KR102050305B1 - Antimicrobial composition against pathogenic bacteria containing Fragaria ananassa extract or its fraction as effective component - Google Patents

Abstract

The present invention relates to an antimicrobial composition for pathogenic bacteria containing a strawberry extract or a fraction thereof as an active ingredient, more specifically, according to the present invention, cultivated strawberry ( Fragariia ananassa leaf extract and fractions thereof are pathogenic bacteria Bacillus cereus , Escherichia coli ), Enterobacter cloacae , Salmonella enterica and Pseudomonas aeruginosa , which have excellent antimicrobial activity, making it very useful for pharmaceuticals, quasi-drugs, food additives or feed additives Can be used.

Description

Antimicrobial composition against pathogenic bacteria containing Fragaria ananassa extract or its fraction as effective component

The present invention relates to a composition for antibacterial against pathogenic bacteria containing a strawberry extract or a fraction thereof as an active ingredient.

Since cosmetics contain various nutrients, including water, they are susceptible to contamination by bacteria and fungi. The negative side of microbial contamination of cosmetics can be summarized in two ways: the first is the risk to the health of consumers, and the second is the fall in commodity value. Since most of the cosmetics have the characteristic of applying to the skin, when the microorganisms are contaminated on the skin, the microorganisms introduced through the wound may cause inflammation or skin trouble. In particular, contamination with pathogenic microorganisms can cause serious health problems. In addition, products that have proliferated with fungi or bacteria cause deterioration of product value by causing odor, destruction of emulsification, and changes in product properties due to changes in product pH. Therefore, various kinds of antimicrobial agents are used to suppress this, so it may be allergic to the skin, so it is important to use a relatively harmless substance. Most of the existing antimicrobial agents used in the past have been manufactured through chemical synthesis, but the emergence of resistant bacteria to such chemical synthetic antimicrobial agents is frequently occurring, and most of them are chemically synthesized antimicrobial agents because of their high toxicity. The use of is being avoided. Therefore, in recent years, efforts have been made to develop materials having antimicrobial activity among natural products.

On the other hand, planting strawberries (Fragaria ananassa Duch.) is a perennial plant belonging to the Rosaceae family, which is a long-growing, labor-intensive crop, but can be grown at low temperatures and harvested from November to May by low-temperature plant cultivation. Strawberry cultivation include flavonoids (flavonid) stream enriched quercetin-7-xylene with the side (quercetin-7-xyloside), plastic covering (fragarin), procyanidin B ₃ (procyanidin B _3), glucose (1-Op days in 1-Op- Kumar coumaroylglucose, 2-heptanol, or ellagic acid. Many studies related to diseases such as anti-inflammatory, anti-cancer, kidney disease using strawberry are in progress. In addition, because of its excellent fragrance and color, it is currently used as a raw material for manufacturing jams, syrups and juices.

On the other hand, Japanese Patent Laid-Open No. 2006-089410 discloses an antimicrobial composition, and Korean Patent No. 0997452 discloses a cosmetic composition using the extract of strawberry faucet as an active ingredient. However, the strawberry of the present invention ( Fragaria Ananassa ) No antimicrobial compositions for pathogenic bacteria containing extracts or fractions thereof as active ingredients have yet been disclosed.

The present invention is derived from the above requirements, the inventors of the present invention, the extracts of cultivated strawberry ( Fragareia ananassa ) leaf extract and its solvent (hexane, ethyl acetate, butanol and water) fractions of the pathogenic bacterium Bacillus cereus ( Bacillus cereus ) , Escherichia coli ), Enterobacter Cloacae ), Salmonella enterica ( Salmonella enterica ) and Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) respectively treated to confirm that effectively inhibit the growth of the pathogenic bacteria, the present invention was completed.

In order to solve the above problems, the present invention is a cultivated strawberry ( Fragaria ananassa ) Provides an antimicrobial composition against pathogenic bacteria containing an extract or a fraction thereof as an active ingredient.

In addition, the present invention is grown strawberry ( Fragaria Ananassa ) provides a cosmetic composition for antimicrobial against pathogenic bacteria containing an extract or a fraction thereof as an active ingredient.

In addition, the present invention is grown strawberry ( Fragaria ananassa ) Provides an antimicrobial food additive for pathogenic bacteria containing the extract or fractions thereof as an active ingredient.

In addition, the present invention is grown strawberry ( Fragaria ananassa ) Provides an antimicrobial feed additive for pathogenic bacteria containing the extract or fractions thereof as an active ingredient.

In addition, the present invention is grown strawberry ( Fragaria Ananassa ) Provides an antimicrobial composition for antibacterial against pathogenic bacteria containing an extract or a fraction thereof as an active ingredient.

According to the invention, cultivated strawberries ( Fragaria ananassa leaf extracts and fractions thereof are pathogenic bacteria Bacillus cereus , Escherichia coli , Enterobacter cloacaea cloacae ), Salmonella enterica and Pseudomonas aeruginosa ) shows excellent antimicrobial activity, it can be very useful for medicines, quasi-drugs, food additives or feed additives.

1 is a cultivated strawberry ( Fragareia) according to an embodiment of the present invention ananassa ) shows a schematic diagram of the preparation process of the fractions of each extract and its solvents (hexane, ethyl acetate, butanol and water).

In order to achieve the object of the present invention, the present invention provides a composition for antibacterial against pathogenic bacteria containing a cultivated strawberry ( Fragareia ananassa ) extract or a fraction thereof as an active ingredient.

By “antimicrobial” is meant the ability to reduce, prevent, inhibit or eliminate the growth or survival of microorganisms at any concentration.

"Antimicrobial composition" of the present invention is a composition having an activity that inhibits the growth of microorganisms, such as bacteria, may mean the same as an antibiotic that is a generic term for an antimicrobial agent, all used in various fields that require antimicrobial effect Forms may be included, for example, in the form of pharmaceuticals, quasi-drugs, food additives or feed additives.

The term “comprising” of the present invention is not limited to including, and does not exclude, for example, other additives, components, indices or steps.

In the antimicrobial composition for pathogenic bacteria according to an embodiment of the present invention, the cultivated strawberry extract may be extracted from strawberry leaves, but is not limited thereto.

In the antimicrobial composition for pathogenic bacteria according to an embodiment of the present invention, the extract may be extracted with water, a lower alcohol of C ₁ ~ C ₄ or a mixture thereof as a solvent, preferably ethanol solvent Extraction may be performed as, but is not limited thereto.

In the antimicrobial composition for pathogenic bacteria according to an embodiment of the present invention, the fraction may be butanol or water fraction of cultivated strawberry ethanol extract, but is not limited thereto.

In the antimicrobial composition for pathogenic bacteria according to an embodiment of the present invention, the pathogenic bacteria are Bacillus cereus , Escherichia coli , Enterobacter cloaca ( Enterobacter) cloacae ), Salmonella enterica and Salmonella enterica and Pseudomonas aeruginosa may be one or more selected from the group consisting of, but is not limited thereto.

In addition, the present invention is grown strawberry ( Fragaria Ananassa ) provides a cosmetic composition for antimicrobial against pathogenic bacteria containing an extract or a fraction thereof as an active ingredient.

The pathogenic bacteria are as described above.

The cultivated strawberry extract or fractions thereof exhibits strong antibacterial activity and low cytotoxicity to human derived cells, so the cultivated strawberry extracts or fractions thereof of the present invention may be usefully used as an active ingredient of a cosmetic composition.

The cosmetic composition of the present invention includes components commonly used in cosmetic compositions in addition to the cultivated strawberry extract or fractions thereof, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers. It includes.

In the cosmetic composition of the present invention, the cultivated strawberry extract of the present invention or a fraction thereof may be added in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight, to the cosmetic composition usually contained.

Cosmetic compositions of the present invention may be prepared in any formulation conventionally prepared in the art and include, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a flexible lotion (skin), nutrition lotion (milk lotion), nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder .

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanta, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, and especially in the case of spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tragacanta and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In addition, the present invention is grown strawberry ( Fragaria ananassa ) Provides an antimicrobial food additive for pathogenic bacteria containing the extract or fractions thereof as an active ingredient.

The cultivated strawberry extract or fractions thereof exhibits strong antibacterial activity and low cytotoxicity to human derived cells, so the cultivated strawberry extracts or fractions thereof may be usefully used as an active ingredient of food additives.

When the cultivated strawberry extract of the present invention or a fraction thereof is used as a food additive, the cultivated strawberry extract or a fraction thereof may be added as it is or used together with other food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient can be appropriately determined according to the purpose of use thereof. In general, the cultivated strawberry extract of the present invention or a fraction thereof is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less with respect to the raw material. However, in the case of long term intake, the amount may be below the above range, and since there is no problem in terms of stability, the active ingredient may be used in an amount above the above range.

There is no particular limitation on the kind of food. Examples of foods to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and all foods in the conventional sense.

In addition, the present invention is grown strawberry ( Fragaria ananassa ) Provides an antimicrobial feed additive for pathogenic bacteria containing the extract or fractions thereof as an active ingredient.

The cultivated strawberry extract or fractions thereof exhibits strong antibacterial activity and low cytotoxicity to human derived cells, so the cultivated strawberry extracts or fractions thereof may be usefully used as an active ingredient of feed additives.

Feed composition of the present invention can be prepared in the form of fermented feed, compound feed, pellet form and silage and the like.

The fermented feed may be prepared by fermenting an organic material by adding various microorganisms or enzymes other than the cultivated strawberry extract or fractions thereof of the present invention, and the compounded feed may include various kinds of general feeds and the cultivated strawberry extract of the present invention or its Fractions can be prepared by mixing. Pellet-type feed can be prepared by applying heat and pressure to the blended feed, etc. in a pellet machine, silage can be prepared by fermenting the green feed to the microorganisms. The wet fermented feed collects and transports organic materials such as food waste, mixes excipients for sterilization and moisture control at a certain ratio, and then ferments at a temperature suitable for fermentation for at least 24 hours, so that the moisture content is about 70%. It can manufacture by adjusting. Fermented dry irrigated feed may be prepared by adjusting the wet fermented feed to contain about 30% to 40% of water by further drying.

In addition, the present invention is grown strawberry ( Fragaria Ananassa ) Provides an antimicrobial composition for antibacterial against pathogenic bacteria containing an extract or a fraction thereof as an active ingredient.

When the composition of the present invention is used as an quasi-drug additive, the cultivated strawberry extract or fraction may be added as it is, or used together with other quasi-drugs or quasi-drug components, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the intended use.

The quasi-drug composition of the present invention is not limited thereto, but may preferably be a disinfectant cleaner, a shower foam, a gagreen, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment, a patch, or a filter filler.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for explaining the present invention in more detail, it is obvious to those skilled in the art that the scope of the present invention is not limited by them.

Materials and methods

1. Material

Experimental planting strawberry leaves ( Fragaria ananassa var. 'Seolhyang') was harvested and washed in May 2016 at Kim Jong-won farm in Nonsan, Chungcheongnam-do, and then shaded and ground to make powder and used for various experiments and analysis.

2. Sample Preparation

The dried cultivated strawberry leaves were extracted three times for 24 hours with 15 times (w / v) 80% ethanol per 200 g of the sample, and the extract was subjected to filtration with filter paper (Whatman No. 2, Maidstone, England). The filtrate was removed using a rotary pressure reducer (EYELA A-1000S, Tokyo Rikakikai co., Tokyo, Japan) in a water bath at 40 ° C, concentrated under reduced pressure, and freeze-dried (SFDSM12-60Hz, Samwon Freezing Engineering Co., Seoul, Korea) and refrigerated below 4 ℃ was used in the experiment. 80% ethanol extract with a separating funnel by solvent fractionation by (Separating funnel) n - hexane (n -hexane), ethyl acetate (ethyl acetate) and n - and then extracted successively with butanol (n- butanol) to rotate the respective fractions under reduced pressure Concentrated under reduced pressure with a concentrator and then lyophilized (Fig. 1).

3. Test strain

Antimicrobial activity was determined by disc diffusion assay using each strain. The strains used for the antibacterial activity is measured in gram-positive Bacillus cereus (Bacillus cereus; B. cereus) KCTC 1012 and Staphylococcus aureus (Staphylococcus aureus; S. aureus) and KCTC 3881, gram-negative bacteria of Enterobacter claw aka Ke (Enterobacter cloacae; E. cloacae) KTCT 1685, Escherichia coli (Escherichia coli; E. coli) KTCT 2441, Salmonella Entebbe Rica (Salmonella enterica; S. enterica) Rouge labor (Pseudomonas aeruginosa in KTCT 1925 and Pseudomonas; P . aeruginosa) KTCT was used under pre-sale a total of six species of 1636 in biological resource Center (Korean collection for Type Culture, Daejeon , Korea) and Korea Culture Center of microorganisms (Korean Culture Center of microorganisms, Seoul , Korea) ( Table 1).

4. Cultivated Strawberry Leaf Extracts and Their Solvents Fraction  Antimicrobial Search

The antibacterial activity of the cultivated strawberry leaf extract and its solvent-specific fractions were grown and cultured for each test strain (Table 1). Specifically, the cultured dormant strains were passaged three times in NB medium (nutrient broth; Becton, Dickinson & Co., MD, USA), and the absorbance value measured at 600 nm was 0.4-0.6 (1 × 10 ⁵ CFU / mL), and used as a test strain for the determination of antimicrobial activity. Each test strain was dispensed with 100 μl of NA (nutrient agar; Becton, Dickinson & Co., MD, USA), an antibacterial test plate, and plated with a sterile swab. Each sample was 1 mg and 5 mg per disc. Absorb as slowly as possible to the paper disk (8mm), volatilize the solvent through the drying process, and incubate at 30 ~ 37 ℃ for 24 hours while keeping it in contact with the plate medium. ) Was measured to compare the antimicrobial activity.

Test strain used in the present invention Test strain badge Temperature (℃) Gram positive
Germ Bacillus cereus NA ¹⁾ / NB ²⁾ 30 Staphylococcus aureus NA / NB 30 Well voice
Germ Escherichia coli (Escherichia coli ) NA / NB 30 Enterobacter cloacae ) NA / NB 30 Salmonella enterica NA / NB 37 Pseudomonas aeruginosa ) NA / NB 37

¹⁾ NA: Nutrient agar

²⁾ NB: Nutrition broth

5. Statistical Processing

All experiments were repeated three times or more, expressed as mean and standard deviation, and the results were analyzed by ANOVA using SPSS 22.0 (Statistical Packege for Social Sciences, SPSS Inc., Chicago IL, USA) software. It was. For items with significant differences, Duncan's multiple range test was used to test the difference at p <0.05.

Example  1. Cultivated Strawberry Leaf Extract and Its Solvents Fraction  Confirmation of antimicrobial activity against pathogenic bacteria following treatment

In Example 1, the cultivated strawberry leaf extract and its solvent-specific fractions were prepared and treated with pathogenic bacteria at a concentration of 5 mg / disc to confirm the antimicrobial activity. As a result, to a Gram-positive bacteria of Bacillus cereus (Bacillus cereus), as shown in Table 1, the gram-negative bacteria Escherichia coli (Escherichia coli ), Enterobacter cloacae ), Salmonella enterica and Pseudomonas aeruginosa ) strain inhibited growth. The 80% ethanol extract showed antimicrobial activity in order of Salmonella enterica (11 mm), Bacillus cereus (10 mm), Enterobacter chlorcaea (10 mm) and Escherichia coli (9.25 mm), and n -butanol. The fraction showed antimicrobial activity in Bacillus cereus (11 mm), and in the water fraction, Escherichia coli (10.25 mm), Bacillus cereus (10 mm), Salmonella enterica (9.5 mm) and Pseudomonas aeruginosa (9.25 mm). It showed growth inhibitory activity in order.

Antimicrobial Effects of Cultivated Strawberry Leaf Extracts and Their Solvent Fractions Against Pathogenic Bacteria Test strain Growth low ring (mm) menstruum Treatment Concentration (5mg / disc) Bacillus cereus 80% ethanol 10 n-hexane ^-1) Ethyl acetate - n-butanol 11 water 10 Staphylococcus aureus 80% ethanol - n-hexane - Ethyl acetate - n-butanol - water - Escherichia coli 80% ethanol 9.25 n-hexane - Ethyl acetate - n-butanol - water 10.25 Enterobacter cloacaea 80% ethanol 10 n-hexane - Ethyl acetate - n-butanol - water 10.5 Salmonella Enterica 80% ethanol 11 n-hexane - Ethyl acetate - n-butanol - water 9.5 Pseudomonas Eruzinosa 80% ethanol - n-hexane - Ethyl acetate - n-butanol - water 9.25

^{1) No} growth deterrent observed

Claims (9)

Antimicrobial composition against Enterobacter cloacae or Pseudomonas aeruginosa containing ethanol extract of cultivated strawberry ( Fragareia ananassa ) leaf or water fraction thereof as an active ingredient. delete delete delete delete An antimicrobial cosmetic composition for Enterobacter cloacae or Pseudomonas aeruginosa , containing an ethanol extract of a cultivated strawberry ( Fragaria ananassa ) leaf or a water fraction thereof as an active ingredient. Antimicrobial food additive for Enterobacter cloacae or Pseudomonas aeruginosa containing ethanol extract of cultivated strawberry ( Fracgaria ananassa ) leaf or water fraction thereof as an active ingredient. Feed additive for antibacterial against Enterobacter cloacae or Pseudomonas aeruginosa containing ethanol extract of cultivated strawberry ( Fragaria ananassa ) or water fraction thereof as an active ingredient. Antimicrobial composition for antimicrobial against Enterobacter cloacae or Pseudomonas aeruginosa containing ethanol extract of cultivated strawberry ( Fragareia ananassa ) leaf or water fraction thereof as an active ingredient.

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