KR102428964B1 - Antimicrobial composition comprising the extracts of hermetia illucens - Google Patents

Abstract

The present invention relates to an antibacterial composition comprising an extract of Hermetia illucens , black soldier fly, and more specifically, a pupa extract after the larva in various growth stages of Dongae, etc. It relates to an antibacterial composition. As an active ingredient of the broad-spectrum antibacterial composition of the present invention, Dongae extract is an antibacterial pharmaceutical composition, health functional food, cosmetic composition, animal feed composition, food, cosmetic or additive for animal feed, as demonstrated through the examples of the present specification. , it has an excellent effect that it can be used for various cleaning agents and various cleaning agents. In addition, the extract of Dongae, etc. of the present invention has excellent thermal stability and no loss of activity even under acidic conditions of pH 2 and plasma, so it can be easily processed into various forms such as liquid, cream, powder, pill, tablet, etc. It can be very usefully used in the pharmaceutical industry, the food industry, the livestock industry, the health industry and the cosmetic industry.

Description

Antibacterial composition comprising extract from Dongae, etc. {ANTIMICROBIAL COMPOSITION COMPRISING THE EXTRACTS OF HERMETIA ILLUCENS}

The present invention relates to an antibacterial composition comprising an extract of Hermetia illucens , black soldier fly, and more specifically, a pupa extract after the larva in various growth stages of Dongae, etc. It relates to an antibacterial composition.

Humans have used various antibacterial agents and preservatives to defend against the invasion of pathogenic microorganisms and to control food spoilage microorganisms. However, in the case of chemical antibacterial agents and preservatives, they show excellent antibacterial activity against various bacteria and fungi, but when used for a long time, many side effects such as redness due to allergy, angioedema, bone marrow suppression, anemia, gastritis, and degenerative brain disease appear. do. In fact, the use of antibiotics with severe side effects, such as chloramphenicol, has been banned in the United States. Therefore, various studies on the development of natural antibacterial agents using natural products that have been used for a long time while ensuring safety are in progress.

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

On the other hand, the black locust (American ferret, Hermetia illucens , Black Soldeir Fly) is an insect in the family Diptera, 13 to 20 mm in length, and the overall color is black. The face is blackish-brown, very short, and the forehead is small and distinctly raised, and it is yellowish. Antennae are blackish brown, shoulder bumps and lateral margins are yellowish brown, and wings are black. The small shield plate is short and the hind margin is obtuse, the wings are large and cloudy, the legs are long and slender, and the base is black, and the 2 basal parts of the middle tarsus and the basal part of the forelimbs are yellowish red. Dongae is mainly distributed in Korea, Japan, and China, and inhabits manure piles and toilets. Adults are slow-moving, so they can be easily spotted, and they do not fly well compared to flies.

Dongae, etc. are recognized worldwide as insects that do not harm humans, and they are omnivorous and are bred for environmental purification and food waste removal. In particular, in the case of larvae, larvae are rapidly increasing in recent years due to their strong digestive ability and rapid growth, and the larvae are widely used for protein feed or for compost production. In general, one larva decomposes 2-3 g of food waste, and attention is being focused around the world for environmental purification. Larvae of Dongae, etc. are known to contain 42% crude protein content, 35% fat, 7% crude fiber, and other ash and water, so they are promising as protein feed. (as of 2019), and is priced between $8 and $40 per kg in the United States. In addition, after 2 to 3 days after mixing the larvae and food waste, it can be used as fecal soil, and in Korea, it is sold at 8,500 won per 20 kg as fecal soil fertilizer.

The larval period of Dongae, etc. varies depending on the breeding environment, but usually takes 15 to 20 days, and it changes to white-off-white-black-brown as it grows. Usually, it takes 2 to 3 days per instar from 1st to 6th instar, then turns into a pupa through 6th instar and emerges as an adult. Adults on the larvae do not feed and mate while flying during the survival period of about 10 days. After that, the female lays about 1,000 very small long oval eggs (average size of 1mm long and 0.2mm short). The total generation period is known to be about 35 to 41 days.

As for the research related to the larvae, a study on the ecology and life patterns of the larvae [Ecological Characteristics of Hermetia illucens in Korea, Jonggil Kim et al., 2008. Journal of the Korean Society of Applied Entomology 47: 337-343; Ecology and larval-derived physiologically active substance properties of Hermetia illucens , Kwan-Ho Park. 2014. Kyungpook National University Ph.D. thesis; Effects of Stress Sound Waves on the Growth of the American Dongae, Jiyoung Park et al., 2013. Journal of the Korean Society for Applied Entomology 52: 227-237], A study on the historical value and practical application of the Chinese dragonfly [ Litopenaeus vannamei ) Effects on the growth and water quality of larvae, Tae-Hoon Kim et al., 2019. Journal of the Korean Society for Environmental Sciences 28: 1-6; Changes in broiler productivity when Hermetia illucens powder is added to feed, Inhak Choi et al., 2018. Journal of Environmental Science 27: 1299-1303; Effect of feeding to broiler birds on the productivity of broilers, Choi, Young-Chul et al., 2013, Journal of the Korean Society of Entomology 51: 30-35; Nutritional value of Dongae as a fish feed, Kwanho Park et al., 2013. Journal of the Korean Society of Entomology 51: 95-98; Effects of adding Hermetia illucens powder to duck feed on duck litter characteristics and livestock environment, 2019. Jeong Tae-ho. Korean Journal of Environmental Science 28: 713-717; Gariglio M et al., 2019. J Anim Sci Biotechnol. doi: 10.1186/s40104-019-0344-7] and a study on fecal soil (Effect of mixed topsoil with fecal soil on the growth and rhizosphere of Korean grass, Lee Sang-beom et al., 2013. Weed & Turfgrass Science 2: 298 -305) is known.

In addition, some studies on the physiological activity of Dongae, etc. have been conducted. Antioxidant activity (Antioxidant activity of American Dongae extract, Park Gwan-ho et al., 2014. Journal of the Korean Society of Entomology 52: 142-146), antibacterial activity (immunity caused by bacteria) Antibacterial activity on induced Dongae et al., Park Gwan-ho et al., 2016. Journal of Life Science 26: 1409-1414; Li Z et al., 2017. Sci Rep. doi: 10.1038/s41598-017-10839-4), Immune enhancing activity in broilers (Lee J. et al., 2018. J Vet Med Sci. 80: 736-740) is reported. In addition, intestinal amylopullulanase has been confirmed (Lee YS et al., 2016. Int J Biol Macromol. 82: 514-521), and the isolation and characterization of intestinal bacteria of the genus Amyelia has been conducted (Yunyeop Lee, 2013, Master's thesis at Kyunghee University). ; Kim Eun-seong et al., 2014. Journal of the Korean Society of Applied Insects, 53: 15-26). However, although antibacterial activity against larvae of Dongae, etc. is known so far, there have been no reports of strong antibacterial activity of extracts of Dongae pupa, dragongak (pupa molting) and adults.

As for the patents related to Dongae, etc., Korean Patent Registration No. 10-1815115 for a spawning ground for Dongae, etc., No. 10-1076686, a feed using larvae for Dongae, etc., No. 10-1759518, for a feed composition for Dongae, etc., and No. 10-2026974 for Dongae, etc. Organic waste treatment apparatus using larvae, antibacterial composition containing larvae extract or its fractions in No. 10-1428157, antibacterial peptide isolated from larvae in Dongae, etc. in No. 10-1726951, Dongae in No. 10-1465096 A composition for controlling plant pathogens comprising a larval extract on the back, an antibacterial composition comprising a peptide fraction isolated from the larvae of the larvae of the larvae of the larvae of the worms immunized in No. 10-1796766, and the larvae of the larvae of the American worms in No. 10-1504879 ( Hermetia illucens ) in the intestine A new useful strain derived from Bacillus genus and its use and No. 10-1491770 discloses a novel mannan degrading gene EM17 with high stability under an organic solvent and a recombinant mannan degrading enzyme produced from its transformant strain, Republic of Korea Patent Publication No. 10 -2017-0068752, egg-laying induction device, No. 10-2017-0111997, No. 10-2017-0111997, protein composition for treatment or prevention of gonorrhea or diseases caused by gonorrhea containing larvae-derived protein or protein fraction in No. 10-2019, No. 10-2019 -0024389 antibacterial composition containing extract from Dongae, etc., and skin care products containing extract from American Dongae, etc. in No. 10-2018-0081554. However, antibacterial patents of larvae in Dongae, etc. have been reported so far, but no patents are known about the strong broad-spectrum antibacterial activity of extracts of pupa, dragon horn and imago of Dongae, etc.

US 10-1428157 B US 10-2019-0024389 A US 10-2017-0111997 A

Park Gwan-ho et al., 2016. Journal of Life Sciences 26: 1409-1414. Antibacterial activity against dongae induced by bacteria Li Z et al., 2017. Sci Rep. doi: 10.1038/s41598-017-10839-4 Alvarez D. et al. 2019. J Insect Sci. 2019 Nov 1;19(6). pii: 17. doi: 10.1093/jisesa/iez120. Prospecting Peptides Isolated From Black Soldier Fly (Diptera: Stratiomyidae) With Antimicrobial Activity Against Helicobacter pylori (Campylobacterales: Helicobacteraceae).

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

In order to solve the above problems, the present invention provides an antibacterial composition comprising an extract of Dongae, etc.

It is preferable that the Dongae etc. extract is an ethanol extract of the pupa of Dongae etc., an ethanol extract of a dragonfly or an ethanol extract of an adult larvae.

It is preferable that the ethanol extract of Dongae et al. Yonggak is a hexene fraction, an ethyl acetate fraction, or a butanol fraction of the ethanol extract of Dongae et al.

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.

As an active ingredient of the broad-spectrum antibacterial composition of the present invention, Dongae extract as an active ingredient, as demonstrated through the examples of the present specification, is an antibacterial pharmaceutical composition, a health functional food, a cosmetic composition, an animal feed composition, food, an additive for cosmetics or animal feed. , it has an excellent effect that it can be used for various cleaning agents and various cleaning agents. In addition, the extract of Dongae, etc. of the present invention has excellent thermal stability and no loss of activity even under acidic conditions of pH 2 and plasma, so it can be easily processed into various forms such as liquid, cream, powder, pill, tablet, etc. It can be very usefully used in the pharmaceutical industry, the food industry, the livestock industry, the health industry and the cosmetic industry.

1 is a photograph of larvae, pupae, dragonflies and adults of Dongae, etc. used in the experiment.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention, in order to test the antibacterial and antifungal efficacy against Dongae, etc., after recovering larvae, pupae, dragonflies and adults by various growth periods, prepare an ethanol extract and evaluate the antibacterial activity of pupae, dragonflies and The adult extract was recovered as an antibacterial active ingredient, and in particular, the hexene fraction, ethyl acetate fraction and butanol fraction of the dragon horn extract showing the strongest antibacterial activity were recovered as broad-spectrum antibacterial active ingredients, and the hexene fraction, ethyl acetate fraction and butanol fraction were treated with heat. By confirming that it has excellent stability and acid stability, it was attempted to utilize extracts and fractions from Dongae, etc. as an antibacterial composition.

In particular, the pupa, dragon horn and adult extracts from the Dongae, and the hexene fraction, ethyl acetate fraction and butanol fraction of the dragon horn extract showed strong antibacterial activity against gram-positive and gram-negative bacteria, and developed as a broad-spectrum antibacterial agent for various bacterial infectious diseases, etc. The present invention was completed by confirming that it can be.

Specifically, the present inventors prepare an ethanol extract for larvae, pupae, dragonflies and adults of various growth stages in order to develop an antibacterial composition using Dongae, etc., which are known to have antioxidant and immune enhancing effects, and target these extracts. As a result of evaluating the antibacterial activity against various bacteria and fungi, strong antibacterial activity not previously reported was confirmed in the ethanol extracts of pupa, dragon dragon and adult larvae, especially in the hexene, ethyl acetate and butanol fractions of the dragon dragon fruit extract from Dongae, etc. By showing strong antibacterial and antifungal activity, it was confirmed that the extracts and fractions can be used as broad-spectrum antibacterial agents.

Accordingly, the present invention provides an antibacterial composition comprising an extract of Dongae, etc.

It is preferable that the Dongae etc. extract is an ethanol extract of the pupa of Dongae etc., an ethanol extract of a dragonfly or an ethanol extract of an adult larvae.

It is preferable that the ethanol extract of Dongae et al. Yonggak is a hexene fraction, an ethyl acetate fraction, or a butanol fraction of the ethanol extract of Dongae et al.

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 experiments of the pupa, dragon horn and imago extracts and antibacterial active fractions of Dongae, etc. of the present invention will be described in more detail.

The present invention comprises the steps of recovering larvae, pupae, dragonflies and adults in various stages of growth and preparing an extract thereof; Evaluating the antibacterial activity of the extract; a step of preparing sequential organic solvent fractions of hexene, ethyl acetate, and butanol from the lysate extract and preparing the water residue obtained thereafter; Evaluating the antimicrobial activity of the fraction; and a step of examining the stability of the active extracts and fractions.

"Dongae extract" contained in the composition of the present invention is obtained by extracting the pupae, dragon larvae and adults of Dongae etc. with an organic solvent, and filtering the extract using a filtration net of 0.06 mm or less, and concentrating it under reduced pressure. can be

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. hot water, or 95% ethanol extraction is most preferred.

As a preferred embodiment, the extract of Dongae and the like of the present invention can be extracted with hot water or ethanol from the pupae, dragonflies and adults of Dongae, etc. In addition, the hot water or ethanol extract may be fractionated sequentially or separately with an organic solvent of hexene, ethyl acetate and butanol to additionally obtain a hexene fraction, an ethyl acetate fraction, a butanol fraction and a water residue.

The hexene fraction, ethyl acetate fraction and butanol fraction of Dongae pupa, dragon horn and imago extract and dragon horn extract of the present invention can be prepared into powder through a conventional powdering process such as drying under reduced pressure, freeze-drying, or spray-drying. The active extracts and fractions are not degraded by various degrading enzymes in plasma, and maintain antibacterial activity even at a heat treatment temperature of 100° C. and a pH in the human stomach of pH 2.

The extracts and fractions of Dongae, etc. of the present invention exhibit strong antibacterial activity, and are related to the prevention or treatment of pathogenic bacterial infections, pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food, cosmetics, additives for animal feed, and various cleaning agents. can be used as a material for

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

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

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, activity of the drug, and the type of disease in the patient; Sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including concomitant drugs, 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 may be administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a 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 age, sex, and weight of the patient, and generally 1 to 5,000 mg per body weight, preferably 100 to 3,000 mg per body weight, is administered daily or every other day Or it can be administered in divided doses 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, disease severity, sex, weight, age, etc., the dosage is not intended to limit 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. Any mode of administration can be envisaged, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intra-cerebroventricular injection. In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the administration route of the pharmaceutical composition of the present invention is oral or parenteral through all general routes as long as it can reach the target tissue. It can be administered orally. In addition, the composition of the present invention may be administered using any device capable of delivering an active ingredient to a target cell. In the present invention, "subject" is not particularly limited, but includes, for example, humans, monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits or guinea pigs. and preferably a mammal, more preferably a human.

In a preferred embodiment, the antibacterial composition of the present invention can be applied as a dietary supplement.

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 with a high risk of infection by pathogenic microorganisms. Foods containing the active ingredient having excellent antibacterial activity of the present invention include, for example, various foods, beverages, gum, tea, vitamin complexes, health supplements, and the like, in powder, granule, tablet, capsule or beverage form. can be used as In general, the active ingredient of the present invention may be added in an amount of 0.01 to 15% by weight of the total food weight, and the health beverage composition may be added in a proportion of 0.02 to 10g, preferably 0.3 to 1g, based on 100 ml. In addition to containing the above compound as an essential ingredient in the proportion indicated, the health functional food of the present invention may contain food pharmaceutically acceptable food supplement additives, such as natural carbohydrates and various flavoring agents, as additional ingredients. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and common sugars such as polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, thaumatin, rebaudioside A, glycyrrhizin, saccharin, aspartame, and the like may be used. The ratio of the flavoring agent is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention includes various nutrients, vitamins, minerals, flavoring agents such as synthetic and natural flavoring agents, colorants and thickeners, 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 natural fruit juice, fruit juice for the production of fruit juice drinks, vegetable drinks, and the like. These components may be used independently or in combination. The proportion of these additives is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the active fraction of the present invention.

In a preferred embodiment, the antimicrobial 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, for example, essence, nourishing cream, body lotion, conditioner, shampoo, body cleanser, lotion, skin, mask pack, etc. It is not limited here. Preparation in the above form can be easily carried out according to various manufacturing processes known to those skilled in the art.

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

The animal is a group of organisms corresponding to plants, mainly ingesting organic matter as nutrients, and has differentiated digestive, embryonic and respiratory organs, specifically echinoderms, crustaceans, molluscs, fish, amphibians, reptiles, birds, 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 snapper, sea bream, cod, flounder , fish such as halibut, birds including poultry such as pheasants or chickens, or mammals such as pigs, cattle, and goats.

The composition for animal feed may further include grains, vegetable protein feed, animal protein feed, sugar or dairy products to the antimicrobial composition comprising the antimicrobial component of the present invention. The grain may specifically be pulverized or crushed wheat, oats, barley, corn and rice, and the vegetable protein feed may specifically include rape, 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 component composed of various kinds of powdered milk and whey powder.

The composition for animal feed may additionally contain ingredients such as nutritional supplements, digestion and absorption enhancers, vocal tract promoters or disease prevention 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, for example, 0.1 g to 100 g of the antimicrobial composition may be included based on 1 kg of the final feed.

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

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

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

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

In addition, the amount of the antimicrobial composition used among the additives may be appropriately used depending on the purpose of use, application form, application site, application target article, etc., for example, it may include 0.01 parts by weight to 50 parts by weight based on the total composition, but 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 as long as the application purpose can be achieved due to the characteristics of the product.

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

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

The detergent may further include an additive selected appropriately according to the purpose and purpose of use, and may be used by mixing with other active ingredients such as a commonly used detergent or additives such as a pigment, surfactant, and preservative. The detergent may be manufactured by powdering, granulating, tableting or liquefying according to the purpose and use thereof, 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, application target article, etc., and may include, for example, 0.01 parts by weight to 50 parts by weight based on the total composition, but The content of the 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 the application purpose can be achieved due to the characteristics of the product.

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

The cleanser may be used as a hand, oral or feminine cleanser in a simple way in order to prevent infection in the body of pathogenic microorganisms that are 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 cleanser.

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

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, etc., for example, it may include 0.01 parts by weight to 50 parts by weight based on the total composition, but the content of the antimicrobial composition is The present invention 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 as long as the application purpose can be achieved due to the characteristics of the product.

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

[Example]

Example 1: Breeding and preparation of larvae, pupae, dragonflies, and adult powders in Dongae, etc.

Dongae, etc. are omnivorous, so breeding is easy, and a mass breeding system has already been established. In Korea, the Rural Development Administration spreads the breeding method to Dongae, etc. and breeds them in many farms, and the larvae and pupae are widely used as feed for various birds, fish, crustaceans, and reptiles. In this example, a feed mixed with sawdust and food is sufficiently supplied in a breeding box at 25° C. for breeding Dongae, etc., and it was bred for more than 1 month as adults. Each of the larvae, pupae, sauropods and adults is shown in FIG. 1, and the size and weight are shown in Table 1. The larvae and adults were collected from the larvae of each growth period, and then dried in a microwave oven for 8 minutes, and then each sample was prepared as a powder using a mortar.

[Table 1] Size and weight of larvae, pupae, dragonfish, and adults

Example 2: Preparation of extracts from Dongae, etc. and analysis of components of the extract

An ethanol extract was prepared from the powder of Dongae etc. prepared in Example 1. At this time, 10 times of ethanol was added to the samples recovered at each growth period, extracted twice at room temperature, the extract was collected, filtered, and concentrated under reduced pressure to prepare a powder. The content of total polyphenols and total flavonoids was measured by analyzing the components of the prepared Dongae extract. For the total polyphenol content, 50 μl of Folin-ciocalteau and 100 μl of Na ₂ CO ₃ saturated solution were added to 400 μl of the extraction sample, left at room temperature for 1 hour, and absorbance was measured at 725 nm. As a standard reagent, tannic acid was used. For the total flavonoid content, each sample was extracted with methanol for 18 hours, and 4 ml of 90% diethylene glycol was added to 400 μl of the filtered extraction sample, 40 μl of 1N NaOH was added again, and the absorbance was measured at 420 nm after reaction at 37° C. for 1 hour. As a standard reagent, rutin was used.

[Table 2] Extraction yield and component analysis of ethanol extracts of larvae, pupae, dragonflies, and adults of Dongae, etc.

As a result, as shown in Table 2, the extraction efficiency of Dongae, etc. increased from 8.2% at the 2nd instar to 13.1% at the 4th instar, which gradually increased as the number of years increased, and then decreased to 1.7% at the 6th instar. appeared low. However, in the case of adults, 18.5% showed the highest extraction yield.

On the other hand, as a result of analysis of the total polyphenol content, it decreased as the number of years increased at 1.8 mg/g at 2 instars, and was hardly detected in the case of 6 years of age. In the case of total flavonoid content, it rather increased as the age increased, and in the case of pupae, it showed the highest 3.4mg/g, and decreased to 2.8mg/g in the sauropod. As a result of these results, it is expected that the extracts of pupa of Dongae et al.

Example 3: Evaluation of antibacterial activity of extracts of larvae, pupae, dragons, and adults such as Dongae

The antibacterial activity of larvae, pupae, dragonflies, and adult extracts of Dongae, etc. prepared in Example 1 was evaluated, and the results are shown in Table 3. As strains used for the evaluation of antimicrobial activity, Escherichia coli KCTC 1682, Proteus vulgaris KCTC 2433, Pseudomonas aeruginosa KACC 10186, Salmonella typhimurium KCTC 1926 were used as Gram-negative bacteria, and Staphylococcus aureus KCTC 1916, Staphylococcus epidermidis ATCC 12228, Staphylococcus epidermidis were used as Gram-positive bacteria. Listeria monocytogenes KACC 10550 and Bacillus subtilis KCTC 1924 were used. On the other hand, for the evaluation of antifungal activity, Saccharomyces cerevisiae IF0 0233 and Candida albicans KCTC 1940, a causative agent of Candida fungal infection, were used.

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

[Table 3] Antibacterial activity of extracts of larvae, pupae, dragonflies, and adults in Dongae, etc.

As a result, as shown in Table 3, ampicillin, which is used as an antibacterial agent in clinical practice, showed a wide range of antibacterial activity against gram-positive and gram-negative bacteria even at 1 μg/disc, and a relatively weak growth inhibitory agent against Pseudomonas aeruginosa and Escherichia coli . (12 mm) was shown. Miconazole, used as an antifungal agent in clinical practice, strongly inhibited the growth of Saccharomyces cerevisiae and Candida albicans even at 1 μg/disc. On the other hand, as a result of evaluation of the ethanol extracts of larvae, pupae, dragons, and adults of Dongae, etc., antibacterial activity did not appear at all in the extracts of 2nd and 3rd instars, and in the extracts of 4th and 6th instars, pupae, dragons and adults at 500μg/disc concentration. It showed excellent antibacterial activity against Gram-positive bacteria. In particular, the extracts of pupae, dragonflies and adults were recognized for antibacterial activity against gram-positive bacteria as well as gram-negative bacteria Escherichia coli and Proteus vulgaris . Therefore, it was confirmed that the extracts of pupa, dragon horn and imago of Dongae, etc. can be used for the control of various bacteria.

Example 4: Sequential preparation of organic solvent fractions of yonggak extract from Dongae et al. and analysis of their useful components

A hexene fraction, an ethyl acetate fraction, a butanol fraction, and a water residue after the fractionation were prepared through sequential organic solvent fractionation for the lychee extract prepared in Example 1. The yield of the prepared fraction and the analysis result of its useful components are shown in Table 4.

[Table 4] Fractionation efficiency and useful component analysis of sequential organic solvent fractions of Dongae et al.

As shown in Table 4, in the case of the fractions of the extract from Dongae et al., the highest fractionation efficiency was shown in the hexene fraction and corresponded to 60% of the ethanol extract. Next, the water residue was 15.9%, the butanol fraction and the ethyl acetate fraction accounted for 10.0 to 10.6% of the lysing ethanol extract.

As a result of measurement of the total polyphenol content of the fractions of the extract from Dongae et al., the ethyl acetate fraction showed the highest content, and hardly contained water residues and hexene fractions. In addition, in the case of the total flavonoid content, the ethyl acetate fraction (5.8 mg/g) showed the highest content, and the butanol fraction showed a content of 4.7 mg/g. Although it is common that total sugar and reducing sugar, which are generally water-soluble, show the highest content in the water residue, the soluble fraction showed the highest total sugar and reducing sugar content in the ethyl acetate fraction. These results mean that most of the hexene fraction of the dragonfish contains fat-soluble lipid components, and the ethyl acetate fraction contains most insect-chemicals.

Example 5: Evaluation of the antibacterial activity of sequential organic solvent fractions of the extract of yonggak extract from Dongae et al.

The antibacterial and antifungal activities of the sequential organic solvent fractions of the Yonggak extract prepared in Example 4 were evaluated in the same manner as in Example 3. The results are shown in Table 5.

[Table 5] Antibacterial activity of organic solvent fractions of yonggak extract from Dongae et al.

As a result, as shown in Table 5, the water residue in the fractions of the lysate extract did not show antibacterial activity, and the hexene fraction, the ethyl acetate fraction, and the butanol fraction showed excellent antibacterial activity against Gram-positive bacteria. In particular, the hexene fraction showed broad antibacterial and antifungal activity against all bacteria and fungi. In addition, antifungal activity, which was not confirmed in the ethanol extract, was confirmed in the hexene fraction and the butanol fraction. These results indicate that the extract of rhubarb contains various antibacterial and antifungal active substances, and in particular, the hexene fraction can be developed as a broad-spectrum antibacterial agent that simultaneously exhibits antibacterial and antifungal activity. Therefore, the above results suggest that, when preparing the extract and active fraction of dragon fruit from Dongae, etc., which are being completely discarded, it can be effectively used for controlling food poisoning-causing bacteria and pathogenic bacteria.

Example 6: Plasma, acid and thermal stability evaluation of the active fractions of pupa, dragon horn and imago extracts of Dongae and the like, and dragon horn extracts

Plasma stability, thermal stability, and acid stability with respect to antibacterial activity were confirmed for the active fractions of the pupa, dragon horn and imago extracts and lystagmus extracts obtained in Examples 1 and 4 above. As a result, the active extract and active fraction showed high stability because no significant loss of antimicrobial activity was observed even after 1 hour heat treatment at 100° C., 1 hour treatment at pH 2 (0.01 M HCl), and 1 hour treatment in plasma. It was. Therefore, it is expected that the pupa, dragon horn and adult extracts of Dongae, and the hexene fraction, ethyl acetate fraction, and butanol fraction of the dragon horn extract will maintain excellent antibacterial activity even under various food processing processes.

Claims (4)

An antifungal composition comprising a hexene fraction or a butanol fraction of an ethanol extract from dragon fruit such as Dongae. delete delete According to claim 1, wherein the antifungal composition is selected from the group consisting of pharmaceutical compositions for the prevention or treatment of fungal infections, health functional foods, cosmetic compositions, animal feed compositions, food additives, cosmetic additives, additives for animal feed, detergents and cleaners. Antifungal composition, characterized in that.

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