KR20050076374A - Utility products comprising soy bean extract fermented by lactic acid bacteria inhibiting ammonia production, allergy, atopy and thereof method - Google Patents

Abstract

The present invention relates to a functional product containing a legume lactobacillus fermentation product having the activity of inhibiting ammonia production of atopy and anti-allergic and enteric harmful bacteria, and a method for producing the same, having an effect on anti-allergic and anti-atopy, In order to suppress ammonia productivity, extracts of alcohol or polyethylglycol and dried and dried soybean or pulse extracts for the treatment of allergies and inflammation are treated with Lactobacillus ashdophilus, Lactobacillus vulgaris, Lactobacillus confusus, At least one of Lactobacillus luteri, Lactobacillus cassia, Bifidobacterium brave, Bifidobacterium infantis, Bifidobacterium bifiderm, Bifidobacterium thermofilum, Streptococcus facecalis, Streptococos thermophilus Atopic dermatitis and anti-allergic and enterotoxic effects obtained by vaccination The invention relates to the functional material and its manufacturing method comprising the lactic acid bacteria fermented bean has an activity to inhibit the production of ammonia.

Description

TECHNICAL PRODUCTS COMPRISING SOY BEAN EXTRACT FERMENTED BY LACTIC ACID BACTERIA INHIBITING AMMONIA PRODUCTION, ALLERGY, ATOPY AND ATOPY AND ANTI-ALLERY THEREOF METHOD}

The present invention, in particular, anti-allergic and anti excellent effect in atopic number of lactic acid bacteria that to suppress intestinal bacteria giinseong ammonia production to cause skin damage 10 ⁸ or more pulses lactic acid fermentation of a functional product containing the lactic acid bacteria fermented product with legumes A functional product containing water.

In modern society, allergic diseases are increasing year by year due to the increase of natural environment pollution, the change of diet, and the stress of stress due to the development of industry and civilization. In 1980, less than 1% of allergic patients, including atopic dermatitis, increased to more than 5% in the 2000s and more than 10% of potential patients.

In other words, allergy is a biochemical phenomenon that shows a specific and modified reaction to an alien substance (antigen, Allergen), and when the released substance causes symptoms, it is called allergen. It is called. It is a pathological process of living organisms resulting from antigen-antibody reactions, which are the source of allergy, and is generally classified into four types from the time required for the reaction and the type of complement involvement.

Type 1 is anaphylatctic (immediate type). The target organs are mainly digestive organs, skin, and lungs. Common symptoms include gastrointestinal allergy, gallbladder, atrophic dermatitis, allergic rhinitis, and bronchial asthma. There is this. Pathology of type 1 causes antigens to come into contact with IgE antibodies attached to the surface of mast cells and basophils, activating target cells to secrete chemical media such as histamine, leukotriene, and PAF to constrict blood vessels and smooth muscle. It is known that it is often combined with type 4 (delay type). In other words, this anaphylaxis and allergic reaction are due to various changes occurring in mast cells and the like. Activation of mast cells to induce degranulation may include antigen to Fc receptors, binding of anti-IgE, lectin, stimulation of anaphylatoxin, calcium ionopore, It is caused by drugs such as synthetic cortical stimulating hormones such as compound K48 / 80 and codeine.

Atopic dermatitis is an immune disease in which genetic and environmental factors are involved. Atopic dermatitis is chronically dry, has a strong pruritus, recurs repeatedly, and dermatitis is easily induced by various stimuli. In many cases, the disease worsens at intervals of 1-2 weeks, and if not properly managed, it progresses to subacute lesions of erythematous rash with exudation and chronic lesions of thickened thyroiditis such as leather. Pathological (Histological) findings of atopic dermatitis Each finding varies with the stage of the lesion, but when it becomes chronic, the epidermis thickens and cells involve in various immune responses. In particular, Langerhans cells responsible for immune response are increased and have abnormally increased antigen presenting ability.

In addition, the number of mast cells is increased by atopic dermatitis, indicating degranulation. In 80% to 90% of patients with atopic dermatitis, serum IgE is elevated, especially when accompanied by respiratory atopy such as allergic rhinitis or asthma. In about 85% of patients with atopic dermatitis, skin tests or RAST (radioallergosorbent test) tests show positive IgE for various foods and inhaled allergens.If serum IgE is elevated, allergic diseases such as atopic dermatitis It is diagnosed with high possibility.

In recent years, research on this has been actively conducted due to the increase of various kinds of allergic diseases, and although there have been many advances in the lifesaving and treatment of allergic diseases due to the development of radical immunology, allergic diseases have yet been sufficiently developed. There is no development of curable drugs. Allergic patients have not been able to cure any of the drugs that have been developed so far, and they are looking forward to improving their symptoms, but have side effects.

On the other hand, legumes are currently used as food ingredients, and easily accessible in Korea contains a large amount of flavonoids and saponins that act as phytoestrogen (phytoestrogen). Isoflavones, a major component of soybeans, are found mainly in soybean embryos, including genistin, genistein, daidzein, daidzin, and puerarin. These are most commonly known phytoestrogens (Anderson et al., Am. J. Clin. Nutr., Vol. 70, No. 3, page 464 1999: Knight et al., Gynecol., Vol. 87, page 897). 1996).

Phytoestrogen is structurally similar to the female hormone steroidal estrogen, which binds to the estrogen receptor and acts as an agonist or antagonist against estrogen. (Messina et al., J Nutr. 131, pages 3095-108S, 2001).

In addition, the isoflavone is present in four forms of aglycone (glycoside) form, sugar-coated glycoside form, acetylglucoside form, and malonylglucoside form, wherein the sugar is separated. The degree depends on the shape of the isoflavones.

The isoflavones are hydrolyzed by beta-glucosidase in the intestine and are absorbed in the form of an active glycoside, that is, aglycone. It is known to be biotransformed.

Conventionally, a technique using an isoflavone having an estrogen action has been introduced in Korean Patent Publication No. 2002-060070 (Hepatocellular Enhancer), and high-purity isoflavone through fermentation using Bacillus licheniformis from soybean embryos. The method for producing aglycone is introduced in Korean Patent Publication No. 2002-032078, but the examples of fermentation with various strains of lactic acid bacteria did not appear.

In addition, in Korean Patent Publication No. 2002-007553, which illustrates a method for producing aglycone isoflavone using microorganisms, an isoflavone glycoside solution is added to an adsorbed resin to convert microorganisms that secrete beta-glucosidase to a column to convert to aglycone. this clearly does not apply to materials, such as the aglycone Korea Patent Publication No. 2001-027341 introduced the method of isoflavones, but using Aspergillus (Aspergillus) or Trichoderma spp (Trichoderma) spp, other lactic acid bacteria It was not used as a strain, nor was it used as a bean.

On the other hand, a method for separating and recovering protein and isoflavones from plant materials was introduced in Korean Patent Publication No. 2002-103921, which prepares protein extracts and contacts them with polar ion exchange to separate isoflavones in the extracts with ion exchange resins. , A method for producing aglycone isoflavones (Korean Patent Publication No. 2001-091098) and the method for separating and purifying isoflavones from soybean embryos (Korean Patent Publication No. 2001-016220). In the flavone extraction method (Korean Patent Publication No. 2001-089863), as shown by using an adsorbent resin, most of them do not use lactic acid bacterium fermentation products and most of them are separated using resins. And acid hydrolysis and ion exchange resin methods. However, there are few products using legume saponin. In addition, most patents are mainly for soybean embryos or soybeans, but none have been developed for the purpose of anti-allergic effects.

On the other hand, lactic acid bacteria are largely classified into the genus Lactobacillus, Bifidobacterium, Streptococcus. When looking at the change of intestinal lactic acid by age, Bifidobacterium (Bifidobacterium) appears at 3-4 days of age, 5 On the first day it becomes the most prevalent bacteria. In addition, the end groups are the reason anaerobic bacteria and Bifidobacterium (Bifidobacterium) is dominant and the other lactic acid bacteria is Lactobacillus bacteria (Lactobacillus), etc. is much less. The balance of intestinal bacteria nearing elderly forces of Bifidobacterium (Bifidobacterium) was dominated by aging is weakening, Lactobacillus bacteria (Lactobacillus) bacteria is greater than the power of Bifidobacterium (Bifidobacterium). Therefore, as the age of the Bifidobacterium bacteria can be taken to be similar to the intestinal flora of infants and young adults. The difference between Lactobacillus and Bifidobacterium is that Lactobacillus has homofermentation and heterofermentation, but Bifidobacterium is heterofermentation ).

Thus, a major feature of the Korean Lactobacillus Bifidobacterium breather probe K-110 (Bifidobacterium breve K- 110) may be effective for the treatment of colon cancer, bladder cancer, multiple cancer (Patent No. 0232639, J. Microbiol. Biotechnol. 9 (3), 368 (1999)), it inhibits the growth of Helicobactor pylori , the causative agent of gastric ulcers, and thus has a protective effect on gastric ulcers and can be used to treat gastric diseases (J. Microbiol. Biotechnol. 10 (4), 532 (2000).

In general, the lactic acid bacteria physiological activity may include inhibiting the growth of harmful bacteria, alleviation of lactose intolerance, lowering of serum cholesterol, anticancer activity, immunity, diarrhea and constipation, vitamin synthesis. Therefore bifidobacteria bacteria (Bifidobacteria) has grown the most in five woosegyun Nikkei started to appear from the age of around 3-4, E. coli found in the original or other lactic acid bacteria is Lactobacillus bacteria (Lactobacillus), Streptococcus Cocos (Sterptococcus), etc. Decreases to one hundredth. This is believed to be due to a factor that promotes the growth of Bifidobacteria existing only in breast milk, and it can be seen that Bifidobacteria is the most important bacteria among the intestinal flora compared to other lactic acid bacteria, Lactobacillus .

Conventional techniques have focused on making lactic acid fermentation products such as yogurt using lactic acid bacteria, and these techniques are most commonly used to make lactic acid bacteria beverages in which lactic acid bacteria are inoculated into milk and added skim milk powder to proliferate lactic acid bacteria. In the case of patents for producing lactic acid bacteria drinks or lactic acid using grains other than milk, Lactobacillus plantarum is used to produce lactic acid fermented beverages derived from vegetables and herbal medicines (Korea Patent Publication No. 2000-0055832) and rice snow drinks ( Application No. 1996-042486), a method for producing lactic acid fermentation product using grain-type rice (Publication No. 92-16030), a method for producing liquid lactic acid fermentation product using grain flour (Publication No. 97-14592), Production method (application number 10-1999-0017375), lactobacillus using lactobacillus asidophilus strains (patent 98-148340), etc., but also using a lactic acid bacteria that inhibits the production of anti-allergic and enterobacterial ammonia. There is little research or patent on fermentation products.

In addition, conventionally, functional products and foods that can improve allergic diseases such as atopic dermatitis have not been developed by using a legume lactobacillus fermentation product.

Accordingly, an object of the present invention is to provide a functional product containing a soybean fermentation product which is effective in anti-allergic and anti-atopy and inhibits the production of intestinal bacterial ammonia by inoculating various lactic acid bacteria into soybean extract or these soybeans. .

After immersing with soybean water, sterilize the finely ground pulverized powder with a mixer or colloidal grinder, or the pulverized crushed powder with 0.5% ascorbic acid added, and inoculate 0.1-1% of the lactic acid bacteria mentioned above after cooling. , fermented bean fermentation, for 24 hours - when was 20 ~ -40 ^o C to freeze 15 to 24 when time has Lyophilisation most number of lactic acid bacteria at least ⁸ X 10, freezing the fermented bean groats drying over ⁸ X 10 An object of the present invention is to provide a method for obtaining a lactic acid bacteria soybean fermentation powder containing lactic acid bacteria.

In addition, an object of the present invention is to improve allergic diseases such as atopic dermatitis using legumes fermented with lactic acid bacteria.

The technical means of the present invention for achieving this object is a functional product containing a pulse or pulse extract for the treatment of allergy and inflammation obtained by extraction and drying in water, alcohol or polyethylene glycol (PG), said pulse or Beans extracts include Lactobacillus acidophillus, Lactobacillus vulgaricus, L. bulgaricus, L. confusus, L. luteri, L. luteri, Lactobacillus casei casei, Bifidobacterium brave (B. breve), Bifidobacterium infantis, B. bifidum, Bifidobacterium thermophyllum ), Streptococcus Facecalis (Str. Facecalis), Streptococcus Thermophilus (Str. Thermophillus) is characterized in that the inoculation.

In addition, another technical solution is to immerse by adding water or ethanol to the beans; Grinding the pulses finely into a mixer or colloid grinder; Sterilizing by adding 0.5% ascorbic acid to the pulse mill; Cooling the pulverized powder added with ascorbic acid to 37 ° C .; Inoculating and culturing at least one or more types of lactic acid bacteria in the pulse powder; Freezing and drying the bean fermentation product; And mixing the frozen, dried bean fermentation product into a functional product.

Yet another technical solution is characterized in that the bean and legume extract is at least one of medicinal beans, red beans, seonbi, white tae, frosted, small beans.

In addition, another technical solution is characterized in that for culturing for 12 hours to 72 hours the pulverized legume inoculated with the lactic acid bacteria, more preferably the culture time is characterized in that the culture for 24 hours.

Hereinafter, the configuration of the present invention will be described in detail with reference to the following examples and accompanying drawings.

In general, lactic acid bacteria are largely Lactobacillus genus, Bifidobacterium, Streptococcus classified as Cocos genus, and mainly the example in order to strengthen them in jangnaegyu layer with Bifidobacterium reduced deumeu Korean lactic acid bacteria of Bifidobacterium breve K-110 and Bacillus subtilis age A soybean fermented product was prepared using seed-1, Lactobacillus del brewier seed-2, and the like.

However, in the present invention, Lactobacillus ashophillus (Lactobacillus acidophillus), Lactobacillus vulgaricus (L. bulgaricus), Lactobacillus confusus (L. confusus), Lactobacillus ruteri (L. luteri), Lactobacillus casei ( L. casei, Bifidobacterium brave (B. breve), Bifidobacterium infantis, B. bifidum, Bifidobacterium thermofilum (B. Soybean lactic acid bacteria fermentation was made by inoculating at least one of thermophyllum, Streptococcus facecalis, and Streptococcus thermophillus into soybeans.

<Example 1>

Preparation of legume lactobacillus fermented product

1 is a schematic diagram showing a method for producing a fermented legume lactic acid bacteria according to the present invention.

In other words, the production method of fermented soybean lactic acid bacteria, first, about 5 times of water or ethanol was added to the dried soybeans and boiled for 5 hours and then filtered to obtain a water extract and ethanol extract. In addition, 2-6 times of water was added to the pulverized bean powder, soaked for 4 hours, sterilized in a 120 ° C. autoclave, followed by lactic acid bacteria, Lactobacillus acidophillus, Lactobacillus acidophillus, and Lactobacillus vulgaris (L. bulgaricus, L. confusus, L. luteri, L. bacillus L. casei, B. breve, Bifidobacterium infantis ( B. infantis, Bifidobacterium B. bifidum, Bifidobacterium thermophyllum, Streptococcus Facecalis, Streptococcus thermophyllus Was inoculated 1% alone or a combination of both and incubated 12% -72 hours (preferably 24 hours) and then lyophilized. The lyophilized soybean fermented product was centrifuged (water extraction fraction) or a fraction obtained by centrifugation with the same amount of ethanol (50% ethanol fraction) was used as a sample.

Yield of Soybean Extract Kinds Extraction rate (%) Water extract Alcohol extract Fermented Water Extract Fermented 50% Alcohol Extract Medicinal beans 5.4 0.6 18.5 12.8 Red bean 5.6 0.6 25.3 13.2 Seonbi 9.5 - 25.9 20.1 various phases 8.7 1.8 24.7 18.2 Seo tae 6.8 6.9 25.7 14.9 Duck egg 1.1 - 15.8 5.4 Shortwave Black Soybean 5.4 0.7 25.2 13.4 kidney bean 6.7 - 23.7 15.4 Black Beans 5.7 1.1 23.9 12.8 Greenpeace 6.8 0.9 22.1 15.7 Gamapin 6.7 - 23.3 13.9 broad bean 5.4 2.0 20.8 12.7 Kidney Beans 7.8 0.5 19.7 15.2 Small beans 7.9 0.9 28.1 21.0

In Table 1, the yield of the extract of soybean, which is the subject of experiment, was compared with that obtained in water or alcohol, compared with Bifidobacterum breve and L. acidophilus as a complex fermentation water. And it can be seen that the yield is significantly increased from the extraction to alcohol.

<Example 2>

Cultivation of RAW 264.7 Cells and Measurement of Nitrite Production Inhibition Effect

The cells were distributed by the Korea Cell Line Bank and cultured in DMEM medium containing 10% FBS and l- glutamine, 5% CO ₂ , and 37 ° C. Passage 2-3 times a week.

RAW 264.7 cells (3 ㅧ 10 ⁵ cells / well) were dispensed into 24well plates and incubated for one day, and then treated with lipoplysaccharide (1µg / ml) and samples. After 24 hours, 0.15 ml of the supernatant was removed and transferred to a 96-well plate, followed by 0.15 ml of Griess reagent (0.2% naphthylethylene diamine in DW: 2% sulfanilamide in 10% H ₃ PO ₄ = 1: 1). Was added and measured at 540 nm of ELISA reader.

Inhibitory Effect of NO Production on RAW264.7 Cells Kinds 50% inhibitory concentration (mg / ml) Water extract Alcohol Extract Fraction Fermented 50% Alcohol Extraction Fraction Medicinal beans 0.05 (induction) 0.04 (derived) 0.08 Red bean 0.15 (induction) 0.17 (induction) 0.18 Seonbi 0.24 (induction) 0.23 (induction) 0.12 various phases 0.48 (induction) 0.52 (induction) 0.23 Seo tae 0.44 (induction) 0.49 (induction) 0.32 Duck egg 0.01 (induction) 0.01 (induction) 0.12 Shortwave Black Soybean 0.04 (derived) 0.05 (induction) 0.24 kidney bean 0.05 (induction) 0.05 (induction) 0.45 Black Beans 0.05 (induction) 0.19 (induction) 0.27 Greenpeace 0.28 (induction) 0.37 (induction) 0.18 Gamapin 0.73 (induction) 0.97 (induction) 0.87 broad bean 0.66 0.95 0.55 Kidney Beans 0.05 (induction) 0.68 (induction) 0.07 Small beans 0.02 (induction) 0.02 (induction) 0.04

* Induction leads to NO production without LPS treatment

Table 2 shows the NO production inhibitory effect in RAW264.7 cells. When water extract or alcohol extraction method is used, NO production is induced in macrophages, which are phagocytic cells, except for silk beans. Rather than treating it, it was likely to cause allergy.

However, fermenting soybeans as described above does not induce the production of NO, thus eliminating allergic potential. Particularly, kidney beans and small beans showed excellent NO inhibitory effect when fermented, resulting in anti-inflammatory effects.

<Example 3>

Cultivation and anti-allergic effect of RBL-2H3 cell line

DBL (Dulbeccos' modified Eagle's medium, Sigma, 22256) containing 10% FBS (fetal bovine serum) and L-glutamine in rat mast cell line (RB-2H3 cell line, Cat Cell No. 22256) (Sigma D-5648) was incubated for 2 hours in a humidified 5% CO ₂ incubator at 37 ° C., and the cells with adherence were suspended using trypsin-EDTA solution. Was used.

RBL-2H3 cells were aliquoted into ⁵ wells of ⁵ × 10 ⁵ cells / well, followed by sensitization with mouse monoclonal IgE 0.5 μg / ml for 12 hours.

The cells were washed with 0.5 ml of siraganian buffer (119 mM NaCl, 5 mM KCl, 0.4 mM MgCl ₂ , 25 mM PIPES, 40 mM NaOH, pH 7.2) and then again 0.16 ml of siraguanian buffer (5.6 mM). Glucose, 1 mM CaCl ₂ , 0.1% BSA was added) and then incubated at 37 ° C. for 10 minutes. After adding 0.04 ml of DSCG as a fermented lactic acid bacterium fermentation product and a control drug prepared in Example 1, respectively, and after 20 minutes, the cells were treated with 0.02 ml of antigen (DNP-BSA 1 μg / ml) at 37 ° C. for 10 minutes. After activation, 0.02 ml of the supernatant was transferred to 96 wells by centrifugation at 2000 rpm for 10 minutes.

To this was added 0.025 ml of substrate solution 1 mM p-NAG (a solution obtained by dissolving p-nitrophenyl-N-acetyl-β-D-glucoamide) in 0.1 M citrate buffer to pH 4.5. After incubation for 10 minutes, 0.2 ml of 0.1 M Na ₂ CO ₃ / NaHCO ₃ was added to stop the reaction, and the respective absorbances were measured with an ELISA analyzer at 405 nm.

Inhibitory Effect of Nucleososaminidase on RBL2H3 Cells Kinds 50% inhibitory concentration (mg / ml) Water extract Alcohol Extract Fraction Fermented 50% Alcohol Extraction Fraction Medicinal beans 0.05 0.05 0.08 Red bean 0.18 0.18 0.18 Seonbi 0.31 0.31 0.12 various phases 0.30 0.30 0.23 Seo tae 0.05 0.05 0.32 Duck egg 0.07 0.07 0.12 Shortwave Black Soybean 0.02 0.02 0.24 kidney bean 0.69 0.69 0.45 Black Beans 0.06 0.06 0.27 Greenpeace 0.48 0.48 0.18 Gamapin 1.10 1.10 0.87 broad bean > 2.00 > 2.00 0.55 Kidney Beans 0.52 0.52 0.07 Small beans  0.25  0.19 0.02

That is, as shown in Table 3, the effect of inhibiting the degranulation of mast cells or basophil cells, which are the cause of allergy, by the effect of inhibiting the secretion of nucleosaminidase of RBL2H3 cells, Was excellent in water extracts such as aek, frost, duck egg, shortwave black soybean, etc., but fermented weak beans, duck egg, green beans, and soybeans. In particular, the effect was even in the mycobacterial fermentation.

<Example 4>

Anti-allergic effect in experimental animals

When carried out according to the method of Inagaki et al. (Int. Arch. Allergy Appl. Immunol., 1988), the dinitrophenol bovine serum protein (Dinitrophenol-bovine serum albumin, hereinafter referred to as DNP-BSA). 10 ug of IgE serum diluted with saline was injected into the back of ether-anesthetized mice and sensitized manually. After 48 hours, 0.2 ml of saline containing 0.2 mg of DNP-BSA and 1.6 mg of Evans blue was tail veined. 30 minutes after the injection into the cervical dislocation is measured by the amount of evans blue pigment leaked on the back. Cut a portion of the back of the mouse, put it in a test tube, add 0.7 ml of IN-KOH and incubate overnight at 37 ℃. 4 ml of 0.6 N phosphoric acid and acetone mixture (5:13) were added to the test tube, followed by shaking and filtration to colorimetrically extract the extracted pigment at 620 nm. Soy water extract, alcohol extract and fermented product are dissolved or suspended in physiological saline and administered orally one hour before antigen administration.

Antiallergic Effect of Soybean Extract and Its Fermentation in Passive Cutaneous Anaphylasis Experimental Mice Composition Dose (mg / kg) Inhibitory activity (%) Water Extract of Beans 200 77.8 ± 8.1 50 39.8 ± 12.9 Alcohol Extract of Pea Beans 200 69.9 ± 7.7 50 35.7 ± 2.2 Water extract obtained by fermenting soybeans with Bifidobacterium breve K-110 100 58.2 ± 7.5 Water extract obtained by fermenting soybeans with Lactobacillus acidophilus 100 69.7 ± 12.1 Water Extracts from Fermented with Streptococcus thermophilus 100 58.4 ± 5.2 A water extract obtained by fermenting soybeans with Bifidobacterium breve K-110 and Lactobacillus acidophilus 100 62.0 ± 3.8 50% alcohol extract from fermented soybeans with Bifidobacterium breve K-110 100 78.2 ± 9.9 50% alcohol extract obtained by fermenting soybeans with Bifidobacterium breve K-110 and Lactobacillus acidophilus 100 76.8 ± 3.1 Disodium cromoglycate 50 37 ± 5.22

That is, as shown in Table 4, the inhibitory effect on passive dermal anaphylaxis, which is a model animal of an atopic allergy, obtained from the soybean extract as it is, is more effective than that of the existing drug, disodium cromoglycate. Excellent effect. In particular, the fermented product of soybean showed a better effect.Bifidobacterium breve K-110 was superior in fermentation alone, but when co-cultured with other Lactobacillus acidophilus It showed a better effect.

Example 5

Anti-allergic Effect of Soybean and Its Fermentation on Contact Dermatitis Model Animals

Fujii Y, Takeuchi H, Tanaka K, Sakuma S, Ohkubo Y, Mutoh S. Effects of FK-506 (tacrolimus hydrate) on chronic oxazolone-induced dermatitis in rats.European Journal of Pharmacology 2002; 456: 115-121 Made a model animal according to.) First, 100ul of oxazolone dissolved in 1.5% alcohol is sensitized on the abdomen of mouse (mouse) of ICR male (weight 25-28 grams). After 1 week, 20ul is applied to both ears with a solution of 1% or 3% dissolved in a mixture of actone: olive oil at a ratio of 4: 1 every 3 days. After 3 days, the thickness of the ear is measured with a Digimatic Micrometer (Mitsutoyo, Japan). Subsequently, oxazolone is applied and the drug is applied again 3 hours later and the thickness of the ear is measured 3 days later. Repeat this three times.

Inhibitory Effect of Soybean Extract on Contact Dermatitis Composition Dose (%) Inhibitory Activity (%)  Water Extract of Beans One 18 ± 1.8 3 34 ± 1.5  Alcohol Extract of Pea Beans One 20 ± 2.5 3 33 ± 2.3  Water extract obtained by fermenting soybeans with Bifidobacterium breve K-110 One 35 ± 2.7  Water extract obtained by fermenting soybeans with Lactobacillus acidophilus One 34 ± 2.1  Water Extracts from Fermented with Streptococcus thermophilus One 28 ± 2.4  A water extract obtained by fermenting soybeans with Bifidobacterium breve K-110 and Lactobacillus acidophilus One 37 ± 3.2  50% alcohol extract from fermented soybeans with Bifidobacterium breve K-110 One 39 ± 2.2  50% alcohol extract obtained by fermenting soybeans with Bifidobacterium breve K-110 and Lactobacillus acidophilus One 42 ± 3.3 Betamethasone (Sigma, USA) One 87 ± 8.5

Therefore, the contact dermatitis inhibitory effect of the extract or fermentation of the soybean as it was, but did not have a lot of side effects than betamethasone showed an excellent effect than the known natural products. In particular, the fermented product of soybean showed a better effect.Bifidobacterium breve K-110 was superior in fermentation alone, but co-cultured with other lactobacillus Lactobacillus acidophilus. Showed better effect.

<Example 6>

Effect of Soybean Extract Fermented with Soybean and Lactic Acid Bacteria on Inhibition of Ammonia Production by Enterobacteriaceae

Soybean water extracts and alcohol extracts and soybean extract fermented extracts and 50% alcohol extracts fermented with B. breve (BB) and Streptococcus thermophyllus (ESTI) were prepared as in Example 1. Thus, the effects of enteric bacteria on the production of ammonia, a harmful substance, are shown in Table 3. Sterilized by adding 1% of soybean and its fermented extract to 10 mL of GAM (general anaerobic medium) medium, and then inoculated with human enterobacteriaceae (liquid diluted to 0.1% by wet weight in persimmon medium), 37 After incubation in anaerobic conditions for 24 hours in a thermostat of ℃ was measured for ammonia production. Ammonia was measured by adding 0.1 M phosphate buffer (pH 7.0), fecal suspension 20 μl and adding 1N H ₂ SO ₄ 100 μl, then mixed solution 1 (1% phenol, 0.005% sodium nitroprusside) and mixed solution 2 (0.1% sodium). 1 ml of hypochlorite, 0.5% sodium hydroxide, and 5.5% sodium phosphate dibasic) were each added, heated at 60 ° C. for 20 minutes, and cooled to room temperature to measure absorbance at 660 nm.

Inhibitory Effects of Ammonia Production by Enteric Bacteria on Soybean and Fermented Soy Kinds Inhibition Rate (%) Water extract Alcohol extract Fermented Water Extract Fermented 50% Alcohol Extract Medicinal beans 67 67 83 86 Red bean 71 75 83 91 Seonbi 78 69 82 90 various phases 67 68 82 82 Seo tae 72 68 83 87 Duck egg 85 74 78 78 Shortwave Black Soybean 76 72 67 87 kidney bean 58 75 87 85 Black Beans 78 77 75 85 Greenpeace 85 77 72 87 Gamapin 83 67 77 88 broad bean 67 69 69 87 Kidney Beans 78 47 78 87 Small beans 77 67 86 90

That is, as shown in Table 6 above, although there are differences depending on the legumes, the water extract of the legumes inhibited the production of ammonia better than the alcohol extracts, and the fermentation extracts and the fermented 50% alcohol extracts inhibited the production of ammonia to a similar extent. In particular, compared to soybeans before fermentation, the production of ammonia, which causes skin troubles, was better suppressed after fermentation. This was superior to any existing natural product.

In conclusion, pulsed Lactobacillus fermentation products had higher yields than normal extracts, and inhibited the production of nitrogen (NO) when fermented by 50% alcohol extraction by Bifidobacterium breb and L. esidophilus combined fermentation. As it shows an anti-inflammatory effect, and also suppresses the degranulation of the cells showed an excellent effect on anti-allergy.

In addition, there is a difference depending on the legumes legumes lactic acid bacteria fermented products are excellent in inhibiting the production of ammonia has the effect of removing or preventing the cause of skin problems.

Therefore, the material according to the present invention can be applied as a food material or various functional cosmetics to suppress skin allergies, atopy, and skin troubles, and can also be used as various functional food materials and food and baby food materials.

As such, those skilled in the art will appreciate that the present invention can be implemented in other specific forms without changing the technical spirit or essential features thereof. Therefore, the above-described embodiments are to be understood as illustrative and not restrictive in all respects, and the scope of the present invention is indicated by the appended claims rather than the foregoing description, and the meaning and scope of the claims and All changes or modifications derived from the equivalent concept should be interpreted as being included in the scope of the present invention.

As described above, in the present invention, by using the fermented legume lactic acid bacteria fermented with the lactic acid bacteria, it is effective for anti-inflammatory and anti-allergic effect, in particular, to suppress skin troubles and to suppress the production of intestinal bacterial total ammonia.

In the present patent, a fermented product is prepared by fermenting soybeans using various lactic acid bacteria, and it is a great feature that the lactic acid bacteria can be obtained by X 10 ⁸ or more by lyophilizing these fermented products. These soybean fermented products can be said to be a technology that can be used as a material for cosmetics (external and dietary), baby foods, baby foods and lactic acid bacteria and functional foods using soybeans.

1 is a flow chart showing a method for producing a legume lactic acid bacteria fermentation according to the present invention.

Claims (9)

A functional product containing allergy and inflammatory pulse or pulse extract, which is obtained by extraction with water, alcohol or polyethylene glycol (PG) and dried, Lactobacillus acidophillus, Lactobacillus vulgaricus, Lactobacillus confusus, L. confusus, L. luteri, Lactobacillus cartose into the pulse or pulse extracts. L. casei, Bifidobacterium brave (B. breve), Bifidobacterium infantis, B. bifidum, Bifidobacterium thermofilum ( B. thermophyllum), Streptococcus Facecalis (Str. Facecalis), Streptococcus thermophillus (Str. Thermophillus) inoculating the activity of inhibiting ammonia production of atopy and anti-allergic and enterotoxic bacteria characterized by Functional product containing legume lactic acid bacteria fermentation having. According to claim 1, wherein the legumes and legumes extract is a soybeans having an activity to inhibit the ammonia production of atopy and anti-allergic and enterotoxic bacteria, characterized in that at least one of medicinal beans, red beans, seonbi, white, frosted, pea beans. Functional product containing lactic acid bacteria fermentation product. Immersing by adding water or ethanol to the beans; Grinding the pulses finely into a mixer or colloid grinder; Sterilizing by adding 0.5% ascorbic acid to the pulse mill; Cooling the pulverized powder added with ascorbic acid to 37 ° C .; Inoculating and culturing at least one or more types of lactic acid bacteria in the pulse powder; Freezing and drying the bean fermentation product; And Preparation of a functional product containing a legic acid lactic acid bacteria fermentation product having an activity that inhibits ammonia production of atopy and anti-allergic and enterotoxic bacteria, characterized in that it comprises the step of mixing the frozen, dried legume fermentation product into a functional product Way. According to claim 3, wherein the legumes and legumes extract is a soybean, which has an activity to inhibit the ammonia production of atopy and anti-allergic and enterotoxic bacteria, characterized in that at least one of medicinal beans, red beans, seonbi, white, frosted, pea beans. Method for producing a functional product containing a lactic acid bacteria fermentation product. According to claim 3, The lactic acid bacteria Lactobacillus acidophillus (Lactobacillus acidophillus), Lactobacillus vulgaricus (L. bulgaricus), Lactobacillus confusus (L. confusus), L. luteri (L. luteri), Lactobacillus L. casei, Bifidobacterium brave, Bifidobacterium infantis, Bifidobacterium bifitum, Bifidobacterium thermophil Activity that inhibits ammonia production of atopy, anti-allergic and enterotoxic bacteria, characterized in that it is at least one of B. thermophyllum, Streptococcus Facecalis, Streptococcus thermophillus Method for producing a functional product containing legume lactic acid bacteria fermentation having. According to claim 3, wherein the legumes inoculated with the lactic acid bacteria pulverized legumes fermented products having an activity to inhibit ammonia production of atopy and anti-allergic and enterotoxic bacteria, characterized in that for 12 hours to 72 hours to incubate Method for producing a functional product containing. The method of claim 6, wherein the incubation time is 24 hours, characterized in that the atopy, anti-allergic and enteric-induced bacterial production method containing a fermented legume lactic acid bacteria fermented product having an activity that inhibits the ammonia production. According to claim 1 or 3, wherein the fermented soybeans and legumes extract is characterized in that the atopy and anti-allergic and characterized in that it is used as a raw material of the soybean lactic acid bacteria fermented products having an activity to inhibit the ammonia production of harmful enteric bacteria Method for producing a functional product containing. According to claim 1 or 3, wherein the fermented soybeans and legumes extract is used as a raw material of the functional food for the treatment of atopy and allergies to inhibit the ammonia production of atopy and anti-allergic and enterotoxic bacteria. Functional product containing legumes lactic acid bacteria fermentation having and a production method thereof.