KR101741165B1

KR101741165B1 - A pharmaceutical or food composition containing the fermentative products of balloon flower converted biologically by Basidiomycota hyphae or lysate of Basidiomycota hyphae with anti-allergenic effect

Info

Publication number: KR101741165B1
Application number: KR1020150165208A
Authority: KR
Inventors: 이상종; 김성필; 허인영; 박선주; 박선옥; 김진만; 남석현
Original assignee: (주)에스티알바이오텍
Priority date: 2014-11-28
Filing date: 2015-11-25
Publication date: 2017-05-30
Also published as: KR20160064992A; KR101784672B1; KR20160064991A; KR20160064993A; KR101773484B1

Abstract

The present invention relates to a therapeutic pharmaceutical composition or food composition having an antiallergic ability comprising a fermentation product of microorganisms (mycelia mycelia) fermented and biologically converted biodiversity through enzymatic treatment, Fermented product of the present invention contains a large amount of antiallergic substance as compared with the non-fermented seedlings.

Description

TECHNICAL FIELD [0001] The present invention relates to a therapeutic pharmaceutical composition or food composition containing fermented microorganisms (bacterium mycelium) or a fermented product of a bellflower which is biologically converted through enzyme treatment, by Basidiomycota hyphae or lysate of Basidiomycota hyphae with anti-allergenic effect}

The present invention relates to a therapeutic pharmaceutical composition or food composition having an antiallergic ability comprising fermented microorganisms (mycelia of mycelia) or further fermented by enzymatic treatment.

Allergic diseases are so widespread that about 25% of all people suffer from one or two allergies. The prevalence of allergic rhinitis in developed countries has increased by 10% every year for the past 30 years. In recent years, the statistics of allergic patients in urban areas show that 15 ~ 20% of rhinitis. Asthma 6 ~ 8%, atopy 10 ~ 15%, eye allergy 3%, and the number of patients with at least one allergy is about 25%. It is estimated that allergic diseases have a very high socioeconomic cost. According to the Korea Asthma Association 2005 report, the socio-economic cost of bronchial asthma reaches 2 trillion won per year. The prevalence of allergic diseases in developed countries is higher than that of underdeveloped and developing countries, and the incidence of allergic diseases is expected to increase in Korea.

Common treatments for allergic diseases include avoidance, drug, surgery and immunotherapy. Avoidance is the most basic and important treatment method to avoid causative antigens. It is a very important method for the management of allergic diseases, but there is a difficulty in completely avoiding causative agents that cause immune reactions. Drug therapy has shown the greatest progress in the treatment of allergic diseases, but current drug therapy is a treatment that inhibits the secretion of chemical messengers. In addition, steroids, antihistamines, sympathomimetics, theophylline, etc. have been used in the selection of therapeutic agents for 20 years or more, with the exception of leukotriene modifiers. Although there is a growing concern about the importance of anti-inflammatory drugs such as steroids for persistent allergic diseases such as chronic asthma, this treatment strategy is not aimed at primary prevention or cure of allergic diseases. Operative therapy includes surgery for cirrhosis, cryosurgery, diathermy, neurosurgery, and laser ablation. However, since the pathology of allergic diseases is not corrected by surgery, , The symptoms often recur. Immunotherapy is a method to induce a change of immune response in the body by continuously and regularly administering the causative antigen when the antigen is revealed through various inspection methods, and as a method to reduce the susceptibility to the causative substance.

As described above, since most of the allergic diseases have no special therapeutic agent or preventative agent other than the symptomatic treatment, there is a need for development of raw materials and products for health functional foods derived from natural materials. Up to now, techniques such as pharmaceutical compositions (KR 0643878 and KR 0375560) for the treatment of bronchial diseases using the bellflower extract as an active ingredient have been developed, but the microorganisms (mycelium mycelium) have been fermented or, further, The development of technologies for the treatment and prevention of allergic diseases using as an active ingredient is insufficient.

Accordingly, the present invention relates to a therapeutic pharmaceutical composition or food composition having antiallergic activity including fermented Dorata fermented by fermentation of mycelia or further biotransformation through enzymatic treatment, and its effect.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide an antiallergic therapeutic pharmaceutical composition or food comprising fermented Dorotus fermented product, &Lt; / RTI > However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

Unless defined otherwise in the specification, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

In one embodiment of the present invention, the basidiomycetes are fungi, which are cells of the fungus as a result of the sexual reproduction of the fungus as a result of the sexual reproduction. There are many things known as mushrooms which are parasitic to other organisms because they can not make their own nutrients. Typical examples are mushroom, mushroom, shiitake, mushroom, mushroom, and mushroom. On the freshly corrugated side of adult mushrooms, a lot of volcanic ash is formed. Most of the basidiomycetes mushrooms live in secondary life in the fluid of higher plants. In particular, they decompose cellulose and lignin to play an important role in the biological circulation of the material. In addition, they form mycorrhiza in the roots of trees, There are many. There are other kinds that are important as edible bacillus or should be cautious with poisoning.

In one embodiment of the present invention, mycelium is a nutritive cell of a filamentous fungus. Functional differentiation such as nutritional function or physiological function can also be seen. Morphologically, the presence or absence of the bulkhead is used for classification, and it develops in germination tube of spore and elongates by tip growth. The group of branched mycelium is called mycelium. Depending on the physiological function, it is distinguished into basal hypha that grows on the surface or inside of the pouch or host, and parasitic bacteria which grows in the air. Parasitoids depend on the transfer of nutrients from basal hyphae, some of which differentiate into spore forming organs and the like. If there are organic matter and suitable environmental conditions such as appropriate humidity and temperature, various mycelial tissues are formed. The mycelial membrane of mycelium of fungus is simple, but it can be seen that it forms a braid connection. In the fungi, it branches into several branches and is entangled like a thread. It does not have chlorophyll and does not have photosynthesis. It attaches to the host organism and absorbs nutrients. In the case of the mushroom-bearing bacterium, the mycelium germinated in the spore has one nucleus, which when fused with another mycelium has two nuclei in one cell. Mycelium with one nucleus is called mycelium, and mycelium with two nuclei is called secondary mycelium. The cell wall of the mycelium is generally composed of chitin or hemicellulose, which is sometimes combined.

In the present specification, the mycelia of mycelia can be selected from mycelium of mushroom, chaga mushroom, shiitake mushroom, mushroom mushroom, mushroom mushroom, The mycelia of these mycelia were collected from the KCCM, KCTC, KCLB, KACC, ATCC, And the like.

In one embodiment of the present invention, Platycodon grandiflorum is a perennial plant having a coarse rootstock. The stem stands straight and grows to the height of 40 ~ 80cm, and almost does not hit the branch. Leaves are alternate to each other or two to three leaves may be in one position. Leaves are elongated egg-like or elliptical in shape, and there is no petiole. Both ends of the leaf are pointed, with sharp sawtooth on the edge, and the back of the leaf appears to be wearing white powder. At the end of the stem, a large bloom of several bell-shaped flowers like a bell is bloomed. The end of the flower is divided into five branches and has five stamens. The diameter of the flower is about 4cm, the color is dark azure, and sometimes there is a flower in white. It is known that Platycodin and Platycodigenin, a kind of saponin contained in the rootstock, are known to cause diadhegmation and vasodilatation.

In the present specification, the bellflower or the bellflower powder means all or part of the bellflower, and a part of the bellflower may be at least one selected from the group consisting of bellflower roots, root hair, stem, branches, leaves, no.

In one embodiment of the present invention, bioconversion refers to the conversion of a substance added using a physiological function of an organism into a chemically modified form.

In one embodiment of the present invention, fermentation refers to a process in which a microorganism decomposes an organic substance using an enzyme contained therein. Fermentation is a phenomenon in which organic matter changes into a simple compound by enzymatic action and releases free energy. In general, microorganisms decompose organic matter and accumulate metabolites. That is, fermentation of alcohol by anaerobic digestion of sugars by anaerobic digestion of yeast, fermentation of lactic acid by lactic acid bacteria decomposes anaerobic sugar into lactic acid by lactic acid bacteria is a typical fermentation, but now, A phenomenon in which alcohol is oxidized to acetic acid by use of oxygen, and a phenomenon in which mold oxidizes glucose to gluconic acid by using oxygen in the air are also referred to as acetic acid fermentation and gluconic acid fermentation. In addition, these anaerobic fermentation can be divided into anaerobic fermentation and aerobic fermentation (oxidation fermentation), and other anaerobic fermentation fermentations include glycerol fermentation, acetone butanol fermentation, 2, 3-butylene glycol fermentation, butyric acid fermentation, Fermentation, and methane fermentation. Examples of aerobic fermentation include citric acid fermentation, itaconic acid fermentation, yeast acid fermentation, 2-keto glutaric acid fermentation, oxalic acid fermentation, fumaric acid fermentation and sorbose fermentation. In addition, production of amino acids, vitamins and antibiotics by microorganisms is also called glutamic acid fermentation, riboflavin fermentation, penicillin fermentation and the like, which is not a good title. The name of the fermentation is usually called the product, but sometimes the name of the substrate is given by cellulosic fermentation, pectin fermentation and so on.

In one embodiment of the present invention, an allergic disease is a disease caused by allergies. Allergy refers to an antigen-antibody reaction that occurs when an IgE antibody is produced against a substance of interest (antigen or allergen), and then the same antigen enters the body. The major diseases are bronchial asthma, allergic rhinitis, urticaria, leukemia, drug allergy, seropositivity, and various types of diseases depending on the type of allergen or tissues causing allergic reactions. Diagnosis of allergic diseases refers to the progress of the family or patient until that time, the time of onset, the living environment, and the contents of the meal consumed. In some cases, it may be necessary to remove the suspicious allergen and observe the improvement of the symptoms, and to induce allergens. Other skin reactions include scratch tests or skin reactions. Therapeutic methods include elimination of allergens, desensitization therapy, and drug therapy to suppress allergic reactions. As a typical allergic disease of the respiratory system, allergic rhinitis shows hypersensitivity to the nasal mucosa of a specific substance. After the nasal mucosa is exposed to the causative substance (allergen), all parts of the nasal mucosa IgE antibody-mediated inflammatory cells are involved in inflammatory reactions caused by various mediators secreted by these cells. It is characterized by three main symptoms: continuous seizure sneezing, clear runny nose, and nasal congestion. Allergic rhinitis can be suspected when two or more of these three symptoms are present. In addition to the characteristic symptoms, it may be accompanied by symptoms such as itching around the nose, headache, loss of smell, and complications such as otitis media, sinusitis and pharyngitis.

In one embodiment of the invention, IgE is an antibody protein, also called immunoglobulin E. It is often called an "allergic antibody" because it plays an important role in allergic reactions. If you are allergic to certain substances (allergens), the immune system mistakenly thinks that harmful substances are harmful to your body. When you are exposed to this particular substance, the immune system begins to produce IgE to protect your body. IgE antibodies remain in the body and then come into contact with allergens again, causing allergic reactions. As a result, people with allergies have elevated serum IgE levels, and IgE has specificity for each allergen.

In one embodiment of the present invention, histamine is a substance produced by the action of histidine decarboxylase, which is an amino acid, histidine. It is mainly stored in mast cells and is released when the cells are damaged due to wound, bacterial infection, Histamine released plays an important role in immune function by expanding the blood vessels and increasing the permeability of the blood vessels, allowing white blood cells and various plasma proteins to move quickly to the site where the bacteria have entered. The excessive secretion of histamine due to excessive immune responses may cause allergic reactions and anaphylaxis. Besides these immune functions, it is one of powerful active agents that regulate the movement of the uterus during pregnancy and secrete gastric juice. It also causes an increase in blood flow due to vasodilation.

In an embodiment of the present invention, the pathogenesis of allergy is described in detail. Allergy refers to a symptom in which an exaggerated immune response to an exogenous substance is exerted. The allergic disease is mainly caused by a Th2 cell mediated immune response Lt; / RTI > Acute and mild allergic diseases such as seasonal allergic rhinitis are characterized by increased allergen-specific IgE antibodies, IgE-dependent activation of mast cells, and invasion of activated Th2 cells and eosinophils into the mucosal surface.

In relation to the onset of allergy, ① allergen-inducing antigen enters our body to activate antigen-specific Th2 cells, ② cytokine secreted by activated Th2 cells (IL-4, etc.) When class switching occurs and IgE antibody production is induced and secreted, our body becomes sensitive to allergens. At this time, when the allergen is reintroduced, the IgE antibody mediated reaction occurs and allergic symptoms appear. That is, when the IgE antibody binds to the IgE receptor of the mast cell and the mast cell is sensitized, the mast cell is activated when the allergic antigen is re-introduced to the mast cell surface, and the degranulation is immediately induced and the intracellular granule such as histamine And inflammatory cytokines, are secreted out of the cell. Allergic reactions are triggered by several chemical allergen mediators secreted. Allergic reactions can be divided into early reaction and late reaction. The initial reaction occurs within a few minutes and secretes mediators of already-produced mediators such as histamine, β-hexosaminidase, and serotonin and newly synthesized lipid components from mast cells And the late response occurs within a few hours. IL-4, IL-5, and IL-13, which are typical cytokines of Th2 immune response, are synthesized and secreted to continue the inflammation reaction. Mediators and cytokines of lipid components, newly produced by mast cells, are the major factors causing allergic acute and chronic inflammatory responses. (4) Eosinophil infiltration and induction of eosinophils in peripheral tissues by cytokines (IL-5, etc.) secreted from cells and mast cells induces eosinophil hyperplasia, which is called eosinophilic allergy. In addition, IL-13 induced hypersensitivity by inducing hypersensitivity. Eosinophilia is a direct cause of induction of allergic hypersensitivity reaction.

In the case of an allergic disease caused by an excessive immune response, it is necessary to control the aggressive immune response. Inhibition of the Th2 immune response by inducing the Th1 immune response and the Treg immune response, (IL-4, IL-5, and IL-13), (3) reduce eosinophils and mast cells in allergic inflammation sites, and Immunomodulating treatments such as reducing hypersensitivity suggest new possibilities for the treatment of allergic diseases.

In one embodiment of the present invention, sepsis refers to a condition in which a microorganism is infected and a serious inflammatory reaction occurs in the whole body. The number of breaths increased more than 24 times per minute (ventilation), heart rate more than 90 times per minute (tachycardia), increase in leukocyte count in blood test, or significant decrease in respiratory rate If you have symptoms that are more than a branch, it is called systemic inflammatory response syndrome (SIRS). This systemic inflammatory response syndrome is called sepsis when it is caused by microbial infection. The source of the infection can be any organs of the body. Pneumonia, pyelonephritis, meningitis, cellulitis, infective endocarditis, peritonitis, pressure ulcer, cholecystitis, and cholangitis may cause sepsis. If such infection occurs, the causative microorganism may invade into the blood and cause sepsis. However, even if microorganisms do not penetrate into the blood, systemic sepsis may occur due to the inflammation reaction of some body parts and the production of inflammatory substances. Early symptoms of sepsis may include respiratory arrest, neurological disturbances such as loss of locomotion (time, place, cognitive ability) or delirium. The skin may appear visually opaque due to lowering of blood pressure and lowering of the amount of blood supplied to the distal end of the body. Bacteremia (a symptom of germs circulating in the blood) can cause germs to move around the bloodstream, to position themselves in certain parts of the body and cause pathological changes in those areas. As the skin changes specific to the causative bacteria, it may help to diagnose the cause of sepsis. Symptoms of the digestive system include nausea, vomiting, diarrhea and intestinal paralysis, and severe gastrointestinal bleeding may also occur.

In one embodiment of the present invention, the pharmaceutical composition means a composition to be administered for a specific purpose. For the purpose of the present invention, the pharmaceutical composition of the present invention is used for enhancing the survival rate or inhibiting the inflammatory active substance in the sepsis-induced group, and comprises a composition for this purpose and a pharmaceutically acceptable carrier, excipient or diluent . The pharmaceutical composition according to the present invention comprises 0.1 to 100% by weight of the active ingredient corresponding to the pharmaceutical composition of the present invention, based on the total weight of the composition. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In one embodiment of the present invention, the food composition is variously used as a food composition for improving allergic diseases. The food composition containing the composition of the present invention as an active ingredient may be used in various foods such as beverages, Tea, vitamin complex, powder, granule, tablet, capsule, confection, rice cakes, bread and the like. Since the food composition of the present invention is composed of a natural food and its fermented product which have little toxicity and side effects, it can be safely used for prolonged use even for prophylactic purposes. When the composition of the present invention is contained in the food composition, the amount thereof may be added in a proportion of 0.1 to 100% of the total weight. Here, when the food composition is prepared in a beverage form, there are no particular limitations other than those containing the food composition at the indicated ratios and may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. That is, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, sucrose and the like and sugar sugars such as polysaccharide, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol can do. Examples of the flavors include natural flavors (such as tau martin, stevia extract (for example, rebaudioside A and glycyrrhizin), and synthetic flavors (for example, saccharin and aspartame). The food composition of the present invention can be used as a food composition containing various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants, pectic acid and its salts, alginic acid and its salts, Stabilizers, antiseptics, glycerin, alcohols, carbonating agents used in carbonated beverages, etc. These components may be used independently or in combination. The ratio of these additives is not so important, Is generally selected in the range of 0.1 to 100 parts by weight per part.

In one embodiment of the present invention, the cosmetic composition is a composition for improving an allergic skin condition or an allergic skin disease. The cosmetic composition comprising the composition of the present invention as an active ingredient can be used as a skin lotion, a nutritional lotion, a nutrition essence, a massage cream, a cosmetic bath additive, a body lotion, a body milk, a bath oil, a baby oil, Skin lotions, skin creams, sunscreen cosmetics, cleansing milks, hair removal products, cosmetics, face and body lotions, face and body creams, skin whitening creams, hand lotions, hair lotions, (Non-medical use), cream soap, facial wash, hair rinse, make-up soap, tooth whitening gel, toothpaste, etc. . &Lt; / RTI > To this end, the compositions of the present invention may further comprise suitable carriers, excipients or diluents conventionally used in the preparation of cosmetic compositions. The carrier, excipient or diluent which may further be added to the cosmetic composition of the present invention include purified water, oil, wax, fatty acid, fatty alcohol, fatty acid ester, surfactant, humectant, thickener, antioxidant, Buffers, lower alcohols, and the like, but are not limited thereto. Further, if necessary, it may contain a whitening agent, a moisturizing agent, a vitamin, an ultraviolet screening agent, a perfume, a dye, an antibiotic, an antibacterial agent, and an antifungal agent. As the oil, hydrogenated vegetable oil, castor oil, cottonseed oil, olive oil, palm oil, jojoba oil and avocado oil may be used. Examples of the wax include wax, wax, carnauba, candelilla, montan, ceresin, liquid paraffin, Can be used. Examples of the fatty acid include stearic acid, linoleic acid, linolenic acid and oleic acid. The fatty acid alcohols include cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol and hexadecanol As the fatty acid ester, isopropyl myristate, isopropyl palmitate, and butyl stearate may be used. As the surfactant, a cationic surfactant, an anionic surfactant and a nonionic surfactant known in the art can be used, and a surfactant derived from a natural material is preferably used. In addition, it may contain a hygroscopic agent, a thickening agent, an antioxidant and the like widely known in the field of cosmetics, and the kind and amount thereof are well known in the art.

In one embodiment of the present invention, there is provided a method for producing a fermentation broth comprising: (a) cultivating a bellflower in a liquid phase; (b) cultivating fermentation broth by inoculating mycelia with a mycelium of a liquid medium, And (c) producing a biotransformation product of the fermentation and enzyme treatment which has improved antiallergic efficacy from the fermentation product produced by the biotransformation fermentation process of the step (b) through the biotransformation enzyme treatment process The present invention also provides a method of producing fermentation products having increased antiallergic activity, comprising the steps of: (a) culturing the seedlings of step (a) in a culture medium, the seedlings obtained by treating the seedlings obtained by pulverizing the seedlings with hydrolytic enzymes, Wherein the enzymatic treatment is a treatment of a cellulolytic enzyme capable of degrading a cell wall component and an enzyme of an amylase family Characterized in that the cellulolytic enzyme is selected from the group consisting of cellulase, hemi-cellulase, pectinase and glucanase in the production of an antiallergic efficacious fermentation product Wherein the mycelia of the bacterium of step (b) are selected from the group consisting of mushroom, chaga mushroom, shiitake mushroom, mushroom mushroom, mushroom mushroom, Wherein the bioconversion product of step (c) inhibits secretion of IgE in IgE-secreting B cells and inhibits the secretion of histamine in mast cells. Wherein the biotransformation product of step (c) is selected from the group consisting of TNF-a, IL-6, IL-10 and IFN- Wherein the biological conversion product of step (c) is selected from the group consisting of TNF-α, IL-1β and nitric oxide in serum, which are indicators of systemic inflammation in sepsis symptoms, And a method for producing the fermented product having an increased anti-inflammatory effect.

In another embodiment of the present invention, there is provided a fermented product having increased antiallergic activity by fermenting culture fermented by inoculation of mycelium of Mycelia into bellflower, wherein the allergen provides an allergic fermented product with increased allergic potency, Antiallergic efficacy reduces the production of IL-2, IL-10, IL-12, and gamma-interferon, as well as reducing IgE production in allergic symptoms and inhibiting the production of IL-4, IL-5 and IL- In a fermented product having increased antiallergic activity.

In another embodiment of the present invention, there is provided a pharmaceutical composition for the treatment of allergy comprising an fermented product having increased antiallergic potency, which is fermented by inoculation with mycelia of Mycelium of phytophagous bacteria, wherein the allergen is an allergic rhinitis Lt; / RTI >

In another embodiment of the present invention, there is provided a pharmaceutical composition for the treatment of inflammation comprising fermented product having increased antiallergic potency, which is fermented by culture inoculated with phytophagous mycelia in a bellflower, To provide a pharmaceutical composition.

In another embodiment of the present invention, there is provided a food composition for anti-allergy comprising a fermented product having increased antiallergic potency, which is fermented by inoculation of mycelium of mycelia with rhizome.

In another embodiment of the present invention, there is provided a cosmetic composition for anti-allergy comprising a fermented product having increased antiallergic potency, which is fermented by culture inoculated with mycelium of Mycobacterium strains in the platycodon.

Hereinafter, the present invention will be described in detail.

Fermented microorganisms (mycelial mycelia) fermented or biologically converted into biotechnology through further enzymatic treatment contain a large amount of antiallergic substances compared to unfermented phytophagous bellflower extract (bellflower) It is anticipated that there will be an effect of reducing the occurrence of allergy.

FIG. 1 is a schematic view showing a culture medium, a bioconversion process, and a fermentation powder obtained from a broth, according to an embodiment of the present invention.
FIG. 2 is a graph showing the activity of inhibiting the IgE antibody production according to the concentration of raw material and fermented product according to one embodiment of the present invention.
FIG. 3 is a graph showing the results of inhibitory activity of mast cell degranulation according to concentration of raw material and fermented product according to one embodiment of the present invention.
FIG. 4 is a graph showing histamine release inhibitory activity according to the concentration of the raw materials and the fermented product, according to an embodiment of the present invention.
FIG. 5 is a graph showing the results of measurement of cytotoxicity according to the concentration of the raw material of the broth and the fermented product, according to an embodiment of the present invention.
FIG. 6 is a graph showing the results of TNF-.alpha. Expression assay according to the concentration of raw materials and fermented products of Doraji according to an embodiment of the present invention.
FIG. 7 is a graph showing the results of evaluating the IL-6 expression-producing ability according to the concentration of the raw material and the fermented product, according to an example of the present invention.
FIG. 8 is a graph showing the results of evaluating the IL-10 expression-producing ability according to the concentration of the raw material and the fermented product, according to an embodiment of the present invention.
FIG. 9 is a graph showing the results of IFN-.beta. Expression assay according to the concentration of the raw Dorago raw material and the fermented product, according to an embodiment of the present invention.
10 is a diagram illustrating a method of producing an experimental animal model of OVA-sensitization (allergic rhinitis / asthma) mouse according to an embodiment of the present invention.
11 is a graph showing the results of measurement of IgE concentration in BALF and blood of an OVA-sensitized animal model according to an embodiment of the present invention.
FIG. 12 is a graph showing the results of measurement of concentrations of representative Th1, Th2 and Treg immunoreactive cytokines in bronchoalveolar lavage fluid (BALF) of an OVA-sensitized animal model according to an embodiment of the present invention.
FIG. 13 is a graph showing the results of measurement of concentrations of representative Th1, Th2, and Treg immunoreactive cytokines in the blood of an OVA-sensitized animal model according to an embodiment of the present invention.
FIG. 14 is a graph showing the results of confirming the effect of BALF on the number of immune cells such as eosinophils in an OVA-sensitized animal model according to an embodiment of the present invention. FIG.
FIG. 15 is a graph showing changes in mortality according to concentration and route of administration of platycodon fermentation products in an animal model in which a sepsis was induced, according to an embodiment of the present invention. FIG.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example 1. Bioconversion process of fermentation and enzyme treatment

Example 1-1. Bellflower preprocessing

For the pre - treatment of the purchased platycodon from the raw material to the pulverized powder, mold contamination degree of the raw material condition was measured, and water washing work was performed to remove foreign matter and contaminants from the raw material. (1) air washing step to remove foreign matters and mold spores, (2) water washing step to remove residual pesticides, (3) alcohol (Et-OH) to remove contaminants such as microorganisms, Washing of the raw materials was carried out through the washing step. After washing and cutting, the raw materials were dried in a hot air drier capable of mass drying, and subjected to pulverization of each of the natural raw materials through crude pulverization and pulverization.

Example 1-2. Obtained Fermented Powder of Bellflower

Platelets with foreign and fungal contamination were treated with enzyme treatment and heat - treatment sterilization for liquid culture medium. The enzymes are hydrolyzed by cellulosic and hemi-cellulosic hydrolytic enzymes capable of decomposing bark, muscle, root, and leaf, and hydrolytic enzymes such as pectinase, glucosidase and amylase hydrolytic enzymes . After the enzyme was added, the reaction was carried out at 60 ° C for 1 hour and sterilized at a high temperature for 30 minutes to culture the broth.

Then, the mycelium of mycelia cultivated separately in the culture (fermentation) medium was added. When the concentration of the residual carbon source is depleted to a certain concentration or less under the condition of inoculating 10% (v / v) culture of mycelium mycelium culture at 28 ~ 30 ° C and pH 4.5 ~ 7, And cultured for 7 to 10 days in a food process. The above mycelium of mycelia may be selected from mycelia such as mushroom, chaga mushroom, shiitake mushroom, mushroom mushroom, mushroom mushroom, snow mushroom mushroom, and skirting mushroom, which are medicinal and edible bacillus species, Ryu Myung-sa is a member of the Korean Society for Microbiology (KCCM), Korea Biotechnology Research Institute (KCTC), Korea Cell Line Bank (KCLB), Korea Agricultural Microbiology Resource Center (KACC) It is possible to use the strain in an institution.

The enzymatic treatment of the biotransformation process for the above fermented product is carried out by adding various enzymes such as cellulose decomposing enzyme, cellulase, hemicellulase, pectinase and glucanase after hot water extraction to an appropriate amount (The optimum amount indicated in the product manual of each enzyme preparation). Examples of the enzymes include commercially available enzymes such as a cellulase (a compound derived from Aspergillus niger), a filtrate (a compound derived from Filtrase (Tricoderma reesei)), a rapidase (a compound derived from a fibrinolytic enzyme, Viscozyme (Complex containing Aspergillus niger) and Sumizyme (a complex containing Aspergillus niger-derived pectinase) was added at the recommended concentration (10% v / v) given in the product manual of each enzyme preparation, The enzyme / substrate reaction was performed by rotating at 250 rpm for 1 to 3 hours.

Platycodon fructus produced by the biotransformation process was lyophilized by enzyme inactivation and sterilization process at 90 ℃ for 1 hour. In addition, the above-mentioned platelet-rich fermented starch powder having an immunostimulatory activity was dissolved in 10 times of water and centrifuged at 10,000 rpm for 30 minutes to remove insoluble residues. The supernatant was lyophilized to obtain a water-soluble powder or 5-fold ethanol Followed by induction of precipitation at 4 DEG C for 24 hours or longer, and the precipitate was lyophilized to obtain a powder of a polymeric polysaccharide fraction having immunological activity. The culture broth, bioconversion process and fermentation powder obtained from the above-mentioned platelets are shown in Fig.

Example 2. Biotransduced Bellflower Fermented Evaluation of in vitro efficacy

Example 2-1. In B cells IgE Evaluation of Antibody Production Inhibitory Activity

Biotransformation was U266B1 cell line (B cells of human origin) for confirming bellflower fermentation water IgE production suppressing effect was purchased from World Cell Line Bank (ATCC), 37 ℃, 5 % CO 2, of 15% under the conditions of saturation humidity The cells were cultured in RPMI-1640 medium supplemented with heat-inactivated fetal bovine serum (FBS) and 100 units of penicillin and 100 μg / ml of streptomycin. After inoculation the U266B1 cells to 1 x 10 ⁶ cells / well density in a 96 well culture plate and cultured overnight. After incubation, B cells were stimulated by treatment with 10 μg / ml of LPS and 5 ng / ml of IL-4. In addition to the stimulation, the seeds and the fermented product were treated at a concentration of 1, 10, and 100 μg / ml, respectively, for 72 hours. After incubation, the supernatant was collected and the amount of IgE secreted from U266B1 cells was measured using a Sandwich ELISA Assay (BETHYL, Montgomery, TX) kit. Doraji was purchased from Yeongcheon (CM) in Gyeongbuk province and Hoengseong (HS) in Kangwon province. Both the raw material and the fermented product were found to have a concentration - dependent inhibitory activity against IgE production. The inhibitory activity of CM raw material and fermented product at 100 ㎍ / ㎖ concentration was 25.3% and 66.9%, respectively. Kangwon - do (HS) acidophilus was inhibited by 27.2% and 66.2% By the effect, there was no difference between two origin. It was found that the activity of inhibiting IgE production was significantly increased in the fermented material compared to the raw material of bellflower. Especially, the inhibitory activity of IgE antibody production was higher at a concentration of 1 ㎍ / ㎖ of the fermented material at a concentration of 100 ㎍ / ㎖ of the raw material will be. The results are shown in Fig.

Example 2-2. In mast cells Degranulation Evaluation of inhibitory activity

Beta-hexyl sosami is a kinase (β-hexosaminidase) inhibition were obtained for (mast cells of rat origin) RBL-2H3 cell line for the activity evaluation at the ATCC, 37 ℃, 5% CO 2, 10% under the conditions of saturation humidity complement Were cultured in DMEM medium containing inactivated FBS (heat-inactivated fetal bovine serum), 100 units of penicillin, and 100 占 퐂 / ml streptomycin. This was dispensed into a 96-well culture dish at 1 × 10 ⁵ cells / well and further cultured overnight. After incubation, the medium was removed and the cells were washed with thyloid buffer (137 mM NaCl, 2.7 mM KCl, 1.8 mM CaCl ₂ , 1.1 mM MgCl ₂ , 11.9 mM NaHCO ₃ , 0.4 mM NaH ₂ PO ₄ , 5.6 mM glucose, pH 7.2), treated with the appropriate concentration of the material, and incubated at 37 ° C for 20 minutes. After three washes with thyloid buffer, 10 μM of A23187 (calcium ionophore) was treated and cultured for 20 minutes to induce degranulation. In the case of inducing degranulation using DNP-BSA, cells were inoculated with 1 ㎍ / ㎖ of IgE, sensitized, and treated with 100 ㎍ / ㎖ of DNP-BSA for 30 min at the appropriate concentration. Respectively. After induction, 100 μl of the supernatant was recovered, and 1 mM p-Nitrophenyl-β-Acetyl-Glucosamide 100 dissolved in 0.1 M citrate buffer (pH 5.0) The reaction was terminated by adding 200 μl of sodium bicarbonate buffer (pH 10.2) in a 37 ° C incubator for 1 hour. The reaction was terminated by using a microplate reader (Bio-Rad, Hercules, CA) The absorbance at 405 nm was measured.

In the beta - hexosaminidase assay, the inhibitory activities of the raw materials and the fermented materials according to their origin were higher in the fermented materials than in the raw materials and fermented material, . β-hexosaminidase assay showed that 41.9% inhibition activity was obtained from 100 g / ㎖ of A23187, and 47.9% inhibition activity of Kwangwon-do, Yeongcheon (CM) Water showed 46.8% and 51.2% inhibitory activity, respectively, and it was confirmed that the hydrolysis activity of the hydrogens (HS) of Gangwon - do was slightly higher than that of Yeongcheon (CM) acid of Gyeongbuk province.

When stimulated with DNP-BSA, the contents and fermented products of Yeongcheon (CM) in Gyeongbuk province showed 27.9% and 31.2% inhibitory activity at 100 ㎍ / ㎖, respectively. The Kangwon province (HS) % And 37.4%, respectively. As in the case of A23187 stimulation, it was confirmed that the HS acid of Kangwon - do had a slightly higher inhibitory activity than that of Yeongcheon (CM) in Gyeongbuk province. However, in both stimulants, The difference was not large. The results are shown in Fig.

Example 2-3. Assessment of histamine inhibitory activity in mast cells

RBL-2H3 cell line (rat origin mast cells) for histamine evaluation was purchased from ATCC and incubated with 10% complement-inactivated fetal bovine serum (FBS) at 37 ° C, 5% CO2, And 100 units of penicillin and 100 [mu] g / ml of streptomycin. After the cells were sensitized with A23187, the raw materials and fermented products of Kangwon-do (HS) acid were treated at the concentrations of 1, 10, and 100 ㎍ / ㎖. To measure histamine, cultured RBL-2H3 cells were treated with 0.25% trypsin and the cells were harvested and suspended in thyloid buffer (137 mM NaCl, 2.7 mM KCl, 1.8 mM CaCl2, 1.1 mM MgCl2, 11.9 mM NaHCO3, 0.4 mM NaH2PO4, 5.6 mM glucose, pH 7.2) at a concentration of 1 × 10 ⁶ cells / well, and then dispensed into a 24-well culture dish. After that, the material was treated at an appropriate concentration and incubated for 15 minutes. Then, 10 μM of A23187 knife The cells were treated with calcium ionophore and cultured again for 20 minutes. After culturing, the reaction was terminated by treatment at 4 ° C for 10 minutes, and the supernatant was recovered. To 1 ml of the recovered supernatant was added 0.2 ml of 1 N NaOH and 0.1 ml of 1% OPT (o-phthalaldehyde), and the mixture was allowed to stand at room temperature for 5 minutes. The reaction was terminated by treatment with 0.2 ml of 1 N HCl and then measured using a fluorescence meter (Molecular Devices, Sunnyvale, Calif.) At excitation wavelength of 360 nm and emission wavelength of 450 nm.

Histamine release inhibition activity was measured by using the raw material and fermented product of Hodongsan (HS) in Kangwon province. As a result, the inhibitory activity of the raw material was 31.6% and the bioconversion fermentation product was 39.9% at the concentration of 100 ㎍ / ㎖, It was confirmed that the enzymatic treatment showed a slight increase in activity depending on the bioconversion process. The results are shown in Fig.

Example 2-4. Cytotoxicity Assessment

The RBL-2H3 cell line cultured by the method of Example 2-2 was dispensed into a 96-well culture dish at a concentration of 1 x 10 ⁴ cells / well, and then the brazen raw material and the fermented product were mixed at a concentration of 1, 10, and 100 μg / And cultured for 48 hours. After the incubation, the supernatant was removed and 200 μl of a 0.5 mg / ml MTT reagent (3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) diluted in the medium was treated and cultured again for 3 hours Respectively. After incubation, the supernatant was removed and 200 μl of DMSO was added to dissolve the intracellular precipitate. The absorbance at 570 nm was measured using a microplate reader (Bio-Rad, Hercules, Calif.).

In the RBL-2H3 mast cell line, the cytotoxicity of the HSCB was not observed at 100 ㎍ / ㎖ of both the raw material and the fermented product. The results are shown in Fig.

Example 2-5. In macrophages TNF Immune response using the expression rate of Activation ability evaluation

In order to evaluate the activation of congenital immune responses of elm fermented products, culture and assay analysis of macrophages (macrophages) were carried out using 5% DMEM medium. The macrophages (macrophages) used for the immunoactivity measurement were RAW 264.7 cells, and the subcultures were subcultured in 0.5 ml each at a concentration of 1 × 10 ⁵ cells / ml in each well of a 24-well culture dish. After culturing for 24 hours, the cell culture was confirmed with a microscope, and the medium was completely removed and then washed 2-3 times with the medium without serum. Each well was filled with 0.5 ml of 5% FBS DMEM medium supplemented with a sample. The concentration of the sample was 1/10, starting at a concentration of 100 μg / ml, and adjusted to 1, 10, and 100 μg / Respectively. In addition, LPS in a final concentration of 1 / / ml was treated with a positive control (200 / / ml stock). 24 wells treated with the sample were incubated for 8-24 hours, and 400 μl of the culture solution of each well was taken. 400 μl of the culture solution was centrifuged at 12,000 rpm for 5 minutes, and the supernatant was used for assay analysis.

The expression of TNF-α was examined in the macrophages to confirm the innate immune response activation effect of the raw material and the fermented product. Experiments were conducted by treating the raw material of the bellflower and the fermented product at a concentration of 10 and 100 μg / ml. The culture supernatant, which had been treated for 18 hours after the sample was treated, was analyzed with an assay of TNF-α (R & D system, MTA00B) according to the manufacturer's protocol.

As a result, it was confirmed that TNF-α expression ratio was higher in the bellflower fermented product than in the bellflower raw material. The results are shown in Fig.

Example 2-6. Immune response using the expression rate of IL-6 in macrophages Activation ability evaluation

The expression of IL-6 was examined in macrophages cultured in the same manner as in Example 2-4 in order to confirm the innate immune response activation effect of the raw material of the bellflower and the fermented product. Experiments were conducted by treating the raw material of the bellflower and the fermented product at a concentration of 10 and 100 μg / ml. The culture supernatant, which had been treated for 18 hours after the sample was treated, was analyzed with an assay of IL-6 (R & D system, M6000B) according to the manufacturer's protocol.

The results showed that IL-6 was almost not expressed in the group treated with the bollard, but the level of IL-6 expression was significantly increased in the group treated with the bellflower fermented product. The results are shown in Fig.

Example 2-7. Immune response using the expression rate of IL-10 in macrophages Activation ability evaluation

The expression of IL-10 was examined in macrophages cultured in the same manner as in Example 2-4 in order to confirm the innate immune response activation effect of the raw material of the bellflower and the fermented product. Experiments were conducted by treating the raw material of the bellflower and the fermented product at a concentration of 10 and 100 μg / ml. The culture supernatant, which was treated for 18 hours after the sample was treated, was analyzed by IL-10 (R & D system, M1000) assay according to the manufacturer's protocol.

As a result, it was confirmed that IL-10 expression was higher in the bellflower fermented product than in the bellflower. The results are shown in Fig.

Example 2-8. In macrophages IFN Immune response using the expression rate of? Activation ability evaluation

In order to confirm the innate immune response activation activity of the raw material and the fermented product, the expression ability of IFN-β was examined in the macrophages cultured in the same manner as in Example 2-4. Experiments were conducted by treating the raw material of the bellflower and the fermented product at a concentration of 10 and 100 μg / ml. The culture supernatant, which had been treated for 18 hours after the sample treatment, was taken and assayed according to the protocol of the manufacturer using an IFN-β (R & D system, 42400) assay.

As a result, it was confirmed that IFN-β expression was higher in the bellflower fermented product than in the bellflower raw material. The results are shown in Fig.

IFN-β is a cytokine that induces a Th1 immune response and is an immunomodulatory cytokine that inhibits the Th1 and Th17 immune responses while inducing a Th1 immune response. Allergy is known to occur as the Th2 immune response increases. Platelet-based fermented products are expected to inhibit the expression of cytokines associated with the Th2 immune response that induces allergy by promoting the expression of IFN-.beta. Associated with the Th1 immune response.

Example 3. In allergic rhinitis models, Fermented In the body (in vivo ) Efficacy evaluation

Example 3-1. Allergic rhinitis Establishment of induction experimental animal model

In order to evaluate the antiallergic activity of the Doraji fermented product, an experimental animal model of OVA-sensitization (allergic rhinitis / asthma) mice was established. Egg white albumin (ovalbumin) was used as an antigen and alumina was used as a reinforcement agent. On the first day of experiment and 14 days, albumin albumin / alumina was sensitized by two intraperitoneal injections. After 2 weeks of elapsed time, allergic reaction was induced by inhalation of albumin albumin for 30 days for 30 days from 30 days. Evaluation of the platycodon fermented product was performed by two ovalbumin / alumina intraperitoneal injections at the beginning and at the 14th day of the experiment. From the 15th day until the end of the experiment, the fermented product was orally administered at a dose of 10 mg / The effects were evaluated. FIG. 10 shows the production method of the OVA-sensitized (allergic rhinitis / asthmatic) mouse experimental animal model.

Example 3-2. BALF And blood IgE produce Control ability evaluation

As shown in FIG. 9, OVA-sensitized (allergic rhinitis / asthma) experimental animal model produced by the method of Example 3-1 induces an allergic reaction by inhalation of albumin albumin daily for 5 days from the 30th day of animal model production. In addition, the bellflower fermented product was sensitized by two ovalbumin / alumina abdominal injections on the day of experiment and on day 14, and daily from day 15 until the end of experiment. On day 35, mice were sacrificed and bronchial alveolar lavage fluid (BALF) was recovered and blood was also collected. The amount of immunoglobulin E (IgE) in the recovered BALF was 45.4% and 80.7%, respectively, and the inhibitory activity against IgE production was significantly increased. Immunoglobulin E in the blood was also inhibited by 25.2% in the fermented broth and 73.6% in the fermented broth. The results are shown in Fig.

Example 3-3. At BALF Cytokine generation Control ability evaluation

In order to investigate the degree of activation of Th2 cells that regulate the proliferation and activity of B cells as the production of IgE was inhibited in OVA-sensitized (allergic rhinitis / asthmatic) experimental animal model due to the administration of the bellflower fermented product, the Th2 immune response IL-5, and IL-13 in the group fed with the bell pepper fermented product by ELISA method as a result of measuring the amount of representative cytokines of Th1 and Treg immunoreactions together with the typical cytokines (OVA only) as compared with the control group (OVA only) and the control group (OVA only), respectively. On the other hand, the production of IL-10, which is a representative cytokine of Treg immunoreactivity with IL-2 and IL-12, which are typical cytokines of Th1 immune response, , And the intake of raw materials was also higher than that of the control group. The results are shown in Fig.

Example 3-4. Evaluation of cytokine concentration in blood

The concentrations of representative cytokines of Th1, Th2 and Treg immunoreactivity in the serum of OVA-sensitized (allergic rhinitis / asthma) experimental animal models were measured using the ELISA method as in Example 3-3. As a result, IL-10, which is a representative cytokine of Treg immunoreactivity, together with IL-2 and IL-12, which are typical cytokines of Th1 immune response, And the intake of raw materials was also increased compared with that of the control group. Gamma interferon (Interferon-γ or IFN-γ), which has been shown to inhibit eosinophil infiltration in allergic inflammation sites, is a representative cytokine secreted by Th1 cells. In contrast to the control and ingestion groups, , And the intake of raw materials was also higher than that of the control group. On the other hand, the production of IL-4, IL-5 and IL-13, which are representative cytokines of Th2 immune response, was reduced in the group fed with the fermented Doraji, compared to the control group (OVA only) And the intake of raw materials was also decreased compared to the control group. The results are shown in Fig.

Example 3-5. At BALF Eosinophilia inhibition and On cell number Check the effect

(BALF) of the experimental animal model of OVA-sensitization (allergic rhinitis / asthma) due to the administration of the fermented Dorago fermented product on the eosinophilia inhibition and the cell number. Experimental results showed that the number of immune cells in the BALF was significantly reduced in the group fed with the platycodon fermented product compared to the group consumed with the raw material. In particular, the number of cells in eosinophils and lymphocytes was significantly reduced. The results are shown in Fig.

Example 4. In the sepsis model, Fermented In the body (in vivo ) Safety evaluation

Example 4-1. Establishment of an animal model for induction of sepsis

Based on the in vivo safety evaluation results of the fermentation of the bellflower, the limit dose and route of administration of the fermentation product in vivo was evaluated. The mice were injected with peritoneal injection of LPS (20 ㎍ / kg) and galactosamine (700 mg / kg) at a concentration of 1, 10 and 100 mg / kg for 2 weeks. Lt; / RTI >

Example 4-2. Comparison of lethal dose by concentration and route of administration

As a result of examining the mortality rate of mice up to 60 hours after the induction of sepsis in the experimental animal model according to the method of Example 4-1, 0 mice and 10 mice in the experimental group administered with 1 mg / kg of Doraji fermented product in 10 mice per experimental group 4 mice were observed to survive 4 mice in the peritoneal injection of 100 mg / kg, and 4 mice were observed in the peritoneal injection of 100 mg / kg. In addition, , And 4 and 2, respectively, survived. That is, as the dose was increased to 1, 10, and 100 mg / kg, the survival rate was improved, and it was confirmed that the borage fermented material was a very safe immune modulating material. The results are shown in Fig.

Example 4-3. Systemic inflammatory response Indicators( TNF -α, IL- 1β , Nitric oxide) evaluation

As shown in Example 4-2, the mortality rate of sepsis at the LPS lethal dose was decreased due to the administration of the platelet fermentation product, so as to confirm the inflammation inhibitory effect of the developmental material in the systemic inflammatory reaction. (10 μg / kg) was administered orally for 2 weeks, followed by sub-lethal dose of LPS (5 μg / kg) and galactosamine (700 μg / kg) depending on the determined dosage and administration route. mg / kg) to induce the systemic inflammatory response. After induction of inflammation for 4 hours, mice were sacrificed and blood was drawn from the heart. Serum was separated and the concentration of TNF-α, IL-1β and nitric oxide in the serum of the systemic inflammatory response was confirmed.

As a result of the experiment, it was confirmed that the mice to which the platelike fermented product was administered inhibited TNF-α production by 40% or more as compared with the control group. IL-1β, a typical inflammatory cytokine, also showed a high inhibitory activity of 45% or more, confirming that the Rhizobium fermented product has an inflammatory response inhibitory activity. The amount of nitric oxide in serum, another indicator of systemic inflammatory reaction caused by LPS, was also measured to show a high production inhibitory activity of 55% or more. The results are shown in Table 1.

Sample TNF -α ( pg / mL) IL- 1β Nitric oxide ( uM ) Vehicle 32.597 ± 1.209 12.174 + 0.896 0.722 + 0.041 Negative control 382.226 ± 15.467 128.462 + - 5.447 19.114 ± 1.005 Bellflower fermentation product 236.904 + 14.414 75.643 ± 3.008 8.524 + 0.566

Claims

(a) culturing the bellflower in a liquid phase;
(b) carrying out a bioconversion fermentation process in which a mycelium of mycelium is inoculated in a liquid medium of a cultured medium of the step (a) to effect culture fermentation; And
(c) producing a bioconverted product of fermentation and enzyme treatment in which the antiallergic effect is improved through the treatment with a fibrinolytic enzyme from the fermentation product produced by the bioconversion fermentation process of the step (b). A method for producing a fermented product with increased allergic potency.

The method according to claim 1,
Wherein the step (a) is performed in a culture medium of the step (d), wherein the step (b) of the step (b) is performed with a hydrolytic enzyme, followed by sterilization to form a liquid Dorado medium.

3. The method of claim 2,
Wherein the hydrolytic enzyme is a cellulolytic enzyme capable of degrading a cell wall component and an amylase, wherein the antiallergic activity is increased.

The method of claim 3,
Wherein said fibrinolytic enzyme is selected from the group consisting of cellulase, hemi-cellulase, pectinase and glucanase.

The method according to claim 1,
Wherein the mycelium of step (b) comprises mycelium of bacterium selected from the group consisting of a mushroom, a mushroom, a shiitake, a mushroom, a mushroom, a mushroom and a mushroom, &Lt; / RTI >

The method according to claim 1,
Wherein the bioconversion product of step (c) inhibits the secretion of IgE in IgE-secreting B cells and inhibits the secretion of histamine in mast cells.

The method according to claim 1,
Wherein the bioconversion product of step (c) promotes the expression of TNF-a, IL-6, IL-10 and IFN-? In macrophages.

The method according to claim 1,
Wherein the biological conversion product of step (c) reduces TNF-α, IL-1β and nitric oxide concentrations in serum, which is an indicator of systemic inflammation, in the symptom of sepsis.

A fermented product with increased antiallergic potency, which is cultured and fermented by inoculating bacillus mycelia with a bellflower.

10. The method of claim 9,
Wherein the allergen is an allergic rhinitis, the fermentation product having increased antiallergic potency.

10. The method of claim 9,
The antiallergic effect reduces the production of IgE in allergic symptoms and inhibits the production of IL-2, IL-10, IL-12 and gamma-interferon in addition to inhibiting the production of IL-4, IL-5 and IL- Wherein the fermentation product has an increased antiallergic activity.

A pharmaceutical composition for the treatment of allergies, comprising fermented product having increased antiallergic potency, which is fermented by culture inoculated with bacillus mycelia in a bellflower.

13. The method of claim 12,
Wherein the allergic is allergic rhinitis.

A pharmaceutical composition for the treatment of inflammation, which comprises fermented product having increased antiallergic potency, which is cultured fermented by inoculation of mycelium of Mycobacterium sp.

15. The method of claim 14,
Wherein said inflammation treatment is a sepsis treatment.

A food composition for antiallergic use, comprising a fermented product having increased antiallergic potency obtained by inoculation of bacillus mycelia in a bellflower and fermented by culture.

A cosmetic composition for antiallergic use, comprising a fermented product having increased antiallergic potency, which is fermented by culture inoculated with bacillus mycelia in a bellflower.