KR20080008491A - Bokbunja Fermentation and Composition Comprising the Same - Google Patents

Abstract

The present invention relates to bokbunja fermentation. Bokbunja fermented product of the present invention is fermented using one or more strains selected from lactic acid bacteria, yeast, Hwangkuk bacteria, Hongguk bacteria, Bacillus subtilis shows the wrinkle improvement efficacy, psoriasis relief efficacy, anti-cancer and flushing efficacy and intestinal efficacy. Therefore, the present invention comprises a cosmetic composition comprising each of the bokbunja fermentation exhibiting such efficacy; Pharmaceutical composition; Food compositions; And feed composition.

Description

Bokbunja fermentation and composition comprising the same {Fermented Rubus coreanus Miq. and the composition containing definitely}

1 is a photograph showing the results of conducting the chorioalantoic membrane assay by dividing the chicken eggs into a control group (Fig. 1a), Example 3 administration group (Fig. 1b), Example 4 administration group (Fig. 1c).

It relates to a bokbunja fermentation and a composition comprising the same.

Academic studies on bokbunja have a lot of antioxidant effects. The antioxidant effect of bokbunja is due to polyphenolic substances, and triterpenosides such as coreanoside F1, suavissimoside, nigaichigoside F1, and F2 were found in bokbunja (Young-Hee Kim, et al., Arch. Pharm. Res . 16: 109- 113 (1993)), and tannins such as epicatechin, catechin, procyanidin B-4, and sanguiin H-4 have been found (Yeon-Ah Lee, et al., Kor. J. Phamacogn . 26: 27-30 ( 1995)) Bokbunja is known to contain various polyphenolic substances as pharmacological substances.

It is known that the components of polyphenolic substances of bokbunja change with maturity of bokbunja, while anthocyanin increases markedly with maturation of bokbunja, while the total amount of polyphenolic substances is hardly changed or decreased (Hwan-Soo Cha, et. al., Korean J. Food Sci. Technol . 33: 409-415 (2001), Kong-Sik Shin, et al ., Korean J. Plant.Res. 16: 109-117 (2003))

It is reported that the pharmacological effect of bokbunja also changes with the change of these components, especially the antioxidant effect is markedly reduced in bokbunja maturation (Kong-Sik Shin, et al ., Korean J. Plant.Res. 16: 109-117 (2003). ))

As described above, studies on the effectiveness of bokbunja have been mainly conducted on antioxidant effects (In Yoon, et al., Korean J. Food Sci. Technol . 35: 499-502 (2003), Jong-Won Lee, et al. ., J. Korean Soc. Food Sci. Nutr . 29: 943-947 (2000), In Yoon, et al., Korean J. Food Sci. Thchnol . 34: 898-904 (2002)), particularly of hydrolyzable tannins. Antioxidant activity has been reported (Kwang-Ho Kim, Et al., Yakhak Heoji 44: 354-357 (2000). For efficacy other than the antioxidant effect, recently angiotensin 1 converting enzyme (ACE), which induces elevated blood pressure, has been reported. Inhibitory effect, report on the inhibitory effect of a-glucosidase activity on blood glucose elevation (Mi-Kyoung Lee, et al., Korean J. Medicinal Crop. Sci . 11: 5-12 (2003)), and antimicrobial effect ( Hwan-Soo Cha, et al., Korean J. Food Sci. Technol . 33: 409-415 (2001)), antihistamine effect (Guang-Zhao Li, et al., Korean J. Phys. Anthrop . 17: 99 -107 (2004)).

Related patents include a method for preparing herbal extracts and extract dried powders having excellent free radical scavenging ability (Korean Patent Office, Publication No. 041650), external topical composition having free radical scavenging ability or protection against intracellular antioxidant system (Korea Patent Office, Application No. 10-2002-0027050) and the most anti-oxidant effect, patents on antimicrobial effects such as the composition for treating periodontal disease containing plant extracts (Korea Patent Office, Registration No. 10-0431170), bokbunja extract effective for diabetic complications (Korea Korean Intellectual Property Office, Registration No. 10-0393908), Cosmetic composition for treating atopic dermatitis (Korea Patent Office, Registration No. 10-0451444), Skin external preparation composition containing raspberry leaf or fruit extract having skin irritation and anti-inflammatory effect (Korea Patent Office, Registration No. 10-0444599), Egg containing mixed extract of Eoseongcho and Bokbun as active ingredients There are patents on skin diseases such as atopy, such as the composition for the prevention and treatment of allergic diseases (Korean Patent Office, Application No. 10-2004-0007404), and the components that inhibit prostaglandins that are increased by ultraviolet rays and those that inhibit melanin synthesis. Various patents such as a whitening cosmetic composition (Korean Patent Office, Registration No. 10-0406124) containing at the same time have been applied or registered. However, research on fermentation of Bokbunja is mostly aimed at improving the flavor of Bokbunja. However, there are only few studies that promote new efficacy with industrial availability by fermenting Bokbunja.

An object of the present invention is to provide a bokbunja fermented product having an industrially high efficacy that is not known in the past through bokbunja fermentation, and further to provide pharmaceuticals, food, cosmetics and feed having such efficacy.

The present invention relates to a bokbunja fermented product obtained by inoculating and fermenting at least one microorganism selected from lactic acid bacteria, yeasts, vinegar fungus, sulfur soup fungus and red yeast fungus.

In the above, bokbunja is applicable in the form of, for example, bokbunja itself or its grinding powder, or extract of bokbunja.

Representative examples of conventional bokbunja fermentation is to enhance the palatability as a liquor by fermenting bokbunja ripened fruit juice using yeast mainly bokbunja. The final purpose of the present invention is to provide a fermented product with high pharmacological efficacy, and thus, bokbunja immature fruit, which has a higher content of polyphenol, which is the main functional substance of plants, is used instead of ripen fruit juice. Therefore, "bokbunja" described in the present invention refers to "unripe fruit unripe fruit" unless otherwise specified. In addition, the bokbunja fermentation product of the present invention is entirely new. Bokbunja immature fruit can be used both as raw unripe fruit and dried bokbunja immature fruit. However, bokbunja immature fruit is generally dried as medicinal ingredients.

Yeasts from the above Saccharomyces ( Saccharomyces) cerevisiae ) CBI-YB41 strain (Accession No. KCCM10763P) is preferred.

In the preparation of the fermentation product, preferably, a nutrient or purified water may be additionally used together with bokbunja, and more preferably, a nutrient and purified water may be further used together to form a mixture before fermentation and then fermented.

The content of the bokbunja in the mixture before fermentation is preferably 0.01 to 10% by weight based on the dry weight of bokbunja. Here, in the case of bokbunja extract as well means 0.01 to 10% by weight equivalent amount on the basis of dry weight of bokbunja.

The nutritional component is preferably at least one component selected from yeast extract, soy protein, malt extract, peptone, tyrosine, cysteine, phytic acid, glucose, maltose, lactose, sucrose and salts.

In addition, the fermentation temperature and time may vary depending on the type and inoculation amount of the fermentation strain, but it is preferable to set at 25 ~ 45 ℃ for 1-7 days in the fermenter.

The fermented product obtained above may preferably be used by sterilizing or disinfecting after removing the cells and insoluble materials by, for example, centrifugation or membrane separation.

Another aspect of the present invention relates to a cosmetic composition for improving wrinkles comprising the bokbunja fermentation.

The present invention also relates to a cosmetic composition for psoriasis relaxation comprising the above bokbunja fermented product.

The present invention also relates to an anti-skin cancer and flushing cosmetic composition comprising the above bokbunja fermentation.

In addition, the present invention relates to a pharmaceutical composition for wrinkle improvement, psoriasis relief, anticancer and flushing or formal dress comprising the above bokbunja fermentation.

The present invention also relates to anti-wrinkle, psoriasis relief, anticancer and flushing or food composition comprising the above bokbunja fermentation.

The present invention also relates to psoriasis alleviation, anticancer and redness relaxation or formal feed composition comprising the bokbunja fermentation.

The present inventors have studied the novel efficacy of the bokbunja fermentation. From bokbunja fermentation, we found (1) excellent collagen production promoting effect related to wrinkle improvement and matrix metalloproteinase-1 (MMP-1) production inhibitory effect. Through continued research, (2) collagen production and psoriasis, etc. Inhibition of Th1 cell activation, which is a major factor in skin diseases, was found to inhibit interferon-gamma production. (3) The inhibitory effect of matrix metalloproteinase-1 on cancer cell proliferation and metastasis and hot flashes. Inhibition of the formation of neovascularization causing the neovascularization was confirmed, and thus the efficacy of inhibiting neovascularization was confirmed. Therefore, (a) wrinkle improvement of bokbunja fermentation, (b) alleviation of diseases caused by T cell overactivation, such as psoriasis, (c) anti-cancer effect against skin cancer and flushing of face, and (d) cosmetics for obtaining formal effects , Medicine, food and feed.

Efficacy of Bokbunja Fruits in Oriental Medicine Literature is as follows.

-Temper is sweet, flat and without poison. Liver, nerves

-Helps with energy, lightens the body and prevents whitening of hair.

-Shows my stomach, improves sexual function, warms up, and strengthens energy.

-Looks at the liver and brightens the eyes.

-Relieves men's lack of energy, semen exhaustion, and fornication. Also, if a woman eats this, she can have children. (Weakness)

-Heals the liver and gods, reduces the amount of urine, and heals the lungs. (Herbaceous greenery)

As can be seen from the above-mentioned research papers as well as the information of the above-mentioned documents, the efficacy of the bokbunja fermentation disclosed in the present invention is completely new efficacy that is not known as the efficacy of bokbunja except antihistamine efficacy. admit.

The bokbunja fermented product for the purpose of alleviating diseases caused by T cell overactivation such as wrinkle improvement, anti-cancer, flushing and psoriasis, and the effect of promoting growth of bifidus bacteria contained in the present invention is not limited to the fermentation product itself, It includes the chemical itself contained in the reprocessed product and bokbunja fermentation to exhibit the above effects.

The bokbunja fermentation products of the present invention, and their compositions are used for the purpose of alleviating diseases caused by T cell overactivation such as wrinkle improvement, anti-cancer, flushing, psoriasis, and promoting the growth of bifidus bacteria, and are specifically defined in the formulation. Without bars, it can be solid, liquid, spray, gel, sol, cream, patch, for example. In addition, the use is not particularly limited and may be a medicine, food, cosmetics or feed. In addition, in each formulation, components other than the above-mentioned essential components may be selected and blended according to the purpose of use. The bokbunja fermentation product of the present invention may be applied as it is, but may be stored and commercialized after drying by a method such as lyophilization or spray drying.

In the case of the pharmaceutical composition comprising the bokbunja fermentation of the present invention, the method of administration is not limited as much as possible by injection as long as it satisfies manufacturing requirements such as oral administration, transdermal administration as well as removal of pyrogenic substances. The bokbunja fermented product of the present invention has no toxicity when administered because it is a fermented strain of bokbunja, which has been widely used as a food for a long time. The dosage may vary greatly depending on the purpose of use, but in order to obtain the efficacy, it is preferable to base the fermentation product of the present invention on the basis of 10 to 10,000 mg per day of 60 kg of adult dry weight.

Hereinafter, the configuration and effect of the present invention will be described in more detail with reference to Examples and Experimental Examples, but the present invention is not limited only to these examples.

Example 1. Bokbunja Extract

 Purified water 1L was added to 10 g of bokbunja crushed powder, and extracted at 110 ° C. for 2 hours under pressurized conditions. After removing the solid material with 400 mesh, the bokbunja extract was sterilized with a 0.4 μm membrane separator.

Example 2. Bokbunja Extract

 1 g of 70% ethanol was added to 10 g of bokbunja grinding powder, followed by extraction for 24 hours. After removing the solid matter with 400 mesh, the bokbunja extract was prepared by sterilization with a 0.4 µm membrane separator.

Example 3 Bokbunja Fermentation

The bokbunja fermentation broth was prepared by adding 2 g of bokbunja extract of Example 1, 10 g of malt extract, and 50 g of glucose to a 5 L fermenter, and sterilizing at 110 ° C. for 30 minutes, and cooling to 37 ° C. Saccharomyces cerevisiae isolated from makgeolli CBI-YB41 strain (Accession No. KCCM10763P) was inoculated and anaerobic fermented at 37 ° C. and 200 rpm for 3 days. After the fermentation was completed, the solid material was removed at 400 mesh and then sterilized with a 0.4 μm membrane separator to prepare a bokbunja fermented product.

Example  4. Bokbunja Fermented products

Bokbunja immature and ground grinding powder 20g, malt extract 10g and glucose 50g was added to 2L of purified water and sterilized for 30 minutes at 110 ℃ to prepare unripe and fermented broth medium and cooled to 37 ℃. Saccharomyces cerevisiae isolated from makgeolli CBI-YB41 strain (Accession No. KCCM10763P) was inoculated and anaerobic fermented at 37 ° C. and 200 rpm for 3 days. After completion of fermentation, the solid material was removed to 400 mesh and then sterilized with a 0.4 μm membrane separator to prepare bokbunja immature and fermented product.

Comparative Example 1. Bokbunja ripening and extract

 1 g of purified water was added to 10 g of dried powder of Bokbunja dried and crushed powder, and extracted at 110 ° C for 2 hours under pressurized condition. After removing solid matters with 400 mesh, it was sterilized with 0.4㎛ membrane separator to extract Bokbunja ripen fruit. Was prepared.

Comparative Example 2. Bokbunja ripening and fermented product

Bokbunja fermentation broth was added to a 5L fermentation tank, and 2g of bokbunja extract of Comparative Example 1, 10g of malt extract, and 50g of glucose were sterilized at 110 ° C for 30 minutes to prepare a bokbunja fermentation medium and cooled to 37 ° C. Saccharomyces cerevisiae isolated from makgeolli CBI-YB41 strain (Accession No. KCCM10763P) was inoculated and anaerobic fermented at 37 ° C. and 200 rpm for 3 days. After completion of fermentation, the solid material was removed by 400 mesh and then sterilized with a 0.4 μm membrane separator to prepare bokbunja and fermented product.

Cosmetics Prescription Example  1 ~ 2.

The composition of the flexible lotion (skin) in the cosmetic containing the resultant of Example 3 or 4 is as follows.

Raw material Content (% by weight)   Result of Example 3 or 4 10.0   glycerin 3.0   Butylene Glycol 2.0   Propylene glycol 2.0   Carboxyvinyl Polymer 0.1   ethanol 10.0   Triethanolamine 0.1   antiseptic 0.4   Pigment 0.001   Spices 0.1   Purified water Remaining amount

Cosmetics Prescription Example  3 ~ 4.

The composition of the nutrient lotion (lotion) in the cosmetics containing the result of Example 3 or 4 is as follows.

Raw material Content (% by weight)   Result of Example 3 or 4 10.0   Beeswax 4.0   Polysorbate 60 1.5   Solbitan Sesquioleate 0.5   Floating paraffin 5.0   Squalane 5.0   Caprylic / Capric Triglycerides 5.0   Butylene glycol 3.0   Propylene glycol 3.0   Carboxyvinyl Polymer 0.1   Triethanolamine 0.2   antiseptic 0.4   Pigment 0.001   Spices 0.1   Purified water Remaining amount

Cosmetics Prescription Example  5 ~ 6.

The composition of the nutrition cream in the cosmetics containing the result of Example 3 or 4 is as follows.

Raw material Content (% by weight)   Result of Example 3 or 4 10.0   Polyoxyethylene sorbitan monostearate 0.7   Sorbitan sesquioleate 0.5   Cetyl alcohol 0.6   Stearic acid 0.75   Glyceryl Monostearate 0.6   Liquid paraffin 15.0   Carboxy Vinyl Polymer 10.0   Triethanolamine 0.2   antiseptic 0.4   Pigment 0.001   Spices 0.1   Purified water Remaining amount

Cosmetics Prescription Example  7-8.

The composition of the pack in the cosmetic product containing the resultant of Example 3 or 4 is as follows.

       Raw material Content (% by weight)   Result of Example 3 or 4 10.0   Polyvinyl alcohol 13.0   Sodium carboxymethylcellulose 0.2   Allantoin 0.1   ethanol 5.0   Nonylphenyl ether 0.3   antiseptic 0.4   Pigment 0.001   Spices 0.1   Purified water Remaining amount

Cosmetics Prescription Example  9-10.

The composition of the massage cream in the cosmetic composition containing the result of Example 3 or 4 is as follows.

     Raw material Content (% by weight)   Result of Example 3 or 4 10.0   Beeswax 10.0   Polysorbate 60 1.5   Solbitan Sesquioleate 0.8   Floating paraffin 40.0   Squalane 5.0   Caprylic / Capric Triglycerides 4.0   Butylene glycol 3.0   Propylene glycol 3.0   Triethanolamine 0.2   antiseptic 0.4   Pigment 0.001   Spices 0.1   Purified water Remaining amount

medicine Formulation example  1 ~ 2: Tablet

Lyophilized 250.0 mg of Example 3 or 4

Lactose BP 250.0 mg

Starch BP 30.0 mg

Pregelatinized Corn Starch BP 15.0 mg

1.0 mg magnesium stearate

The lyophilisate of Example 3 or 4 was sieved and mixed with lactose, starch and pregelatinized corn starch, then a suitable volume of purified water was added and granulated into a powder. The granules were dried, mixed with magnesium stearate and compressed to prepare tablets, respectively.

medicine Formulation example  3 ~ 4: Capsule

Lyophilized 250.0 mg of Example 3 or 4

Starch 1500 100.0 mg

Magnesium Stearate BP 1.0 mg

The lyophilisate of Example 3 or 4 was sieved and mixed with starch and magnesium stearate, and then filled into gelatin capsules to prepare capsules, respectively.

medicine Formulation example  5 ~ 6: Liquid

200.0 g of the result of Example 3 or 4

637.5 g per bag

2.0 g of sodium carboxymethylcellulose

0.28 g of methyl paraben

0.12 g of propylparaben

20 ml of ethanol

A solution in which sodium carboxymethylcellulose was dissolved in 400 ml of purified water was added to a solution of white sugar in 500 ml of purified water, followed by mixing. Methyl paraben and propyl paraben were added and dissolved therein, followed by ethanol and purified water to make 1000 ml. The resultant of Example 3 or 4 was mixed here, and the liquid formulation was respectively prepared.

medicine Formulation example  7-8: Injection

200.0 mg of the result of Example 3 or 4

Dilute sodium hydroxide pH 7.5 ~ 8.5

Sodium Chloride BP for Injection Up to 2ml

The result of Example 3 or 4 was dissolved in a suitable volume of dilute sodium hydroxide solution to adjust to pH 7.5-8.5, then volume was adjusted using sodium chloride BP for injection and thoroughly mixed. The solutions were filled in 2 ml type ampoules of clear glass, encapsulated under an upper grid of air by dissolving the glass and then sterilized in an autoclave to prepare injections respectively.

food Prescription Example  1 ~ 2: Drink

After diluting 10 ml of the resultant of Example 3 or 4 to distilled water to 80 ml, 7 ml to 10 wt% of liquid fructose and 0.1 to 0.2 wt% of citric acid were added thereto to prepare 100 ml of the mixed beverage.

feed Prescription Example  1 ~ 2: Powder feed

The lyophilisate of Example 3 or 4 was mixed in a general powdery feed in a 1: 100 weight ratio to be used as a powder feed.

Feed Formulation Examples 3-4: Liquid Feed

The resultant of Example 3 or 4 is mixed with general liquid feed or drinking water at 1: 100 weight ratio, and used as a liquid feed.

Experimental Example 1. Collagen production promoting effect

To evaluate the collagen production promoting effect of bokbunja fermentation, primary cultured human fibroblasts were passaged in FGM-2 (Clonetics) medium, treated with trypsin-EDTA, detached from the bottom of the culture vessel, and used as a test medium (0.2). % FBS, 0.1% BSA) medium was replaced with 5 Ｘ 10 ⁴ cells / ml, and then 100 μl of the 96-hole microplate was incubated at 37 ° C. and 5% CO ₂ for 24 hours. After replacing with 135 μl of fresh test medium, 15 μl of bokbunja extract and bokbunja fermented product properly diluted with PBS were added and further incubated for 3 days. ELISA was used to measure the collagen produced from the cells. Specifically, 50 μl of the cell culture solution was transferred to a new 96-hole microplate, coated and washed at 37 ° C. for 2 hours, and then blocked with 0.5% BSA. The secondary antibody was treated with an anti-type 1 procollagen antibody (TaKaRa) to bind to the antigen, washed again and then an anti-IgG antibody conjugate (phosphatase) as a secondary antibody [anti-IgG antibody conjugate (phosphatase): treated with sigma] to bind to the primary antibody, washed, and added with pNPP solution, a substrate solution, reacted for 30 minutes, and then measured for absorbance at 405 nm. Collagen production rate was calculated in comparison with.

Collagen production promoting effect (concentration of the sample by volume) Experimental substance Collagen production rate (%) Comparative Example 2 (0.5%) 145.3 Example 3 (0.5%) 307.3 Example 4 (0.5%) 306.9 TGF-β1 (5 ng / ml) 159.2

From Table 1, it can be seen that bokbunja ripening and fermented products (Comparative Example 2), bokbunja fermented products (Examples 3 and 4) all have collagen production promoting effect, but bokbunja fermented products of the present invention (Examples 3 and 4) ) Showed significantly higher collagen production promoting effect, which is significantly higher than the collagen production promoting effect at the maximum effective concentration of TGF-β1, which is known as a major cytokine for the regulation of collagen production. Wrinkle improvement cosmetics was found.

Experimental Example 2. Inhibitory Effect of MMP-1 Production

To evaluate the inhibitory efficacy of matrix metalloproteinase-1 (MMP-1) production, primary cultured human fibroblasts were passaged in FGM-2 (Clonetics) medium and treated with trypsin-EDTA and removed from the bottom of the culture vessel. After replacing with DMEM (2% FBS, 0.1% BSA) medium used as a test medium to make 2 Ｘ 10 ⁴ cells / ml, dispense 100 μl into a 96-hole microplate at 37 ° C and 5% CO ₂ . Incubated for 24 hours. After replacing with 135 μl of fresh test medium, 15 μl of bokbunja extract and bokbunja fermented samples diluted appropriately with PBS were added and further incubated for 3 days. ELISA was used to measure the MMP-1 produced from the cells. Specifically, 50 µl of the cell culture solution was transferred to a new 96-hole microplate, coated and washed at 37 ° C. for 2 hours, and then blocked with 0.5% BSA. The antibody was treated with an anti-matrix metalloproteinase-1 antibody (Sigma) to bind the antigen and washed again, followed by an anti-IgG antibody conjugate (phosphatase) as a secondary antibody. anti-IgG antibody conjugate (phosphatase): treated with Sigma] to bind to the primary antibody, washed, added pNPP solution, a substrate solution, reacted for 30 minutes, and then absorbed at 405 nm. The yield was calculated by comparing the absorbance of the culture.

Inhibitory Effect on MMP-1 Production Experimental substance MMP-1 production rate (%) Experimental substance MMP-1 production rate (%) Example 3 (1.000 v / v%) 23.1 Example 4 (1.000 v / v%) 24.3 Example 3 (0.100 v / v%) 25.7 Example 4 (0.100 v / v%) 25.1 Example 3 (0.010 v / v%) 49.1 Example 4 (0.010 v / v%) 48.7 Example 3 (0.001 v / v%) 76.5 Example 4 (0.001 v / v%) 77.1

It can be seen from Table 2 that the bokbunja fermentation products of the present invention (Examples 3 and 4) were effective in inhibiting MMP-1 production even at very low concentrations (0.001 v / v%). MMP-1 is an enzyme that breaks down collagen. Inhibition of MMP-1 is used as an indicator of the efficacy of wrinkle improvement materials in the cosmetic industry. In addition, MMP-1 is an essential process for cancer metastasis. It has been shown to induce vasodilation, the cause of flushing. Therefore, it can be seen that the bokbunja fermented product of the present invention has not only anti-wrinkle effect but also cancer metastasis suppression and hot flashes relief effect.

Experimental Example 3. Angiogenesis inhibitory effect

CAM (chorioallantoic membrane) assay was used to evaluate the neovascularization inhibitory effect. In detail, chicken eggs were incubated at 37 ° C. for 3 days. Samples were added to the incubated CAMs and dried coverslips were raised and further incubated for 2 days to evaluate angiogenesis inhibition efficacy.

Inhibition of neovascularization sample Test amount Number of eggs showing inhibitory activity / number of eggs tested Control (PBS) 10 μl 0/10 10-fold dilution of Example 3 10 μl 9/10 10-fold dilution of Example 4 10 μl 9/10

From the results of Table 3, it can be seen that the bokbunja fermented product of the present invention (Examples 3 and 4) has an angiogenic inhibitory effect.

Experimental Example 4. Interferon-gamma production inhibitory effect

To assess the efficacy of INF-γ inhibition, the spleen cells of mice were suspended in RPMI (10% FBS) medium at 1 × 10 ⁶ cells / ml and in a concentration of 0.5 ug / ml to induce INF-γ. After adding PHA (Phytohemaglutinin) and 25ng / ml PMA (phorbol 12-myristate 13-acetate), 150 μl were dispensed into a 96-hole microplate and 15 μl of the sample was added thereto, followed by incubation for 24 hours. INF-γ was measured in the culture medium using a Biosource ELISA kit.

Interferon-gamma production inhibitory effect Experimental substance INF-γ production rate (%) Experimental substance INF-γ production rate (%) Example 3 (2.27 v / v%) 21.12 Example 4 (2.27 v / v%) 22.18 Example 3 (0.57 v / v%) 31.86 Example 4 (0.57 v / v%) 29.25 Example 3 (0.14 v / v%) 43.21 Example 4 (0.14 v / v%) 44.54

Table 4 confirmed that the bokbunja fermentation of the present invention (Examples 3 and 4) has an interferon-gamma production inhibitory effect. Since interferon gamma is known as a factor that inhibits collagen production in vivo, it can be seen that the inhibitor of the production of interferon gamma has an effect of improving wrinkles. In addition, since interferon gamma is known as a major factor of psoriasis, it can be seen that the bokbunja fermentation product of the present invention also has psoriasis relaxing effect.

Experimental Example 5. Antihistamine Effect

To evaluate the antihistamine efficacy, mast cells ATCC TIB-64 was precultured in modified DMEM (10% FBS) medium for 48 hours, washed three times with buffer solution, and then washed to 5 X 10 ⁵ cells / ml. Dispense the amount of released histamine using a histamine quantitative kit (Oxford Biomedical Research) after dispensing on the microplate, adding Bokbunja extract and Bokbunja fermentation, reacting for 30 minutes, and adding C48 / 80 to induce histamine release. It was.

Antihistamine effect Experimental substance Histamine release rate (%) Experimental substance Histamine release rate (%) Experimental substance Histamine release rate (%) Example 1 (4.17 v / v%) 35.2 Example 3 (4.17 v / v%) 29.9 Example 4 (4.17 v / v%) 30.2 Example 1 (1.04 v / v%) 31.6 Example 3 (1.04 v / v%) 28.2 Example 4 (1.04 v / v%) 31.3 Example 1 (0.52 v / v%) 65.6 Example 3 (0.52 v / v%) 36.1 Example 4 (0.52 v / v%) 34.6

 Table 5 shows that both bokbunja extract (Example 1) and bokbunja fermentation (Examples 3 and 4) have an anti-histamine effect and anti-histamine efficacy is improved by yeast fermentation of bokbunja. It is known that Bokbunja extract has antihistamine efficacy, and the Bokbunja fermentation product of the present invention enhances the efficacy of Bokbunja extract.

Experimental Example 6. Bifidus growth promoting effect

Bifidobacterium adolescentis was precultured in brain heart infusion broth and then inoculated in the same medium in 1% volume to evaluate the effect of Bifidobacterium growth promotion. Diluted solution was added and cultured at 37 ° C. for 16 hours, and then absorbance was measured at 630 nm to evaluate bifidus growth promoting effect.

Bifidus growth promoting effect Experimental substance Absorbance Experimental substance Absorbance Example 3 (5.000 v / v%) 0.206 Example 4 (5.000 v / v%) 0.212 Example 3 (2.500 v / v%) 0.171 Example 4 (2.500 v / v%) 0.169 Example 3 (1.250 v / v%) 0.133 Example 4 (1.250 v / v%) 0.135 Example 3 (0.625 v / v%) 0.118 Example 4 (0.625 v / v%) 0.121 Example 3 (0.313 v / v%) 0.115 Example 4 (0.313 v / v%) 0.113 Example 3 (0.156 v / v%) 0.109 Example 4 (0.156 v / v%) 0.111 Example 3 (0.078 v / v%) 0.108 Example 4 (0.078 v / v%) 0.108 Example 3 (0.039 v / v%) 0.105 Example 4 (0.039 v / v%) 0.106 Example 3 (0.020 v / v%) 0.104 Example 4 (0.020 v / v%) 0.104 Example 3 (0.000 v / v%) 0.101 Example 4 (0.000 v / v%) 0.102

 Table 6 shows that bokbunja fermentation (Examples 3 and 4) has a bifidus growth promoting effect.

Each of the various compositions, i.e., each cosmetic composition exhibiting anti-wrinkle efficacy, psoriasis relief, anti-cancer and flushing efficacy and formal efficacy, including bokbunja fermented products as a non-toxic natural ingredient by the present invention; Pharmaceutical composition; Food compositions; And feed composition.

Claims (13)

Bokbunja fermented product obtained by inoculating and fermenting at least one microorganism selected from lactic acid bacteria, yeasts, Bacillus subtilis, Hwangkuk fungi and red yeast fungi. The bokbunja fermentation according to claim 1, wherein the bokbunja is bokbunja itself or its grinding powder, or extract of bokbunja. The bokbunja fermentation according to claim 1, wherein the yeast is Saccharomyces cerevisiae CBI-YB41 strain (Accession No. KCCM10763P). The bokbunja fermentation according to claim 1, which is obtained after forming a mixture before fermentation using nutrients or purified water together with bokbun or further using both nutrients and purified water. The bokbunja fermentation according to claim 4, wherein the content of bokbunja in the mixture before fermentation is 0.01 to 10% by weight based on dry weight of bokbunja. The bokbunja fermentation according to claim 4, wherein the nutrient is at least one component selected from yeast extract, soy protein, malt extract, peptone, tyrosine, cysteine, phytic acid, glucose, maltose, lactose, sucrose and salts. . The bokbunja fermentation according to claim 1, which is further obtained by centrifugation or membrane separation to remove the cells and insoluble materials, followed by sterilization or sterilization. Claim 1 cosmetic composition for improving wrinkles comprising the bokbunja fermentation. Cosmetic composition for psoriasis relaxation comprising the bokbunja fermentation of claim 1. Claim 1 anti-skin cancer and flushing cosmetic composition comprising the bokbunja fermentation. Wrinkle improvement, psoriasis relief, anticancer and flushing or formal medicine composition comprising the bokbunja fermentation of claim 1. Wrinkle improvement, psoriasis relief, anticancer and redness relief or formal food composition comprising the bokbunja fermentation of claim 1. Psoriasis alleviation, anticancer and flushing or formal feed composition comprising the bokbunja fermentation of claim 1.

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