KR20130080172A - Novel antifungal peptide isolated from solanum tuberosum l cv. jopung and use therof - Google Patents

Abstract

PURPOSE: A novel antifungal peptide isolated from Solanum tuberosum L cv. Jopung is provided to have an antifungal activity against Candida albicans and to be used in pharmaceutical compositions, cosmetic compositions, agrochemicals, preservatives for foods, cosmetics, and pharmaceutical products. CONSTITUTION: An antifungal peptide has an amino acid sequence of sequence number 1 and is isolated from Solanum tuberosum L cv. Jopung. The molecular weight of the antifungal peptide is 1.0831 kDa. A method for preparing the peptide comprises the steps of: freeze-drying and pulverizing bulbs of Solanum tuberosum L cv. Jopung; extracting the pulverized bulbs and preparing an extract; centrifuging the extract and collecting a supernatant fraction; and isolating and purifying the supernatant fraction by chromatography. An antifungal pharmaceutical composition contains the peptide as an active ingredient.

Description

Novel antifungal peptide isolated from potato and its use {Novel antifungal peptide isolated from Solanum tuberosum L cv. Jopung and use therof}

The present invention is a potato ( Solanum tuberosum L cv. A novel antifungal peptide isolated from Jopung) and an antifungal pharmaceutical composition containing the same as an active ingredient.

Functional antibiotics, such as pesticides, food preservatives, cosmetics, and pharmaceuticals, are broadly used as additives to prevent the functions of crops as pesticides, deterioration, decay, discoloration, and chemical changes of foods and medicines. Included in a narrow sense is to inhibit the growth of microorganisms such as bacteria, fungi, yeast to inhibit the development or disinfection of decay microorganisms in pesticides, food and pharmaceuticals. Ideal conditions such as pesticides, food preservatives, cosmetics, and pharmaceuticals should be nontoxic and effective in trace amounts.

The plants and animals producing the peptide antibiotic produced naturally (Bevins et al., Ann. Rev. Biochem., 59, 395-414, 1990). The peptide antibiotic is structurally divided into three groups, the first being a cysteine-rich β-sheet peptide, the second being an α-helical amphiphilic molecule, and the third 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 present among these structures is a structure that does not have a cysteine residue and forms an amphiphilic alpha helical, and is an antimicrobial peptide found in insects, cecropin. ) Has this structure.

Natural antimicrobial agents are antimicrobial agents that selectively control viruses, bacteria, yeasts, molds, etc. by maximizing selectivity, biochemical, and physiological differences from natural or microbial resources, but are harmless to humans or minimize toxicity. Currently reported natural antimicrobial agents have been developed, such as chynochithiol from cypress, megnonole from magnolia, grapefruit seed extract DF-100, etc., but the scale is currently very small. Currently, natural herb extracts are known to have a high bactericidal effect against microorganisms by inhibiting cell proliferation mechanisms, and have been reported as safe natural antibacterial agents that are nontoxic, noncorrosive and harmless to the human body. However, plants currently used as natural antimicrobial agents are not grown easily because they are small in cultivation scale or are not cultivated a lot in Korea and are difficult to cultivate. However, potato has a large growing area, and its quantity is also high, so it is easy to secure the material and lower the raw material cost. The potato peels are dried and powdered and placed in the bathroom or in a humid place to prevent insects.When hot potatoes are cultivated in hot water, they can be ground or sliced and glued to the affected area. It acts as an antibacterial agent.

Thus, the inventors of the potato ( Solanum tuberosum L cv. Jopung) extract proteins and isolate and purify peptides to obtain Potide-J, which has antifungal activity against Candida albicans , against human red blood cells. The present invention was completed by confirming that hemolytic activity was not found to be cytotoxic and confirming its function as a natural antifungal agent.

The object of the present invention is potato ( Solanum) tuberosum L cv. It is to provide an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung).

Still another object of the present invention is to provide a method for preparing the antifungal peptide.

Still another object of the present invention is to provide an antifungal composition using the antifungal peptide.

In order to achieve the above object, the present invention is a potato ( Solanum tuberosum L cv. An antifungal peptide consisting of the amino acid sequence set forth in SEQ ID NO: 1 isolated from Jopung) is provided.

In addition,

1) Potato ( Solanum tuberosum L cv. Lyophilizing the bulb of Jopung) and pulverizing it into powder;

2) preparing an extract by extracting the powder of step 1) with a buffer solution;

3) recovering the supernatant fraction of the extract by centrifuging the extract of step 2); And

4) It provides a method for producing the antifungal peptide comprising the step of separating and purifying the supernatant fraction of step 3) using chromatography.

The present invention also provides an antifungal pharmaceutical composition comprising the antifungal peptide as an active ingredient.

The present invention also provides an antifungal cosmetic composition containing the antifungal peptide as an active ingredient.

The present invention also provides a non-toxic pesticide containing the antifungal peptide as an active ingredient.

In addition, the present invention provides a preservative for preserving medicines, cosmetics, food or feed containing the antifungal peptide as an active ingredient.

Potato of the invention ( Solanum tuberosum L cv. Four of Tide antifungal peptides consisting of the amino acid sequence represented by SEQ ID NO: 1 isolated from Jopung) - J (Potide-J) are Candida albicans (Candida albicans ) has excellent antifungal activity and does not show cytotoxicity, so it can be usefully used as pharmaceutical composition, cosmetic composition, pesticide, food preservative, cosmetic preservative and pharmaceutical preservative.

1 is a potato ( Solanum tuberosum L cv. Figure shows a series of steps to isolate and purify the antibiotic functional peptide Potide-J from Jopung.
Figure 2 is a diagram showing the results of the electrophoresis of the proteins of AFP-J (AFP-J).
Figure 3 is a diagram showing the separation of the disulfide-dissolved peptide obtained by dissolving the protein extracted in order to obtain Potide-J (Potide-J) in HCl.
FIG. 4A shows the anti-bioactivity of purified peptides digested with HCl by Candida albicans ) is a diagram showing the measurement using cells.
4B is a diagram showing the antibiosis of Potide-J was measured using Candida albicans cells.
FIG. 5 is a diagram showing the antimicrobial activity of synthesized Potide-J using Candida albicans cells.
A: Candida albicans untreated with Potide-J; And
B: Candida albicans treated with 60 μg / ml of Potide-J.

Hereinafter, the present invention will be described in detail.

The present invention is a potato ( Solanum tuberosum L cv. An antifungal peptide consisting of the amino acid sequence set forth in SEQ ID NO: 1 isolated from Jopung) is provided.

The potatoes may be used without limitation, those grown specifically, commercially available.

The antifungal peptide specifically has a molecular weight of 1.0831 kDa, has antifungal activity against pathogenic fungi, and more specifically, the pathogenic fungus is Candida albicans ( Candida). albicans ), C. catenulate , C. intermidia , C. rugosa , C. glabrata and C. melibiosica Is one selected from the group consisting of, and most specifically, the pathogenic fungi is Candida albicans , but is not limited thereto.

In addition, the antifungal peptide does not specifically exhibit cytotoxicity, but is composed of an amino acid size of 10-mer (mer), but is not limited thereto.

The present inventors, in order to prepare a peptide having antibiotic activity, potato ( Solanum) tuberosum L cv. The protein was extracted from Jopung) to obtain AFP-J using Fast Protein Liquid Chromatography (FPLC) (see FIG. 1), which was separated using SDS-PAGE. It was confirmed that the specific protein of kDa (see Figure 2). In addition, the AFP-J was isolated and purified peptides using C18 Reverse Phase (RP) -High Performance Liquid Chromatography (HPLC) (see Figure 3), Candida albicans ( Candyda albicans ) Potide-J with antifungal activity against was obtained (see FIGS. 4A and 4B).

In addition, the inventors of the present invention confirmed the amino acid sequence and molecular weight of the potide-J, using an amino acid analyzer and MALDI-MS (Matrix Assisted Laser Desorption Ionization-Mass Spectrometry), the potide-J is SEQ ID NO: 1 It consists of an amino acid sequence described as and confirmed that the molecular weight is 1.0831 kDa.

In addition, the present inventors confirmed that the potide-J did not exhibit cytotoxicity in order to confirm that the potide-J showed cytotoxicity, and the red blood cell destruction ability of the potide-J was examined (see Table 1). .

In addition, the present inventors synthesized the peptide by the solid phase method using Fmoc (N- (9-fluorenyl) methoxycarbonyl) -protected amino acid in order to determine the antifungal activity of the synthesized peptide, and antifungal against Candida albicans As a result of the activity, it was confirmed that only the linear peptide, Potide-J, had antifungal activity (see FIG. 5).

Therefore, the potato of the present invention ( Solanum tuberosum L cv. Potide-J, an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung), is useful as a safe antibacterial agent to the human body because it not only has antifungal activity against Candida albicans but also exhibits no cytotoxicity. Can be used.

In addition,

1) Potato ( Solanum tuberosum L cv. Lyophilizing the bulb of Jopung) and pulverizing it into powder;

2) preparing an extract by extracting the powder of step 1) with a buffer solution;

3) recovering the supernatant fraction of the extract by centrifuging the extract of step 2); And

4) It provides a method for producing the antifungal peptide comprising the step of separating and purifying the supernatant fraction of step 3) using chromatography.

In the above method, the potato of step 1) can be used without limitation, specifically grown and commercially available.

In the above method, the buffer solution of step 2) is specifically 50 mM Tris-HCl pH 7.5, 10 mM EDTA, 150 mM NaCl, 1% dimethyl sulfoxide (DMSO) and 0.1% β-mercaptoethanol (β-mercaptoethanol), but is not limited thereto.

In the above method, the chromatography of step 4) is specifically Fast Protein Liquid Chromatography (FPLC) or Reverse Phase (RP) -High Performance Liquid Chromatography (HPLC), More specifically, separation by high-performance protein chromatography, purification by C18 reverse-phase high performance liquid chromatography, but is not limited thereto.

In the method, the antifungal peptide of step 4) specifically has a molecular weight of 1.0831 kDa, has antifungal activity against pathogenic fungi, and more specifically, the pathogenic fungus is Candida albicans ( Candida). albicans ), C. catenulate , C. intermidia , C. rugosa , C. glabrata and C. melibiosica Is one selected from the group consisting of, and most specifically, the pathogen fungus Candida albicans ( Candida albicans ), but is not limited thereto.

In addition, the antifungal peptide does not specifically exhibit cytotoxicity, but is composed of an amino acid size of 10-mer (mer), but is not limited thereto.

In addition, the present invention potato ( Solanum) of the present invention tuberosum L cv. It provides an antifungal pharmaceutical composition comprising an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung).

The pharmaceutical composition is specifically to have antifungal activity against pathogenic fungi, more specifically Candida albicans ( Candida albicans ), C. catenulate , C. intermidia , C. rugosa , C. glabrata and C. melibiosica It has antifungal activity against any one selected from the group consisting of), and most specifically, it has antifungal activity against Candida albicans , but is not limited thereto.

Potato of the invention ( Solanum tuberosum L cv. Fortide-J, an antifungal peptide consisting of the amino acid sequence of SEQ ID NO. 1 isolated from Jopung), has antifungal activity against Candida albicans and does not show cytotoxicity. It can be usefully used.

The novel 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.

In other words, the novel peptides of the present invention can be administered in a variety of parenteral formulations, which, when formulated, are prepared using diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. that are commonly used. do. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As a suppository base, witepsol, macrogol, tween 61, cacao paper, Liu ling, glycerogelatin and the like can be used.

In addition, the novel peptides of the present invention can be used in admixture with a variety of carriers that are accepted as pharmaceuticals, such as physiological saline or organic solvents, and carbohydrates such as glucose, sucrose or dextran to increase stability or absorption. Antioxidants such as ascorbic acid or glutathione, chelating agents, small molecule proteins or other stabilizers can be used as medicaments.

The effective dose of the novel 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.

In the pharmaceutical composition of the present invention, the total effective amount of the novel peptide of the present invention may be administered to the patient in a single dose by bolus form or by infusion for a relatively short period of time. Multiple doses may be administered by a fractionated treatment protocol with long term administration. 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 > of < / RTI > novel peptide as a pharmaceutical composition.

In addition, the present invention potato ( Solanum) of the present invention tuberosum L cv. It provides an antifungal cosmetic composition containing an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung) as an active ingredient.

Potato of the invention ( Solanum tuberosum L cv. Potide-J, an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung), is useful as an antifungal cosmetic composition because it not only has antifungal activity against Candida albicans but also exhibits no cytotoxicity. Can be used.

The cosmetic composition of the present invention contains components commonly used in cosmetic compositions in addition to the novel peptide as an active ingredient, and includes conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers. Include.

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 may be prepared in the form of a flexible lotion (skin), nutrition lotion (milk lotion), nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder .

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanta, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components. Can be.

In the case where 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. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / 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.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, and microcrystals are used as carrier components. Soluble cellulose, aluminum metahydroxy, bentonite, agar or tragacanta and the like can be used.

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

In addition, the present invention potato ( Solanum) of the present invention tuberosum L cv. It provides a non-toxic pesticide containing an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung) as an active ingredient.

Potato of the invention ( Solanum tuberosum L cv. Potide-J, an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung), has not only antifungal activity against Candida albicans but also does not show cytotoxicity and thus is useful as a nontoxic pesticide. Can be.

In addition, the present invention potato ( Solanum) of the present invention tuberosum L cv. The present invention provides a preservative for preserving pharmaceuticals, cosmetics, food or feed containing an antifungal peptide consisting of the amino acid sequence of SEQ ID NO: 1 isolated from Jopung) as an active ingredient.

Potide-J, an antifungal peptide consisting of the amino acid sequence set forth in SEQ ID NO: 1 isolated from the potato of the present invention ( Solanum tuberosum L cv. Jopung), not only has antifungal activity against Candida albicans but also cells It does not show toxicity and can be useful as a preservative for preserving medicines, cosmetics, food or feed.

Food preservatives, cosmetic preservatives and pharmaceutical preservatives are additives used to prevent the deterioration, decay, discoloration and chemical change of foods and medicines. These include fungicides and antioxidants. Functional antibiotics, such as inhibiting the development or disinfection of decay microorganisms in food and pharmaceuticals, are included. The ideal conditions for these food preservatives, cosmetics and pharmaceutical preservatives should be nontoxic and effective in trace amounts. Pesticides to eradicate pests of crops must also inhibit the growth of harmful microorganisms and harm the human body so that humans can safely consume pesticide-spread crops.

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

However, the following Examples are only for illustrating the present invention and do not limit the content of the present invention.

< Example  1> Protein Extraction from Potatoes

Ana Segura et al. (Ana Segura et al, 1999) and Quang Khai Huynh et al. (Quang Khai Huynh et al, 1992) were used to extract proteins using potato bulbs. Potato bulbs were ground to powder using mortar and pestle in the presence of liquid nitrogen, followed by protein extraction buffer (50 mM Tris-HCl). pH 7.5), 10 mM EDTA, 150 mM NaCl, 1% dimethyl sulfoxide (DMSO) and 0.1% β-mercaptoethanol (β-mercaptoethanol) were extracted.

< Example  2> Peptide  detach

After centrifuging the protein extract of <Example 1> at 6,000 × g for 30 minutes, the supernatant was chromatographed using a Sephacryl S-100 column (2.5 × 95 cm). In the ammonium bicarbonate buffer (pH 8.0), an antiadsorbent fraction (fraction I and fraction II) without antifungal activity and an active adsorption fraction (fraction III) were separated. Fraction III shown was isolated by Fast Protein Liquid Chromatography (FPLC) using a Superdex 200 prep grade column.

As a result, as shown in Fig. 1, a large adsorption peak was named AFP-J (Fig. 1).

< Example  3> Peptide  Detach Confirm

Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) was used to confirm AFP-J separation of <Example 2> by Laemmli (Laemmli et al. 1972). According to the method of) was performed using a flat gel (slab gel), and the sample and the standard protein was loaded (loaded) to 15% acrylamide gel (acrylamide gel). After electrophoresis, the gel was stained with Coomassie brilliant blue R-250, and destaining buffer (MeOH: glacial acetic acid: distilled water) 30: 10: 60) to decolorize the protein (band) was confirmed. Molecular weight standard proteins include Low Molecular Weight Range Bovine Serum Albumin (66 kDa), Chicken Egg Ovalbumin (45 kDa), Rabbit Muscle GAPDH (Glyceraldehyde) -3-phosphate Dehydrogenase) (36 kDa), Bovine Erythrocytes Carbonic Anhydrase (29 kDa), Bovine Pancreas Trypsinogen (24 kDa), Soybean Trypsin Inhibitor (20 kDa), Bovine Milk a-Lactalbumin (14.2 kDa), Bovine Lung Aprotinin (6.5 kDa) to determine the molecular weight of the protein.

As a result, as shown in FIG. 2, AFP-J (AFP-J) was confirmed to include a specific protein having a molecular weight of 15 kDa (FIG. 2).

< Example  4> Peptide  refine

AFP-J of <Example 2> was dissolved in HCl and treated at 60 ° C. for 24 hours. Then add loading buffer to terminate the reaction and place all samples in a Shim-pack VP-ODS, C18 column (4.6 × 250 mm; 4.6 ± 0.3 im particles; 12.0 ± 1.0 nm pores; Shimadzu Purification by Reverse Phase (RP) -High Performance Liquid Chromatography (HPLC). And eluted by a linear gradient (40% to 80%, 1% / min) of the buffer solution (B of Acetonitrile with 0.1% TFA) (Lee et al., 1999).

As a result, as shown in Figure 3 it was confirmed that divided into three peaks (①, ② and ③) (Fig. 3).

< Example  5> Potide -second( Potide -J) antifungal activity measurement

<Example 4> peak fractions Candida albicans , a pathogenic fungus, was used as a strain to measure antifungal activity. The fungal water was diluted to 2 × 10 ³ cells per well in the agar-hole, and the fungi were dispensed. Then, only the Candida albicans was dispensed in the negative control group and inoculated with the potide-jay in the other hole of the experimental group. And incubated for 24 hours in a 28 ℃ incubator.

As a result, as shown in Figures 4a and 4b, ① and ③ peaks did not show antifungal activity, but ② peaks prevented the aggregation of fungal cells after 4 hours, and after 24 hours, the aggregation of fungal cells was very strongly inhibited. It was confirmed (Fig. 4a and 4b). Therefore, the peak ② was named Potide-J.

< Example  6> Potide -second( Potide -J) amino acid sequence and molecular weight confirmation

Potide-J, which has antimicrobial activity derived from potato bulbs, was dissolved in 6N-HCl, hydrolyzed and dried at 110 ° C for 22 hours, and then amino acid sequence was confirmed with an amino acid analyzer, followed by MALDI-MS (Matrix). The molecular weight was confirmed by Assisted Laser Desorption Ionization-Mass Spectrometry (Kratos Analytical Ins).

As a result, as shown in SEQ ID NO: 1, the N-amino terminal sequence of the potide-J is NH ₂ -Ala-Val-Cys-Glu-Asn-Asp-Leu-Asn-Cys-Cys, and the molecular weight is 1.0831 kDa. It was confirmed (SEQ ID NO: 1).

< Example  7> Potide -second( Potide -J) cytotoxicity measurement

To determine if Potide-J is cytotoxic, the red blood cell destruction capacity of the peptide was examined. Dilute human red blood cells with phosphate buffer (PBS, pH 7.0) to a concentration of 8% and serially dilute the peptides at a concentration of 1/2 from 200 μM / well to 1 hour at 37 ° C. After reacting for a while, the amount of hemoglobin contained in the supernatant by centrifugation at 1,000 g was investigated by measuring the absorbance at a wavelength of 414 nm. In order to investigate the degree of cell destruction, 1% Triton X-100 was added to human erythrocytes, and the absorbance of the supernatant was measured. The cell destruction ability of 1% Triton X-100 was 100%, which resulted in erythrocyte destruction of the peptide. It was calculated according to the following Equation 1 in comparison with the performance.

In the above formula, absorbance A is the absorbance of the peptide solution at 414 nm wavelength, absorbance B is the absorbance of PBS at 414 nm wavelength and absorbance C is the absorbance of 1% Triton X-100 at 414 nm wavelength.

As a result, since red blood cells were not destroyed as shown in Table 1, it was confirmed that Potide-J did not show cytotoxicity (Table 1).

Cytotoxicity Findings Peptides
( Peptie ) % Red blood cell destruction Hemolysis ) 40 20 10 5 2.5 1.25 0.62 0.31 Potide -second( Potide -J) 0 0 0 0 0 0 0 0 Malitine ( Melittin ) 100 100 100 95 73 36 19 0

(Melittin is an antibiotic peptide known to be highly cytotoxic and tested as a control.)

< Example  7> Potide -second( Potide -J) synthesized and synthesized Potide Determination of Jay's Antifungal Activity

Linear peptides, peptides with disulfide bonds between 3 and 9 Cys, and peptides with disulfide bonds between 3 and 10 Cys, in solid phase using Fmoc (N- (9-fluorenyl) methoxycarbonyl) -protected amino acids Three synthetic peptides were made. The resulting peptides were separated using Reverse Phase (RP) -High Performance Liquid Chromatography (HPLC) using a Waters 15-μm Deltapak C18 column (19 × 300 mm). Elution by linear gradient (0% to 60%) of buffer (A of acetonitrile with 0.1% TFA) followed by Vydac C18 column (4.6 × 250 mm, 300 μs, 5 nm) Purity was determined by reverse phase-high performance liquid chromatography using) and molecular weight was confirmed by Matrix Assisted Laser Desorption Ionization-Mass Spectrometry (MALDI-MS). Antibiotic activity is Candida albicans (Candida a plate medium albicans ) and potide-jay were measured by smearing.

As a result, as shown in FIG. 5, it was confirmed that potide-jay, which is a linear peptide, exhibited antifungal activity by inhibiting the growth of Candida albicans (FIG. 5).

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

< Manufacturing example  1> Preparation of Pharmaceutical Formulations

<1-1> Sanje  Produce

2g peptide of the present invention

Lactose 1g

The above components were mixed and packed in airtight bags to prepare powders.

<1-2> Preparation of tablets

100 mg of peptide of the present invention

100 mg of corn starch

100 mg of milk

2 mg 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

100 mg of peptide of the present invention

100 mg of corn starch

100 mg of milk

2 mg magnesium stearate

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

&Lt; 1-4 >

1 g of the peptide of the present invention

Lactose 1.5 g

1 g of glycerin

Xylitol 0.5 g

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

<1-5> Preparation of granules

150 mg of the peptide of the present invention

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

<110> Industry-Academic Cooperation Foundation, Chosun University <120> Novel antifungal peptide isolated from Solanum tuberosum L cv.          Jopung and use therof <130> 11p-11-36 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Potide-J <400> 1 Ala Val Cys Glu Asn Asp Leu Asn Cys Cys   1 5 10

Claims (12)

Potatoes ( Solanum tuberosum L cv. An antifungal peptide consisting of the amino acid sequence set forth in SEQ ID NO: 1 isolated from Jopung).
The antifungal peptide of claim 1, wherein the antifungal peptide has a molecular weight of 1.0831 kDa.
The antifungal peptide of claim 1, wherein the antifungal peptide has antifungal activity against pathogenic fungi.
4. The method of claim 3 wherein the pathogenic fungus Candida albicans (Candida albicans ), C. catenulate , C. intermidia , C. rugosa , C. glabrata and C. melibiosica Antifungal peptides, characterized in that any one selected from the group consisting of.
1) Potato ( Solanum tuberosum L cv. Lyophilizing the bulb of Jopung) and pulverizing it into powder;
2) preparing an extract by extracting the powder of step 1) with a buffer solution;
3) recovering the supernatant fraction of the extract by centrifuging the extract of step 2); And
4) A method for producing a peptide according to claim 1, comprising the step of separating and purifying the supernatant fraction of step 3) using chromatography.
The method of claim 5, wherein the buffer solution of step 2) is 50 mM Tris-HCl pH 7.5, 10 mM EDTA, 150 mM NaCl, 1% dimethyl sulfoxide (DMSO) and 0.1% β-mercaptoethanol A method for producing a peptide according to claim 1, comprising (β-mercaptoethanol).
An antifungal pharmaceutical composition comprising the peptide of claim 1 as an active ingredient.
The pharmaceutical composition of claim 7, wherein the pharmaceutical composition has antifungal activity against pathogenic fungi.
The method of claim 8, wherein the pathogenic fungus Candida albicans (Candida albicans ), C. catenulate , C. intermidia , C. rugosa , C. glabrata and C. melibiosica The pharmaceutical composition for antifungal, characterized in that any one selected from the group consisting of).
An antifungal cosmetic composition containing the peptide of claim 1 as an active ingredient.
Non-toxic pesticide containing the peptide of claim 1 as an active ingredient.
A preservative for preserving medicines, cosmetics, food or feed containing the peptide of claim 1 as an active ingredient.

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