KR20200058353A - Antimicrobial composition comprising the hexane and butanol fractions of teleogryllus emma extract - Google Patents

Abstract

The present invention relates to an antibacterial composition comprising a hexane fraction or butanol fraction of an extract of Teleogryllus emma as an effective component, and more specifically, to an antibacterial composition for a wide range of gram-positive bacteria, which comprises, as an effective component, a hexane fraction or butanol fraction collected by sequential fractionation of an ethanol extract of Teleogryllus emma, which is a native species of grasshopper in South Korea. The hexane fraction or butanol fraction of an extract of Teleogryllus emma as an effective component of the antibacterial composition of the present invention, as demonstrated in examples in the present specification, exhibits excellent antibacterial activity against gram-positive bacteria and food-poisoning bacteria causing allergic food poisoning, such as Proteus vulgaris, and thus may be utilized for a pharmaceutical composition, a health functional food, a cosmetic composition, an animal feed composition, food, cosmetics, or an animal feed additive, a variety of detergents and cleansers, and the like. Moreover, the hexane fraction or butanol fraction of an extract of Teleogryllus emma of the present invention has excellent thermal stability and shows no loss in activity even in an acidic condition of pH 2, and thus can be easily processed into various forms including liquid, cream, powder, tablets, pills, and the like, and may be extremely usefully used in drug industry, food industry, livestock industry, health industry, and cosmetics industry.

Description

Antimicrobial composition comprising hexene fraction or butanol fraction of the king cricket extract as an active ingredient {ANTIMICROBIAL COMPOSITION COMPRISING THE HEXANE AND BUTANOL FRACTIONS OF TELEOGRYLLUS EMMA EXTRACT}

The present invention relates to an antimicrobial composition containing a hexene fraction or a butanol fraction of an extract of a king cricket ( Teleogryllus emma ) as an active ingredient, and more specifically, a hexene recovered by sequentially ethanol extracting a ethanol extract of a king cricket, a native species of domestic cricket It relates to a wide range of antibacterial compositions against Gram-positive bacteria containing a fraction or a butanol fraction as an active ingredient.

According to the U.S. Center for Disease Control and Prevention, in-hospital blood flow infection is one of the leading causes of death in the United States. In addition, infection of pathogenic bacteria and fungi through 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 have different treatment methods depending on the severity of the infection, but oral antimicrobial agents and topical cleanliness enable successful treatment. However, more severe or complex infections that are common in patients with underlying risk factors (e.g., vascular blood flow abnormalities, diabetes) and / or infections caused by difficult or multi-resistant bacteria and fungi are difficult to treat. Anti-microbial treatment and invasive surgical tissue removal may be necessary. Illustratively, the glycopeptide antibiotic vancomycin has been successfully used for severe in-hospital infections caused by multi-drug-resistant Gram-positive bacteria, particularly MRSA, coagulase-negative Staphylococcus and Enterococcus. However, recently, it has been reported that the separation of strains resistant to vancomycin is increasing, and it is true that current antibiotics have very limited treatment for severe infections caused by some of the pathogens. Therefore, there is an increasing need to develop new antibiotics that are more effective against resistant bacteria.

On the other hand, cricket belongs to the cricket family of arthropod grasshoppers, and there are about 1,200 species worldwide, and about 30 are known in Korea. Among them, the king cricket ( Teleogryllus emma ) is a cricket native to East Asia, including Korea. Unlike the Gryllus bimaculatus , which has been imported from abroad, it is a domestic indigenous species that adapts to the climate of Korea, which has four distinct seasons and winter. to be. The body is shiny brown or dark brown, and the parietal becomes narrower toward the front. The inside of compound eyes is yellow, connected to a white horizontal band with a round face, and the antenna is thin and long and looks like a thread. The front breast back is slightly narrower than the head, and the edges of the front and rear edges are slightly narrower and bent toward the back, the center is slightly thicker, the surface is wider, and the front edge of the next piece is round and has a light yellow pattern. In Korea, it is commonly found in gardens and surrounding grasslands, in humid, dark warehouses and toilets, and in accreditation. It is the most common type of cricket. On the other hand, a paired cricket registered as edible in Korea in 2016 is an insect of the grasshopper family Cricket that inhabits subtropical regions. It is an insect that has been imported into Korea and raised for food, medicinal, pet, feed, and educational purposes. In particular, the body is black overall, and there is a yellow spot on the base of the fore wing, so it is called 'two-spotted cricket' in English, and in Korean, it is called a double-headed cricket.

Currently, domestic and foreign research is concentrated on twin-star crickets, and research on king crickets is very poor. Studies on the king cricket include artificial artificial egg larvae and storage of the eggs of the Teleogryillus emma (Kim Nam-Jung et al., 2005. Journal of the Korean Society of Applied Entomology, 44, 61-65). The Korean Society of Applied Entomology 2003 Fall Conference p. 68-68) and the excellent anti-inflammatory activity of the king cricket (Hwang Jae-sam et al., 2015. Skin-friendly and anti-inflammatory mechanism of insect useful substances, National Academy of Agricultural Science) , Influence of the exotic species Gryllus bimaculatus on the survival rate of the native species Teleogryllus emma (Lee Eun-a et al., 2013. Ecology and environment, 46, 67-74), two crickets ( Gryllus bimaculatus , Teleogryllus emma ) Immune activity comparison of extracts (Dong-Hwan Seo et al., 2003. The 46th Spring Conference of the Korean Society of Proceedings, p.79, 2003) is known. There is no situation.

As a research on twin crickets, the effect of the breeding density of twin crickets on feeding, adult mortality and hatching larvae has been reported (Park Young-gyu et al., 2013. Korean Society of Sericultural Insects 51, 89-94), and crickets are added for feed and edible use. Effects of feed on the composition of chicken meat and eggs (Ahn Mi-young et al., 2000. Korean Poultry Association 27, 197-202), Physicochemical properties of edible crickets by mountain area (Kim Eun-mi et al., 2015. Korean Journal of Food Preservation, 22, 831- 837) and in the case of repeated administration of cricket extract in rats, no toxicity was reported (Hwang Seok-yeon et al., 2004. Korean Society for Experimental Zoology 20, 194-199). Useful physiological activities of twin-brick crickets include the effects of cricket water and methanol extracts on alcohol metabolism (Lee Yong-Woo et al., 2004. Korean Society for Pharmacy Medicine 35, 175-178), water-soluble cricket crickets and liver protective effects of alcohol extracts (Ahn Mi-Young et al., 2002 , Korean Journal of Food Science and Technology 34, 684-687), TCDD (2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin) prevents the effect of cricket extract on hepatotoxicity of rats (Park Jong-bae, 2004. Master of Daejeon University) Thesis), when the cricket is added as a feed, it is known to increase the antioxidant enzyme activity and growth rate of African catfish (Taufek et al., 2016. Fish Physiol Biochem. 42, 1143-1155), and prevent DNA oxidation when administered to rats for 1 month There is a report (Ahn MY et al., 2015. Toxicol Res. 31, 173-180), but until now, the antibacterial effect of the twin-brick cricket extract itself has not been known.

The patent for the king cricket is registered as the Republic of Korea Patent No. 10-1790018 in 2016 [Composition for the prevention or treatment of inflammatory bowel disease containing the extract of the king cricket], 2018 Korea Patent No. 10-1854658 [Contain the extract of the king cricket as an active ingredient The pharmaceutical composition and health functional food for prevention or treatment of thrombosis to be described], and No. 10-1856602 [Antioxidant composition containing Wanggwi-Turami extract as an active ingredient] are known, and strong antibacterial activity of the Wanggwi-Turami extract has not been reported. . On the other hand, with respect to the paired cricket, Republic of Korea Registered Patent No. 10-1661176 [Method of manufacturing cricket for food and cricket raw material for food prepared accordingly], No. 10-1493916 [Insect edible method, its edible insect , Method for preparing food dough using such edible insects], No. 10-1686179, [Cricket with enhanced arginine content to produce nitric oxide] is known to be effective in preventing hypertension, and with regard to the use of twin cricket, [No. 10-1429679] [Cosmetic composition for skin whitening containing cricket extract], No. 10-1660381 has an anti-angiogenic ability or a therapeutic effect for diseases caused by dysfunction of vasodilation reaction [Insect N-glycan and Its use] is registered, and in Patent Publication No. 10-2013-0134097 [Composition for preventing or treating dermatitis containing cricket extract], Patent Publication No. 10-2016-0096548 is fed with a feed of specific composition. [Antioxidant, anti-aging and skin beauty improving effect and cosmetic composition containing the same] for the use of a paired cricket with increased antioxidant active ingredient, [10-2014-0036369] [Feed feed using dried cricket Manufacturing method], [10-2016-0128113] [Method of manufacturing processed foods using edible insects] and [10-2016-0118860] [powder mixture and insect noodles] are disclosed. In addition, [Cricket Scattering and Incubation Mechanism] is registered in Korean Utility Model No. 20-0477102, and [Cricket Bulk Collector] is registered in No. 20-0477103. However, there have been no reports of crickets, especially the antimicrobial composition of crickets.

KR 10-1790018 B KR 10-1854658 B KR 10-1856602 B

 2017 Korean Life Science Society 58th General Assembly and International Academic Conference Poster P2-32.  2017 Korea Microbiological Biotechnology Society Yeungnam Branch Conference P013.

The present invention has been devised 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 a hexene fraction or a butanol fraction of a king cricket extract as an active ingredient.

In order to solve the above problems, the present invention provides an antimicrobial composition containing a hexene fraction or a butanol fraction of an extract of a king cricket ( Teleogryllus emma ) as an active ingredient.

It is preferable that the king cricket extract is a king cricket ethanol extract.

It is preferable that the antibacterial composition has antibacterial activity against Gram positive bacteria.

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

Hexene fraction or butanol fraction of wanggwitturami extract as an active ingredient of the antimicrobial composition of the present invention, poisoning that causes allergic food poisoning, such as, as demonstrated in Examples of the present specification, the gram-positive bacteria and Proteus vulgaris (Proteus vulgaris) It exhibits excellent antibacterial activity against bacteria, and has excellent effects that can be used for pharmaceutical compositions, dietary supplements, cosmetic compositions, animal feed compositions, additives for food, cosmetics or animal feed, various cleaners, and various cleaners. In addition, the hexene fraction or butanol fraction of the king cricket extract of the present invention has excellent thermal stability and does not exhibit loss of activity even under acidic conditions of pH 2, so it is easily processed into various forms such as liquid, cream, powder, pill, tablet, etc. It can be used in the pharmaceutical industry, food industry, livestock industry, health industry and cosmetics industry.

1 is a photograph of a king cricket used in an embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention, to test the antibacterial activity against the king cricket, first feed the mixture of wheat bran and fishmeal in a breeding box at 25 ° C, breed the king cricket as an adult, remove the wings, and fast for 24 hours. Inducing bowel movements, and then rapidly freezing to -20 ° C to kill, then freeze-dried at -50 ° C, and then, using a pestle, to prepare a king cricket powder. Next, an ethanol extract was prepared using the manufactured powder, and the sequential organic solvent fraction, a hexene fraction, an ethyl acetate fraction, a butanol fraction, and a water residue were recovered from the ethanol extract, and the hexene fraction and butanol fraction were antibacterial active ingredients. Was confirmed, and the active ingredient was intended to utilize the hexene fraction or the butanol fraction of the king cricket extract as an antibacterial composition by confirming that it has excellent thermal and acid stability characteristics.

Specifically, the present inventors prepared an ethanol extract targeting a king cricket to develop an antibacterial composition using a king cricket known to have anti-inflammatory, hepatoprotective, and antihypertensive effects, and prepared a sequential organic solvent fraction targeting these extracts. then, by checking the result of test a wide range of antimicrobial activity thereof, shows excellent antibacterial activity against Gram-negative bacteria, such as positive bacteria and Proteus vulgaris gram in hexene fraction and the butanol fraction (Proteus vulgaris), can be developed with these broad antibacterial agents such as It was confirmed that the present invention was completed.

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

It is preferable that the king cricket extract is a king cricket ethanol extract.

In addition, the antimicrobial composition of the present invention is preferably antibacterial activity, more specifically, antibacterial activity against Proteus vulgaris among Gram positive bacteria and Gram negative bacteria.

Hereinafter, a method and an efficacy test of the active fraction of the extract of the king cricket of the present invention will be described in more detail.

Inventors of the present invention to achieve the object of the present invention, breeding a king cricket; Pre-treatment and lyophilization of adult; Powdering the adult; Preparing a cricket extract; Sequential organic solvent fraction preparation of hexene, ethyl acetate and butanol from the extract of the king cricket and preparation of water residues obtained afterwards; The extracts and fractions were subjected to experiments of evaluating the antibacterial activity and investigating the stability of the active substance.

As a preferred embodiment, the king cricket breeding of the present invention uses the same method as the normal insect breeding, and can feed the king cricket as an adult by supplying a feed mixed with wheat bran and fish meal in a breeding box at 25 ° C. Thereafter, after removing the wing of the adult, 24 hours fasting and inducing natural bowel movements, rapidly freezing to -20 ° C and killing, and then freeze-dried at -50 ° C, and the freeze-dried adult is made into a powder using a mortar. . Thereafter, the step of extracting with a solvent and filtering the extract using a filter network of 0.06 mm or less, and concentrating it under reduced pressure can obtain the king cricket extract of the present invention, additionally, hexene, ethyl acetate and The organic solvent fraction and water residue of butanol can be recovered.

The solvent used in the present invention is water (cold water, hot water), alcohol, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, butanol, etc.), mixed solvents of the lower alcohol and water, etc. Can be used, hot water, or 95% ethanol extraction is most preferred. Wangguturami hot water or ethanol extract may be sequentially or separately fractionated with an organic solvent of hexene, ethyl acetate and butanol to additionally obtain a hexene fraction, ethyl acetate fraction, butanol fraction and water residue.

The hexene fraction or butanol fraction of the king cricket extract of the present invention may be prepared as a powder through a conventional powdering process such as vacuum drying and lyophilization, or spray drying. They are not degraded by various degrading enzymes in plasma, and they maintain activity even at a heat treatment of 100 ° C and a pH in the human stomach of pH 2.

The hexene fraction or butanol fraction of the king cricket extract of the present invention exhibits strong antimicrobial activity, pharmaceutical composition, health functional food, cosmetic composition, animal feed composition, food, cosmetic, animal feed additives related to prevention or treatment of pathogenic bacteria infection And various cleaning agents.

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, injections of sterile injectable solutions, etc. It can be used orally or by various routes including intravenous, intraperitoneal, subcutaneous, rectal and topical administration. The pharmaceutical composition may further include a carrier, excipient, or diluent, and examples of suitable carrier, excipient, or diluent that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, acacia rubber, 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 include a filler, an anti-coagulant, 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, "a pharmaceutically effective amount" means 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 patient's disease, severity, activity of the drug, Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs and other factors well known in the medical arts. 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 a conventional therapeutic agent, and may be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those 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 patient's age, sex, and body weight, and generally 1 to 5,000 mg per body weight, preferably 100 to 3,000 mg per day or every other day Alternatively, it may be administered in 1 to 3 times a day. However, the dosage may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, etc., so that the dosage does not limit the scope of the present invention in any way. The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracerebroventricular injection. In the present invention, "administration" means providing a predetermined substance to a patient in any suitable way, 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 the active ingredient to target cells. “Subject” in the present invention is not particularly limited, and 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.

In 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, which can lower the likelihood of infection from a subject with high risk of infection of pathogenic microorganisms. Foods containing the active ingredient having excellent antimicrobial activity of the present invention include, for example, various foods, beverages, gums, teas, 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 can generally be added at 0.01 to 15% by weight of the total food weight, and the health drink composition can be added at a rate of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml. The dietary supplement of the present invention may contain, as an essential ingredient in the indicated proportions, the above compound as an essential ingredient, and may contain, as an additional ingredient, food additives, such as natural carbohydrates and various flavoring additives. Examples of the natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and sugars such as xylitol, sorbitol, and erythritol, and common sugars such as dextrin and cyclodextrin. As the flavoring agent, taumatin, rebaudioside A, glycyrrhizine, saccharin, aspartame, or the like can be used. The flavoring agent is generally used in an amount of about 1 to 20 g, preferably about 5 to 12 g per 100 ml of health functional food of the present invention. In addition to the above, the health functional food of the present invention includes various nutrients, vitamins, minerals, synthetic flavoring agents, flavoring agents such as natural flavoring agents, coloring agents and neutralizing agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, and protective colloids It may contain a thickening agent, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonic acid used in carbonated beverages and the like. In addition, the health functional food of the present invention may contain natural fruit juice and fruit flesh for the production of fruit juice drinks and vegetable drinks. These ingredients can be used independently or in combination. The proportion of these 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 the use of a cosmetic composition.

The cosmetic composition of the present invention can be prepared in the form of, for example, essence, nourishing cream, body lotion, rinse, shampoo, body cleanser, lotion, skin, mask pack, etc. with excellent antibacterial activity of the present invention. It is not limited to. Manufacturing in this form may be readily 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 the plant, mainly consumes organic matter as nutrients, refers to digestion, excretion, and respiratory system differentiation, specifically, echinoderm, crustacean, mollusk, fish, amphibians, reptiles, birds, It may be a mammal, and preferably, arthropods including crustaceans such as sea urchins or sea cucumbers, crabs, shrimp, and crustaceans, mollusks such as cephalopods, gastropods, or bivalve shellfish, red snapper, sea bream, cod, flounder , Birds including fish such as flounder, poultry such as pheasants or chickens, or mammals such as pigs, cows, and goats.

The composition for animal feed may further include grains, vegetable protein feed, animal protein feed, sugar or dairy products in addition to the antimicrobial composition comprising 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 be specifically composed of rapeseed, soybean, and sunflower as main components, and the animal protein feed may be specifically May be blood powder, meat powder, bone powder, and fish powder, and the sugar or dairy product may be a dry component composed of various powdered milk and whey powder.

The animal feed composition may additionally use components such as nutritional supplements, digestion and absorption enhancers, stimulators, or disease prevention agents.

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

In addition, the feed composition can be made of granular or granular material with a viscosity depending on the degree of grinding of the ingredients, the composition can be supplied as a mesh or formed into a desired separated shape for further processing and packaging, and storage In order to facilitate the pelletization, expansion or extrusion process, for ease of storage, excess water may be desiccated and dried.

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

The additive is added to impart preservability to the food, cosmetics, or animal feed 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 additives may further include additional additives selected appropriately according to the purpose and use of the additives, and may be mixed 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 depending on the purpose and use of the additive, and a conventional method may be used for packaging for productization.

In addition, the amount of the antimicrobial composition used among the additives can be appropriately used depending on the purpose of use and application type, application site, and the object to be applied, for example, may include 0.01 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 as long as it can achieve the application purpose due to the characteristics of the product.

In a preferred embodiment, the present invention can be applied to the antibacterial composition as a cleaning agent.

The cleaning agent may be preferably a detergent for clothes, a dishwashing agent, a food cleaning agent, a household appliance cleaning agent. Since the antimicrobial composition is a natural component and is not harmful to the human body, more preferably, it can be used for the purpose of washing or imparting antimicrobial properties to various household products including kitchenware or food washing.

The cleaning agent may further include an additive selected appropriately according to the purpose and use of the cleaning agent, and may be used by mixing with other active ingredients such as commonly used cleaning agents, 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, and a conventional method for packaging may be used for productization.

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 site, and object to be applied, and may include, for example, 0.01 to 50 parts by weight based on the total composition, but the antibacterial property 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 application purpose due to the characteristics of the product.

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

The cleaning agent may be used as a hand, mouth, or female cleaning agent in a simple manner in order to prevent infection of a pathogenic microorganism, which is commonly susceptible to everyday life, in a preventive manner. Since the antibacterial composition is a natural ingredient and is not harmful to the human body, it can be safely applied even when used as a cleanser for hands, mouths, and women.

The cleaning agent may further include additional additives appropriately selected according to the purpose and purpose of use, and may be mixed with other active ingredients such as commonly used additives, additives such as pigments, surfactants, and preservatives. . The cleaning agent may be prepared by powdering, granulating, tableting or liquefying according to the purpose and purpose of use, but in the case of a liquid type, it may be conveniently used.

In addition, the amount of the antimicrobial composition used in the cleaning agent may be appropriately used depending on the purpose of use, the application form, and the application site, and may include, for example, 0.01 to 50 parts by weight based on the total composition, but the content of the antibacterial composition is It is not limited to this. 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 application purpose due to the characteristics of the product.

Hereinafter, the present invention will be described in more detail through specific examples. The following examples describe one preferred embodiment of the present invention, and the scope of the present invention is not limited by the matters described in the following examples.

[Example]

Example 1: Breeding, fasting, lyophilization, and powder preparation

Crickets are omnivorous and easy to breed, and mass breeding systems have already been established. Since 2005, the Rural Development Administration has spread the pet insect king cricket breeding method and has become a pet insect (March 28, 2005, press release), and has been widely used as a food feed for various reptiles. In this embodiment, for feeding the king cricket, a feed mixture of wheat bran and fish meal is sufficiently supplied in a breeding box at 25 ° C., and raised for 8 months or more and raised as a king cricket adult (FIG. 1). After removing the wings of the adult, 24 hours fasting and inducing natural bowel movements, and rapidly frozen to -20 ℃ to kill, then freeze-dried at -50 ℃, lyophilisate was prepared as a powder using a mortar.

On the other hand, commercially available twin-pair crickets were prepared as fasting, defecation induction, lyophilization, and powder as controls, and freeze-dried powders of brown beetle (millworm), slug, and longevity beetle larvae were purchased and used as controls. The insects used are shown in Table 1.

[Table 1] Information of king cricket, twin-star cricket, brown pelar, slug and scarab beetle

Example 2: Preparation of various insect extracts and component analysis of the extract

Ethanol extract was prepared for the powder of the king cricket, twin-star cricket and control insects prepared in Example 1. At this time, 10 times ethanol was added to each insect sample, and the extract was collected twice, filtered at room temperature, filtered, and concentrated under reduced pressure to prepare a powder. Total polyphenols, total flavonoids, total sugars, and reducing sugars were measured by analyzing the components of various prepared insect extracts. The total polyphenol content was 50 μl of Folin-ciocalteau, 100 μl of Na ₂ CO ₃ saturated solution in 400 μl of the extracted sample solution, and allowed to stand at room temperature for 1 hour, and absorbance was measured at 725 nm. As a standard reagent, tannic acid was used. The total flavonoid content was extracted by stirring each sample with methanol for 18 hours, adding 4 ml of 90% diethylene glycol to 400 μl of the filtered extraction sample, adding 40 μl of 1N NaOH again, reacting at 37 ° C. for 1 hour, and measuring absorbance at 420 nm. Rutin was used as a standard reagent. The reducing sugar was quantified using the DNS method and the total sugar was phenol-sulfuric acid method.

[Table 2] Extraction yield and component analysis of extracts from ethanol extracts of king crickets, twin-star crickets, brown beetles, slugs, and beetle larvae

As a result, as shown in Table 2, the extraction efficiency of the king cricket was 1.53 times higher than that of the twin-star cricket, and 0.74 times lower than that of the brown legs. This is believed to correlate with the fat-soluble crude fat content in the insect powder. On the other hand, longevity scarab larvae and slugs showed extraction efficiency of 24.9 ~ 28.9%. In the case of the analysis of the total polyphenol content of the insect extract, the king cricket and twin-star cricket extracts were 15.5-15.6 mg / g, which was significantly higher than that of the brown larvae 2.6 mg / g, and the total flavonoid content was 47 times that of the cricket larvae. It was high. In the case of total polyphenols and total flavonoids, the longevity scarab larvae showed 18.9 and 4.6 mg / g, showing a high content. On the other hand, the total sugar content and reducing sugar content were similar to those of the king and two-star crickets, about 7.5 to 8.5 mg / g. On the other hand, slugs showed high total sugar and reduced sugar content. These results indicate that the Wang Cricket and Ssangbyong Cricket extracts exhibit very similar component contents.

Example 3: Evaluation of antimicrobial activity of the king cricket extract

The antibacterial activity of the king cricket, twin-star cricket and control insect extract prepared in Example 2 was evaluated, and the results are shown in Table 3. Rating bacteria bacteria are gram Escherichia coli KCTC 1682, Proteus vulgaris KCTC 2433, Pseudomonas aeruginosa KACC 10186, as were used for Salmonella typhimurium KCTC 1926, Gram positive bacteria are Staphylococcus aureus KCTC 1916, Staphylococcus epidermidis ATCC 12228, Listeria monocytogenes KACC 10550, Bacillus subtilis KCTC 1924 was used. Meanwhile, for evaluation of antifungal activity, Saccharomyces cerevisiae IF0 0233 and Candida albicans KCTC 1940, a causative agent of candidiasis fungal infection, were used.

First, in the case of antibacterial activity evaluation, Nutrient broth (Difco Co., USA) was inoculated with each bacterium and cultured at 37 ° C. for 24 hours, and then each strain was adjusted to OD ₆₀₀ 0.1 for Nutrient agar (Difco Co. , USA) 100 μl was plated on a sterile petri dish (90 × 15 mm, green cross, Korea) containing medium, and 5 μl of each sample was added to sterile disc-paper (diameter 6.5 mm, Whatsman No. 2) at 37 ° C. Incubation was carried out for 24 hours, and in the case of fungi, after incubation for 24 hours at 30 ° C 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 miconazole (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 three or more evaluations.

[Table 3] Antibacterial activity of ethanol extract of king cricket, double-star cricket, brown pelar, slug, and longevity beetle larvae

As shown in Table 3, in the case of ampicillin (1 μg / disc), which was first used as an antibacterial active control, it exhibited a growth inhibition ring of 8.0 to 28.0 mm for Gram-positive bacteria and Gram-negative bacteria, thereby providing broad and powerful antibacterial bacteria. The activity was confirmed, and the antifungal agent myconazole (1 μg / disc) showed growth inhibition rings of 20 to 26 mm for C. albicans and S. cerevisiae , respectively. On the other hand, the antibacterial activity was not shown at all up to 500 μg / disc concentration in the five insect extracts. To date, no antibacterial activity of these edible insects has been reported.

Example 4: Preparation of sequential organic solvent fractions of ethanol extracts of king crickets, twin-star crickets, jerky, and beetle larvae and their component analysis

Sequential organic solvent fractions of hexene, butanol, and butanol were targeted for the extracts of king crickets, twin-star crickets, jerks, and beetle larvae prepared in Example 2, and finally water residues were respectively recovered. At this time, the extraction efficiency, the organic solvent fraction efficiency, extract / fraction analysis results are shown in Table 4 to Table 8.

[Table 4] Preparation of sequential organic solvent fractions of ethanol extract of Wangguiturami and analysis of components of fractions

As shown in Table 4, 86.3% of the ethanol extract of the king cricket was transferred to the hexene fraction, and it was found that most of the extract is a lipid component. 3.4%. As a result of measuring the total polyphenol and total flavonoid content of the fractions, the highest 27.2mg / g and 3.4mg / g were found in the butanol fraction, and the total sugar and reducing sugar contents were the highest 215.1mg / g and 207.8mg / g in the butanol fraction. Shown.

[Table 5] Preparation of sequential organic solvent fractions of ethanol extract of Ssangyeol cricket and analysis of the components of the fractions

As shown in Table 5, 75.8% of the ethanol extract of the twin-brick cricket was transferred to the hexene fraction, and it was found that most of the extract was a lipid component, water residue in the extract was 16.4%, butanol fraction was 2.7%, and ethyl acetate fraction Accounted for 1.6%. As a result of measuring the total polyphenol and total flavonoid contents of the fractions, the highest was 48.5mg / g and 2.5mg / g in the butanol fraction, and the total sugar and reducing sugar contents were also the highest in the butanol fraction, 21.4mg / g and 19.7mg / g. Shown. Although the extracts of the king cricket and the paired cricket showed a similar composition and component content, it was found that there was a significant difference in the constituent components as a result of fractionation.

[Table 6] Preparation of sequential organic solvent fractions of brown ethanol extract and component analysis of fractions

As shown in Table 6, it was found that 84.4% of the brownish ethanol extract was transferred to the hexene fraction, so that most of the extract was a lipid component, water residue of the extract was 6.5%, ethyl acetate fraction was 4.9%, butanol fraction Silver occupies 2.8%, and it is confirmed that the brown meal contains a significant amount of lipid components. As a result of measuring the total polyphenol and total flavonoid content of the fractions, the butanol fraction showed the highest 29.4mg / g and 2.6mg / g, and the total sugar and reducing sugar content was the highest in the ethyl acetate fraction, 42.1mg / g and 40.9mg / g. It was shown.

[Table 7] Preparation of sequential organic solvent fractions of ethanol extract of beetle larvae and component analysis of fractions

As shown in Table 7, 30.4% of the longevity beetle larvae ethanol extract is 30.4% of the hexene fraction, 28.1% of the ethyl acetate fraction, 20% of the butanol fraction, and 16% of the water residues, so the extract contains substances of various polarity It was confirmed that the fraction yield for each solvent showed a large difference from the existing crickets and brown pebbles. As a result of measuring the total polyphenol content of the fraction, a high content of 6.6 mg / g was found in the butanol fraction and water residue, and a total flavonoid content measurement result showed the highest content of 1.8 mg / g in the butanol fraction. The total sugar and reducing sugar contents were the highest in the butanol fraction, 22.9 mg / g and 2.7 mg / g.

Example 5: Evaluation of antimicrobial activity of sequential organic solvent fractions of king cricket extract

Antibacterial activity was measured in the same manner as in the antibacterial and antifungal activity evaluation of Example 3, targeting the fractions of the king cricket extract and various insect extracts prepared in Example 4. As a result, it is shown in Tables 8-11.

[Table 8] Antibacterial activity evaluation of sequential organic solvent fractions of ethanol extracts

[Table 9] Evaluation of antibacterial activity of sequential organic solvent fractions of ethanol extracts from Ssangyeol cricket

[Table 10] Antibacterial activity evaluation of sequential organic solvent fractions of brown ethanol extract

[Table 11] Antibacterial activity evaluation of sequential organic solvent fractions of ethanol extract of Jang beetle larvae

As shown in Table 8 to Table 11, it was confirmed that the hexene fraction and butanol fraction of the king cricket extract showed a wide range of strong antibacterial activity against the Gram-positive bacteria and the Gram-negative bacteria Proteus vulgaris . The fraction of the extract showed very weak antibacterial activity and showed selective antibacterial activity only for specific bacteria. The above results suggest that the hexene fraction and butanol fraction of the king cricket extract can be effectively used to control food poisoning-causing bacteria and pathogenic bacteria.

Example 6: Evaluation of acid and thermal stability of the hexene fraction and butanol fraction of the king cricket extract

The hexene fraction and butanol fraction of the ethanol extract of the king cricket obtained in Example 4 were checked for acid and thermal stability against antibacterial activity. The active fraction maintained excellent activity without serious loss of antibacterial activity even after 1 hour heat treatment at 100 ° C and 1 hour treatment at pH 2 (0.01M HCl). Therefore, it is expected that the hexene fraction and butanol fraction of the ethanol extract of the king cricket will maintain excellent antibacterial activity even under various food, cosmetic, and pharmaceutical processing processes.

Claims (4)

An antimicrobial composition containing hexene fraction or butanol fraction of a king cricket ( Teleogryllus emma ) extract as an active ingredient. The antibacterial composition according to claim 1, wherein the king cricket extract is an ethanol extract of king cricket. The antimicrobial composition of claim 1, wherein the antimicrobial composition has antibacterial activity against Gram-positive bacteria. The antimicrobial composition according to any one of claims 1 to 3, wherein the antimicrobial composition is selected from the group consisting of pharmaceutical compositions, dietary supplements, cosmetic compositions, animal feed compositions, food additives, cosmetic additives, animal feed additives, detergents and cleaners. Antibacterial composition, characterized in that.

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