KR20200129788A - Antimicrobial composition comprising the extract of sageretia thea - Google Patents

Abstract

The present invention relates to an antibacterial composition containing a Sageretia theezans extract as an active component. More specifically, the present invention relates to a broad-spectrum antibacterial composition containing an extract of Sageretia theezans leaves or stems as an active component. The Sageretia theezans extract, as the active component of the broad-spectrum antibacterial composition of the present invention, as demonstrated through the examples of the present specificiation, has excellent antibacterial activity that can be used for an antibacterial pharmaceutical composition, health functional food, a cosmetic composition, an animal feed composition, food, cosmetic or an animal feed additive, various detergents, and various cleaning agents. In addition, the Sageretia theezans extract of the present invention has excellent thermal stability, and does not exhibit any loss of activity in an acidic condition of pH 2 and plasma, so that it is possible to be easily processed into various forms such as liquid, cream, powder, pills, tablets, etc., thereby being able to be very usefully used in the pharmaceutical industry, food industry, livestock industry, health industry, and cosmetics industry.

Description

Antimicrobial composition containing Sangdong tree extract as an active ingredient {ANTIMICROBIAL COMPOSITION COMPRISING THE EXTRACT OF SAGERETIA THEA}

The present invention relates to an antibacterial composition containing an extract of Sangdong trees ( Sageretia thea or Sageretia theezans ) as an active ingredient, and more particularly, to a broad-spectrum antibacterial composition containing an extract of a leaf or stem of a tree of Sangdong as an active ingredient.

Humans have used various antibacterial agents and preservatives to prevent invasion of pathogenic microorganisms and control food spoilage microorganisms. While these chemical antimicrobial agents and preservatives show excellent antibacterial activity against various bacteria and fungi, when used for a long time, many side effects such as redness due to allergy, vascular edema, bone marrow suppression, anemia, gastritis, and degenerative brain disease appear. In fact, the use of antibiotics with severe side effects such as chloramphenicol has been banned in the United States. Therefore, various studies on the development of natural antibacterial agents using natural products with long-term use experience while securing safety are in progress.

In addition, according to the U.S. Center for Disease Control and Prevention, bloodstream infections in hospitals are one of the leading causes of death in the United States. In addition, infection of pathogenic bacteria and fungi through skin and soft tissues is recognized as a universal medical disease, and is understood to be mainly caused by trauma or surgery. Many pathogenic bacteria and fungi infections are treated differently depending on the severity of the infection, but successful treatment is possible with oral antimicrobial agents and topical cleansing. However, more severe or complex infections commonly occurring in patients with infections caused by underlying risk factors (e.g., vascular blood flow, diabetes) and/or difficult to treat or multi-resistant bacteria and fungi are venous Measures such as antimicrobial treatment and invasive surgical tissue removal may be necessary. For example, vancomycin, a glycopeptide-based antibiotic, has been successfully used against severe nosocomial infections caused by multi-drug-resistant Gram-positive bacteria, especially MRSA, coagulase-negative Staphylococcus and Enterococcus. However, it has been reported that the isolation of strains resistant to vancomycin is increasing recently, and it is true that treatment for severe infections caused by some of the pathogens is very limited with current antibiotics. Therefore, there is an increasing need to develop new antibiotics that are more effective against the increasing resistance bacteria.

On the other hand, Sangdong tree ( Sageretia thea or Sageretia theezans ) is an evergreen broad-leaved shrub belonging to the Rhamnaceae genus Sageretia, and is distributed in Asia and North America, including Korea, and is mainly distributed in Jeju Island and the south coast of Korea. Sangdong trees have small leaves and thorns, so they are also called barberry. The leaves turn yellow in autumn, some of them winter, and flowers bloom from October to November. Fruits mature in black purple from April to May of the following year, and are similar in color to blueberries, but small in size. In the private sector, the leaves of Sangdong trees have been used to treat colds, fever, hepatitis, and skin diseases, and the fruits of Sangdong trees have been used for food.

As for the study on Sangdong trees, the hypertrophy and seed germination characteristics of fruits of Sangdong trees (Song Cheol et al., 2015, Korean J. Medicinal Crop Science 23: 311-318; Sang Cheol Song et al., 2015, Korean J. Medicinal Crop Science 23: 8- 12), and cultivation studies such as investigation of the growing environment of native sites (Song Cheol et al., 2014, Korean J. Medicinal Crop Science 22: 301-305) have been mainly conducted in the past, but recently, studies on the antioxidant activity of fruit extracts (Lei, Kechan, etc.) , 2016, International J. Food Properties 19: 210-221), whitening activity (Kyung-ah Ko, 2015, Master's thesis, Jeju National University), cancer cell growth inhibitory activity (Kyung-ah Ko, 2017, Journal of Applied Biological Chemistry 60: 161-171) is known. In the case of the leaves of the sapling tree, the antioxidant activity of the flavonoid-concentrated components (Chung SK, 2009, J Med Food. 12:1310-1315), review of the hydrolysis conditions for the analysis of the flavonoid components of the leaves (Jin-soon Park et al., 2010, Korean Food Journal of Storage and Distribution (Journal of the Korean Journal of Storage and Distribution 17: 261-266), HIV-1 protease inhibitory activity of leaf extracts (Joo-kwon Park, 2001, Master's thesis at Sunchon National University; Park JC, 2002, Phytother Res. 16: 422-426) and cancer cell growth of leaf extracts. Inhibitory activity (Hana Kim et al., 2018. The Plant Resources Society Publication 108) and anti-inflammatory activity (Hana Kim et al., 2018 The Plant Resources Resources Society Publication 98) are known. Studies on the stem of a sapling tree have been relatively insignificant, and the protective effect of the extract of sapling tree trunks (twigs) on apoptosis (Kang KB et al., J Nat Prod. 2018, 81: 1819-1828) and inhibiting the growth of cancer cells of the stem extract (Hana Kim et al., 2018. The Plant Resources Society of Korea Publication 108) and anti-inflammatory activity (Hana Kim et al., 2018. The Plant Resources Resources Society Publication 98) are known. Unknown.

In addition, as for patents related to Sangdong trees, Korean Patent Registration No. 10-1854446 [cosmetic composition containing Sangdong tree extract and fine needle], 10-1716489 [cosmetic compositions containing Sangdong tree extract], No. 10-1743200 [A composition for skin whitening containing a fraction of the fruit extract of Sangdong tree as an active ingredient] is known, so the use of the tree fruit extract in the cosmetic industry is well reported. No. 10-0969134 A method for separating a fraction containing a large amount of useful flavonoids and a novel flavonoid substance from the above-ground part] is disclosed, and No. 10-1831981 discloses [a composition for preventing, improving or treating cancer containing an extract or a fraction thereof as an active ingredient] Although known, until now, no patent has been known for the broad-spectrum antibacterial activity of the extract of the sacrum.

KR 10-1828584 B

Hana Kim et al., 2018. The Plant Resources Society of Korea Presentation 108: Anti-cancer cell growth inhibitory activity of leaf and stem extracts Hana Kim et al., 2018. The Plant Resources Society of Korea Presentation 98: Anti-inflammatory activity of leaf and stem extracts

The present invention has been conceived to solve the problems of the prior art as described above, and the problem to be solved in the present invention is to provide a broad spectrum antibacterial composition containing a sacrum extract as an active ingredient.

In order to solve the above problems, the present invention provides an antibacterial composition containing an extract of Sageretia thea or Sageretia theezans as an active ingredient.

It is preferable that the Sangdong tree is a leaf or a stem of a Sangdong tree.

It is preferable that the above-mentioned homologous tree extract has antibacterial activity against Gram-positive bacteria and Gram-negative bacteria.

The antimicrobial composition is preferably selected from the group consisting of pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food additives, cosmetic additives, animal feed additives, detergents and cleaners.

Sangdong tree extract as an active ingredient of the broad-spectrum antibacterial composition of the present invention, as demonstrated through the examples of the present specification, antibacterial pharmaceutical composition, health functional food, cosmetic composition, animal feed composition, food, cosmetic or animal feed additive , It has excellent antibacterial activity that can be used for various cleaning agents and various cleaning agents. In addition, the Sangdong tree extract of the present invention has excellent thermal stability, does not show any loss of activity even in the acidic condition of pH 2 and plasma, and can be easily processed into various forms such as liquid, cream, powder, pills, tablets, etc. It can be very useful in the pharmaceutical industry, food industry, livestock industry, health industry and cosmetics industry.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention prepared extracts of leaves, stems, and fruits of Sangdong tree in a certain way in order to test the antibacterial and antifungal effects on the tree, and recovered an extract having antibacterial activity, and the extract was human red blood cells. Although it did not show any hemolytic activity against, it was to be used as an antibacterial composition by confirming that it has excellent characteristics of thermal stability and acid stability. In particular, the present invention was completed by confirming that the extract exhibits a strong antibacterial activity against Gram-positive bacteria and Gram-negative bacteria, and thus can be developed as a broad spectrum antibacterial agent against various bacterial infectious diseases.

Specifically, the present inventors prepared an ethanol extract for the leaves, stems, and fruits of the sacrum tree in order to develop an antibacterial composition using a sapling tree known to have anti-inflammatory, hepatoprotective, and antihypertensive effects, and various bacteria As a result of evaluating the antimicrobial activity against, it was confirmed that strong antibacterial activity against Gram-positive bacteria was confirmed in the extracts of the leaves and stems of the gram-negative tree, and strong against Escerichia coli and Proteus vulgaris , known as food poisoning bacteria among Gram-negative bacteria. By confirming the antibacterial activity, it was confirmed that the extract can be used as a broad spectrum antibacterial agent.

Accordingly, the present invention provides an antimicrobial composition containing the extract of Sangdong tree as an active ingredient.

It is preferable that the Sangdong tree is a leaf or a stem of a Sangdong tree.

It is preferable that the above-mentioned homologous tree extract has antibacterial activity against Gram-positive bacteria and Gram-negative bacteria.

The antibacterial composition may be applied to a use selected from the group consisting of pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food additives, cosmetic additives, animal feed additives, detergents and cleaning agents.

Hereinafter, the method for preparing the extract of the present invention and the efficacy test will be described in more detail.

In order to achieve the object of the present invention, the inventors of the present invention carried out a specific manufacturing method and efficacy experiment as follows, and the present invention comprises the steps of preparing an extract of leaves, stems, and fruits of a sapling tree; Evaluating the antibacterial activity of the extracts; And a stability investigation step.

The "Sangdong tree extract" included in the composition of the present invention may be obtained by extracting the Sangdong tree with an organic solvent and filtering the extract using a filter net of 0.06mm or less, and concentrating it under reduced pressure.

The solvent used in the present invention includes water (cold water, hot water), alcohol, anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, butanol, etc.), and a mixed solvent of the lower alcohol and water. Available, 70% ethanol extraction is most preferred.

As a preferred embodiment, the homologous tree of the present invention can be extracted with ethanol. In addition, the ethanol extract may be sequentially or fractionated with an organic solvent of hexene, ethyl acetate and butanol to additionally obtain a hexene fraction, an ethyl acetate fraction, a butanol fraction, and a water residue.

The Sangdong tree extract of the present invention may be prepared into a powder through a conventional powdering process such as vacuum drying, freeze drying, or spray drying. Sangdong tree extract is not decomposed by various degrading enzymes in plasma, and maintains antibacterial activity even at a heat treatment of 100℃ and a pH of 2 in the stomach.

The extract of the present invention has a strong antibacterial activity, and thus the material for pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, foods, cosmetics, animal feed additives, and various cleaning agents related to the prevention or treatment of pathogenic bacteria infection. Can be used as

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as a pharmaceutical composition.

The pharmaceutical composition is formulated in various forms such as oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and injections of sterile injectable solutions according to the usual method for each purpose of use. It may be used, and may be administered orally, or administered through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and topical administration. Such pharmaceutical compositions may further include a carrier, excipient, or diluent, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil And the like. In addition, the pharmaceutical composition of the present invention may further contain a filler, an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, a preservative, and the like.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, drug activity, Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

The effective amount of the component having antimicrobial activity in the pharmaceutical composition of the present invention may vary depending on the age, sex, and body weight of the patient, and generally 1 to 5,000 mg, preferably 100 to 3,000 mg per body weight is administered daily or every other day. Alternatively, it can be administered in 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, the severity of the disease, sex, weight, age, etc., the dosage amount is not limited by any method. The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration can be expected and can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intra-cerebroventricular injection. In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the route of administration of the pharmaceutical composition of the present invention is oral or parenteral through all general routes as long as it can reach the target tissue. It can be administered orally. In addition, the composition of the present invention may be administered using any device capable of delivering an active ingredient to target cells. In the present invention, the "object" is not particularly limited, but includes, for example, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, mouse, rabbit, or guinea pig And, preferably, a mammal, more preferably a human.

As a preferred embodiment, the antimicrobial composition of the present invention can be applied as a health functional food.

The health functional food of the present invention can be used in various ways such as effective foods and beverages that can lower the likelihood of infection from a subject having a high risk of infection by pathogenic microorganisms. Foods containing an active ingredient having excellent antibacterial activity of the present invention include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplement foods, etc., in the form of powders, granules, tablets, capsules or beverages Can be used as In general, the active ingredient of the present invention may be added in an amount of 0.01 to 15% by weight of the total food weight, and the health beverage composition may be added in an amount of 0.02 to 10g, preferably 0.3 to 1g, based on 100 ml. The health functional food of the present invention may contain the compound as an essential component in the indicated ratio as an additional component, as well as food supplementary additives acceptable for food, such as natural carbohydrates and various flavoring agents. Examples of the natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and conventional sugars such as polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, taumatin, rebaudioside A, glycyrrhizin, saccharin, aspartame, and the like may be used. The proportion of the flavoring agent is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention includes a variety of nutrients, vitamins, minerals, synthetic flavors, and flavoring agents such as natural flavors, colorants and thickeners, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloids. It may contain thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the health functional food of the present invention may contain pulp for the production of natural fruit juices, fruit juice drinks, and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is generally selected from 0.01 to about 20 parts by weight per 100 parts by weight of the active fraction of the present invention.

As a preferred embodiment, the antimicrobial composition of the present invention can be applied as a use of a cosmetic composition.

The cosmetic composition of the present invention may be prepared in the form of ingredients having excellent antibacterial activity of the present invention, for example, essence, nutritional cream, body lotion, conditioner, shampoo, body cleanser, lotion, skin, mask pack, etc. It is not limited to this. Manufacturing in this form may be easily performed according to various manufacturing processes known to those skilled in the art.

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as an animal feed composition.

The animal is a group of organisms corresponding to plants, mainly ingesting organic matter as nutrients, and refers to a differentiated digestion, embryonic and respiratory organs, specifically echinoderm, crustacean, molluscs, fish, amphibians, reptiles, birds, It may be a mammal, and preferably, echinoderm animals such as sea urchins or sea cucumbers, arthropods including crustaceans such as crabs, shrimp, and shrimp, cephalopods, gastropods or mollusks such as bivalve, red snapper, bream, cod, flounder , Fish such as halibut, birds including poultry such as pheasants or chickens, or mammals such as pigs, cows, and goats.

The composition for animal feed may further include grain, vegetable protein feed, animal protein feed, sugar or dairy product to the antibacterial composition containing the antibacterial component of the present invention. The grain may be specifically crushed or crushed wheat, oats, barley, corn, and rice, and the vegetable protein feed may specifically include rapeseed, soybean and sunflower as main components, and the animal protein feed may be specifically May be blood meal, meat meal, bone meal and fish meal, and the sugar or dairy product may specifically be a dry component composed of various milk powder and whey powder.

The composition for animal feed may further include ingredients such as nutritional supplements, digestion and absorption enhancers, sexual ulcer stimulators or disease prevention agents.

The antimicrobial composition contained in the composition for animal feed of the present invention may vary depending on the purpose of use and conditions of use of the feed, for example, 0.1g to 100g of the antimicrobial composition may be included based on 1 kg of the final produced feed.

In addition, the feed composition may be prepared as a hard-viscous granulated or granular material depending on the degree of pulverization of the ingredients, and the composition may be supplied as a mesh or may be formed into a desired separated shape for further processing and packaging, and storage For ease of storage, it may be subjected to a process of pelletizing, swelling or extrusion, and preferably excess water may be removed by drying.

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as an additive for food, cosmetics or animal feed.

The additive is added to impart preservability to food, cosmetics, or animal feed to be used, and the antibacterial composition is a natural component and is not harmful to the human body, so it can be safely applied to food, cosmetics or animal feed.

The additive may further include additional additives appropriately selected according to the purpose and use of the additive, and may be used in combination with other active ingredients such as commonly used additives or additives such as pigments, surfactants, and preservatives. . The additive may be prepared by powdering, granulating, tableting, or liquefying according to the purpose and use thereof, and a conventional method may be used for packaging for commercialization.

In addition, the amount of the antimicrobial composition among the additives may be appropriately used depending on the purpose of use, application form, application site, and application object, and may include, for example, 0.01 parts by weight to 50 parts by weight based on the total composition. The content of the composition is not limited thereto. Since the antimicrobial composition of the present invention is a natural component and is not harmful to the human body, it can be used in various amounts if it can achieve the purpose of application due to the characteristics of the product.

As a preferred embodiment, the antimicrobial composition of the present invention can be applied as a cleaning agent.

The detergent may be preferably a detergent for clothes, a dishwasher, a food detergent, or a home appliance detergent. Since the antibacterial composition is a natural component and is not harmful to the human body, more preferably, it can be used for the purpose of washing various household items including kitchen utensils or food washing or for imparting antibacterial properties.

The detergent may additionally contain selected additives according to its purpose and use, and may be used by mixing with other active ingredients such as commonly used detergents or additives such as pigments, surfactants, and preservatives. The detergent may be prepared by powdering, granulating, tableting, or liquefying depending on the purpose and use thereof, and a conventional method may be used for packaging for commercialization.

In addition, the amount of the antimicrobial composition among the detergents may be appropriately used depending on the purpose of use, application type, application site, and the product to be applied, and may include, for example, 0.01 parts by weight to 50 parts by weight based on the total composition, but the antimicrobial properties The content of the composition is not limited thereto. Since the antimicrobial composition of the present invention is a natural component and is not harmful to the human body, it can be used in various amounts as long as it can achieve the purpose of application due to the nature of the product.

As a preferred embodiment, the antimicrobial composition of the present invention can be applied as a cleaning agent.

The cleaning agent may be used as a hand, oral, or feminine cleaning agent in a simple manner in order to prevent infection in the body of pathogenic microorganisms that are often susceptible to infection in daily life. Since the antibacterial composition is a natural component and is not harmful to the human body, it can be safely applied even when used as a cleanser for hands, mouth, and women.

The cleaning agent may further contain additional additives appropriately selected according to the purpose and use of the cleaning agent, and may be used by mixing with other active ingredients such as commonly used additives or additives such as pigments, surfactants, and preservatives. . The cleaning agent may be prepared by powdering, granulating, tableting, or liquefying depending on the purpose and use of the cleaning agent, but in the case of a liquid type, convenient use will be possible.

In addition, the amount of the antimicrobial composition among the cleaning agents may be appropriately used depending on the purpose of use, application form, and application site, and may include, for example, 0.01 parts by weight to 50 parts by weight based on the total composition, but the content of the antibacterial composition is It is not limited thereto. Since the antimicrobial composition of the present invention is a natural component and is not harmful to the human body, it can be used in various amounts if it can achieve the purpose of application due to the characteristics of the product.

Hereinafter, the present invention will be described in more detail through examples. The following examples are only one preferred specific example of the present invention, and the scope of the present invention is not limited to the scope of the following examples.

[Example]

Example 1: Preparation of extracts for each part of Sangdong tree and analysis of useful components thereof

Sangdong trees harvested in Jeju Island in 2018 were purchased and used to prepare ethanol extracts. Specifically, after dividing the sapling tree into leaves, stems, and fruits, 10 times the amount of ethanol was added to each sample, extracted twice at room temperature, collected and filtered, concentrated under reduced pressure, and prepared as a powder. The extract was prepared.

The contents of total polyphenols, total flavonoids, total sugars and reducing sugars were measured by analysis of the components of the extract of the sacrum. For the total polyphenol content, 50 μl of Folin-ciocalteau and 100 μl of Na ₂ CO ₃ saturated solution were added to 400 μl of the extraction sample, and allowed to stand at room temperature for 1 hour, and the absorbance was measured at 725 nm. Tannic acid was used as a standard reagent. For the total flavonoid content, each sample was extracted with methanol for 18 hours, and then 4 ml of 90% diethylene glycol was added to 400 μl of the filtered extraction sample solution, 40 μl of 1 N NaOH was added, and the absorbance was measured at 420 nm after reaction at 37° C. for 1 hour. Rutin was used as a standard reagent. Reducing sugar was quantified by DNS method and total sugar was quantified by phenol-sulfuric acid method.

[Table 1] Analysis of Extraction Efficiency and Useful Components of Extracts by Part of Sangdong Tree

As shown in Table 1, the ethanol extraction efficiency of the sapling tree was found in the order of fruit> leaves> stem, and showed a yield of 15.5-20%. As a result of analyzing the useful components of the extracts by site, the highest content of 307.5mg/g was found in the stem extract, followed by leaf extract and fruit extract. On the other hand, the flavonoid content measurement results were in the order of leaf extract (77.0mg/g)> stem extract (63.9mg.g)> fruit extract (28.2mg/g). The total sugar and reducing sugar contents were similarly in the order of fruit> stem> leaf. Therefore, it is expected that leaf and stem extracts will exhibit more potent physiological activity than the fruit of the Sangdong tree, which has been studied.

Example 2: Evaluation of antibacterial activity of extracts for each part of Sangdong tree

The antimicrobial activity of the extracts of leaves, stems and fruits of Sangdong tree prepared in Example 1 were evaluated, and the results are shown in Table 2. As the strain used in the antibacterial activity evaluation, a Gram-negative bacteria Escherichia coli KCTC 1682, Proteus vulgaris KCTC 2433, Pseudomonas aeruginosa KACC 10186, was used Salmonella typhimurium KCTC 1926, a gram-positive Staphylococcus aureus KCTC 1916, Staphylococcus epidermidis ATCC 12228, Listeria monocytogenes KACC 10550, Bacillus subtilis KCTC 1924 were used. Meanwhile, Saccharomyces cerevisiae IF0 0233 and Candida albicans KCTC 1940, the causative agent of candidiasis fungal infection, were used to evaluate antifungal activity.

First, in the case of antibacterial activity evaluation, each bacteria was inoculated into Nutrient broth (Difco Co., USA) and cultured at 37° C. for 24 hours, and then each strain was adjusted to OD ₆₀₀ 0.1 to obtain Nutrient agar (Difco Co., USA). , USA) A sterile petri dish (90mm, Green Cross, Korea) containing the medium was plated at 100μl each, and 5μl of each sample was added to a sterile disc-paper (diameter 6.5mm, Whatsman No. 2), and 37°C for 24 hours. In the case of fungi, in the case of fungi, after incubation at 30° C. for 24 hours in the same manner using Sabouraud dextrose (Difco Co. USA), the size of the growth inhibition ring was measured to evaluate the antibacterial activity. As a control, the antibacterial agent ampicillin and the antifungal agent myconazole (miconazole, Sigma Co., USA) were used at a concentration of 1 μg/disc, respectively, and the size of the growth inhibition ring was the diameter of the part where growth did not appear with the naked eye. It was measured in mm and expressed as a representative result after three or more evaluations.

[Table 2] Antibacterial activity of extracts from leaves, stems and fruits of Sangdong tree

As a result, as shown in Table 2, ampicillin, which is used as an antibacterial agent in clinical practice, exhibited broad antibacterial activity at 1 μg/disc, but showed relatively weak growth inhibition rings (9-12 mm) for Pseudomonas aeruginosa and Escherichia coli . Done. Miconazole, used clinically as an antifungal agent, strongly inhibited the growth of Saccharomyces cerevisiae and Candida albicans even at 1 μg/disc. On the other hand, the extracts of the leaves and stems of gram-positive trees exhibited excellent growth inhibition against Gram-positive bacteria Listeria monocytogenes , Bacillus subtilis , Staphylococcus epidermidis , Staphylococcus aureus, and Gram-negative bacteria Proteus vulgaris and Escherichia coli at a concentration of 250 μg/disc. On the other hand, at 250 μg/disc concentration, the fruit extract of Sangdong tree showed a growth inhibition ring of 8.0 mm against Listeria monocytogenes , a Gram-positive bacteria, but did not show antibacterial activity against other bacteria. Therefore, it was expected that the extracts of leaves and stems of the sakura tree could be used as a broad spectrum antibiotic.

Example 3: Human Erythrocyte Hemolytic Activity of Extracts from Leaves and Stems

Since the fruit of the Sangdong tree is used for food, and the leaves and stems are used as tea, it is a safe food raw material. Therefore, it is judged that the extract of the sapling tree does not cause a separate acute toxicity. In order to evaluate the possibility of acute toxicity of the extracts from the extract of sacrum, the hemolytic activity of human red blood cells was evaluated, and the results are shown in Table 3. At this time, the hemolytic activity was evaluated according to the previous report (Ho-Yong Son, 2014 ㆍKorean J. Microbiol. Biotechnol. 42: 285-292), and simply 100 μl of human red blood cells washed three times with PBS was placed in a 96-well microplate. Then, 100 μl of sample solution of various concentrations was added and reacted for 30 minutes at 37°C. After that, the reaction solution was centrifuged for 10 minutes (1,500 rpm), and 100 μl of the supernatant was transferred to a new microtiter plate. It was measured at. DMSO (2%) was used as a solvent control for the sample, and Triton X-100 (1 mg/ml) was used as an experimental control for hemolysis of red blood cells. Hemolytic activity was calculated using the following formula.

[Table 3] Hemolytic activity of human red blood cells of Sangdong tree extract

First, it was confirmed that DMSO and water used as control cells had no hemolytic activity, and triton X-100 hemolytic 100% red blood cells at a concentration of 1 mg/ml. In addition, it was confirmed that amphotericin B, which is used as an anticancer agent and antifungal agent, hemolyzes more than 50% of red blood cells at a concentration of 0.025mg/ml. It was confirmed that the extract of the present invention did not show any red blood cell hemolysis up to the concentration of 1 mg/ml, and thus had no acute toxicity and red blood cell hemolysis activity.

Example 4: Plasma, acid, and heat stability evaluation of leaf and stem extracts

Plasma stability, thermal stability, and acid stability against antimicrobial activity were confirmed for the ethanol extracts of the leaves and stems of Sangdong trees obtained in Example 1 above. As a result, the active extract exhibited high stability without significant loss of antimicrobial activity even after 1 hour heat treatment at 100°C, 1 hour treatment at pH 2 (0.01M HCl), and 1 hour treatment in plasma. Therefore, the above mentioned Sangdong tree leaf and stem extract is expected to maintain excellent antibacterial activity even under various food processing processes.

Claims (4)

Sangdong tree ( Sageretia thea ) antibacterial composition containing the extract as an active ingredient. The antibacterial composition according to claim 1, wherein the Sangdong tree is a leaf or stem of a Sangdong tree. The antimicrobial composition according to claim 1, wherein the extract of the sapling tree has antibacterial activity against Gram-positive bacteria and Gram-negative bacteria. The method of any one of claims 1 to 3, wherein the antimicrobial composition is selected from the group consisting of pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food additives, cosmetic additives, animal feed additives, detergents and cleaners. Antibacterial composition, characterized in that the.