KR100962587B1 - A method for fermentation of natural plants and herbal medicines, a fermented product prepared therefrom and a paharmaceutical composition, a cosmetic compositon and a food composition comprising the product - Google Patents

Abstract

Provided are fermentation methods of natural plant materials and herbal medicines using Nuruk and lactic acid bacteria, fermented products prepared by the above methods, and cosmetic compositions and food compositions containing the same.

According to the present invention, it is possible to increase the fermentation efficiency of lactic acid bacteria without the addition of a carbon source by fermenting a natural plant material or a physiological activity that is easily ingested in daily life and using medicinal herbs used in oriental medicine using Nuruk and lactic acid bacteria. Since water is very stable and safe and has excellent efficacy of anti-aging, anticancer, anti-inflammatory, and immunity, it may be usefully used in pharmaceutical compositions, cosmetic compositions and food compositions, fermentation methods of natural plant materials and herbal medicines, the method It can be provided a fermentation product prepared by and a pharmaceutical composition, a cosmetic composition and a food composition containing the same.

Description

Method for fermentation of natural plants and herbal medicines, a fermented product prepared there from and a paharmaceutical composition , a cosmetic compositon and a food composition comprising the product}

The present invention relates to a fermentation method of natural plant materials and herbal medicines using Nuruk and lactic acid bacteria, fermented products prepared by the above method, and cosmetic compositions and food compositions containing the same, more specifically, natural foods that are easily ingested in daily life. By fermenting herbal medicine used in oriental medicine with yeast and lactic acid bacteria because of strong vegetable material or physiological activity, lactic acid bacteria fermentation efficiency can be increased without adding carbon source, and the prepared fermented products have excellent stability and safety, and have excellent anti-aging and anticancer activity. , Anti-inflammatory, immunity improvement, etc., which can be usefully included in pharmaceutical compositions, cosmetic compositions and food compositions, fermentation methods of natural plant materials and herbal medicines, fermented products prepared by the method and pharmaceutical compositions containing the same, Of cosmetic compositions and food compositions to be.

With the advent of an aging society in recent years, well-being trends are taking root throughout the society, which has amplified the interest in herbal medicines that have been introduced to natural vegetable materials and traditional folk remedies in all fields such as food, cosmetics, and residential environment.

In the cosmetics industry, where natural plant materials are relatively easy to commercialize, there have been attempts to use them as raw materials for a long time. In the fields of functional foods, preventive medicine and therapeutic medicine, oriental herbal medicine has been used in the US and Europe. As licensing and commercialization began, interest in the human utility of natural plant materials and herbal medicines increased.

Natural plant material or herbal medicine contains various biologically active substances and antioxidants, so it shows various effects such as anti-aging, anti-cancer, anti-inflammatory and immune function improvement, but it is unstable and sometimes irritating after extraction, so it is toxic to human body. Often. Therefore, in recent years, many attempts have been made to stabilize the natural extracts, reduce their toxicity, or convert them into stable derivatives to increase their effectiveness. As a part, biological transformation methods using microorganisms or enzymes have been developed. Representative methods include fermentation as an example.

Fermentation is a process of producing a substance that is useful for human life by decomposing organic substances using enzymes of microorganisms. Representative fermentation microorganisms that produce substances useful for the human body include lactic acid bacteria, yeast, and various foods. Produced by fermentation process. In addition to producing useful substances, fermentation also plays a role in immunity and detoxification by the lactic acid bacteria involved in fermentation. Furthermore, the components ingested through fermentation by the intestinal microorganisms are decomposed and converted into low-molecular substances that are easily absorbed by human cells, or converted into unstable or inactive forms in the active form, which is absorbed by the intestinal microorganisms. This shows the importance of biotransformation.

Accordingly, attempts to maximize the efficacy of natural plant materials and herbal medicines or solve safety problems by microbial fermentation such as lactic acid bacteria and yeast have important meanings in the development of various foods, cosmetics, medicines and the like. In particular, the microorganisms that propagate in traditional Korean foods such as Kimchi and Doenjang, which are reported to have excellent anticancer effect, anti-inflammatory effect, anti-aging effect, and immunity enhancing effect, are reported to have excellent biological activity and are isolated from traditional fermented foods. Many studies are underway to exploit a microbe.

Nuruk, a kind of microorganism that propagates in Korean traditional food, has been used to make alcohol in Korea since ancient times. Nuruk is a fungus with alcohol-producing fungi that grows in cereals. The main role of nuruk is to saccharify starch-like raw materials such as rice. In the primitive age, salivary amylase was used to make sake, but later fungal mycelium was applied. The use of yeast or malt that has sprouted by dipping barley has become popular. Currently, Nuruk is widely used to increase citric acid and glycosylation by cultivating bacteria such as the main strains Rhyzopus, Usami and Oryzae.

Conventional natural plant materials and herbal medicine techniques are mostly using simple extracts, supercritical extracts, extracts using a variety of solvents, but some of these natural plant materials and herbal medicines are carried out fermentation process using yeast fungi or lactic acid bacteria Techniques have been disclosed. For example, Korean Patent Laid-Open Publication No. 2005-64688 discloses an invention for fermenting and extracting soybean by allowing yeast mold to be propagated using rice straw in preparing a health beverage. No. 6960 discloses a cosmetic composition prepared by fermenting mung beans with lactic acid bacteria.

However, natural plant materials and herbal medicines contain various components according to their types, and among them, materials containing a lot of antimicrobial ingredients or materials containing no carbon source in a form easily used by microorganisms may contain lactic acid bacteria. Even if the fermentation is performed by inoculation, the reaction is not performed well. In order to solve this problem, a method of supplying a separate carbon source or using a purified enzyme has been developed. However, since several enzymes have to be added separately, the process is complicated and economically disadvantageous.

Therefore, the first problem to be solved by the present invention, by effectively linking the saccharification step using the yeast and fermentation step by lactic acid bacteria, fermentation method of natural plant material and herbal medicine that can maximize the lactic acid bacteria fermentation efficiency without the addition of additional carbon source To provide.

The second problem to be solved by the present invention is to provide a fermentation product prepared by the above method, which is very excellent in stability and safety and has excellent efficacy of anti-aging, anticancer, anti-inflammatory, immunity and the like.

The third object of the present invention is to provide a pharmaceutical composition containing the fermented product.

The fourth problem to be solved by the present invention is to provide a cosmetic composition containing the fermented product.

The fifth problem to be solved by the present invention is to provide a food composition containing the fermented product.

In order to achieve the first object of the present invention,

Saccharification step using yeast;

Fermentation step through lactic acid bacteria culture; And

Obtaining a supernatant from the culture result

It provides a natural plant material and fermentation method of the herbal medicine.

According to a preferred embodiment of the present invention, the saccharification step may include the step of pulverizing natural plant material and herbal medicine, adding the yeast, and immersing.

According to another preferred embodiment of the present invention, the pulverized product of the grinding step may have an average particle diameter of 10㎛ to 1mm.

According to another preferred embodiment of the present invention, the natural plant material and herbal medicine is ginger, brown root, licorice, green tea, kelp, Angelica, Magmundong, Bokbunja, Cornus, Sangbaekpi, Schisandra chinensis, Ginkgo biloba, Licorice, Astronomical dong, Gardenia, Chamomile, Shiitake mushroom, persimmon leaf, kidney bean, cinnamon, red ginseng, gold and silver acorn, acorn, bark, pogongyoung, situation mushroom, pomegranate, matsutake, horse riding, wormwood, ganoderma lucidum, gall bladder, burdock, yulpi, golden, rhubarb, yellow white, chrysanthemum, gilkyung, Bamboo, cactus, jade juk, baekryeok, white paper, fueryeong, loquat, shiho, aloe, motherwort, chajeoncho, changza, cheongung, astronomical dong, earth and sand, sesao, passerby, brown rice, safflower, gamgu, deficiency, oats, mung bean, jujube, deodeok , Sweet pumpkin, cabbage, machi, quince, black bark, thyme, sanda flower, eoseongcho, lotus root, hard bones, gwahyang, rhubarb, article, white weight, baekbuja, white paper, casualty, ginseng, red citrus fruit, cabbage, cabbage, ginseng, red ginseng, cheonhwabun, Tomatoes, papayas, mutters, almonds, bran Flesh, Oat, Deer Antler, Cordyceps Sinensis, Grape Seed, White Nymph, Agaricus Mushroom, Chaga Mushroom, Lotus Root, Turmeric, Red Pepper, Garlic, Wolfberry, Guava, Gujeolcho, Golden Pine, Root, Elm, Arbor, Tortilla, Perilla, Rooibos, Hemp, Ephedra , Plum, plum, white, thyme, centella, golgoji, borage, bokbunja, non-recovery, visa, loquat, betel nut, mountain mulberry, cornus, sancho, fresh vinegar, acerola, acacia, wormwood, olive leaf, wasong, dew Garden, nutmeg, ginkgo biloba, root, purple leaf, bamboo shoot, earth, jimo, jisil, jihwang, chajeoncho, changja, irises, irises, caviar, chlorella, spirulina, earthenware, parsley, pogongyoung, mutter, pedestrian, hamcho, walnut , Fennel or mixtures thereof.

According to another preferred embodiment of the present invention, the amount of the yeast added may be 1 to 10 parts by weight based on 100 parts by weight of the pulverized product.

According to another preferred embodiment of the present invention, the amount of the yeast added may be 2 to 3 parts by weight based on 100 parts by weight of the pulverized product.

According to another preferred embodiment of the present invention, the immersion step may be performed for 18 to 30 hours at a temperature of 20 to 40 ℃.

According to another preferred embodiment of the present invention, the method may further include sterilizing the resultant after the immersion step.

According to another preferred embodiment of the present invention, the strain used to prepare the yeast is spider web fungus ( rhyzopus nigricans ), yeast fungus ( Aspergillus oryzae ) or a mixed strain thereof.

According to another preferred embodiment of the present invention, the lactic acid bacteria is Lactobacillus Sakei ( Lactobacillus sakei B2-16), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus flange tareom (Lactobacillus plantarum P23 ), Lactococcus lactis A164 ), Bacillus coagulum coagulance ) or a mixed strain thereof.

According to another preferred embodiment of the present invention, during the fermentation step through the lactic acid bacteria culture Saccharomyces yeast Saccharomyces ( Saccharomyces cerevisiae ) strain may be added further.

According to another preferred embodiment of the present invention, the inoculation amount of the lactic acid bacteria or yeast may be 10 ⁵ to 10 ⁸ cfu / g or 10 ⁵ to 10 ⁸ cfu / ㎖ initial inoculation.

According to another preferred embodiment of the present invention, the lactic acid bacteria is a strain isolated from kimchi or miso, the yeast may be a strain used in the production of sake.

According to another preferred embodiment of the present invention, the fermentation step through the lactic acid bacteria culture may be performed for 18 to 120 hours at a temperature of 15 to 40 ℃.

According to another preferred embodiment of the present invention, the supernatant obtaining step may be performed by filtration or centrifugation.

In order to achieve the second object of the present invention,

It provides a fermented product prepared by the fermentation method of the natural plant material and herbal medicine.

The present invention to achieve the third object,

It provides a pharmaceutical composition containing the fermented product.

In order to achieve the fourth object of the present invention,

It provides a cosmetic composition containing the fermented product.

In addition, the present invention to achieve the fifth object,

It provides a food composition containing the fermented product.

Hereinafter, the present invention will be described in more detail.

The present invention intends to use the fermentation process as a biological conversion method for such natural plant material or herbal medicine, in order to use more physiological activity of natural plant material or herbal medicine more stable, less toxic, and effective, and saccharification using yeast By effectively linking the step and the fermentation step by lactic acid bacteria, it was intended to maximize the lactic acid bacteria fermentation efficiency without the addition of a separate carbon source.

As a biological conversion method for natural plant material or herbal medicine, the most economical and proven method of safety is fermentation method. However, in order to perform the fermentation process using microorganisms smoothly, it is necessary to provide conditions for growth of the microorganisms, but most natural plant materials and herbal medicines cannot provide such an environment. This is because glucose is easy to use as a carbon source in order to grow microorganisms well, but most natural plant materials and herbal medicines do not have many carbon sources of this type. Therefore, culturing fermented microorganisms on various natural plant materials or herbal medicines known to have good physiological activity makes it difficult to induce the desired biological conversion.

In the present invention, Nuruk (or grain) and lactic acid bacteria were used to solve this problem. In general, yeast means that molds containing alcohol-producing enzymes are propagated in grains such as rice. The main role of yeast is to saccharify starch raw materials present in grains. Therefore, by using the result of the saccharifying action of yeast as a growth nutrient of lactic acid bacteria, it is possible to perform fermentation on natural plant material and herbal medicine in an effective and convenient way.

Specifically, the fermentation method of natural plant material and herbal medicine according to the present invention, the saccharification step using Nuruk; Fermentation step through lactic acid bacteria culture; And obtaining a supernatant from the culture result, in particular, the saccharification step may include pulverizing natural plant material and herbal medicine, adding yeast, and immersing.

1 schematically shows a process diagram for a fermentation method of natural plant material and herbal medicine according to the present invention, hereinafter, the present invention will be described in more detail with reference to FIG.

First, the grinding step is to facilitate the saccharification process by the yeast, the grinding material obtained from the grinding step may have an average particle diameter of 10㎛ to 1mm.

Natural plant materials and herbal medicines to which the present invention can be applied include, but are not limited to, various substances having various physiological activities useful for the human body, but are not limited thereto. Ginger, brown root, licorice, green tea, kelp, Angelica, and Macmundong , Bokbunja, Cornus, White peach, Schisandra chinensis, Ginkgo biloba, Korean herb, Astronomical dong, Gardenia, Chamomile, Shiitake, Persimmon leaf, Kidney bean, Cinnamon, Red ginseng, Gold silver coin, Acorn, Peony, Pogongyoung, Situation mushroom, Pomegranate, Pine mushroom, Horse riding, Mugwort, Ganoderma lucidum, gall bladder, burdock, yulpi, golden, yellow lotus, yellow white, chrysanthemum, gilgyeong, bamboo, cabbage, jade porridge, count, white paper, fueryeong, loquat, siho, aloe, motherwort, chajeoncho, changja, cheongung, cheonmundong, tosa, Shou, pedestrian, brown rice, safflower, gums, glitches, oats, mung beans, jujube, deodeok, sweet pumpkin, cabbage, machi, quince, bark skin, thyme, sanda flower, eoseongcho, lotus root, kobon, kwak hyang, rhubarb, article, salary, baekbuja, Blank paper , Prickly pear, tangerine, cabbage, tofu, ginseng, red ginseng, natural flower, tomato, papaya, pine rapeseed, almond, mistletoe, oat, antler, cordyceps, grape seed, white goose, agaricus mushroom, chaga, lotus root, turmeric, red pepper, garlic, Gugija, guava, gujeolcho, calendula, root root, elm, arthropod, tofu, perilla, rooibos, hemp, ephedra, plum, plum, white, thyme, centella, golboji, borage, bokbunja, non-repair, visa, loquat leaf, betel nut Purple, mountain mulberry, Cornus, Sancho, Sinseoncho, acerola, acacia, wormwood, olive leaf, hawthorn, dew, grass, nutmeg, ginkgo biloba, root, purple leaf, bamboo shoot, earthly, jimo, jisil, jihwang, chajeoncho, changja Examples include irises, nectars, caviar, chlorella, spirulina, earthenware, parsley, pogongyoung, sea stalks, almonds, seaweeds, walnuts, fennel or mixtures thereof.

On the other hand, as a raw grain for producing yeast, a wheat flour or a wheat flour is used as a raw material, and mung bean, glutinous rice or barley may be used together. In addition, yeasts include dokmari, barley straw, bark straw, mulberry, wormwood, resident, lotus, young leaves of pine needles, pine needles and the like. Raw grains can be cooked or roasted, steamed, steamed, or raw wheat. The grain size can range from 12% to fine powder. Types of main ingredients include mulberry leaves, mugwort, lactating juice, peach seed powder, apricot seed powder and melon powder.

The amount of yeast added for the saccharification process may be 1 to 10 parts by weight, preferably 2 to 3 parts by weight, based on 100 parts by weight of the pulverized product. If the amount of yeast is more than 10 parts by weight, there is a concern such as the occurrence of a mold odor, if less than 1 part by weight does not cause saccharification enough, there is a problem that the subsequent lactic acid bacteria fermentation efficiency is not preferable.

The yeast fungus is generally classified into yellow yeast, black yeast or red yeast fungus according to its color, and is also classified into rice yeast (sick soup) or scattered yeast (new soup). A fungus that inhabits the byeongguk is geared as a fermented type from the inside, there is a cobweb mold (rhyzopus nigricans), yeast, mold and hair, a fungus that lives in the new national is a breathing that breed only on the surface, koji mold (Aspergillus oryzae ).

In manufacturing yeast in the present invention, the rice cake yeast or scattered yeast may be used evenly, in particular, spider web mold ( rhyzopus nigricans ), yeast fungi ( Aspergillus oryzae ) or mixed strains thereof are preferably used.

The soaking step after the addition of the yeast is preferably carried out at a temperature of 20 to 40 ° C. for 18 to 30 hours, since this is the optimum temperature that indicates the activity of the yeast fungus derived enzyme. Organic acids produced by the yeast fungus at this temperature include kojic acid and gluconic acid, and the enzymes induced include amylase, maltase, invertase, cellulase, inulinase and protease.

In addition, after the immersion step, in order to increase the purity of the fermentation product obtained from the fermentation step through the subsequent lactic acid bacteria culture, sterilization step may be further performed, such sterilization step is for example 20 minutes at 121 ℃, 1.5 atmosphere By heat treatment for from 40 minutes.

Next, when the saccharification step is completed as described above, the fermentation step through the lactic acid bacteria culture is performed.

Lactic acid bacteria that can be used in the present invention, but is not limited thereto, Lactobacillus sakei B2-16), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus flange tareom (Lactobacillus plantarum P23 ), Lactococcus lactis A164 ), For example, Bacillus coagulance or a mixed strain thereof, including the yeast Saccharomyces ( Saccharomyces) cerevisiae ) strain may be added.

The inoculation amount of the lactic acid bacteria or yeast is preferably 10 ⁵ to 10 ⁸ cfu / g or 10 ⁵ to 10 ⁸ cfu / ㎖ initial inoculation, when the initial inoculation bacteria is less than 10 ⁵ cfu / g or 10 ⁵ cfu / ㎖ There is a problem in that the fermentation time is long, and when the amount exceeds 10 ⁸ cfu / g or 10 ⁸ cfu / ㎖ there is a problem of excessive inoculation amount even if the increase of the effect is not large.

The lactic acid bacteria can be easily separated from kimchi or doenjang, which is a traditional Korean food, and the yeast may also use strains used in the production of sake.

On the other hand, the fermentation step through the lactic acid bacteria culture, when considering the optimum lactic acid bacteria fermentation conditions, may be performed for 18 to 120 hours at a temperature of 15 to 40 ℃.

Finally, a step of obtaining a supernatant from the fermentation product through the lactic acid bacteria culture is performed, which may be performed by various methods such as filtration or centrifugation.

The second object of the present invention is to provide a fermented product prepared by the fermentation method of the natural plant material and herbal medicine, the fermentation product according to the present invention, as described in the following examples, collagen synthesis promoting effect, skin whitening Excellent effect, antioxidant effect, etc., to be included as an active ingredient in various pharmaceutical compositions, cosmetic compositions or food compositions having the effects of preventing and improving skin aging, assisting skin turnover, preventing and improving skin damage, antioxidant, anti-inflammatory, etc. Can be.

Accordingly, the third, fourth and fifth task of the present invention is to provide a pharmaceutical composition, a cosmetic composition and a food composition containing the fermentation product, and the compositions according to the present invention provide the main effects aimed at by the present invention. It may contain other ingredients that can give a synergistic effect to the main effect within the range of not being damaged, and for example, fragrances, pigments, fungicides, antioxidants, preservatives, humectants, thickeners, inorganic salts, In addition to an additive such as an emulsifier and a synthetic polymer, it may further include auxiliary ingredients such as water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, and seaweed extracts. The components may be appropriately selected and blended by those skilled in the art according to the formulation or purpose of use, and the amount may be selected within a range that does not impair the object and effect of the present invention, but 0.01 to 5 weight based on the total weight of the composition. %, Preferably 0.01 to 3% by weight.

In addition, the formulation of the composition according to the present invention may also be in various forms such as solution, emulsion, viscous mixture, tablet, powder, etc., which may be administered by various methods such as simple drinking, injection, spray or squeeze method. .

According to the present invention, it is possible to increase the fermentation efficiency of lactic acid bacteria without the addition of a carbon source by fermenting a natural plant material or a physiological activity that is easily ingested in daily life and using medicinal herbs used in oriental medicine using Nuruk and lactic acid bacteria. Since water is very stable and safe and has excellent efficacy of anti-aging, anticancer, anti-inflammatory, and immunity, it may be usefully used in pharmaceutical compositions, cosmetic compositions and food compositions, fermentation methods of natural plant materials and herbal medicines, the method It can be provided a fermentation product prepared by and a pharmaceutical composition, a cosmetic composition and a food composition containing the same.

Hereinafter, the present invention will be described in more detail with reference to Examples and Evaluation Examples. However, these examples and evaluation examples are intended to explain the present invention in more detail, and the scope of the present invention is not limited by these examples and evaluation examples according to the gist of the present invention is common knowledge in the art. It will be self-evident for those who have.

Example  1. Natural plant material or herbal medicine according to the present invention Fermented product  Produce

100 g of brown root was washed with cold water of 30 ° C. or less for about 30 minutes, pulverized with a pulverizer, and then water was added so that the pulverized powder was 3% by weight based on solids. . Nuruk is mixed with wheat grass and water at a ratio of 5: 1 and mixed well, and then inoculated with cultured spider fungus ( rhyzopus nigricans ), yeast fungus ( Aspergillus oryzae ) or a mixed strain thereof, and inoculated with 3∼3 at 25 ° C to 30 ° C. The culture was used for 5 days. At this time, the amount of Nuruk added was 2.5 parts by weight based on 100 parts by weight of the pulverized product. After immersion, the immersion product was sterilized by heat treatment at 121 ° C. for 30 minutes. Subsequently, Lactobacillus , previously cultured plantarum P23 was added to a concentration of 10 ⁷ cfu / ml, then incubated at 37 ° C. for 3-5 days, and the supernatant was collected using a continuous centrifuge.

Example  2 to 32. Natural plant material or herbal medicine according to the present invention Fermented product  Produce

Instead of brown root, licorice (Example 2), green tea (Example 3), kelp (Example 4), Angelica (Example 5), Macmundong (Example 6), Bokbunja (Example 7), Cornus milk (Example) Example 8) Ginger (Example 9), Morus bark (Example 10), Schisandra chinensis (Example 11), Ginkgo biloba (Example 12), Licorice (Example 13), Astronomical dong (Example 14), Gardenia (Example) 15), chamomile (Example 16), shiitake mushroom (Example 17), situation mushroom (Example 18), red ginseng (Example 19), pomegranate (Example 20), wormwood (Example 21), sewage Example 22) Casualty (Example 23), Pedestrian (Example 24), Aloe (Example 25), Muduck (Example 26), March (Example 27), Eochocho (Example 28), Archery (Example) 29), fermentation according to Examples 2 to 32 by the same method as in Example 1, except that poongyoung (Example 30), thorn stem (Example 31), and worms (Example 32) were used. Waters were prepared.

Comparative example  1 to 32. Natural plant material or herbal medicine according to the prior art Fermented product  Produce

A fermentation product was obtained in the same manner as in Example 1, except that the fermentation step was performed directly through lactic acid bacteria culture without performing the saccharification step using Nuruk.

Evaluation example  1. Evaluation of fermentation efficiency

PH was used as a measure for comparing the fermentation degree of the fermentation products according to Examples 1 to 32 and Comparative Examples 1 to 32. In the case of lactic acid bacteria, the pH showed a tendency to decrease in proportion to the growth of the cells, and the fermentation efficiency was shown by comparing the degree of drop in pH. The results are shown in Table 1 below. Referring to Table 1, the fermentation efficiency (% reduction in pH) is a percentage of the pH which was further reduced in the examples when compared with the comparative example. For example, in the case of Example 1 for rooting, the pH was further reduced by 15% compared to Comparative Example 1.

Evaluation example  2. Cytotoxicity Test

After inoculating the skin cells (Fibroblast) in 96-well plate, the medium to which the fermentation products according to Examples 1 to 32 were added for each concentration was incubated for 1 to 2 days. After the medium was removed, MTT solution was added and reacted for 2 to 3 hours. Then, the MTT solution was removed, DMSO was added, and the formazone crystal was mixed. Absorbance was measured to compare the degree of cell death (toxic concentration), and compared with the extract of the natural plant or herbal medicine used in Examples 1 to 32, the result of expressing the concentration of the fermentation product added to show the same cell death in multiples. Shown in For example, in the case of Example 1 for the root of the roots compared to the extract of the root of the root itself, the concentration added to show the same apoptosis can be seen that the cytotoxicity is significantly reduced in the case of the fermentation according to the present invention.

Evaluation example  3. Fermented product  Whitening effect

The whitening effect of the fermented products according to Examples 1 to 32 was evaluated by the following three methods, the results are shown in Table 1 below. The whitening effect of the fermentation product is based on the melanin synthesis inhibitory effect and tyrosinase expression inhibition effect in melanin tumor cells as follows-(no whitening effect), + (shows a slight whitening effect), ++ (whitening effect Excellent), +++ (the whitening effect is very good), and the results of Example 17 for which the whitening effect is most excellent are shown in FIGS. 2 and 3.

Inhibitory Effects of Melanin Synthesis on Melanin Tumor Cells

In order to determine the effect of inhibiting melanin synthesis in melanin tumor cells of the fermentations according to Examples 1 to 32 were carried out the following experiment.

First, the mouse-derived malignant melanin tumor cell line B16F1 (KCLB 8007) was added to DMEM (Dulbecco's Modified Eagle's Medium) medium containing penicillin (100 IU / mL), streptomycin (100 g / mL), and 10% FBS (fetal bovine serum). Incubated in an incubator containing 5% carbon dioxide while maintaining 37 ° C. The obtained melanin tumor cells were seeded in 1 × 10 ⁴ cells / well in a 24-well plate and cultured for 1 day. After incubation, the medium was removed, and then, in order to induce melanin synthesis, melanin synthesis was induced by adding 50 nM of alpha-MSH (melanocyte stimulating hormone; melanocyte stimulating hormone), except for the untreated group. The fermentation products obtained through the 32 to 32 were incubated for 3 days by adding DMEM medium added 0.1%, 0.5%, 1% and 800 μM kojic acid (positive control), respectively. The amount of synthesized melanin secreted by melanin tumor cells by each treatment group was visually checked and the change was photographed by a digital camera. The cells of each treatment group were collected and 100 μl of 10% DMSO (dimethyl sulfoxide) and 1N NaOH were collected. Each was added to dissolve the melanin, the absorbance at 405nm was measured to measure the amount of melanin and the results for Example 17 are shown in FIG.

2) Tyrosinase  Expression inhibitory effect

In order to determine the effect of inhibiting tyrosinase expression of the fermented products according to Examples 1 to 3, the following experiment was performed.

First, the mouse-derived malignant melanin tumor cell line B16F1 (KCLB 8007) was replaced with DMEM (Dulbecco's Modified Eagle's Medium) containing penicillin (100 IU / mL), streptomycin (100 g / mL Hyclone), and 10% FBS (fetal bovine serum Hyclone). Hyclone) medium and maintained at 37 ℃ incubated in an incubator containing 5% carbon dioxide. The obtained melanin tumor cells were seeded in 1 × 10 ⁵ cells / well in 6-well plates and cultured for 1 day. After incubation, the medium was removed, and then, in order to induce melanin synthesis, melanin synthesis was induced by adding 50 nM of alpha-MSH (melanocyte stimulating hormone) except for the untreated group, and at the same time, lactic acid bacteria lysate 0.5% and 1% were added to the DMEM medium and incubated for 1 day. After collecting the cells of each treatment group, lysates were obtained by lysing the cells with SDS cell lysis buffer (2% SDS addition), and the total protein amount was quantified by Brad-ford method. Western samples were prepared by correcting the total protein amount in each treatment group. The prepared samples were separated according to size on 10% SDS PAGE, and then the proteins transferred to PVDF membranes were visualized on X-ray films using antityrosinase antibodies as primary antibodies. An X-ray photograph of the fermentation product according to Example 17 is shown in FIG. 3. 3, in the case of shiitake mushroom fermentation product according to the present invention, no band is visible. Therefore, it can be seen that no tyrosinase is produced, which means that the fermentation product according to the present invention is excellent in inhibiting the expression of tyrosinase.

Evaluation example  4. Fermented product  Collagen synthesis promoting effect (anti-aging effect)

In order to measure the anti-aging effect of the fermented products according to Examples 1 to 32, the effect of promoting collagen synthesis in human fibroblasts was examined.

First, normal human fibroblasts were charged with DMEM (Dulbecco's Modified Eagle's Medium) medium containing penicillin (100 IU / mL), streptomycin (100 g / mL), and 10% FBS (fetal bovine serum). Culture was maintained in an incubator containing 5% carbon dioxide while maintaining the ℃. The obtained fibroblasts were dispensed into 6-well plates at 3 × 10 ⁵ cells per well and incubated for 24 hours under cell culture conditions. After incubation, the medium was removed, and each fermented product obtained in Examples 1 to 32 and Comparative Examples 1 to 32 and 5, 10, and 20 nM of vitamin C (positive control group) at 0.05, 0.1, and 0.2% concentration were serum-free ( cultured in serum-free medium (DMEM) for 24 hours. The culture solution of each treatment group was collected and subjected to protein electrophoresis. Subsequently, the protein transferred to the PVDF membrane was visualized on an X-ray film using an anti-procollagen type 1 antibody as the primary antibody, and the densities of the collagen bands visualized above were quantified and compared. In the anti-aging efficacy column of Table 1, it is indicated as-(no anti-aging effect), + (slightly anti-aging effect), ++ (excellent anti-aging effect), + + + (excellent anti-aging effect) It was. Meanwhile, the X-ray photographs of Examples 1 to 17 of the present invention are illustrated in FIGS. 4 to 6.

Evaluation example  5. Fermented product  Antioxidative effect

Superoxide dismutase (SOD) -like activity was measured to determine the antioxidant effect according to Examples 1 to 32.

First, lactic acid bacteria lysate was prepared at a concentration of 0.083, 0.83, 8.3%, and SOD (Sigma, S-2515) was prepared at a concentration of 0.1, 1, 10 U (unit) / ml to be used as a positive control. SOD assay kit (Dojindo molecular technologies, INC.) Was used to measure SOD activity. SOD is harmful free radicals (O ^{2 -)} inde representative antioxidant enzyme that converts hydrogen peroxide (Hydrogen peroxide) harmless, as the xanthine oxidase (xanthine oxidase) that is included in the kit been radicals created by action, the sample is held The higher the SOD activity is, the more active oxygen does not change the substrate of WST-1 to WSH-1 formazan. On the other hand, if there is no SOD activity, the change in color is increased because WSH-1 easily changes to WSH-1 formazan by the produced active oxygen.

The results of SOD-like activity measured by observation of color change are shown in Table 1 as antioxidant efficacy,-(no antioxidant effect), + (shows some antioxidant effect), ++ (excellent antioxidant effect), ++ It is indicated as + (an excellent antioxidant effect). On the other hand, SOD-like activity results for the fermentations of Examples 1 to 17 are shown in Figures 7 and 8.

Hereinafter, according to the following composition, formulations of soap, lotion, cream, pack and cosmetic liquid forms were prepared, respectively.

Production Example  One. Soap

Fermented product of Example 1 1.00

Maintenance 75.00

Sodium Hydroxide 5.00

Spices 10.00

Purified water level

(Unit: weight%)

Production Example  2. Lotion Type

Fermentation of Example 1 3.00

L-ascorbic acid-2-magnesium phosphate salt 1.00

Water Soluble Collagen (1% Aqueous Solution) 1.00

Sodium Citrate 0.10

Citric Acid 0.05

Licorice Extract 0.20

1,3-butylene glycol 3.00

Purified water level

(Unit: weight%)

Production Example  3. Cream

Fermented product of Example 1 1.00

Polyethylene Glycol Monostearate 2.00

Self-emulsifying glycerin monostearate 5.00

Cetyl Alcohol 4.00

Squalene 6.00

Tri2-ethylhexaneglyceryl 6.00

Sphingolipids 1.00

1.3-butylene glycol 7.00

Purified water level

(Unit: weight%)

Production Example  4. Packed

Fermentation of Example 1 5.00

Polyvinyl Alcohol 13.00

L-ascorbic acid-2-magnesium phosphate salt 1.00

Lauroylhydroxyproline 1.00

Water Soluble Collagen (1% Aqueous Solution) 2.00

1,3-butylene glycol 3.00

Ethanol 5.00

Purified water level

(Unit: weight%)

Production Example  5. Cosmetic liquid type

Fermentation Example 2 2.00

Hydroxyethylene Cellulose (2% Aqueous Solution) 12.00

Xanthan gum (2% aqueous solution) 2.00

1,3-butylene glycol 6.00

Dark Glycerin 4.00

Sodium Hyaluronate (1% aqueous solution) 5.00

Purified water level

The natural plant material and the herbal fermented product prepared according to the present invention are widely used in various fields such as pharmaceutical industry, beauty industry and food industry because they have excellent stability and safety and have excellent anti-aging, anti-cancer, anti-inflammatory and immunity improvement effects. Can be applied.

1 is a view schematically showing a process diagram for the fermentation method of natural plant material and herbal medicine according to the present invention.

Figure 2 is a graph showing the melanin synthesis inhibitory effect of the fermentation Example 17.

3 is an X-ray photograph showing the effect of inhibiting the tyrosinase expression of the fermentation product of Example 17.

Figure 4 is an x-ray picture to confirm the collagen synthesis effect of the fermentation according to Examples 1 to 5 of the present invention.

Figure 5 is an x-ray picture to confirm the collagen synthesis effect of the fermentation according to Examples 6 to 11 of the present invention.

Figure 6 is an x-ray picture to confirm the collagen synthesis effect of the fermentation according to Examples 12 to 17 of the present invention.

7 is a graph showing the results of SOD-like activity for fermentations according to Examples 1 to 8. FIG.

8 is a graph showing the results of SOD-like activity for the fermentations according to Examples 9 to 17.

Claims (19)

Saccharification step using yeast; Fermentation step through lactic acid bacteria culture; And Obtaining a supernatant from the culture result, wherein the saccharification step is a step of pulverizing natural plant material and herbal medicine, adding the yeast, and immersing for 18 to 30 hours at a temperature of 20 to 40 ℃ Fermentation method of natural plant material and herbal medicine comprising a. delete The method of fermenting natural plant material and herbal medicine according to claim 1, wherein the pulverized product of the grinding step has an average particle diameter of 10 µm to 1 mm. According to claim 1, The natural plant material and herbal medicine is ginger, brown root, licorice, green tea, kelp, Angelica, ganmundong, bokbunja, cornus, baekbaekpi, Schisandra chinensis, ginkgo biloba, licorice, cheonmundong, gardenia, chamomile, shiitake mushroom, persimmon leaf, Kidney beans, cinnamon, red ginseng, gold and silver coins, acorns, bark, pogongyoung, situation mushrooms, pomegranates, pine mushrooms, horse riding, wormwood, ganoderma lucidum, gall bladder, burdock, yulpi, golden, yellow lotus, yellow white, chrysanthemum, gilyeong, bamboo, cabbage, jade porridge , Count, White paper, Fuling, Pipa, Siho, Aloe, Motherwort, Chajeoncho, Changzaza, Cheongung, Astronomical dong, Tosaja, Shouao, Pedestrian rice, Brown rice, Safflower, Gums, Deductor, Oats, Green beans, Jujube, Deodeok, Sweet pumpkin, Doin, Machi, Chinese quince, Peel, Thyme, Sanda, Eoseongcho, lotus root, Gobon, Gwakyang, Rhubarb, Freshness, Baekbu, Baekbuja, White paper, Casualties, Kashiwapi, Tangerine, Doin, Tofu, Ginseng, Red ginseng, Cheonhwa, Tomato, Papaya , Passersby, mistletoe, oats, antler, cordyceps, Dose, White Zap, Agaricus Mushroom, Chaga Mushroom, Lotus Root, Turmeric, Red Pepper, Garlic, Wolfberry, Guava, Gujeolcho, Geumsong, Root, Elm, Arsenal, Tofu, Perilla, Rooibos, Hemp, Ephedra, Plum, Plum, Grab, Thyme, Centella, Borgoji, Boraji, Bokbunja, Non-Suri, Visa, Loquat leaf, Betel nut, Mountain mulberry, Cornus, Sancho, Sinchocho, Acerola, Acacia, Mugwort, Olive leaf, Wasong, Emulsion, Raw grass, Nutmeg, Ginkgo leaf, From the group consisting of roots, barley, bamboo shoots, earth, jimo, jishil, jihwang, chajeoncho, changja, irises, Japanese fern, caviar, chlorella, spirulina, earthenware, parsley, pogongyoung, mongrel, pedestrian, seaweed, walnut and fennel Fermentation method of natural plant material and herbal medicine, characterized in that any one or more selected. The method of fermenting natural plant material and herbal medicine according to claim 1, wherein the amount of the yeast is added in an amount of 1 to 10 parts by weight based on 100 parts by weight of the ground product. The fermentation method of natural plant material and herbal medicine according to claim 1, wherein the amount of the yeast is added in an amount of 2 to 3 parts by weight based on 100 parts by weight of the ground product. delete The method of fermenting natural plant material and herbal medicine according to claim 1, further comprising sterilizing the resultant after the dipping step. According to claim 1, wherein the strain used to prepare the yeast are rhyzopus nigricans , yeast fungus ( Aspergillus) oryzae ) or a fermentation method of natural plant material and herbal medicine characterized in that the mixed strains. According to claim 1, The lactic acid bacteria Lactobacillus sakei ( Lactobacillus sakei B2-16 ), Lactobacillus brevis ( Lactobacillus brevis), Lactobacillus plantarum P23 , Lactococcus lactis ( Lactococcus lactis A164 ) and Bacillus Fermentation method of natural plant material and herbal medicine, characterized in that any one or more selected from the group consisting of Bacillus coagulance . According to claim 1, wherein the fermentation step through the lactic acid bacteria culture Saccharomyces yeast Saccharomyces ( Saccharomyces cerevisiae ) Fermentation method of natural plant material and herbal medicines, characterized in that further adding strains. The fermentation method of natural plant material and herbal medicine according to claim 11, wherein the inoculation amount of lactic acid bacteria or yeast is 10 ⁵ to 10 ⁸ cfu / g or 10 ⁵ to 10 ⁸ cfu / ml. According to claim 11, The lactic acid bacteria are fermentation method of natural plant material and herbal medicine characterized in that the strain isolated from kimchi or miso. The fermentation method of natural plant material and herbal medicine according to claim 1, wherein the fermentation step through the lactic acid bacteria culture is performed for 18 to 120 hours at a temperature of 15 to 40 ℃. The method of claim 1, wherein the step of obtaining the supernatant is characterized in that the fermentation method of natural plant material and herbal medicines are carried out by filtration or centrifugation. A fermentation product prepared by the method according to any one of claims 1, 3 to 6 and 8 to 15. A pharmaceutical composition containing a fermentation product according to claim 16. Cosmetic composition containing a fermentation product according to claim 16. Food composition containing a fermentation product according to claim 16.

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