KR20210009480A - Antimicrobial composition comprising the extract of immature fruit extract of magnolia denudata - Google Patents

Abstract

The present invention relates to an antibacterial composition including an immature fruit extract of yulan (Magnolia denudata) as an active ingredient, and more particularly, to a broad-spectrum antibacterial composition including an ethanol extract of an immature fruit of yulan immediately after a yulan seed is produced as an active ingredient. As demonstrated through an embodiment of the present disclosure, the immature fruit extract of the yulan as the active ingredient of the broad-spectrum antibacterial composition according to the present invention is used for an antibacterial pharmaceutical composition, a health functional food, a cosmetic material composition, an animal feed composition, an additive for a food, a cosmetic product, or an animal feed, various detergents, various cleaning agents, and the like. In addition, since the immature fruit extract of yulan according to the present invention has excellent thermal stability and no loss of activity even under acidic conditions of pH 2 and within plasma, the immature fruit extract is easily processed into various forms such as a liquid, cream, powder, a pill, and a tablet, so that the immature fruit extract is very usefully used in a pharmaceutical industry, a food industry, a livestock industry, a health industry, and a cosmetic industry.

Description

Antibacterial composition containing extract of immature white magnolia as an active ingredient {ANTIMICROBIAL COMPOSITION COMPRISING THE EXTRACT OF IMMATURE FRUIT EXTRACT OF MAGNOLIA DENUDATA}

The present invention relates to an antibacterial composition containing an extract of Magnolia denudata immature fruit as an active ingredient, and more particularly, a broad-spectrum antibacterial composition containing an ethanol extract of immature fruit immediately after the seed of white magnolia is produced as an active ingredient It is about.

Humans have used various antibacterial agents and preservatives to prevent invasion of pathogenic microorganisms and control food spoilage microorganisms. However, in the case of chemical antimicrobial agents and preservatives, they exhibit excellent antibacterial activity against various bacteria and fungi, but when used for a long time, many side effects such as redness due to allergy, vascular edema, bone marrow suppression, anemia, gastritis, and degenerative brain disease appear. 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 with long-term use experience while securing safety are in progress.

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

On the other hand, Magnolia denudata is a deciduous tall tree of the magnolia family, reaching 10-15m in height, and its stem bark is grayish-white, with many hairs pressed by young branches and winter snow. Petiole is 1~2cm long, wide obovate, 8~15cm long, 4~11cm wide, there are hairs on both sides, but gradually disappears. Flowers bloom before leaves, bell-shaped, and white. Sepals and petals are not distinguished, and they are narrow obovate, 7-8cm long, 3-4cm wide, and do not spread out. There are many surgeries, and they are arranged in a spiral shape. It is planted for ornamental purposes in Korea, and flower buds are used for medicinal purposes. Magnolias are classified into various types according to flower color and shape. Representatively, white magnolia, purple magnolia, and star magnolia are known, and most of the domestic landscape trees use white magnolia native to China with excellent pollution resistance and cold resistance.

The buds of magnolia are known as shinyi (辛夷) and are used as a very valuable medicinal material in oriental medicine, and shinyi is known for its astringent, capillary dilatation, anti-inflammatory, blood pressure lowering, pain relief, sedation, and antibacterial properties. Magnolias have a large amount of fruit after harvesting the buds and after the flowers have faded. Until now, this fruit has no special purpose and has been discarded.

To date, studies on magnolia trees have been reported on antibacterial and antioxidant activities of various magnolia trees (Sungsook Lee et al., 2010. Wood Engineering 38: 579~586). Most studies on flower buds and leaves have been reported. , Research on white magnolia, which is the largest number of landscape trees in Korea, is insignificant. Studies related to white magnolia include the antioxidant activity, anticancer activity and anti-inflammatory activity of white magnolia flower extract (Jin-Woo No, 2009, Journal of Food and Nutrition Science 38: 1478-1484), and isolation of melanin production inhibitors from Shin-I (辛夷) ( Heo, Gwang-Hwa et al., 2004, Journal of the Korean Society of Herbal Medicine 35: 152~156), Shin-I's Allergy Inhibition Effect (Seungmin Hong, 2013, Master's Thesis at Seoul National University) and anti-obesity effect (Changsuk Kong et al., 2011, J. Agricultural and Food Chemistry 59: 5665~ 5670), Antioxidant and Anticancer Effects of Magnolia Calyx Extract (Park, Hye et al., 2015. Molecules J. Synthetic Chemistry Natural Product Chemistry 20: 12154~12165), Inflammatory cytokines and NO in LPS-activated peritoneal macrophages. Inhibitory effects (Kim Do-yoon et al., 2007. Journal of Dong-Eui Physiological Pathology 21: 916-920), analysis of the components of white magnolia leaves (Du, Jiang et al., 2001, J. Asian Natural Products Res. 3: 313-319) are known. In addition, studies related to the white magnolia fruit include analysis of the lignan component of the white magnolia fruit (Lee Seong-suk, 2010, Wood Engineering 38: 579-586), and the purification of cytotoxic (apoptotic) phenolic compounds from the white magnolia fruit (Noshita Toshiro et al., 2009. Biotechnology Biochemistry 73: 726-728), larval activity of lignan obtained from white magnolia seeds (Wang, Zhang-qian et al., 2016, Pest Management Science 72: 897-906), IL-4 and IL-13 in immune T cells The expression inhibitory activity of (Jin Mirim et al., 2013, In vitro Cellular Developmental Biology. Animal 49: 805-814), etc., have been known, but to date, no report on the potent antibacterial activity of the immature fruit extract of Magnolia denudata has been known. .

Patents related to magnolia trees include Korean Patent Registration No. 10-1397188, [Method of manufacturing magnolia flower tea], No. 10-1079980, [Lignan components isolated from Shinyi and its use], and No. 10-0795512. And Skin Protection Composition Containing Magnolia Extract], No. 10-1563802, [Bola branch made using magnolia flower tea, and its manufacturing method], No. 10-1560367, [New neorignan derived from Hwang magnolia and brain containing the same] Cell protection composition], No. 10-1024043 [(Japan) method of sesamin separation from magnolia] and No. 10-0909572 (separated from Japanese magnolia leaves and bark) [Obobatol and its synthetic derivatives as active ingredients Containing a composition for preventing and treating diabetes and metabolic syndrome diseases] is disclosed, and in Korean Patent Publication No. 10-2009-0025645 [An antiseptic cosmetic composition containing magnolia extract], [Magnolia] in No. 10-2018-0093238 Petal tea and its manufacturing method], No. 10-2015-0065341 [fragrance composition that reproduces the scent of magnolia], No. 10-2012-0000243 [Anti-cancer agent composition containing Japanese magnolia fruit extract], No. 10- 2008-0044612 [C-kit protein activity inhibitory composition containing Japanese magnolia extract], 10-2016-0070986 [Pharmaceutical composition having antioxidant effect containing magnolia flowers fermented by microorganisms] , No. 10-2016-0070972 [Functional food composition having antioxidant effect containing magnolia flowers fermented by microorganisms] and No. 10-2005-0111257 [Magnolia extract for improvement of symptoms of degenerative central nervous system disease] Functional food containing a] is disclosed. However, until now, there is no known patent for antibacterial activity of the immature white magnolia fruit extract.

KR 10-2012-0000243 A

Sung-Suk Lee et al., 2010. Wood Engineering 38: 579-586

The present invention has been conceived 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 immature white magnolia fruit as an active ingredient.

In order to solve the above problems, the present invention provides an antibacterial composition containing an extract of immature fruits of Magnolia denudata as an active ingredient.

The white magnolia immature fruit extract is preferably an ethanol extract of the white magnolia immature fruit immediately after the seed is generated.

It is preferable that the antibacterial is antibacterial and antifungal activity.

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

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

1 shows the fruit of white magnolia. 1: immature fruit before seed generation, 2: immature fruit immediately after seed generation, 3: mature fruit with seed fully formed and fruit open.

Hereinafter, the present invention will be described in detail.

In order to test the antibacterial and antifungal efficacy of the white magnolia fruit, the inventors of the present invention prepare the white magnolia immature fruit and mature fruit extract by a certain method, and the immature fruit extract immediately after the seed is produced among the fruits of various periods is antibacterial. The extract was recovered as an active ingredient, and it was confirmed that the extract exhibited almost no hemolytic activity against human red blood cells, but had excellent thermal stability and acid stability, so that the immature white magnolia fruit extract was used as an antibacterial composition.

In particular, the active extract showed a strong antibacterial activity against not only Gram-positive and Gram-negative bacteria, but also fungi, and it was confirmed that it can be developed as a broad spectrum antibacterial agent against various bacteria and fungal infectious diseases, thereby completing the present invention.

Specifically, in order to develop an antibacterial composition using white magnolia fruit known to have anticancer, anti-inflammatory, and antioxidant effects, the present inventors prepared an ethanol extract for immature white magnolia fruit, and evaluated its antimicrobial activity against various bacteria and fungi. As a result, the immature white magnolia fruit extract showed strong antibacterial activity against Gram-positive bacteria, Gram-negative bacteria, and fungi, and it was confirmed that the extract can be used as a broad spectrum antibacterial agent.

Accordingly, the present invention provides an antibacterial composition containing an extract of immature white magnolia fruit as an active ingredient.

The white magnolia "immature fruit" may mean the fruit immediately after the seed is generated.

The white magnolia immature fruit extract is preferably extracted with a solvent selected from the group consisting of water, alcohol, and anhydrous or hydrous alcohol having 1 to 4 carbon atoms, and ethanol extraction is most preferred.

The white magnolia immature fruit extract may be sequentially or fractionated with an organic solvent of hexene, ethyl acetate, and butanol to additionally obtain a hexene fraction, an ethyl acetate fraction, a butanol fraction, and a water residue.

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

Hereinafter, a method for preparing the immature white magnolia fruit extract of the present invention and an efficacy experiment will be described in more detail.

The inventors of the present invention, in order to achieve the object of the present invention, the steps of preparing an immature white magnolia fruit extract; Evaluating the antibacterial activity of the extract; And it includes the step of investigating the stability of the immature white magnolia fruit extract.

The "white magnolia immature fruit extract" included in the composition of the present invention may be obtained by extracting the immature white magnolia fruit with an organic solvent and 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 includes water (cold water, hot water), alcohol, anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, butanol, etc.), and a mixed solvent of the lower alcohol and water. Hot water, or 70% ethanol extraction is most preferred.

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

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

Baek magnolia immature fruit extract of the present invention exhibits strong antibacterial activity, and is used in pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, foods, cosmetics, animal feed additives, and various cleaning agents related to the prevention or treatment of infection of pathogenic bacteria. Can be used as a material.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[Example]

Example 1: Preparation of white magnolia fruit extract and analysis of useful components thereof

In 2018, white magnolia fruits from Andong, Gyeongbuk were collected and used to prepare ethanol extracts. Specifically, immature fruits before the seeds of white magnolia are produced (recovered on June 10, 2018: immature fruits (represented as 1)), and immature fruits immediately after the seeds are generated (recovered on July 20, 2018: immature fruits) (Represented as (2)), after dividing into mature fruits (recovered on September 10, 2018: marked as mature fruits (3)) with open pulp and exposed mature seeds (Fig. 1), 10 for each sample After adding pear of ethanol, extracting twice at room temperature, collecting the extract, filtering, and then concentrating under reduced pressure to prepare an ethanol extract.

The content of total polyphenols, total flavonoids, total sugars and reducing sugars was measured by component analysis of the white magnolia fruit 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, and the absorbance was measured at 725 nm after standing at room temperature for 1 hour. Tannic acid was used as a standard reagent. 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 solution, 40 μl of 1 N NaOH was added, and the absorbance was measured at 420 nm after 1 hour reaction at 37°C. . Rutin was used as a standard reagent. Reducing sugar was quantified by DNS method and total sugar was quantified by phenol-sulfuric acid method.

[Table 1] Analysis of extraction efficiency and useful components of fruit extract by maturation period of Baek magnolia

As shown in Table 1, the ethanol extraction efficiency of the white magnolia fruit was higher as the maturity increased, and the extraction efficiency of the immature fruit (1) before the seed was generated was only 1.2%, but the immature fruit (2) was It increased to 2.7%, and the yield of mature fruit (3) was 4.3%. As a result of analysis of total polyphenol content in fruit extract, the highest content of 232.3mg/g in the extract of immature fruit (1) before seed formation was shown, and it gradually decreased as the fruit matured, and 97.8 in the case of mature fruit (3). It was expressed as mg/g, which was 42% level compared to the initial immature fruit (1). The flavonoid content measurement result also showed the highest 78.2mg/g in the early immature fruit (1) extract, and then 27.2~31.0mg/g in the immature fruit (2) and mature fruit (3) extracts. ) Compared to 40%. On the other hand, the total sugar content increased as the fruit matured, showing 117.7 mg/g in the initial immature fruit (1) extract, while 153.7 mg/g and 307.2 mg/g in the immature fruit (2) and mature fruit (3) extract, respectively. g. As a result of evaluating the reducing sugar content, the content of the extract of the immature fruit (2) was increased 1.37 times compared to the initial immature fruit (1), but the extract of the mature fruit showed 27.6 mg/g, which was 42% higher than that of the immature fruit (1). Indicated. Therefore, it is expected that changes in useful physiological functions due to changes in components during the maturation process of white magnolia fruit are expected, and in the actual antioxidant activity evaluation results, immature fruit extract showed stronger activity than mature fruit extract.

Example 2: Evaluation of antibacterial activity of fruit extracts according to maturation period of white magnolia

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

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

[Table 2] Antibacterial activity of fruit extracts by maturation period of white magnolia

As a result, as shown in Table 2, ampicillin, which is used as an antibacterial agent in clinical practice, exhibited broad antimicrobial activity even at 1 μg/disc, and showed relatively weak growth inhibition rings (9 to 15 mm) for Pseudomonas aeruginosa and Salmonella typhimurium . Done. Miconazole, used clinically as an antifungal agent, strongly inhibited the growth of Saccharomyces cerevisiae and Candida albicans even at 1 μg/disc. On the other hand, Magnolia Denudata immature fruit extract was characterized by the Gram-positive bacteria is Listeria monocytogenes, Bacillus subtilis, Staphylococcus epidermidis, Staphylococcus aureus, gram-negative bacteria of Proteus vulgaris and Pseudomonas aeruginosa and fungal of Saccharomyces cerevisiae and Candida albicans for the growth inhibition at 500μg / disc density.

Example 3: Human Red Blood Cell Hemolytic Activity of Fruit Extracts by Maturation Period of White Magnolia

The buds of white magnolia are used as a medicinal herb called god, but their fruits are being discarded for no special purpose. In addition, it has not been used for food until now. Therefore, in order to evaluate the possibility of acute toxicity of the white magnolia fruit extract, human red blood cell hemolysis activity was evaluated, and the results are shown in Table 3. At this time, the hemolytic activity was evaluated according to the previous report (Ho-Yong Son, 2014 ㆍKorean J. Microbiol. Biotechnol. 42: 285-292), and simply 100 μl of human red blood cells washed three times with PBS was placed in a 96-well microplate. Then, 100 μl of sample solution of various concentrations was added and reacted for 30 minutes at 37°C. After that, the reaction solution was centrifuged for 10 minutes (1,500 rpm) to transfer 100 μl of the supernatant to a new microtiter plate, and the degree of hemoglobin outflow due to hemolysis was 414 nm. It was measured at. DMSO (2%) was used as a solvent control for the sample, and Triton X-100 (1 mg/ml) was used as an experimental control for hemolysis of red blood cells. Hemolytic activity was calculated using the following formula.

[Table 3] Human Red Blood Cell Hemolytic Activity of Fruit Extracts by Maturation Period of Baek Magnolia

First, it was confirmed that DMSO and water used as control cells had no hemolytic activity, and triton X-100 hemolytic 100% red blood cells at a concentration of 1 mg/ml. In addition, it was confirmed that amphotericin B, which is used as an anticancer agent and antifungal agent, hemolyzes more than 50% of red blood cells at a concentration of 0.025mg/ml. On the other hand, white magnolia mature fruit (3) extract and immature fruit (1) extract did not show any red blood cell hemolysis up to the concentration of 1 mg/ml, and immature fruit (2) extract showed 82.0% hemolysis at 1 mg/ml concentration, 0.5 At mg/ml concentration, 63.8% of red blood cell hemolysis was observed. As a result of calculating HC ₅₀ (concentration representing 50% hemolysis) from the erythrocyte hemolytic activity evaluation result, amphotericin B was 0.01449 mg/ml, and in the case of immature fruit (2) extract, it was found to be 0.4253 mg/ml, and immature fruit ( 2) The hemolytic activity of the extract was found to be insignificant at the level of 1/30~1/40 of amphotericin B currently used as an antibacterial agent.

Example 4: Plasma, acid and heat stability evaluation of immature white magnolia fruit extract

Plasma stability, thermal stability, and acid stability against antimicrobial activity were confirmed for the extract of immature white magnolia fruit (2) obtained in Example 1 above. Baek magnolia immature fruit (2) extract did not show any significant loss of antibacterial activity even after 1 hour heat treatment at 100°C, 1 hour treatment at pH 2 (0.01M HCl), and 1 hour treatment in plasma. Therefore, the extract of the immature white magnolia fruit (2) is expected to maintain excellent antibacterial activity even under various food processing processes.

Claims (4)

Baek magnolia ( Magnolia denudata ) Antibacterial composition containing the immature fruit extract as an active ingredient. The antibacterial composition according to claim 1, wherein the extract of immature white magnolia fruit is an ethanol extract of immature white magnolia fruit immediately after seed is generated. The antibacterial composition according to claim 1, wherein the antibacterial is antibacterial and antifungal activity. The method of any one of claims 1 or 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. Antibacterial composition, characterized in that the.

Images