KR20230077110A - Antimicrobial composition comprising the extract of spent mushroom substrate for lentinus edodes - Google Patents

Abstract

The present invention relates to an antibacterial composition containing an extract of spent mushroom substrate (SMS) after harvesting shiitake mushrooms, which is generated as a by-product during the cultivation of shiitake mushrooms (Lentinus edodes). More specifically, the present invention relates to an antibacterial composition with excellent antibacterial and antifungal activity, containing the ethyl acetate fraction of a hot water extract of an SMS remaining after harvesting shiitake mushrooms by inoculating and culturing shiitake mushroom spawn in a medium for artificial cultivation of shiitake mushrooms. As an active ingredient in the broad-spectrum antibacterial composition of the present invention, the ethyl acetate fraction of the hot water extract of the SMS generated as a by-product during artificial cultivation of shiitake mushrooms exhibits excellent antibacterial activity, thereby having excellent effects of being used for purposes such as antibacterial pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, additives for food, cosmetics or animal feed, various detergents, and various cleaning agents. In addition, the ethyl acetate fraction of the hot water extract of the SMS generated as a by-product during artificial cultivation of shiitake mushrooms of the present invention has excellent thermal stability and does not show loss of activity even under acidic conditions of pH 2 and in plasma, thereby being easily processed into various forms such as liquid, cream, powder, pills, and tablets, so that the same can be very useful in the pharmaceutical industry, food industry, livestock industry, health industry, and cosmetics industry.

Description

Antibacterial composition containing mushroom waste medium extract after harvesting shiitake mushrooms

The present invention relates to an antibacterial composition comprising an extract of spent mushroom substrate (SMS) after harvesting shiitake mushrooms, which is generated as a by-product during cultivation of shiitake mushrooms ( Lentinus edodes ), and more specifically, for artificial cultivation of shiitake mushrooms The present invention relates to an antibacterial composition containing an ethyl acetate fraction of a hot-water extract of a mushroom waste medium remaining after harvesting shiitake mushrooms by inoculating and culturing shiitake mushroom spawn in a medium, and having excellent antibacterial and antifungal activity.

Humans have been using various antibacterial agents and preservatives to defend against the invasion of pathogenic microorganisms and control food spoilage microorganisms. However, in the case of chemical antibacterial agents and preservatives, while showing excellent antibacterial activity against various bacteria and fungi, long-term use causes many side effects such as allergy-induced flare-ups, vascular edema, bone marrow suppression, anemia, gastritis, and degenerative brain disease. do. In fact, the United States has banned the use of antibiotics with severe side effects, such as chloramphenicol. Therefore, various studies on the development of natural antibacterial agents using natural products with long-term experience of use while ensuring safety are in progress.

According to the U.S. Center for Disease Control and Prevention, hospitalized bloodstream infections are one of the leading causes of death in the United States. In addition, infection of pathogenic bacteria and fungi through the skin and soft tissue is recognized as a universal medical disease, and is understood to be mainly caused by trauma or surgery. Many pathogenic bacterial and fungal infections can be successfully treated with oral antimicrobial agents and topical cleansing, although treatment methods vary depending on the severity of the infection. However, more severe or complex infections, which often occur in patients with underlying risk factors (e.g., abnormal blood flow, diabetes) and/or infections caused by difficult-to-treat or multi-resistant bacteria and fungi, are Treatment with antimicrobial agents and invasive surgical tissue removal may be necessary. For example, vancomycin, a glycopeptide antibiotic, has been successfully used against severe nosocomial infections caused by multi-drug-resistant Gram-positive bacteria, particularly MRSA, coagulase-negative Staphylococcus and Enterococcus. However, it has recently been reported that the isolation of strains resistant to vancomycin is increasing, and it is true that current antibiotics are very limited in treating severe infections caused by some of the pathogens. Therefore, there is an increasing need for the development of new antibiotics that are more effective against the gradually increasing resistant bacteria.

On the other hand, shiitake mushrooms are edible mushrooms belonging to the genus Pleurotus and Pine nut, and have the highest content of vitamin C among mushrooms, and contain a large amount of various amino acids and ergosterol. In terms of health functionalities, anticancer, lung and gastrointestinal disease prevention, immunity enhancement against viruses, promotion of antibody production, lowering blood pressure, prevention of arteriosclerosis, blood lipid concentration control function, antidiabetic effect, etc. are known.

Patents related to shiitake mushrooms include Korean Registered Patent No. 10-1839507 [Method of manufacturing seasoned salted fish marinated with shiitake mushrooms and fermented shiitake mushrooms], No. 10-1830763 [Method of manufacturing rice doenjang containing shiitake mushroom extract] and rice miso paste prepared therefrom], No. 10-1823460 [Method for producing semi-dried rockfish using shiitake mushroom powder], No. 10-1759312 [Method for producing shiitake mushroom powder, production of nutritional bar using shiitake mushroom powder] Method and nutritional bar produced thereby], and No. 10-1673573 [Method for manufacturing shiitake mushroom tea and shiitake mushroom tea produced thereby] are disclosed, and Korean Patent Publication No. 10-2004-0038957 [shiitake mushroom powder Foods with the subject of laver production] are disclosed. Most of the patents known to date relate to shiitake mushroom cultivation and various food preparations using shiitake mushrooms, and patents on useful physiological activities of shiitake mushrooms are very limited.

In addition, patents related to useful physiological activity of shiitake mushrooms include Korean Patent Registration No. 10-1487742 [Food composition having antioxidant effect containing shiitake mushroom and cinnamon extract], No. 10-1806808 [fermentation using shiitake mushroom mycelium] [Composition for prevention, improvement, or treatment of female menopausal symptoms containing fermented saline fermented soybeans as an active ingredient], No. 10-1611947 [Health food composition for preventing and improving vascular disease and manufacturing method thereof], No. 10-1266528 [A cosmetic composition for moisturizing and/or whitening containing a polysaccharide extracted from shiitake mushroom mycelium culture as an active ingredient] is disclosed, and Patent Publication No. 10-2017-0036912 [Atopy containing shiitake mushroom extract as an active ingredient] and composition for preventing and improving contact dermatitis], Patent Publication No. 10-2003-0056753 [Health food composition for controlling obesity containing shiitake mushroom mycelium, Agaricus mycelium, and water-soluble chitosan], No. 10-2008-0110212 [shiitake mushroom A composition that helps bone growth using a hot water extract] has been disclosed.

The total annual production of edible mushrooms in Korea is reported to be 162,292 tons in 2017 from 173,354 tons in 2013. In 2017, oyster mushrooms were 58,784 tons, king oyster mushrooms 48,588 tons, enoki mushrooms 38,092 tons, and button mushrooms 10,173 tons. production, and shiitake mushroom production is estimated to be 25,000 tons (Ministry of Agriculture, Food and Rural Affairs, 2016. Production of special crops). These mushroom cultivations are gradually changing from natural media to using specific artificial media. As the artificial cultivation and production of mushrooms increase, the emission of mushroom waste media is also rapidly increasing. Considering that 5 kg of mushroom medium is generally consumed for the production of 1 kg of mushrooms, it is estimated that more than 2 million tons of waste mushroom media are emitted annually in Korea. In particular, in the case of shiitake mushrooms, sawdust medium from various trees is used. In 2015, sawdust medium imported from China was 36,285 tons, more than doubled in three years compared to 11,825 tons in 2012 (Export and import statistics of the Customs Service; Weekly Chosun, 2016. 4. 11).

It is known that 20-25% of nutrients are required for mushroom cultivation when artificial media are used (Williams et al, 2001. Biores Technol. 79:227-230.; Bae et al, 2006. J Anim Sci & Technol. 48:237-246). Therefore, a significant amount of nutrients remain in the mushroom waste medium. In addition, since the mushroom mycelium is already mass-cultured in the mushroom waste medium, various enzymes such as β-glucosidase and cellulase derived from mushroom fruiting bodies and mycelia, as well as various physiological activities such as plant growth promoting active ingredients, antibacterial substances, and disease resistance derivatives, etc. (Hautzel and Anke., 1990, Z Naturforsch. 45: 1093-1098; Lim et al., 2012. Mycobiology. 41: 160-166; Parada et al., 2012 Crop Protection 30: 443-450; Suess, 2006, Agriculture and Lands. pp. 1-101). Therefore, the mushroom waste medium can be used as a nutritionally or physiologically active material. However, to date, research on the development of high value-added functional materials using waste mushroom medium has been very limited, and a strong antibacterial composition of post-harvest mushroom waste medium extracts generated after shiitake mushroom ash has not been known.

In Korea, some of the waste mushroom medium was used for feed (Jeong Jong-cheon et al., 2012. J. Mushroom Sci. Prod. 10: 174-178), and 10 to 30% was added to mushroom cultivation and reused as an artificial medium (Lee Nam-gil et al., 2015. J. Mushrooms, 13: 310-313), but when the sawdust content is high in the artificial medium, it is not reused for feed or mushroom cultivation and is almost used as compost. Recently, studies have been actively conducted in foreign countries to efficiently purify barn waste using various microorganisms and enzymes of mushroom waste medium (Luo X et al., 2018, Bioresour. Technol. 250: 611-620; Meng L et al., 2018. Bioresour. Technol. 253: 197-203; Meng X et al., 2018. Bioresour. Technol. 251: 22-30), suggesting the possibility of using mushroom waste medium as a new environmental cleaning agent in the future.

The content of useful substances in the waste mushroom medium varies depending on the composition of the medium before mushroom inoculation and the type and condition of the cultured mushrooms. Until now, most studies have been focused on the waste medium of mushrooms for enoki mushrooms (Jeong Jong-cheon et al., 2012. J. Mushroom Sci. Prod. 10: 174-178), but recently, the waste medium for ericium erinaceus (Lee Sang-yeop, 2015, The Korean Journal of Mycology 43: 185-190) and beta-glucan content of shiitake sawdust medium are also being studied (Park Young-ae, 2016. Korean Journal of Mycology, 44: 296-299). To date, the known useful physiological activities of mushroom waste media include the growth promotion and pest control effects of pepper growth promotion and pestilence inhibition effects of water extracts of mushroom waste media of oyster mushrooms, shiitake mushrooms, and ericium erinaceus at room temperature (Kwak Amin et al., 2015. J. Mushrooms , 13: 16-20; Kang DS et al., 2017. Plant Pathology J. 33: 264-275) and the anti-inflammatory effect of the water extract of oyster mushroom mushroom waste medium is known (Rivero-Perez N. et al., 2016. Indian J Pharmacol. 48: 141-144). In addition, water-soluble polysaccharides have been reported as the main active ingredient of mushroom waste medium, and recently, antioxidant activity of polysaccharides has been reported in shiitake mushroom mushroom waste medium (Zhu H et al., 2018, Int. J Biol. Macromol. 112: 976-978), acidic polysaccharides with cancer cell growth inhibitory activity (Zhang Y et al., 2017. Int. J. Med. Mushrooms. 19: 395-403), and polysaccharides with antibacterial activity have also been identified ( Zhu H, 2012, Int. J. Biol. Macromol. 50: 840-843). However, no strong antibacterial activity has been reported in shiitake mushroom waste medium.

In addition, all patents related to mushroom waste medium are manufacturing fuel pellets using mushroom waste medium (Korean Registered Patent No. 10-1728233, method of manufacturing fuel pellets using waste medium or waste wood after mushroom cultivation), feed manufacturing (Korea Patent Publication No. 10-2014-0066036, feed using mushroom waste medium and its manufacturing method), and saccharification enzyme preparation (Korean Patent No. 10-1458628, method for producing saccharification enzyme from mushroom waste medium), section Antithrombotic active composition (Republic of Korea Patent No. 10-2128625, pharmaceutical composition and health functional food for preventing or treating thrombosis containing medium extract after harvesting shiitake mushroom as an active ingredient) and pepper blight containing oxalic acid as an active ingredient / Tomato green blight control composition (Republic of Korea Patent No. 10-1873978, composition for controlling plant diseases including medium extract after shiitake mushroom harvest and method for producing the same) is known, but obtained from hot water extract of shiitake mushroom waste medium There have been no reports of potent broad-spectrum antibacterial and antifungal activity of ethyl acetate fractions.

KR 10-1611947 B KR 10-1873978 B

Zhu H, 2012, Int. J. Biol. Macromol. 50: 840-843 Amin Kwak et al., 2015. J. Mushrooms, 13: 16-20 Kang DS et al., 2017. Plant Pathology J. 33: 264-275 Hyerin Yu et al., 2018. Journal of the Korean Mushroom Society 16: 170-174

The present invention has been made to solve the problems of the prior art as described above, and the problem to be solved in the present invention is to provide an antimicrobial composition containing an extract of mushroom waste medium generated as a by-product during artificial cultivation of shiitake mushrooms. .

In order to solve the above problems, the present invention provides an antibacterial composition comprising a mushroom waste medium (Spent Mushroom Substrate, SMS) extract after harvesting shiitake mushrooms ( Lentinus edodes ).

The waste mushroom medium is preferably a waste mushroom medium after harvesting shiitake mushrooms 1 to 3 times.

The extract is preferably extracted with hot water and then fractionated with ethyl acetate.

The antibacterial 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.

The ethyl acetate fraction of the hot-water extract of the mushroom waste medium generated as a by-product during the artificial cultivation of shiitake mushrooms as an active ingredient of the broad-spectrum antibacterial composition of the present invention exhibits excellent antibacterial activity, making it an antibacterial pharmaceutical composition, health functional food, cosmetic composition, and animal feed It has excellent effects that can be used for applications such as compositions, food, cosmetics, additives for animal feed, various detergents, and various cleaning agents. In addition, the ethyl acetate fraction of the hot water extract of the mushroom waste medium generated as a by-product during the artificial cultivation of shiitake mushrooms of the present invention has excellent heat stability and does not show loss of activity even in acidic conditions of pH 2 and plasma, so it can be used in liquid, cream , It can be easily processed into various forms such as powder, pill, tablet, etc., so it can be used very usefully in the pharmaceutical industry, food industry, livestock industry, health industry and cosmetics industry.

Figure 1 shows, from left to right, a sterilization medium for growing shiitake mushrooms used in the experiment, a waste mushroom medium after harvesting shiitake mushrooms once by inoculating and culturing shiitake spawn in the sterilization medium, and a waste mushroom medium after harvesting three times. , and photographs of harvested shiitake mushrooms.
Figure 2 is a sterilization medium for shiitake mushroom cultivation used in the experiment from left to right, a waste mushroom medium after harvesting shiitake mushrooms once by inoculating and culturing shiitake spawn in a sterilization medium, and a waste mushroom medium after harvesting three times. , and a photograph showing the hot water extract of each of the harvested shiitake mushrooms.

Hereinafter, the present invention will be described in detail.

In order to verify the antibacterial efficacy of the shiitake mushroom waste medium, the inventors of the present invention inoculated and cultured the shiitake mushroom spawn in a sterilization medium for shiitake mushroom cultivation (new medium before inoculation) and a sterilization medium to harvest the shiitake mushrooms once. Waste mushroom medium after harvesting, waste mushroom medium after harvesting three times, and dry powder of each harvested shiitake mushroom were prepared. Thereafter, each of the hot water extracts was prepared in a certain way, and as a result of evaluating their antibacterial and antifungal activities, no significant antibacterial activity was found in all of the extracts. This meant that, as in the previous report (Korean Patent No. 10-1873978), when the oxalic acid of the mushroom waste medium was not concentrated, the antimicrobial activity was very weak in the shiitake mushroom powder, sterilized medium, and post-harvest medium.

Then, hexene fractions, ethyl acetate fractions, butanol fractions, and water residues after organic solvent fractionation were prepared from each of the extracts, and as a result of evaluating the antibacterial activity thereof, the ethyl acetate fraction of the hot water extract of the mushroom waste medium after cultivation of shiitake mushrooms was recovered as a broad-spectrum antibacterial active component, and it was confirmed that the fraction had excellent heat stability and acid stability while not exhibiting any hemolytic activity against human red blood cells, and thus the ethyl acetate fraction was classified into gram-positive bacteria, gram-negative bacteria and The present invention was completed by confirming that it could be developed as a broad-spectrum antibacterial agent for fungal infections.

Specifically, the inventors of the present invention, in order to develop an antibacterial composition in mushroom waste medium after mass cultivation of shiitake mycelium and immature fruiting bodies, sterilized medium, mushroom waste medium after one harvest of shiitake mushrooms, mushroom waste after three shiitake mushrooms Following analysis of the nutrient components and physicochemical properties of the culture medium and shiitake mushrooms, hot water extracts were prepared for each of the above samples and antibacterial activity was evaluated, and then sequential organic solvent fractions were prepared for each extract, and After harvesting three times, strong antibacterial and antifungal activities were confirmed in the ethyl acetate fraction of the mushroom waste medium hot water extract. In addition, it was confirmed that the ethyl acetate fraction of the mushroom waste medium hot water extract after harvesting the shiitake mushroom did not exhibit hemolytic activity on human red blood cells.

Therefore, the present invention provides an antibacterial composition comprising an extract of shiitake mushroom ( Lentinus edodes ) after harvesting mushroom waste medium (Spent Mushroom Substrate, SMS).

The waste mushroom medium is preferably a waste mushroom medium after harvesting shiitake mushrooms 1 to 3 times.

The extract is preferably extracted with hot water and then fractionated with ethyl acetate.

The antibacterial 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.

Hereinafter, the production method and efficacy test of the ethyl acetate active fraction of the mushroom waste medium ethanol extract after harvest during artificial cultivation of shiitake mushrooms of the present invention will be described in more detail.

The present invention is a sterilization medium for artificial cultivation of shiitake mushrooms, a waste mushroom medium after harvesting shiitake mushrooms once by inoculating and culturing shiitake spawn in a sterilization medium, a waste mushroom medium after harvesting three times, and harvested shiitake Preparing each powder of mushrooms; preparing an extract from the four samples; Evaluating the antibacterial and antifungal activity of the extract; preparing sequential organic solvent fractions of hexene, ethyl acetate and butanol from the hot water extracts and preparing a water residue obtained thereafter; and evaluating the antibacterial and antifungal activities of the fractions and examining the stability of the active substances of the extracts and active fractions and the hemolytic activity of human red blood cells of the active fractions.

"Post-harvest mushroom waste medium extract during artificial cultivation of shiitake mushrooms" included in the composition of the present invention collects and pulverizes discarded mushroom waste media after harvesting shiitake mushrooms 1 to 3 times after inoculation and cultivation of shiitake mushrooms. doing; It can be obtained by extracting the pulverized mushroom waste medium with a solvent, filtering the extract using a filter net of 0.06 mm or less, and concentrating it under reduced pressure.

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

The extract, preferably a hot water extract, may be sequentially or individually fractionated with organic solvents of hexene, ethyl acetate and butanol to additionally obtain a hexene fraction, an ethyl acetate fraction, a butanol fraction, and a water residue after organic solvent fractionation.

After mushroom harvesting in artificial cultivation of shiitake mushrooms of the present invention, the mushroom waste medium extract and its active fraction may be prepared into powder through a conventional powdering process such as reduced pressure drying, freeze drying, or spray drying. They are not degraded by various degrading enzymes in plasma, and maintain their activity even in heat treatment at 100 ° C and pH in the human stomach of pH 2.

The ethyl acetate fraction of the hot water extract of the mushroom waste medium after cultivation of the shiitake mushroom of the present invention exhibits strong antibacterial activity, and is suitable for pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, and foods related to the prevention or treatment of pathogenic bacterial infections. , it can be used as a material for cosmetics, additives for animal feed, and various cleaners.

As a preferred embodiment, the antibacterial 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 injection solutions according to conventional methods according to each purpose of use. It can be used and can be administered orally or through various routes including intravenous, intraperitoneal, subcutaneous, rectal, topical, and the like. These pharmaceutical compositions may further include carriers, excipients, or diluents, and examples of suitable carriers, excipients, or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, gum acacia, alginates, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. etc. can be mentioned. In addition, the pharmaceutical composition of the present invention may further include fillers, anti-agglomerating agents, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, and the like.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type, severity, drug activity, It may be determined according to factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, 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 in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

The effective amount of the component having antibacterial activity in the pharmaceutical composition of the present invention may vary depending on the patient's age, sex, and weight, and is generally administered in an amount of 1 to 5,000 mg per body weight, preferably 100 to 3,000 mg per day or every other day. Alternatively, it may be divided into 1 to 3 doses per day. However, since it may increase or decrease according to the route of administration, severity of disease, sex, weight, age, etc., the dosage is not limited to the scope of the present invention in any way. The pharmaceutical composition of the present invention may be administered to a subject through various routes. All modes of administration can be envisaged, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine intrathecal 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 common routes as long as it can reach the target tissue. can be administered orally. In addition, the composition of the present invention may be administered using any device capable of delivering active ingredients to target cells. In the present invention, "subject" includes, but is not particularly limited to, for example, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig. and, preferably, a mammal, more preferably a human.

As a preferred embodiment, the antibacterial 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 possibility of infection from a subject at high risk of infection by pathogenic microorganisms. Foods containing the active ingredient with excellent antibacterial activity of the present invention include, for example, various foods, beverages, gum, tea, vitamin complexes, health supplements, etc., in the form of powders, granules, tablets, capsules or beverages can be used as The active ingredient of the present invention may generally be added in an amount of 0.01 to 15% by weight of the total food weight, and the health drink composition may be added in an amount of 0.02 to 10g, preferably 0.3 to 1g, based on 100 ml. In addition to containing the above compound as an essential component in the indicated ratio, the health functional food of the present invention may contain food additives such as natural carbohydrates and various flavors as additional components. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Thaumatin, rebaudioside A, glycyrrhizin, saccharin, aspartame, etc. may be used as the flavoring agent. The proportion of the flavoring agent is generally about 1 to 20g, preferably about 5 to 12g, per 100ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention is various nutrients, vitamins, minerals, flavors such as synthetic flavors and natural flavors, colorants and enhancers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloids It may contain thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like. In addition, the health functional food of the present invention may contain fruit flesh for production of natural fruit juice, fruit juice beverages, and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is generally selected from the range of 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 antibacterial composition of the present invention can be applied as a cosmetic composition.

The cosmetic composition of the present invention can be prepared in the form of an ingredient having excellent antibacterial activity of the present invention, for example, essence, nutrient cream, body lotion, rinse, shampoo, body cleanser, lotion, toner, mask pack, etc. It is not limited here. Preparation in the form can be easily carried out according to various manufacturing processes known to those skilled in the art.

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

The animals are a group of organisms corresponding to plants and mainly consume organic matter as nutrients, and have differentiated digestive, embryonic, and respiratory organs. Specifically, echinoderms, crustaceans, mollusks, fish, amphibians, reptiles, birds It may be mammals, preferably echinoderms such as sea urchins or sea cucumbers, arthropods including crustaceans such as crabs, shrimps and lobsters, molluscs such as cephalopods, gastropods or bivalves, red sea bream, sea 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 products to the antibacterial composition containing the antibacterial component of the present invention. The grain may be specifically ground or crushed wheat, oats, barley, corn and rice, the vegetable protein feed may be specifically rapeseed, soybean and sunflower as main components, and the animal protein feed is specifically may be blood meal, meat meal, bone meal, and fish meal, and the sugar or dairy product may be a dry ingredient composed of various powdered milk and whey powder.

The composition for animal feed may additionally use ingredients such as nutritional supplements, digestion and absorption enhancers, growth promoters, or disease preventive agents.

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

In addition, the feed composition may be made of a hard granular or granular material depending on the degree of grinding of the ingredients, and the composition may be supplied as a mesh or formed into a desired separated shape for further processing and packaging, and may be stored It may be pelletized, expanded, or extruded for storage, and excess water may preferably be removed by drying for ease of storage.

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

The additive is added to impart preservation to food, cosmetics, or animal feed, 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 other additives appropriately selected depending on the purpose and purpose of use, and may be mixed with other active ingredients such as commonly used additives or additives such as pigments, surfactants, and preservatives. . The additives may be prepared by powdering, granulating, tableting or liquefying according to the purpose and purpose of use, 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 area, application target product, etc., for example, may include 0.01 part by weight to 50 parts by weight based on the total composition, but the antibacterial The content of the composition is not limited thereto. Since the antibacterial 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 the purpose of application can be achieved due to the nature of the product.

As a preferred embodiment, the antibacterial composition of the present invention can be applied for use as a detergent.

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

The detergent may further include appropriately selected additives depending on the purpose and purpose of use, and may be mixed 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 purpose of use, and a conventional method may be used for packaging for commercialization.

In addition, the amount of the antimicrobial composition used in the cleaning agent may be appropriately used depending on the purpose of use, application form, application area, application target product, etc., for example, may include 0.01 part by weight to 50 parts by weight based on the total composition, but the antibacterial 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 the purpose of application can be achieved due to the nature of the product.

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

The cleaning agent can be used as a hand, oral, or feminine cleaner in a simple way to prevent infection in the body of pathogenic microorganisms that are often easily infected 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 hand, oral, and feminine cleaner.

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

In addition, the amount of the antimicrobial composition used in the cleaner may be appropriately used depending on the purpose of use, application form, application area, etc., for example, may include 0.01 part by weight to 50 parts by weight based on the total composition, but the content of the antibacterial composition It is not limited to this. Since the antibacterial 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 the purpose of application can be achieved due to the nature of the product.

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

[Example]

Example 1: Physicochemical Characteristics Analysis and Color Analysis of Sterilized Medium for Shiitake Mushroom Cultivation, Wasted Mushroom Medium after 1st Harvest, Wasted Mushroom Medium after 3rd Harvest and Shiitake Mushrooms

A medium for growing shiitake mushrooms (composition: oak 80%, bran 19%, edible gypsum 1%) manufactured in Jinzhen, China in 2017 was pressed and sealed, sterilized at 100 ° C for 8 hours to prepare an aseptic medium, and then shiitake mushroom spawn 808 was inoculated and cultured in a culture room for about 120 days, and then cultured again in a cultivation house at 23 ° C and 70% humidity for 40 days, and then generation work was performed. Afterwards, the generated shiitake mushrooms were harvested once (1st cycle harvest), and after the first harvest, a mushroom waste medium was prepared, and then the shiitake mushrooms were regenerated and harvested 2 times (2nd cycle harvest) and 3rd harvest (3rd cycle harvest) After harvesting the remaining three times, the mushroom waste medium was recovered (FIG. 1).

Photographs of the sterilization medium, the waste mushroom medium after the first harvest, the waste mushroom medium after the third harvest, and the harvested shiitake mushrooms are shown in Figure 1, and their crude protein, crude carbohydrates, crude lipid, ash and moisture content are shown in food Table 1 shows the amount of heat measured according to revolution.

[Table 1] Nutrient composition analysis of sterilization medium, waste mushroom medium after 1st harvest, waste mushroom medium after 3rd harvest and shiitake mushrooms

Moisture content changed from 46.8% at the beginning to 52.6% after the first harvest and 54.0% after the third harvest. The ash content was also 6.55% in the sterilized medium, but increased rapidly to 14.77~14.93% in the mushroom waste medium after harvest. The ash content of shiitake mushrooms was 0.8%. On the other hand, the change in the crude lipid content was insignificant, and the crude carbohydrate decreased rapidly from 43.3% to 25.7% after the first harvest, and then showed 23.2% after the third harvest. This is judged to be the result of the medium after the 1st and 3rd harvests containing a large amount of shiitake mushroom mycelium and fruiting bodies.

On the other hand, as a result of color analysis of each sample after grinding the shiitake mushroom medium and shiitake mushrooms, the sterilized medium showed relatively low brightness, redness, and yellowness, and the color difference was 73.3. After one harvest, the mushroom waste medium and 3 After harvesting, the mushroom waste medium showed 46.3 and 44.75, respectively, showing similar brightness, redness, and yellowness (Table 2). At this time, color difference analysis was measured using a Hunter Color Difference meter (Super color SP-80 Colormeter, Tokyo Denshoku Co., Japan), brightness (white 100 ~ 0 black), redness (red 100 ~ -80 green), Yellowness (yellow 70 to -80 black) was measured. At this time, the chromaticity of the standard white board was set as L value of 92.44, a value of -0.06, and b value of 1.35, and was measured three times per sample to obtain an average value. It was calculated using

[Table 2] Color difference analysis of sterilization medium, waste mushroom medium after 1st harvest, waste mushroom medium after 3rd harvest and shiitake mushroom powder

Example 2: Sterilization medium, mushroom waste medium after 1st harvest, mushroom waste medium after 3rd harvest and shiitake mushroom hot water extract preparation and physicochemical characterization analysis thereof

Hot water extracts were prepared from the sterilization medium prepared in Example 1, the waste mushroom medium after the first harvest, the waste mushroom medium after the third harvest, and dried shiitake mushroom powder. After adding 10 times distilled water to each sample and extracting at 100 ° C. for 1 hour, the pH, brix and color difference of the obtained extract were measured, and the results are shown in Table 3.

[Table 3] Physicochemical properties of sterilization medium, mushroom waste medium after 1st harvest, mushroom waste medium after 3rd harvest, and shiitake mushroom powder hot water extract

The initial pH of the sterilization medium was 5.6, but it was reduced to 4.6 in the waste mushroom medium after the first and third harvests. In addition, the initial brix of the sterilization medium increased from 6.0 to 16.0 and 20.0 for the mushroom waste medium after the first and third harvests, and the acidity also increased from 0 initially to 0.83 and 1.40 after the first and third harvests. . On the other hand, in the color analysis of the hot water extract, the brightness and redness decreased and the yellowness increased as the number of harvests increased, and the color difference increased from 76.8 in the initial sterilization medium to 78.2 in the mushroom waste medium after the third harvest. That is, the longer the culture of shiitake mushrooms, the higher the acidity, brix, and yellowness of the mushroom waste medium (FIG. 2).

Example 3: Component analysis of sterilization medium, mushroom waste medium after 1 harvest, mushroom waste medium after 3 harvest, and hot water extract of shiitake mushroom

After filtering the hot water extract prepared in Example 2, it was concentrated under reduced pressure to prepare a powder, and the extraction efficiency and component analysis results of each extract are shown in Table 4. Component analysis determined total polyphenol, total flavonoid, total sugar and reducing sugar content. For the total polyphenol content, 50 μl of Folin-ciocalteau and 100 μl of a saturated solution of Na ₂ CO ₃ were added to 400 μl of the extraction sample solution, and the absorbance was measured at 725 nm after being allowed to stand at room temperature for 1 hour. As a standard reagent, tannic acid was used. For the total flavonoid content, each sample was stirred and extracted with methanol for 18 hours, 4ml of 90% diethylene glycol was added to 400μl of the filtered extraction sample solution, 40μl of 1N NaOH was added, and the absorbance was measured at 420nm after reaction at 37℃ for 1 hour. . Rutin was used as a standard reagent. Reducing sugar was quantified using the DNS method, and total sugar was quantified using the phenol-sulfuric acid method.

[Table 4] Component analysis of sterilization medium, waste mushroom medium after 1 harvest, waste mushroom medium after 3 harvests, and hot water extract of shiitake mushroom

As shown in Table 4, the extraction efficiency was very low at 1% in the sterile medium, but the mushroom waste medium showed a high yield of 5.1 to 9.2%, and the third harvest was higher than the mushroom waste medium (5.1%) after the first harvest. After the mushroom waste medium (9.2%), the extraction yield was about 1.8 times higher. The extraction efficiency of the hot water extract of dried shiitake mushroom powder was very high at 34.7%. This is considered to be due to the increase in soluble components due to the decomposition of the sterilization medium as the cultivation of shiitake mushrooms progresses.

As a result of total polyphenol analysis, the sterilization medium showed 20.5mg/g, which was 2.5 times higher than that of shiitake mushrooms. It showed a significantly higher content than harvested shiitake mushrooms. This pattern was also found in the total flavonoid content analysis (Table 4). Therefore, it was predicted that the mushroom waste medium after the first and third harvests would show better physiological activity than the sterilized medium for mushroom cultivation and the harvested shiitake mushrooms.

On the other hand, in the case of total sugar, the highest content of 144.9mg / g was shown in the sterilized medium, and in the case of reduced sugar, the highest content of 53.4 ~ 54.0mg / g was shown in the mushroom waste medium after the 1st and 3rd harvest. On the other hand, shiitake mushroom extract showed very low total sugar (21.6 mg/g) and reducing sugar (19.7 mg/g) content, unlike other samples.

Example 4: Antibacterial activity of sterilization medium, mushroom waste medium after 1 harvest, mushroom waste medium after 3 harvest and hot water extract of shiitake mushroom

Antibacterial activity was evaluated for various extracts prepared in Example 2, and the results are shown in Table 5. At this time, the disc diffusion method was used for antibacterial activity, and the sample concentration per disc was adjusted to 0.5 mg.

Escherichia coli KCTC 1682, Proteus vulgaris KCTC 2433, Pseudomonas aeruginosa KACC 10186, and Salmonella typhimurium KCTC 1926 were used as strains for antibacterial activity evaluation, and Staphylococcus aureus KCTC 1916 and Staphylococcus epidermidis ATCC 12228 were used as gram-positive bacteria. , Listeria monocytogenes KACC 10550 and Bacillus subtilis KCTC 1924 were used. On the other hand, for evaluation of antifungal activity, Saccharomyces cerevisiae IF0 0233 and Candida albicans KCTC 1940, the causative agent of candidiasis fungal infection, were used.

First, in the case of antibacterial activity evaluation, each bacterium was inoculated into Nutrient broth (Difco Co., USA) and cultured at 37° C. for 24 hours, and each strain was adjusted to an OD ₆₀₀ of 0.1 to Nutrient agar (Difco Co. , USA) 100 μl each was spread on a sterile petri dish (90 mm, Green Cross, Korea) containing the medium, and 5 μl of each sample was applied to a sterile disc-paper (diameter 6.5 mm, Whatsman No. 2) for 24 hours at 37 ° C. In the case of fungi, after incubation at 30 ° C. for 24 hours in the same manner using Sabouraud dextrose (Difco Co. USA), the antibacterial activity was evaluated by measuring the size of the growth inhibition ring. As a control, antibacterial agent ampicillin and antifungal agent miconazole (Sigma Co., USA) were used at a concentration of 1 μg/disc, respectively. It was measured in mm, and it was shown as a representative result after 3 or more evaluations.

[Table 5] Sterilization medium, mushroom waste medium after 1 harvest, mushroom waste medium after 3 harvest, antibacterial activity of harvested shiitake mushroom powder hot water extract

As shown in Table 5, in the case of ampicillin (1μg/disc) used as an antibacterial activity control, growth inhibitory rings of 20.0 to 30.0 mm were applied to all bacteria except Escherichia coli ( E. coli ) and P. aeruginosa ( P. aeruginosa ). In the case of myconazole (1 μg/disc), which is an antifungal agent, growth inhibitory rings of 10.5 mm and 9.5 mm were shown for C. albicans and S. cerevisiae , respectively. On the other hand, the sterilization medium for shiitake mushroom cultivation, the waste mushroom medium after the 1st harvest, the waste mushroom medium after the 3rd harvest, and the hot water extract of dried shiitake mushroom powder showed no growth inhibition for all of the experimental bacteria and fungi at a concentration of 500 μg/disc. Didn't show up. In the case of shiitake mushroom extract, antibacterial activity has been reported at a very high concentration of 43.1mg/disc (Kim Yong-doo et al., 2003. Journal of the Korean Food Storage and Distribution Society 10: 89-93), but this high concentration is beyond the practically applicable part, so it is meaningless. There is no report (assuming that human blood is 4 liters, 43mg/disc concentration is the concentration at which an antimicrobial agent should be ingested close to 50g). Normally, activity must be recognized at a concentration of 1mg/disc or less to be used as an actual antibacterial agent. Therefore, these results mean that the sterilization medium for shiitake mushroom cultivation, the waste mushroom medium after the first harvest, the waste mushroom medium after the third harvest, and the dried shiitake mushroom dry powder extract have no antibacterial and antifungal activity.

Example 5: Sterilization medium for shiitake mushroom cultivation, preparation of sequential organic solvent fractions of waste mushroom medium after 1 harvest and hot water extract of mushroom waste medium after 3 harvest and analysis of components

In order to confirm the active ingredients of the sterilization medium for shiitake mushroom cultivation prepared in Example 2, the mushroom waste medium after the first harvest and the hot water extract of the mushroom waste medium after the third harvest, hexene, ethyl acetate, Sequential organic solvent fractionation was performed using butanol, and finally a water residue was recovered. Components of each recovered fraction were analyzed in the same manner as in Example 3, and the results are shown in Table 6.

[Table 6] Sterilization medium, preparation and component analysis of sequential organic solvent fractions of mushroom waste medium after 1 harvest and hot water extract of mushroom waste medium after 3 harvest

As shown in Table 6, in the case of the hot-water extract of the sterilization medium for shiitake mushroom cultivation, 90.7% was converted to water residue, indicating that the hot-water extract contained a significant amount of water-soluble components. On the other hand, the water residues of the mushroom waste medium after the first harvest and the extract of the mushroom waste medium after the third harvest were 69.3% and 78.3%, respectively, indicating a relatively reduced water-soluble component. The fat-soluble hexene fraction of the hot-water extract of the sterilization medium for shiitake cultivation, the waste mushroom medium after the first harvest, and the waste mushroom medium after the third harvest accounted for 0.1, 0.2, and 0.1% of the hot-water extract, respectively, and were negligible. On the other hand, the yields of ethyl acetate fractions containing high concentrations of polyphenols and flavonoids accounted for 2.7, 5.7, and 8.1% of each hot water extract, and gradually increased as the cultivation period increased. In the case of the butanol fraction, it accounted for 10.8, 24.3, and 17.2% in the hot water extract of the sterilization medium, the waste mushroom medium after the first harvest, and the waste mushroom medium after the third harvest, respectively, showing the highest fraction yield in the waste mushroom medium after the first harvest. was

On the other hand, in the case of the ethyl acetate fraction containing polyphenols and flavonoids at high concentrations, the total polyphenol and flavonoid content was highest in the sterilized medium, and the total sugar content was also the highest in the ethyl acetate fraction of the sterilized medium. . However, the reducing sugar content was relatively high in the ethyl acetate fraction of the mushroom waste medium after the first and third harvests. These changes are understood to be the changes that occurred due to the decomposition and use of oak sawdust and wheat bran, which are the components of the medium, as the shiitake mushrooms inoculated in the sterilization medium grew.

In particular, as a result of analyzing the total polyphenol content of each fraction, in the case of the sterilization medium, the ethyl acetate fraction showed the highest value of 185.7 mg/g, followed by the hexene fraction > butanol fraction > water residue. However, in the case of the mushroom waste medium after the first harvest, the hexene fraction showed the highest 54.8mg / g, followed by the ethyl acetate fraction > butanol fraction > water residue, and in the case of the mushroom waste medium after the third harvest, ethyl The acetate fraction showed the highest value of 95.4 mg/g, followed by the water residue > butanol fraction > hexene fraction. These results mean that the components of the sterilization medium and the mushroom waste medium extract after harvest are completely different, and the components of the mushroom waste medium extract after one harvest and the mushroom waste medium extract after three harvests are different.

Example 6: Antibacterial Activity of Sequential Organic Solvent Fractions of Sterilization Medium for Shiitake Mushroom Cultivation, Wasted Mushroom Medium after 1st Harvest and Hot Water Extract of Wasted Mushroom Medium after 3rd Harvest

The antibacterial and antifungal activities of the various fractions obtained in Example 5 were evaluated in the same manner as in Example 4. As shown in Table 7, antibacterial activity was not shown in the case of the sterilization medium for shiitake mushroom cultivation, the mushroom waste medium after the first harvest, and the hot water extract of the mushroom waste medium after the third harvest, but the ethyl acetate fraction of each extract had very strong antibacterial activity. In particular, the ethyl acetate fraction of the mushroom waste medium after the first harvest and the hot water extract of the mushroom waste medium after the third harvest showed excellent antibacterial and antibacterial activity with a broad antibacterial spectrum against gram-positive bacteria, gram-negative bacteria and fungi. showed antifungal activity. In the case of the ethyl acetate fraction prepared from the mushroom waste medium after the first harvest, the overall antibacterial activity was superior to the ethyl acetate fraction prepared from the mushroom waste medium after the third harvest, and the growth inhibitory activity against Gram-positive bacteria was more powerful.

[Table 7] Sterilization medium for shiitake mushroom cultivation, antibacterial activity of the sequential organic solvent fractions of the waste mushroom medium after the first harvest and the hot water extract of the mushroom waste medium after the third harvest

Example 7: Human erythrocyte hemolytic activity of mushroom waste medium after the first harvest and hot-water extract of the mushroom waste medium after the third harvest and active fractions thereof

Shiitake mushrooms have been used for food and medicine for a long time, and mushroom waste medium after harvesting shiitake mushrooms has been used as feed. In the present invention, human erythrocyte hemolytic activity was tested to evaluate the acute toxicity of mushroom lung medium extracts and their active fractions, and the results are shown in Table 8. At this time, the hemolytic activity was evaluated according to the previous report (Jung In-chang, Son Ho-yong, 2014, Korean J. Microbiol. Biotechnol. 42: 285-292), and simply 100 μl of human red blood cells washed three times with PBS was added to a 96-well It was added to a microplate, 100μl of sample solution of various concentrations was added, reacted at 37 ° C for 30 minutes, and then the reaction solution was centrifuged for 10 minutes (1,500 rpm), and 100μl of the supernatant was transferred to a new microtiter plate, and hemoglobin flow due to hemolysis The degree was measured at 414 nm. DMSO (2%) was used as a solvent control for the sample, and Triton X-100 (1 mg/ml) was used as a control for red blood cell hemolysis. Hemolytic activity was calculated using the following formula.

[Table 8] Human erythrocyte hemolytic activity of the mushroom waste medium after the first harvest and the hot water extract of the mushroom waste medium after the third harvest and active fractions thereof

First, it was confirmed that DMSO and water used as controls had no erythrocyte hemolytic activity, and triton X-100 100% hemolyzed erythrocytes at a concentration of 1 mg/ml. On the other hand, after the first and third harvests, the mushroom waste medium extract and its active fractions showed no hemolytic activity even at 1 mg/ml. Considering that amphoteracin B, an anticancer drug known to have hemolytic activity, exhibits hemolytic activity of 55.5% at a concentration of 0.00625mg/ml and 93.7% at a concentration of 0.05mg/ml, mushroom waste medium extract and its active fraction after harvesting shiitake mushrooms are considered to have no hemolytic activity.

Example 8: Evaluation of Plasma, Acid and Thermal Stability of Mushroom Waste Medium After 1st Harvest and Hot-Water Extract and Active Fractions of Mushroom Waste Medium After 3rd Harvest

Thermal stability and acid stability for antibacterial activity were confirmed for the ethyl acetate active fraction of the mushroom waste medium after the first harvest and the mushroom waste medium extract after the third harvest obtained in Examples 2 and 5. The ethyl acetate active fractions showed no decrease in antibacterial and antifungal activity even when heat treated at 100° C. for 1 hour and treated at pH 2 (0.01M HCl) for 1 hour. Therefore, the active fraction is expected to maintain antibacterial and antifungal activity during the manufacturing process of various foods, cosmetics, and household products.

[R&D task]

[Assignment identification number] 2021-0357

[Name of department in charge] Ministry of SMEs and Startups

[Research management institution] Andong National University Industry-University Cooperation Foundation

[Research project name] R&D research on fostering star companies at the Gyeongbuk Regional Center for Value Creation

[Name of research project] Development of high value-added products using local agricultural products

[Contribution rate] 100%

[Research period] 2021. 09.27.-2021.12.25.

[Organizer] Corporate Support Center for the Disabled Gyeongbuk Regional Center

[Research Institution] Department of Food and Nutrition, Andong National University

Claims (4)

Shiitake mushroom ( Lentinus edodes ) Mushroom waste medium after harvesting (Spent Mushroom Substrate, SMS) antibacterial composition containing an extract. The antimicrobial composition according to claim 1, wherein the waste mushroom medium is a waste mushroom medium after harvesting shiitake mushrooms 1 to 3 times. The antimicrobial composition according to claim 1, wherein the extract is extracted with hot water and then fractionated with ethyl acetate. The method according to any one of claims 1 to 3, wherein the antibacterial 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. Antimicrobial composition, characterized in that being.

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