substances derived from various organisms. Many researches have been carried out on a number of biologically active substances, among which antimicrobial peptides isolated from various organisms are known to act in various ways, ranging from bacteria, fungi and viruses. Antimicrobial peptides are also known to play an important role in host defense and the innate immune system.

Allergy refers to a phenomenon in which a living organism in contact with an exotic substance reacts differently to the substance. When an organism comes into contact with a certain exogenous substance, an antigenic antibody reaction causes a rapid change in the response ability in vivo. . For a heterologous substance, a living organism produces antibodies and lymphocytes that specifically react with the antigen (antigen), and when the antigen is contacted again, various immune responses are produced. This immune response or immune response is one of the important defense mechanisms for the self-preservation of living organisms. It usually acts on the living body in a protective manner, but sometimes this mechanism causes disability to the living body. Allergy means "hypersensitive", meaning that the Greek word allos is the etymology, which means "modified." The term allergy was first used by the French scholar von Pirke in 1906. Allergies or allergies are pronounced in English, and allergies are pronounced in German. Antigens that cause allergic reactions are called allergens. Typical allergens include pollen, drugs, vegetable fibers, bacteria, food, dyes, and chemicals. There are several defense mechanisms in the immune system to protect the body against antigens. The most abundant of these are lymphocytes, which are specific for reacting to specific antigens, including B cells and T cells. B cells bind to an antigen to produce an antibody that is a protein that destroys and neutralizes the antigen. Instead of producing antibodies, T cells directly bind to the antigen and stimulate the attack. An allergic reaction occurs as an immediate allergic or delayed allergy, depending on which of the B or T cells the antigen reacts to. The diseases caused by allergies include various diseases including autoimmune diseases and collagen diseases. Generally, allergic diseases are classified into classical allergic diseases such as anaphylactic shock, food allergy, allergic rhinitis, hay fever, bronchial asthma , Drug allergies, plant allergies, urticaria, eczema, and allergic contact dermatitis. They are allergic diseases that cause disease, but there are cases where other biochemical conditions are necessary for their development. The same symptoms may also be caused by non-allergic mechanisms.

Accordingly, the inventors of the present invention have been studying various physiological activities using a peptide derived from a rice plant ( Oxya chinensis sinuosa ), and confirmed that the novel peptide oxyacin-5 has an antibacterial, antifungal or antiallergic potency, there was.

Korean Patent No. 10-0625875 (Novel peptide isolated from Aspergillus nidulans and a pharmaceutical composition containing the same as an active ingredient, applicant: Chosun University, registered on September 12, 2006) Korean Patent No. 10-0964136 entitled " Antibacterial or Antifungal Peptide Gene Isolated from Ulmus japonicus Larvae and Synthetic Peptide Having Antibacterial or Antifungal Activity, Applicant: Korea, Date of Registration: June 09, 2010) Korean Patent No. 10-1743113, entitled: Antimicrobial Peptide Oxyacyline-1 Derived from a Rice Mullet and Its Composition, Applicant: Korea, Registered on May 29, 2017) Korean Patent No. 10-1740551 (entitled "Antimicrobial Peptide Oxyacyline-2 Derived from a Rice Mullet and Its Composition, Applicant: Korea, Registration Date: May 22, 2017) Korean Patent No. 10-1697179 (Title of the Invention: Composition for prevention and treatment of atopic dermatitis comprising scolopendrazine-1 peptide as an active ingredient, applicant: Korea, registered on January 11, 2017)

It is an object of the present invention to provide an oxyacyl -5 peptide having antibacterial, antifungal or antiallergic potency separated from a rice plant ( Oxya chinensis sinuosa ) and a composition containing the same.

The present invention relates to a peptide represented by the following SEQ ID NO: 1 synthesized from a gene of an antibacterial, antifungal or antiallergic peptide isolated from a rice plant ( Oxya chinensis sinuosa ).

SEQ ID NO: 1: RRVGRRQR-NH ₂

The peptide may exhibit an antimicrobial activity against at least one of Staphylococcus aureus and Escherichia coli 0-157, and the antifungal activity against Candida albicans Lt; / RTI > In addition, the peptide has an effect of inhibiting secretion of? -Hexosaminidase, histamine, and TNF-alpha (Tumor necrosis factor-?).

Accordingly, the present invention provides an antibacterial, antifungal or antiallergic composition characterized by containing the peptide as an active ingredient. The antiallergic composition is characterized in that it has an effect of preventing or treating an allergic disease selected from anaphylactic shock, food allergy, allergic rhinitis, hay fever, bronchial asthma, drug allergy, plant allergy, urticaria, eczema and allergic contact dermatitis do.

In addition, the present invention can provide a composition for improving or treating food poisoning or enteritis containing the peptide as an active ingredient. The food poisoning or enteritis is characterized by being caused by one or more causative bacteria selected from Staphylococcus aureus and Escherichia coli 0-157.

Due to the antimicrobial and antifungal effects, the present invention may provide a pharmaceutical composition for antimicrobial or antifungal preparation containing the peptide as an active ingredient, an antibiotic, a food preservative, a cosmetic preservative or a pharmaceutical preservative.

Hereinafter, the present invention will be described in detail.

The present invention relates to a peptide represented by SEQ ID NO: 1 synthesized from a gene of an antibacterial, antifungal or antiallergic peptide isolated from a rice plant ( Oxya chinensis sinuosa ). Accordingly, the present invention may relate to an antibacterial, antifungal or antiallergic peptide directly isolated from a rice plant ( Oxya chinensis sinuosa ) having the same sequence as SEQ ID NO: 1.

Accordingly, the present invention may relate to an antibacterial, antifungal or antiallergic peptide directly isolated from a rice plant ( Oxya chinensis sinuosa ) having the same sequence as SEQ ID NO: 1.

The peptide of the present invention may have a carboxyl end in the form of -NH ₂ or -OH.

In the present invention, in order to identify a peptide gene derived from the rice plant ( Oxya chinensis sinuosa ), firstly, the rice grasshopper is rapidly frozen with liquid nitrogen to isolate the total RNA, and the gene information about the rice hatchlot transcripts is confirmed using the RNA seq analysis method Based on the sequence of the translated amino acids (peptides) from each transcript, it is possible to use a known unicin, antifungal or antiallergic peptide, Can be selected.

The amino acid sequence of the peptide is RRVGRRQR-NH ₂ , which is composed of 8 amino acids and is named Oxyasin-5.

The present invention also provides an antibacterial, antifungal or antiallergic composition comprising the peptide as an active ingredient, a composition for preventing / treating enteritis, an antibiotic, a food preservative, a cosmetic preservative or a pharmaceutical preservative.

More specifically, the oxyacin-5 peptide is useful as an antibiotic activity candidate, such as Staphylococcus aureus , Escherichia coli 0-157 ( Escherichia coli 0-157), Candida albicans Antimicrobial or antifungal activity is shown by RDA assay (radial diffusion assay) against Candida albicans . In addition, the oxyacyl-5 peptide has an effect of suppressing secretion of β-hexosaminidase, histamine and TNF-α, which are typical proteins expressed in allergic diseases . Therefore, the oxyacyl-5 peptide of the present invention can be usefully used as an antibacterial, antifungal or antiallergic composition.

In addition, since the composition containing the oxyacyl-5 peptide of the present invention as an active ingredient shows little cytotoxicity, it can be usefully used as a safe antimicrobial, antifungal or antiallergic composition.

 The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The pharmaceutical composition containing the oxyacyl-5 peptide of the present invention can also be administered parenterally at the time of clinical administration and can be used in the form of a general pharmaceutical preparation. The peptide or the composition containing the peptide may be administered in various forms of parenteral administration. In the case of formulation, the peptide may be prepared using a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, 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 non-aqueous solvent and suspension agent. As the base of the suppository, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

In addition, the oxyacyl-5 peptide of the present invention can be used in combination with various carriers permitted as medicaments such as physiological saline or an organic solvent, and can be used as glucose, sucrose or dextran such as sucrose, Antioxidants such as carbohydrates, ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers can be used as pharmaceuticals.

The total effective amount of the oxyacyl-5 peptide of the present invention or a peptide having more than 95% homology thereto in the above pharmaceutical composition may be in the form of a bolus or a single dose by infusion for a relatively short period of time, , And may be administered by a fractionated treatment protocol in which multiple doses are administered over a prolonged period of time. The concentration is determined in consideration of various factors such as the route of administration and the number of treatments of the pharmaceutical composition as well as the age and health condition of the patient. Thus, in view of this point, It will be possible to determine the appropriate effective dose depending on the particular use as the pharmaceutical composition containing the novel peptide of the invention.

In addition, the present invention provides antibiotics, food preservatives, cosmetic preservatives, and pharmaceutical preservatives containing the oxysacin-5 peptide as an active ingredient. In this case, food preservative, cosmetic preservative or pharmaceutical preservative is an additive used to prevent deterioration, decay, discoloration and chemical change of food or medicine. It includes antiseptic and antioxidant and inhibits the growth of microorganisms such as bacteria, fungi and yeast And also includes functional antibiotics such as inhibiting the growth of microorganisms or sterilizing the microorganisms in foods or medicines. Ideal conditions for these food preservatives, cosmetics or pharmaceutical preservatives should not be toxic and should be effective in trace amounts.

In addition, the present invention can provide a composition for improving or treating food poisoning or enteritis containing the peptide as an active ingredient. The food poisoning or enteritis is caused by at least one of Staphylococcus aureus and Escherichia coli 0-157 ( Escherichia coli 0-157).

The present invention can provide a composition for preventing or treating an allergic disease containing the peptide as an active ingredient. The allergic disease is characterized by being a disease selected from anaphylactic shock, food allergy, allergic rhinitis, hay fever, bronchial asthma, drug allergy, plant allergies, urticaria, eczema and allergic contact dermatitis.

The present invention relates to an oxyacyl-5 peptide derived from a rice-grasshopper and an antibacterial, antifungal or antiallergic composition containing the same. Since the oxyacin-5 peptide exhibits excellent antibacterial, antifungal and antiallergic activity with little cytotoxicity, it can be provided as a composition for treating antibiotics, food preservatives, cosmetic preservatives, pharmaceutical preservatives and various allergic diseases. No research has been conducted on peptides having the same amino acid sequence as the present invention in antimicrobial peptides related to the rice-grasshopper, which have antibacterial, antifungal or antiallergic properties.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a photograph showing antibiotic activity or antifungal activity of the oxyacin-5 peptide of the present invention. FIG.
FIG. 2 is a graph showing the results that the oxyacin-5 peptide of the present invention has no cytotoxic effect.
FIG. 3 is a graph showing the effect of the oxyacyl-5 peptide of the present invention on the secretion of β-hexosaminidase. FIG.
FIG. 4 is a graph showing the results of confirming the effect of the oxyacin-5 peptide of the present invention on the secretion of histamine.
5 is a graph showing the inhibitory effect of the oxyacyl-5 peptide of the present invention on TNF-a secretion.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are for illustrative purposes only and that the scope of the invention is not limited by these examples.

≪ Example 1 > Selection of antimicrobial peptides derived from rice hulls &

Example 1-1. Immunization and RNA collection of rice-grasshoppers

In order to identify the peptide gene showing antimicrobial, antifungal or antiallergic activity from the rice hermit, the whole rice was firstly isolated by rapid freezing of the rice hermit with liquid nitrogen. More specifically, rice grasshoppers were immunized and RNA was collected through the following procedure.

The female insect adults were supplied from Jeonnam Agricultural Research and Extension Service and used for the experiment. For immunization, E. coli was first cultured in a TSB (Tryptic Soy Broth, Difco, USA) liquid medium at 37 ° C and 200 rpm for 18 hours in a shaking incubator, and then incubated again for 2 hours and 30 minutes under the same conditions , The cell number was taken to be 2 x 10 ⁶ colony forming units (CFU), injected into the body cavity using a microsyringe and incubated at 25 ° C for 18 hours to induce immunization.

Next, to maximize the expression of the physiologically active substance, RNA-seq was performed using total rice of the rice-grasshopper immunized with normal rice and E. coli. First, prepare a normal rice seedlings and an immunized rice seedlings. To maintain the freshness of the tissues as much as possible, they are rapidly frozen with liquid nitrogen and ground in a mortar and then mixed with 800 μl of TRIZOL and reacted at room temperature for 5 minutes. Add 160 μl of chroloform and mix. The mixture is centrifuged at 12,000 g for 15 minutes at 4 ° C. Then, the supernatant is taken, and 500 μl of isopropanol is added. The mixture is reacted at room temperature for 10 minutes and then centrifuged at 12,000 g for 10 minutes at 4 ° C. The precipitated RNA was washed with 800 μl of 75% ethanol, dried and dissolved in sterilized water to prepare RNA. Prepared total RNA was checked for RNA purity and quality using an Agilent 2100 Bioanalyzer instrument.

Examples 1-2. Selection of antimicrobial, antifungal or antiallergic peptides

Using the RNA seq analysis method, gene information on rice transporter transcripts was confirmed. Based on the sequence of the translated amino acids from each transcript and the existing antibacterial, antifungal or antiallergic peptide properties, filtration. As a result, new antimicrobial, antifungal or antiallergic peptide presumed genes were selected. The selected amino acid sequence was RRVGRRQR-NH ₂ , which was composed of 8 amino acids and named Oxyasin-5 (Table 1).

Peptides Amino acid sequence Oxyasin-5 RRVGRRQR-NH ₂

≪ Example 2: Synthesis and separation of oxyacin-5 >

The oxyacyl-5 peptide derived from the rice-grasshopper identified in Example 1 was synthesized and isolated and purified.

First, in order to synthesize oxyacin-5 peptide, the present inventors used Merrifield's liquid phase method (Merrifield, RB., J. Am. Chem. Soc., 85, 2149, 1963 ) Was used to prepare the peptide. Method of the peptide synthesis the Fmoc (9-fluorenylmethoxycarbonyl) amino acids of the N _α were synthesized and used as a protecting group (protecting group) of the amino group (amino group) to the conventional method (solid phase method).

Concretely, the peptide having carboxyl terminal in the form of -NH ₂ used Rink Amide MBHA-Resin as the starting material, and the peptide having carboxyl terminal in the form of -OH used Fmoc-amino acid-Wang Resin as a starting material. The elongation of the peptide chain by coupling of the Fmoc-amino acid was performed by DCC ( N -hydroxybenzo-triazole (HOBt) -dicyclo-hexylcar-bodiimide) method.

After the Fmoc-amino acid of the amino terminal of each peptide was coupled, the Fmoc group was removed with 20% piperidine / N-methylpyrrolidone (NMP) solution, washed several times with NMP and dichloromethane (DCM) It was dried with gas. A solution of TFA (trifluoroacetic acid) -phenol-thioanisole-H ₂ O-triisopropylsilane (85: 5: 5: 2.5: 2.5, vol./vol.) Was added and the reaction was carried out for 3 hours. The peptides were separated and the peptide was precipitated with diethyl ether. The crude peptide thus obtained was subjected to a reverse phase-HPLC column (Delta Pak, C ₁₈ 300 Å, 15 μm, 19.0 mm × 30 mm) using an acetonitrile gradient containing 0.1% TFA , Waters).

After the synthetic peptide was hydrolyzed with 6 N HCl at 110 ° C for 24 hours, the resulting residue was concentrated under reduced pressure, dissolved in 0.02 N HCl, and the amino acid composition was measured with an amino acid analyzer (Hitachi 8500 A). The molecular weight was calculated based on the sequence of the synthesized peptides, and the exact molecular weight was measured using a MALDI mass spectrometer (MALDI).

As a result, it was confirmed that the antimicrobial, antifungal or antiallergic peptide having the correct amino acid sequence was synthesized because the calculated molecular weight and the calculated molecular weight were identical.

≪ Example 3: Investigation of antimicrobial or antifungal activity against pathogenic microorganisms of oxyacin-5 antimicrobial peptide >

RDA (radial diffusion assay) was performed to investigate the antimicrobial activity against oxyacin-5 peptide. Melittin, a powerful antimicrobial peptide isolated from bees, was used as a positive control in the present invention. In the present invention, the oxyacin-5 peptide was stored at -20 ° C and dissolved in sterilized distilled water.

Staphylococcus aureus , Escherichia coli 0-157 ( Escherichia coli 0-157), 3% (w / v) trypticase soy broth (TSB) was used to measure the antimicrobial activity of the peptides. At 37 ° C and 200 rpm for 18 hours, and then cultured for 2 hours and 30 minutes at a concentration of ⁴ × 10 ⁶ CFU (colony forming units) / ml under the same conditions.

Thereafter, a sterilized underlay gel consisting of citrate phosphate buffer (9 mM sodium phosphate, 1 mM sodium citrate, pH 7.4) and 1% (w / v) type I (low electroendosmosis) agarose and 0.03% TSB ( ⁴ × 10 ⁶ colony forming units / ㎖) in an underlay gel and mix. After pouring on a square plate, if the underlay gel is hardened, a hole with a diameter of 3 mm is pored and 5 μl of peptide per concentration .

10 ml of overlay gel (6% TSB, 1% agarose) was poured and cultured at 37 ° C again to confirm the formation of a clear zone (CLEAR ZONE) after culturing at 37 ° C for 3 hours to allow the peptide to diffuse. The results are shown in Fig.

Candida albicans , a pathogenic fungus, was grown overnight at 30 ° C and 200 rpm in YPD medium (1% yeast extract, 2% peptone, 2% dextrose) They were inoculated for 2 hours to become algebra.

Then Citrate phosphate buffer (9 mM sodium phosphate , 1 mM sodium citrate, pH 7.4) and 1% (w / v) type (low electroendosmosis) agarose, 0.03% of bacterial culture eda sterilized underlay gel consisting of TSB (4x10 ⁶ colony forming units / ㎖). After mixing, the mixture was poured into a square plate. When the underlay gel was hardened, a hole with a diameter of 3 mm was punched out and 5 μl of peptide was added per concentration.

The peptide was incubated at 37 ° C for 3 hours to allow the peptide to diffuse, and 10 ml of overlay gel (6% TSB, 1% agarose) was poured thereon and cultured again at 37 ° C to confirm formation of a clear zone. 1.

Referring to FIG. 1, it was confirmed that the oxyacin-5 peptide has activity against Gram-negative bacteria and positive bacteria, which is increased in a concentration-dependent manner. This shows that it can be used as a therapeutic agent against existing harmful strains. In addition, as a result of the measurement of antifungal activity, it was confirmed that a clear zone (CLEAR ZONE) was formed and the activity was increased in a concentration-dependent manner as in the case of Gram-negative bacteria / positive bacteria.

<Example 4: Cytotoxicity of oxyacin-5>

Experiments were conducted using RBL-2h3 cells known as inflammation and allergy-related cells. The cells were cultured in DMEM medium containing antibiotics (100 units / ml penicillin, 100 μg / ml streptomycin) and 10% fetal bovine serum (FBS) in a 5% CO ² incubator at 37 ° C.

AQueous One Solution Reagent (MTS) (Promega, USA)를 첨가하고 3 ~ 4 시간 반응시켜 색의 변화를 확인하였고 이를 multi detecter (Beckman DTX8800)를 이용하여 450nm 파장에서 흡광도 측정을 통해 확인하였다. Cytotoxicity was confirmed by MTS assay. To this end, 100 μl of 2 × 10 ⁴ cells / well cells were dispensed into a 96-well culture vessel and treated with oxyacyl-5 in a concentration-dependent manner for 24 hours. Afterwards, 10 [mu] l CellTiter 96

AQueous One Solution Reagent (MTS) (Promega, USA) was added and reacted for 3 ~ 4 hours to confirm the change of color. It was confirmed by absorbance measurement at 450nm wavelength using multi detector (Beckman DTX8800).

The results are shown in FIG. 2. Referring to FIG. 2, it is confirmed that the cell survival rate of oxyjacin-5-treated group is not significantly different from that of the untreated group. Therefore, it can be seen that oxy-5 is not cytotoxic at a maximum concentration of 200 占 퐂 / ml.

Example 5 Identification of antiallergic activity of oxyacin-5 [

Example 5-1. β-hexosaminidase release assay

As another method for measuring the antiallergic activity of oxyacyl-5 peptide, the amount of β-hexosaminidase secreted in RBL-2h3 cells was confirmed. Cells were prepared at 10 ⁶ cells / ml, and then 200 μl of each was added to 24 wells, treated with IgE and then incubated overnight. The prepared cells were treated with oxyacin-5 and allowed to react for 30 minutes. After that, DNP was applied to each well for 3 hours at a concentration of 10 μg / ml to induce allergy. The supernatant was collected and measured using the β-hexosaminidase release assay kit. The results are shown in FIG.

Referring to FIG. 3, it was confirmed that the oxyacin-5 peptide decreased the secretion amount of? -Hexosaminidase.

Example 5-2. Histamine release assay

The amount of histamine secreted in RBL-2h3 cells was determined to measure antiallergic activity of oxyacyl-5 peptide. Cells were prepared at 10 ⁶ cells / ml, and then 200 μl of each was added to 24 wells. The cells were treated with IgE and then incubated overnight. The prepared cells were treated with oxyacin-5 and allowed to react for 30 minutes. Then, to induce allergy, DNP was applied for 3 hours at a concentration of 10 μg / ml for each well. The supernatant was collected and measured using a histamine release assay kit. The results are shown in FIG.

Referring to FIG. 4, it was confirmed that the oxyacin-5 peptide reduced histamine secretion amount.

Example 5-3. Confirm TNF-α inhibition

In order to elucidate the mechanism of antiallergic activity of oxyacin-5 peptide, the secretion inhibition of TNF-α, which is known to be secreted in the course of allergy induction, was confirmed through experiments. As a method, 10 ⁶ cells / ㎖ of cells were prepared, and then 200 μl of each was added to 24 wells, followed by treatment with IgE. The prepared cells were treated with oxyacin-5 and allowed to react for 30 minutes. After that, DNP was used to induce allergy. The supernatant was collected at a concentration of 10 μg / ml for each well, and the amount of TNF-α secretion was measured by ELISA. The results are shown in FIG.

Referring to FIG. 5, it can be seen that the oxyacin-5 peptide reduces the secretion of TNF-? From mast cells.

These results indicate that the oxyacyl-5 peptide of the present invention is not only cytotoxic, but also exhibits antibacterial, antifungal or antiallergic activity and thus can be used as a composition for the treatment of safe antibiotics, food preservatives, cosmetic preservatives, medicines and allergic diseases .

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Oxyasin-5 peptide isolated from Oxya chinensis sinuosa and          antimicrobial, antimycotic and antiallergic composition          comprising it <130> 240 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 8 <212> PRT <213> Unknown <220> <223> Oxya chinensis sinuosa <400> 1 Arg Arg Val Gly Arg Arg Gln Arg   1 5

Claims (9)

The peptide represented by SEQ ID NO: 1, which is synthesized in a gene of an antibacterial, antifungal or antiallergic peptide isolated from a rice plant ( Oxya chinensis sinuosa ).
SEQ ID NO: 1: RRVGRRQR-NH ₂ The method according to claim 1,
Wherein said peptide exhibits an antimicrobial activity against any one or more of Staphylococcus aureus and Escherichia coli 0-157 ( Escherichia coli 0-157). The method according to claim 1,
Wherein said peptide exhibits antifungal activity against Candida albicans . The method according to claim 1,
Wherein said peptide has an effect of inhibiting the secretion of? -Hexosaminidase, histamine and TNF-alpha. An antimicrobial, antifungal or antiallergic composition comprising the peptide of any one of claims 1 to 4 as an active ingredient. 6. The method of claim 5,
Wherein the composition is effective for the prevention or treatment of an allergic disease selected from anaphylactic shock, food allergy, allergic rhinitis, hay fever, bronchial asthma, drug allergy, plant allergy, urticaria, eczema and allergic contact dermatitis. , An antifungal or antiallergic composition. A composition for improving or treating food poisoning or enteritis, which comprises the peptide of any one of claims 1 to 4 as an active ingredient. An antibiotic comprising the peptide of any one of claims 1 to 4 as an active ingredient. A preservative for foods, cosmetics or pharmaceuticals containing the peptide of any one of claims 1 to 4 as an active ingredient.

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