KR102550348B1 - Antimicrobial composition comprising fermented product of Chrysanthemum extract by lactic acid bacteria as effective component - Google Patents

Abstract

Since the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention has antibacterial activity against gram-negative and gram-positive bacteria, it can be usefully used as an active ingredient in antibiotics, cosmetic compositions, food additives, feed additives, biological pesticides and quasi-drugs.

Description

Antimicrobial composition comprising fermented product of Chrysanthemum extract by lactic acid bacteria as effective component}

The present invention relates to an antibacterial composition containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In general, direct or indirect damage caused by pathogenic microorganisms causes many economic, environmental and medical problems. In particular, loss due to spoilage during the food distribution process, harm to the human body and environmental pollution due to the use of excessive chemical pesticides on crops, and the emergence of antibiotic-resistant strains due to misuse and abuse of antibiotics are already chronic problems throughout society. there is. In addition, most of the currently used antibiotics are manufactured through chemical synthesis, and if synthetic antibiotics are used excessively for a long period of time, they may remain in the human body and cause toxicity or create new resistant bacteria. In particular, benzoic acid, paraben, imidazolidinyl urea, and chorphenesin, which are used in external preparations, have excellent stability and antiseptic efficacy, but cause toxicity and skin problems. The downside is that it can cause irritation and allergies. Therefore, in recent years, it is necessary to develop environmentally friendly and harmless natural substances that can replace the function of antibiotics through research on natural substances having antibiotic activity in a wide range of pathogenic bacteria and fungi and new antibacterial substances isolated therefrom. am.

On the other hand, chrysanthemum is a plant with the longest history among ornamental plants cultivated in the Orient, and mainly uses flower parts as food or medicine. Chrysanthemum contains flavonoids, phenols, etc., so it has an antioxidant effect, and has recently been reported to have pharmacological effects such as anti-inflammatory, lowering blood pressure, and diuresis. In the private sector, chrysanthemum flowers are known to be effective in relieving symptoms of headache, dizziness, ringing in the ears, and blood circulation.

On the other hand, Korean Patent Registration No. 2242755 discloses 'skin anti-inflammatory, anti-wrinkle, whitening and antibacterial functional cosmetic composition containing chrysanthemum flower extract from Udo, Jeju', and Korean Patent Publication No. 2011-0120394 discloses 'extract from chrysanthemum flower 'antibacterial composition' is disclosed, but there is no description about the antibacterial composition containing the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention as an active ingredient.

The present invention has been derived from the above needs, and the present inventors inoculated and fermented Lactobacillus acidophilus KCTC3140 strain in a chrysanthemum extract to prepare a lactic acid bacteria fermentation product of the chrysanthemum extract, By confirming that the fermentation product of lactic acid bacteria has antibacterial activity against Bacillus cereus , Listeria monocytogenes , Escherichia coli or Salmonella enterica , the present invention completed.

In order to solve the above problems, the present invention provides an antibacterial composition containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In addition, the present invention provides an antibiotic containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In addition, the present invention provides an antibacterial cosmetic composition containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In addition, the present invention provides an antibacterial food additive containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In addition, the present invention provides an antibacterial feed additive containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In addition, the present invention provides an antibacterial biological pesticide containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In addition, the present invention provides an antibacterial quasi-drug composition containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In addition, the present invention provides an antibacterial method in a non-human subject comprising the step of administering a pharmaceutically effective amount of a lactic acid bacteria fermented product of a chrysanthemum extract to a non-human subject.

Since the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention has antibacterial activity against gram-negative and gram-positive bacteria, it can be usefully used as an active ingredient in antibiotics, cosmetic compositions, food additives, feed additives, biological pesticides and quasi-drugs.

In order to achieve the object of the present invention, the present invention provides an antibacterial composition containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient.

In the antibacterial composition of the present invention, the chrysanthemum may preferably be a flower part of small flowered chrysanthemum, but is not limited thereto.

In addition, the extraction solvent of the chrysanthemum extract may be water, C ₁ ~ C ₄ lower alcohol, or a mixture thereof, preferably C ₁ ~ C ₄ lower alcohol, more preferably ethanol. , but not limited thereto.

In addition, in the antibacterial composition of the present invention, the lactic acid bacteria fermented product of the chrysanthemum extract may be prepared by inoculating and fermenting the chrysanthemum extract with a strain of the genus Lactobacillus , preferably Lactobacillus acidophilus ( Lactobacillus acidophilus ) KCTC3140 strain may be inoculated and prepared by fermentation, but is not limited thereto.

The lactic acid bacteria fermented product of the chrysanthemum extract of the present invention may be prepared by fermentation at 32 to 42° C. for 40 to 56 hours, and preferably may be prepared by fermentation at 37° C. for 48 hours, but is not limited thereto.

In addition, the lactic acid bacteria fermented product of the chrysanthemum extract shall include any one of a fermentation broth obtained by extraction and fermentation, a diluted or concentrated fermentation broth, a dried product obtained by drying the fermentation solution, a crude product, or a purified product.

The lactic acid bacteria fermented product of the chrysanthemum extract of the present invention preferably has antibacterial activity against gram-negative bacteria or gram-positive bacteria, but is not limited thereto.

The gram-negative bacteria are all gram-negative bacteria known in the art as gram-negative bacteria including Escherichia genus, Salmonella genus, Pseudomonas genus, Leptospira genus and Rickettsia genus It is preferable, specifically Escherichia ( Escherichia ) Or Salmonella ( Salmonella ) Genus It is more preferable that any one or more selected from the group consisting of, more specifically Escherichia coli ( Escherichia coli ) or Salmonella Enterica ( Salmonella enterica ) is most preferred, but is not limited thereto.

The gram-positive bacteria are known in the art as gram-positive bacteria including genus Bacillus , genus Listeria , genus Staphylococcus , genus Corynebacterium and genus Lactobacillus . It is preferably all Gram-positive bacteria, more preferably Bacillus or Listeria , more preferably Bacillus cereus or Listeria monocytogenes . , but not limited thereto.

The present invention also provides an antibiotic containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient. The lactic acid bacteria fermented product of the chrysanthemum extract is as described above.

Since the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention exhibits antimicrobial activity against gram-negative or gram-positive bacteria, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention can be usefully used as an active ingredient of antibiotics.

The lactic acid bacteria fermented product of the chrysanthemum extract of the present invention can be administered parenterally during clinical administration and can be used in the form of general pharmaceutical preparations. Parenteral administration may refer to administration through an administration route other than oral administration, such as rectal, intravenous, peritoneal, intramuscular, arterial, transdermal, nasal, inhalational, ocular and subcutaneous administration. When the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention is used as a medicine, it may additionally contain one or more active ingredients exhibiting the same or similar functions.

That is, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention can be administered in various actual parenteral formulations. prepared using Formulations for parenteral administration include sterilized aqueous solutions, water-insoluble agents, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, liurine fat, glycerogeratin, and the like may be used.

In addition, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention may be mixed with various carriers accepted as pharmaceuticals such as physiological saline or organic solvents, and may be mixed with glucose, sucrose or dextran to increase stability or absorption. Antioxidants such as carbohydrates, ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers can be used as pharmaceuticals.

In the antibiotics of the present invention, the effective dosage of the patient is determined in consideration of various factors such as the age and health condition of the patient as well as the route of administration and number of treatments as well as the administration route and the number of treatments. At this time, those skilled in the art will be able to determine an appropriate effective dose according to the specific use as an antibiotic of the lactic acid bacteria fermentation of the chrysanthemum extract of the present invention.

In addition, the present invention provides an antibacterial cosmetic composition containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient. The lactic acid bacteria fermented product of the chrysanthemum extract is as described above.

Since the lactic acid bacteria fermented product of the chrysanthemum extract exhibits antibacterial activity against gram-negative bacteria or gram-positive bacteria, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention can be usefully used as an active ingredient in a cosmetic composition.

The cosmetic composition of the present invention includes components commonly used in cosmetic compositions in addition to the lactic acid bacteria fermented product of the chrysanthemum extract, for example, antioxidants, stabilizers, solubilizers, conventional adjuvants such as vitamins, pigments and flavors, and carriers. includes

In the cosmetic composition of the present invention, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention may be added in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight, in the cosmetic composition usually contained.

The cosmetic composition of the present invention can be prepared in any formulation conventionally prepared in the art, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , It may be formulated as an oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be formulated into a formulation of softening lotion (skin), nourishing lotion (milk lotion), nourishing 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, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. are used as carrier components. It can be.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol oil, fatty acid esters of glycerol, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar or tragacantha and the like may be used.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, 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.

The present invention also provides an antibacterial food additive containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient. The lactic acid bacteria fermented product of the chrysanthemum extract is as described above.

Since the lactic acid bacteria fermented product of the chrysanthemum extract exhibits antibacterial activity against Gram-negative bacteria or Gram-positive bacteria, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention can be usefully used as an active ingredient of an antibacterial food additive.

When the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention is used as a food additive, the lactic acid bacteria fermented product of the chrysanthemum extract can be added as it is or used together with other food ingredients, and can be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of its use. In general, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake, the amount may be less than the above range, and since there is no problem in terms of stability, the active ingredient may be used in an amount greater than the above range.

There is no particular limitation on the type of food. Examples of foods to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all foods in a conventional sense.

The present invention also provides an antibacterial feed additive containing a lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient. The lactic acid bacteria fermented product of the chrysanthemum extract is as described above.

Since the lactic acid bacteria fermented product of the chrysanthemum extract exhibits antibacterial activity against Gram-negative bacteria or Gram-positive bacteria, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention can be usefully used as an active ingredient of an antibacterial feed additive.

The feed additive of the present invention replaces conventional antibiotics and suppresses the growth of harmful food pathogens to improve the health of animals, improve weight gain and quality of livestock, and increase milk production and immunity.

The present invention also provides an antiseptic composition, an antibacterial biological pesticide and an antibacterial quasi-drug composition containing the lactic acid bacteria fermented product of chrysanthemum extract as an active ingredient. The lactic acid bacteria fermented product of the chrysanthemum extract is as described above.

Since the lactic acid bacteria fermented product of the chrysanthemum extract exhibits antibacterial activity against gram-negative bacteria or gram-positive bacteria, the lactic acid bacteria fermented product of the chrysanthemum extract of the present invention can be usefully used as an active ingredient in antibacterial biological pesticides, antiseptic compositions, and quasi-drug compositions. .

The preservative composition includes a cosmetic preservative or a pharmaceutical preservative. The food preservatives, cosmetic preservatives, and pharmaceutical preservatives are additives used to prevent deterioration, decay, discoloration, and chemical change of medicines, and include bactericides and antioxidants, and inhibit the growth of microorganisms such as bacteria, molds, and yeasts, thereby inhibiting the growth of food products. And functional antibiotics such as inhibiting the growth of spoilage microorganisms or sterilizing action in pharmaceuticals are also included. As an ideal condition for such an antiseptic composition, it should not be toxic and should be effective even in a small amount.

When the composition of the present invention is used as a quasi-drug additive, the lactic acid bacteria fermented product of the chrysanthemum extract can be added as it is or used together with other quasi-drugs or quasi-drug ingredients, and can be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use.

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

The present invention also provides an antibacterial method in a non-human subject comprising the step of administering a pharmaceutically effective amount of the lactic acid bacteria fermented product of the chrysanthemum extract to the non-human subject. The lactic acid bacteria fermented product of the chrysanthemum extract is as described above. The non-human subject may preferably be a non-human mammal, but is not limited thereto.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are only to illustrate the present invention, and the content of the present invention is not limited to the following examples.

Example 1. Preparation of lactic acid bacteria fermented product of chrysanthemum extract

250 ml of 70% (v/v) ethanol was added to 50 g of the flower part of the dried small flowered chrysanthemum, followed by repeated stirring and extraction at room temperature for 5 hours twice, followed by concentration under reduced pressure to prepare a chrysanthemum extract. 10 ml of the chrysanthemum extract and 5 g of glucose were added to a 250 ml Erlenmeyer flask, and the final 50 ml was added with distilled water, followed by sterilization at 121° C. for 15 minutes. Thereafter, the culture solution of Lactobacillus acidophilus KCTC3140 strain pre-cultured in MRS medium for 12 to 16 hours was inoculated with 1% (v / v) of the sterilized chrysanthemum extract, and 48 °C at 150 rpm at 37 ° C. Lactobacillus fermented product of chrysanthemum extract was prepared by culturing for a period of time, and after centrifugation at 4° C. and 5,000 g for 10 minutes, the supernatant was collected and used as a sample for measuring antibacterial activity.

The culture medium of Lactobacillus acidophilus KCTC3140 strain cultured in MRS medium was centrifuged at 4° C. and 5,000 g for 10 minutes, and then the supernatant was collected and used as a control (lactic acid bacteria culture medium) sample.

Example 2. Measurement of antibacterial activity

The strains used for measuring the antibacterial activity of the present invention were distributed from KCTC, Bacillus cereus with resource number KCTC3624, Listeria monocytogenes with resource number KCTC3710, Escheri with resource number KCCM11835 Escherichia coli , Salmonella enterica whose resource number is KCTC1925, was cultured and used under the conditions shown in Table 1 below.

Culture conditions of strains used to measure antibacterial activity strain culture medium Incubation temperature (℃) Gram
positive bacteria Bacillus cereus TSB, NB 30 Listeria monocytogenes BHI 30 Gram
Negative bacteria Escherichia coli TSB, NB 37 Salmonella enterica NB 37

(TSB: Tryptic Soy Broth, NB: Nutrient Broth, BHI: Brain Heart Infusion)

The culture medium of the strain used for the measurement of antibacterial activity (OD ₆₀₀ =0.4~0.6) was added at 1% (v/v) to 10 ml of 0.8% (w/v) NB soft agar medium and 0.5% (w/v) yeast A plate for measuring antibacterial activity was prepared by layering the extract on a solid medium. After spotting 50 μl of the lactic acid bacteria fermented product or lactic acid bacteria culture medium of the chrysanthemum extract of Example 1 on a plate for measuring antibacterial activity, and then incubating at 37 ° C. for more than 12 hours, the clear zone The diameter of was measured.

As a result, it was confirmed that the lactic acid bacteria fermented product of the chrysanthemum extract had antibacterial activity against Bacillus cereus, Listeria monocytogenes, Escherichia coli, and Salmonella enterica. ) or Listeria monocytogenes (KCTC3710), it was confirmed that the antibacterial activity was increased compared to the lactic acid bacteria culture medium alone (Table 2).

Antibacterial activity measurement result strain Lactobacillus culture medium of chrysanthemum extract
Lactobacillus fermented product Gram
positive bacteria Bacillus cereus (KCTC3624) + ++ Listeria monocytogenes (KCTC3710) - + Gram
Negative bacteria Escherichia coli (KCCM11835) + + Salmonella enterica (KCTC1925) + +

(-: No activity, +: Inhibiting ring less than 10 mm, ++: Inhibiting ring more than 10 mm)

Claims (14)

An antibacterial composition for Escherichia coli containing a fermented product prepared by inoculating and fermenting a strain of Lactobacillus acidophilus in an ethanol extract of chrysanthemum as an active ingredient. The antibacterial composition for Escherichia coli according to claim 1, wherein the chrysanthemum is small flowered chrysanthemum. delete delete delete delete delete An antibiotic for Escherichia coli containing a fermented product prepared by inoculating and fermenting a strain of Lactobacillus acidophilus in an ethanol extract of chrysanthemum as an active ingredient. Chrysanthemum ethanol extract Lactobacillus acidophilus ( Lactobacillus acidophilus ) Strain inoculated and fermented, containing a fermented product prepared by fermentation as an active ingredient Escherichia coli ( Escherichia coli ) Antibacterial cosmetic composition for. An antibacterial food additive for Escherichia coli containing a fermented product prepared by inoculating and fermenting a strain of Lactobacillus acidophilus in ethanol extract of chrysanthemum as an active ingredient. An antibacterial feed additive for Escherichia coli containing a fermented product prepared by inoculating and fermenting a strain of Lactobacillus acidophilus in an ethanol extract of chrysanthemum as an active ingredient. An antibacterial biological pesticide for Escherichia coli containing a fermented product prepared by inoculating and fermenting a strain of Lactobacillus acidophilus in an ethanol extract of chrysanthemum as an active ingredient. An antibacterial quasi-drug composition for Escherichia coli containing, as an active ingredient, a fermented product prepared by inoculating and fermenting a strain of Lactobacillus acidophilus in an ethanol extract of chrysanthemum. Escherichia in a non-human subject comprising administering a fermented product prepared by inoculating a pharmaceutically effective amount of chrysanthemum ethanol extract with a Lactobacillus acidophilus strain and fermenting it to a non-human subject Antibacterial method against Escherichia coli .