KR101853701B1 - Compositions with bioconversion Sesamum indicum for improvement of immunity, diabetes, hyperlipemia or protection against liver damage - Google Patents

Abstract

The present invention relates to a composition containing a sesame bioconversion product for enhancing immunity, alleviating diabetes, alleviating hyperlipemia or protecting liver. The composition containing the sesame bioconversion product of the present invention can increase the immunity regarding microbial infection through activation of macrophages, alleviate diabetes, alleviate damage of the liver or damage of Langerhans islet of pancreas, and inhibit production of triglycerides. Moreover, alcoholic fatty liver or alcoholic fatty liver diseases expressed by alcohol can be inhibited.

Description

[0001] The present invention relates to a composition for improving immunity, for improving diabetes, for improving hyperlipidemia, or for protecting liver, which comprises sesame bioconversion. [0002] The present invention relates to a composition for preventing or alleviating diabetes, hyperlipemia or protection against liver damage,

The present invention relates to a composition for improving immunity, for improving diabetes, for improving hyperlipemia or for protecting liver containing a sesame bioconversion product.

Immunity is largely divided into innate immunity (aka natural immune) and acquired immunity (aka acquired immunity). The three Nobel laureates of the 2011 Nobel Prize in Physiology awarded the Nobel Prize for medicine for discovering the core principles of the immune system against bacteria, viruses and fungi.

According to these studies, foreign invaders such as pathogenic microorganisms enter our bodies and are first associated with specific receptors on the cell surface of immune cells responsible for innate immunity. At this time, our body discovers external antigens, which are external intruders, and responds quickly. As a result, an inflammatory reaction occurs in cells and tissues, and the body feels fever or body fatigue.

This early immune response occurs immediately, regardless of the identity of the foreign antigen. Congenital immunity reacts nonspecifically to the antigen. In congenital immunity, macrophage, polymorphonuclear leucocyte, and NK cell, which can kill infected cells, are the most common. In fact, most infections are congenital It is defended by immune.

However, when immunity in the human body is weakened for various reasons, many immune enhancers have been developed and used to enhance immunity. Recently, immune activities such as anti-complement activity and lymphocyte division induction activity of electric polysaccharides extracted from fungi including fungi and mushroom have attracted attention. Therefore, in order to develop a new immunomodulating substance, polysaccharides produced in fungi have been intensively studied.

On the other hand, the risk of diabetes is emphasized because it can lead to various complications as well as death itself. The prevalence of diabetes is not only high in the world but also in Korea, the prevalence of diabetes and diabetes complications is higher than the OECD average. Therefore, the social cost is gradually increasing, which is attracting attention as a task to be solved.

Diabetes mellitus is classified into two types. First, the pancreatic beta cell function is broken and insulin is not properly secreted. Therefore, the first type of insulin is not controlled, and there is no problem with insulin secretion, but insulin sensitivity is low. Type 2 diabetes mellitus does not control blood sugar properly because the signal is not properly transmitted.

Type 1 diabetes is mostly congenital, and because insulin secretion is poor, it can be managed by simple treatment with insulin.

On the other hand, type 2 diabetes occurs mainly as an acquired factor, and it is known that type 2 diabetes occurs mainly due to insulin resistance. In the case of normal people, when the blood glucose concentration is increased through digestion and absorption after food intake, insulin secretion is smoothly performed, and cells of each tissue to which glucose is to be absorbed normally recognize insulin and absorb glucose. Absorbed sugars are used for a variety of uses, such as energy production or glycogen storage, thereby lowering blood sugar. However, in a person with insulin resistance, the cells in each tissue are less sensitive to insulin, failing to recognize secreted insulin normally, and not having proper glucose uptake and utilization.

If insulin does not play a role in insulin resistance, glucose can not be absorbed as an energy source by each tissue, or glucose can not be properly decomposed into glycogen, Eventually leading to type 2 diabetes. Such diabetes increases triglycerides, cholesterol, and may also cause liver and pancreatic tissue damage.

On the other hand, alcohol can cause liver damage, which is called alcoholic fatty liver. Currently, the number of chronic liver disease patients in the world reaches about 600 ~ 800 million, and the first cause of death in Korea is the liver disease. However, the currently marketed medicines are not effective in treating liver fibrosis or cirrhosis, and have been making efforts to develop them at home and abroad. However, the results are still insufficient.

On the other hand, sesame (Sesamum indicum ) is a dicotyledonous plant, also called homa, 麻麻, and 麻麻. Sesame seeds are capillary, cylindrical, and contain about 80 seeds. Sesame seeds contain 45-55% oil and 36% protein. Sesame seeds are used for various foods and seasonings, and are especially used in Asia including Korea. It also uses squeezed oil from the seeds and leaves the oil, which is then used as feed and fertilizer.

Because sesame contains a lot of lecithin ingredient, it not only prevents spots or freckles, it also has antioxidant effect because it is rich in vitamin E, and it prevents the aging of skin. It is said that it shows good efficacy for dental health. In addition, sesame contains essential fatty acids, and it is known that it steadily consumes sesame seeds, it helps activation of brain function, improves memory and concentration, and shows good efficacy against dementia.

However, up to now, it has been confirmed that a composition exhibiting both immunological enhancement, diabetic improvement, triglyceride and cholesterol suppression, and hepatoprotective effect, including a sesame bioconversion bioconverted into a superficial hyphae, has not been studied.

Korean Patent No. 10-1467903 (entitled "Immune Enhancer Produced Through Mycelial Fermentation and Bioconversion Process of Mycelia of Mycelia of Black Rice)" discloses a method for producing an immunological active substance produced by mycelia fermentation and biotransformation process (From mycelium of bacillus mycelium, mushroom, chaga mushroom, shiitake mushroom, thrips mushroom, and snow blossom mushroom) on a liquid black rice meadow prepared by cultivating black rice into a fermentation broth from a culture produced by the culturing process A method for producing a bioconversion product that dramatically improves immunity through a bioconversion process and a biotransformation product are described as a composition characterized by a polymeric polysaccharide as an active ingredient having an immunostimulating effect. Korean Patent Registration No. 10-0517899 entitled "Biodegradable Ginseng Composition and Method for Producing the Same", the ginseng composition is prepared by suspending a raw material of ginseng in water and adding lactic acid bacteria for about 24 hours to 72 hours To obtain a biotransformed ginseng liquid; And a concentrating step of concentrating or freezing / drying the biotransformed ginseng solution which has undergone the above process as it is, concentrating the biotransformed ginseng solution by centrifugation, and filtering only the supernatant to obtain a biotransformed ginseng concentrate Of the present invention.

In the present invention, it is intended to develop and provide a novel composition exhibiting an immunity enhancing effect, diabetic improvement, hyperlipidemia improvement or liver protecting effect.

To achieve the above object, the present invention sesame (Sesamum indicum ), comprising the steps of: (a) preparing a liquid medium; (B) preparing a fermented product by inoculating shiitake mycelia into the liquid medium after the step (a) and culturing the same; (C) a step of adding a fibrinolytic enzyme to the fermentation product after the step (b); And a sesame bioconversion product prepared from a process comprising the steps of:

In the immunoenhancing food or pharmaceutical composition of the present invention, the sesame bioconverting agent is preferably subjected to 'enzyme inactivation and sterilization step (d)' in which the step (c) is followed by treatment at 80 to 100 ° C for 0.5 to 2 hours, May be prepared from the process further comprising. Further, after the 'enzyme inactivation and sterilization step (d)', the 'drying and pulverization step (e)' may be further included.

In the immunoconjugation food or pharmaceutical composition of the present invention, the liquid medium of step (a) is preferably prepared by adding at least one selected from amylase or cellulase to sesame, reacting, . At this time, the amylase or cellulase enzyme is preferably added in an amount of 0.1 to 0.5% (w / v) and reacted for 0.5 to 2 hours. When the amylase or cellulase enzyme is treated, sesame starch and cellulose can be decomposed, which facilitates preparation into a liquid medium.

In the immunoconjugation food or pharmaceutical composition of the present invention, the mushroom mycelium of step (b) may be produced by culturing the cellulolytic enzyme and discharging it to the outside. The released cellulose decomposing enzyme can hydrolyze sesame seeds.

In the immunoconjugation food or pharmaceutical composition of the present invention, the fibrinolytic enzyme of step (c) may be, for example, a cellulase, a hemicellulase, a pectinase, a glucanase, , Beta-glucanase, beta-glucosidase, lysozyme, viscozyme, filtrase, rapidase and sumizyme ). ≪ / RTI >

In the immunomodulating food or pharmaceutical composition of the present invention, the sesame bioconversion preferably has immunity against microbial infection (activation) through activation of macrophages. The immunity (resistance) refers to resistance to disease.

In the immunomodulating food or pharmaceutical composition of the present invention, the sesame bioconversion may preferably be a polysaccharide. In this case, the polysaccharide is preferably obtained by adding distilled water to the sesame bioconversion product obtained through step (c) to extract a water-soluble substance; Removing the residue through centrifugation after extracting the water-soluble substance, and separating the extracted supernatant (B); Adding EtOH to the extracted supernatant, mixing and maintaining (c); Separating the resulting precipitate after said holding (d); Adding distilled water to the separated precipitate to dissolve the precipitate, and then removing undissolved material by centrifugation or filtration; And performing dialysis against the sediment lysis solution after the centrifugation or filtration step (f).

In the pharmaceutical composition for enhancing immunity of the present invention, the pharmaceutical composition preferably contains 0.00001 to 100% by weight of a sesame bioconversion product relative to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the pharmaceutical composition for immunostaining according to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, diluent or excipient in addition to the active ingredient. Examples of usable carriers, excipients or diluents 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, and one or more selected from among them may be used. When the pharmaceutical composition is a pharmaceutical, it may further include at least one selected from a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent or an antiseptic agent.

In the pharmaceutical composition for enhancing immunity of the present invention, the formulation of the pharmaceutical composition may be in a desired form depending on the method of use, and may be in the form of a pharmaceutical composition, To be formulated by employing a method known in the art. Examples of specific formulations include, but are not limited to, PLASTERS, GRANULES, LOTIONS, LINIMENTS, LEMONADES, AROMATIC WATERS, POWDERS, SYRUPS, OPHTHALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS, ), Suspensions, DECOCTIONS, INFUSIONS, OPHTHALMIC SOLUTIONS, TABLETS, SUPPOSITORIES, INJECTIONS, SPIRITS, CATAPLSMA, ), Capsules, CREAMS, TROCHES, TINCTURES, PASTES, PILLS, soft or hard gelatin capsules.

In the pharmaceutical composition for enhancing immunity of the present invention, the dosage of the pharmaceutical composition is preferably determined in consideration of the administration method, the age, sex, weight, and severity of the disease. For example, the pharmaceutical composition of the present invention can be administered at least once per day, 0.00001 to 100 mg / kg (body weight) based on the active ingredient. However, the dosage is only an example and may be changed by a doctor's prescription depending on the condition of the recipient.

In the food composition for immunostaining according to the present invention, the food composition preferably contains 0.00001 to 100% by weight of sesame bioconversion to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the food composition for enhancing immunity of the present invention, the food composition may be, for example, meat, cereal, caffeinated beverages, ordinary beverages, chocolate, bread, snacks, confectionery, candy, pizza, jelly, noodles, Alcoholic beverages, alcoholic beverages, vitamin complexes, and other health supplement foods, but the present invention is not limited thereto.

On the other hand, the present invention relates to a process for producing a liquid medium comprising sesame (a); (B) preparing a fermented product by inoculating shiitake mycelia into the liquid medium after the step (a) and culturing the same; (C) a step of adding a fibrinolytic enzyme to the fermentation product after the step (b); A diabetic improvement food composition or a pharmaceutical composition for the prevention or treatment of diabetes, which comprises a sesame bioconversion product prepared from a process comprising the steps of:

In the 'diabetic food composition for improvement of diabetes mellitus' or 'pharmaceutical composition for the prevention or treatment of diabetes' of the present invention, the sesame bioconversion product may be prepared by mixing the enzymatic fire Activation and sterilization step (d) 'are further included. Further, after the 'enzyme inactivation and sterilization step (d)', the 'drying and pulverization step (e)' may be further included.

In the 'diabetic food composition for improving diabetes' or the 'pharmaceutical composition for the prevention or treatment of diabetes' of the present invention, the liquid medium of step (a) is preferably at least one selected from the group consisting of amylase or cellulase And then sterilization treatment is carried out. At this time, the amylase or cellulase enzyme is preferably added in an amount of 0.1 to 0.5% (w / v) and reacted for 0.5 to 2 hours. When the amylase or cellulase enzyme is treated, sesame starch and cellulose can be decomposed, which facilitates preparation into a liquid medium.

In the 'diabetic food composition for improving diabetes' or the 'pharmaceutical composition for the prevention or treatment of diabetes' of the present invention, the mushroom mycelium of step (b) may be produced by culturing and then releasing the cellulolytic enzyme to the outside. The released cellulose decomposing enzyme can hydrolyze sesame seeds.

In the 'diabetic food composition for improving diabetes' or the 'pharmaceutical composition for the prevention or treatment of diabetes' of the present invention, the fibrotic enzyme of step (c) includes, for example, cellulase, hemicellulase, pectinase, glucanase, beta-glucanase, beta-glucosidase, lysozyme, viscozyme, filtrase, Rapidase and Sumizyme may be used.

In the 'diabetic food composition for improving diabetes' or 'the pharmaceutical composition for preventing or treating diabetes' of the present invention, the sesame bioconversion can improve liver damage or damage of the pancreas to Langerhans is caused by diabetes mellitus.

In the 'diabetic food composition for improving diabetes' or the 'pharmaceutical composition for the prevention or treatment of diabetes' of the present invention, the sesame bioconversion may preferably be a polysaccharide. In this case, the polysaccharide is preferably obtained by adding distilled water to the sesame bioconversion product obtained through step (c) to extract a water-soluble substance; Removing the residue through centrifugation after extracting the water-soluble substance, and separating the extracted supernatant (B); Adding EtOH to the extracted supernatant, mixing and maintaining (c); Separating the resulting precipitate after said holding (d); Adding distilled water to the separated precipitate to dissolve the precipitate, and then removing undissolved material by centrifugation or filtration; And performing dialysis against the sediment lysis solution after the centrifugation or filtration step (f).

In the pharmaceutical composition for preventing or treating diabetes according to the present invention, the pharmaceutical composition preferably contains 0.00001 to 100% by weight of a sesame bioconversion product relative to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the pharmaceutical composition for preventing or treating diabetes according to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, diluent or excipient in addition to the active ingredient. Examples of usable carriers, excipients or diluents 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, and one or more selected from among them may be used. When the pharmaceutical composition is a pharmaceutical, it may further include at least one selected from a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent or an antiseptic agent.

In the pharmaceutical composition for the prevention or treatment of diabetes according to the present invention, the formulation of the pharmaceutical composition may be in a desired form depending on the method of use, and in particular, may be formulated so as to provide rapid, sustained or delayed release of the active ingredient It is preferable to employ a method known in the art to formulate it. Examples of specific formulations include, but are not limited to, PLASTERS, GRANULES, LOTIONS, LINIMENTS, LEMONADES, AROMATIC WATERS, POWDERS, SYRUPS, OPHTHALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS, ), Suspensions, DECOCTIONS, INFUSIONS, OPHTHALMIC SOLUTIONS, TABLETS, SUPPOSITORIES, INJECTIONS, SPIRITS, CATAPLSMA, ), Capsules, CREAMS, TROCHES, TINCTURES, PASTES, PILLS, soft or hard gelatin capsules.

In the pharmaceutical composition for preventing or treating diabetes according to the present invention, the dose of the pharmaceutical composition is preferably determined in consideration of the administration method, the age, sex, weight, and severity of the disease. For example, the pharmaceutical composition of the present invention can be administered at least once per day, 0.00001 to 100 mg / kg (body weight) based on the active ingredient. However, the dosage is only an example and may be changed by a doctor's prescription depending on the condition of the recipient.

In the food composition for improving diabetes mellitus of the present invention, the food composition preferably contains 0.00001 to 100% by weight of sesame bioconversion to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the food composition for improving diabetes mellitus of the present invention, the food composition may be at least one selected from the group consisting of meat, cereal, caffeinated beverages, general beverages, chocolate, bread, snacks, confectionery, candy, pizza, jelly, Alcoholic beverages, alcoholic beverages, vitamin complexes, and other health supplement foods, but the present invention is not limited thereto.

The present invention also relates to a process for producing a liquid medium comprising (a) preparing a liquid medium containing sesame seeds, (B) preparing a fermented product by inoculating shiitake mycelia into the liquid medium after the step (a) and culturing the same; (C) a step of adding a fibrinolytic enzyme to the fermentation product after the step (b); Or a sesame bioconversion product produced from a process comprising the steps of: (a) adding a sesame bioconversion product prepared by a process comprising the steps of: At this time, the hyperlipemia may be preferably caused by 'diabetes'. In the present invention, the term 'hyperlipidemia' is meant to include hyperlipidemia caused by a rise in triglyceride levels in the body. That is, the hyperlipemia of the present invention is a concept including a disease caused by an increase in the content of triglycerides in the body, including the concept of hypertriglyceridemia.

In the 'food composition for improving hyperlipidemia' or 'pharmaceutical composition for the prevention or treatment of hyperlipidemia' according to the present invention, the sesame bioconversion product may be prepared by mixing the enzymatic fire, which is treated at 80 to 100 ° C for 0.5 to 2 hours, Activation and sterilization step (d) 'are further included. Further, after the 'enzyme inactivation and sterilization step (d)', the 'drying and pulverization step (e)' may be further included.

In the 'food composition for improving hyperlipidemia' or the 'pharmaceutical composition for the prevention or treatment of hyperlipidemia' according to the present invention, the liquid medium of step (a) is preferably at least one selected from the group consisting of amylase or cellulase And then sterilization treatment is carried out. At this time, the amylase or cellulase enzyme is preferably added in an amount of 0.1 to 0.5% (w / v) and reacted for 0.5 to 2 hours. When the amylase or cellulase enzyme is treated, sesame starch and cellulose can be decomposed, which facilitates preparation into a liquid medium.

In the 'food composition for the improvement of hyperlipemia' or 'the pharmaceutical composition for the prevention or treatment of hyperlipidemia' according to the present invention, the mushroom mycelium of step (b) may be cultured to produce a fibrolysis enzyme and be discharged to the outside. The released cellulose decomposing enzyme can hydrolyze sesame seeds.

In the 'food composition for improving hyperlipidemia' or the 'pharmaceutical composition for the prevention or treatment of hyperlipidemia' according to the present invention, the fibrotic enzymes of step (c) include, for example, cellulase, hemicellulase, pectinase, glucanase, beta-glucanase, beta-glucosidase, lysozyme, viscozyme, filtrase, Rapidase and Sumizyme may be used.

In the 'food composition for improving hyperlipidemia' or the 'pharmaceutical composition for the prevention or treatment of hyperlipidemia' according to the present invention, the sesame bioconversion inhibits the production of triglyceride and LDL-cholesterol and the production of high-density cholesterol (HDL- cholesterol. < / RTI >

In the 'food composition for improving hyperlipemia' or the 'pharmaceutical composition for the prevention or treatment of hyperlipidemia' according to the present invention, the sesame bioconversion may preferably be a polysaccharide. In this case, the polysaccharide is preferably obtained by adding distilled water to the sesame bioconversion product obtained through step (c) to extract a water-soluble substance; Removing the residue through centrifugation after extracting the water-soluble substance, and separating the extracted supernatant (B); Adding EtOH to the extracted supernatant, mixing and maintaining (c); Separating the resulting precipitate after said holding (d); Adding distilled water to the separated precipitate to dissolve the precipitate, and then removing undissolved material by centrifugation or filtration; And performing dialysis against the sediment lysis solution after the centrifugation or filtration step (f).

Meanwhile, in the pharmaceutical composition for preventing or treating hyperlipidemia of the present invention, the pharmaceutical composition preferably contains 0.00001 to 100% by weight of sesame bioconversion to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the pharmaceutical composition for preventing or treating hyperlipidemia of the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, diluent or excipient in addition to the active ingredient. Examples of usable carriers, excipients or diluents 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, and one or more selected from among them may be used. When the pharmaceutical composition is a pharmaceutical, it may further include at least one selected from a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent or an antiseptic agent.

In the pharmaceutical composition for the prevention or treatment of hyperlipidemia according to the present invention, the formulation of the pharmaceutical composition may be in a desired form depending on the method of use, and in particular, may be formulated so as to provide rapid, sustained or delayed release of the active ingredient It is preferable to employ a method known in the art to formulate it. Examples of specific formulations include, but are not limited to, PLASTERS, GRANULES, LOTIONS, LINIMENTS, LEMONADES, AROMATIC WATERS, POWDERS, SYRUPS, OPHTHALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS, ), Suspensions, DECOCTIONS, INFUSIONS, OPHTHALMIC SOLUTIONS, TABLETS, SUPPOSITORIES, INJECTIONS, SPIRITS, CATAPLSMA, ), Capsules, CREAMS, TROCHES, TINCTURES, PASTES, PILLS, soft or hard gelatin capsules.

In the pharmaceutical composition for preventing or treating hyperlipidemia of the present invention, the dose of the pharmaceutical composition is preferably determined in consideration of the administration method, the age, sex, weight, and severity of the disease. For example, the pharmaceutical composition of the present invention can be administered at least once per day, 0.00001 to 100 mg / kg (body weight) based on the active ingredient. However, the dosage is only an example and may be changed by a doctor's prescription depending on the condition of the recipient.

On the other hand, in the food composition for improving hyperlipemia of the present invention, the food composition preferably contains 0.00001 to 100% by weight of sesame bioconversion to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the food composition for improving hyperlipidemia according to the present invention, the food composition may be at least one selected from the group consisting of meat, cereal, caffeinated beverages, ordinary beverages, chocolate, bread, snacks, confectionery, candy, pizza, jelly, noodles, Alcoholic beverages, alcoholic beverages, vitamin complexes, and other health supplement foods, but the present invention is not limited thereto.

On the other hand, the present invention relates to a process for producing a liquid medium comprising sesame (a); (B) preparing a fermented product by inoculating shiitake mycelia into the liquid medium after the step (a) and culturing the same; (C) a step of adding a fibrinolytic enzyme to the fermentation product after the step (b); Or a sesame bioconversion product produced from a process comprising the steps of: (a) providing a liver-protecting composition for preventing or treating liver disease;

In the 'liver protection food composition' or the 'pharmaceutical composition for preventing or treating liver disease' of the present invention, the sesame bioconversion product may be treated with an enzyme fire which is treated at 80 to 100 ° C for 0.5 to 2 hours after step (c) Activation and sterilization step (d) 'are further included. Further, after the 'enzyme inactivation and sterilization step (d)', the 'drying and pulverization step (e)' may be further included.

In the 'liver protecting food composition' or the 'pharmaceutical composition for preventing or treating liver disease' according to the present invention, the liquid medium of step (a) is preferably at least one selected from the group consisting of amylase or cellulase And then sterilization treatment is carried out. At this time, the amylase or cellulase enzyme is preferably added in an amount of 0.1 to 0.5% (w / v) and reacted for 0.5 to 2 hours. When the amylase or cellulase enzyme is treated, sesame starch and cellulose can be decomposed, which facilitates preparation into a liquid medium.

In the 'liver protecting food composition' or the 'pharmaceutical composition for preventing or treating liver disease' of the present invention, the mushroom mycelium of step (b) may be produced through the cultivation to produce a cellulolytic enzyme and be discharged to the outside. The released cellulose decomposing enzyme can hydrolyze sesame seeds.

In the 'liver protecting food composition' or the 'pharmaceutical composition for the prevention or treatment of liver disease' of the present invention, the fibrinolytic enzyme of step (c) includes, for example, cellulase, hemicellulase, pectinase, glucanase, beta-glucanase, beta-glucosidase, lysozyme, viscozyme, filtrase, Rapidase and Sumizyme may be used.

In the 'liver protecting food composition' or the 'pharmaceutical composition for preventing or treating liver disease' of the present invention, the sesame bioconduct may preferably inhibit alcoholic fatty liver or alcoholic fatty liver hepatitis expressed by alcohol.

In the 'liver protecting food composition' or the 'pharmaceutical composition for the prevention or treatment of liver disease' of the present invention, the sesame bioconversion may preferably be a polysaccharide. In this case, the polysaccharide is preferably obtained by adding distilled water to the sesame bioconversion product obtained through step (c) to extract a water-soluble substance; Removing the residue through centrifugation after extracting the water-soluble substance, and separating the extracted supernatant (B); Adding EtOH to the extracted supernatant, mixing and maintaining (c); Separating the resulting precipitate after said holding (d); Adding distilled water to the separated precipitate to dissolve the precipitate, and then removing undissolved material by centrifugation or filtration; And performing dialysis against the sediment lysis solution after the centrifugation or filtration step (f).

In the pharmaceutical composition for preventing or treating liver diseases according to the present invention, the pharmaceutical composition preferably contains 0.00001 to 100% by weight of a sesame bioconversion product relative to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the pharmaceutical composition for preventing or treating liver disease according to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, diluent or excipient in addition to the active ingredient. Examples of usable carriers, excipients or diluents 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, and one or more selected from among them may be used. When the pharmaceutical composition is a pharmaceutical, it may further include at least one selected from a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent or an antiseptic agent.

In the pharmaceutical composition for the prevention or treatment of liver disease according to the present invention, the formulation of the pharmaceutical composition may be in a desired form depending on the method of use, and can be used to provide rapid, sustained or delayed release of the active ingredient It is preferable to employ a method known in the art. Examples of specific formulations include, but are not limited to, PLASTERS, GRANULES, LOTIONS, LINIMENTS, LEMONADES, AROMATIC WATERS, POWDERS, SYRUPS, OPHTHALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS, ), Suspensions, DECOCTIONS, INFUSIONS, OPHTHALMIC SOLUTIONS, TABLETS, SUPPOSITORIES, INJECTIONS, SPIRITS, CATAPLSMA, ), Capsules, CREAMS, TROCHES, TINCTURES, PASTES, PILLS, soft or hard gelatin capsules.

In the pharmaceutical composition for preventing or treating liver disease according to the present invention, the dose of the pharmaceutical composition is preferably determined in consideration of the administration method, the age, sex, weight, and severity of the disease. For example, the pharmaceutical composition of the present invention can be administered at least once per day, 0.00001 to 100 mg / kg (body weight) based on the active ingredient. However, the dosage is only an example and may be changed by a doctor's prescription depending on the condition of the recipient.

On the other hand, in the food composition for liver protection of the present invention, the food composition preferably contains 0.00001 to 100% by weight of sesame bioconversion to the whole composition. When the content is less than 0.00001% by weight, the effect is insignificant.

In the food composition for liver protection of the present invention, the food composition may be, for example, meat, cereal, caffeinated beverages, general beverages, chocolate, bread, snacks, confectionery, candy, pizza, jelly, noodles, , An alcoholic beverage, a drink, a vitamin complex, and other health supplement foods, but the present invention is not limited thereto.

The sesame bioconversion product of the present invention can increase immunity against microbial infection through the activation of macrophages, improve diabetes, improve liver damage or damage of the pancreas to Langerhans islet, and inhibit the production of triglycerides I will exert. It also has the effect of inhibiting alcoholic fatty liver or alcoholic fatty liver disease expressed by alcohol.

BRIEF DESCRIPTION OF THE DRAWINGS FIG.
Fig. 2 is a process chart of the post-treatment and recovery operation of the sesame biological conversion product of the present invention.
FIG. 3 is a graph showing the results of NO production of macrophages which are remarkably improved by the sesame bioconversion of the present invention and which are increased in NO production ability and concentration-dependent manner.
4 is a graph showing the cytokine expression potential of sesame and the sesame bioconversion of the present invention.
5 is a graph showing the antimicrobial activity of Salmonella in the sesame bioconversion product of the present invention.
FIG. 6 is a graph showing the effect of the sesame bioconversion of the present invention on the salmonellosis of macrophages by concentration.
7 is a graph showing the effect of the sesame bioconversion of the present invention on inhibition of salmonellosis in macrophages by concentration.
8 is a graph showing the delayed effect of the sesame biotransformation according to the present invention in the mortality rate caused by Salmonella infection of a mouse.
FIG. 9 is a graph showing the inhibitory effect of the sesame biotransformation of the present invention on intraabdominal salmonella.
10 is a graph showing the penetration inhibiting activity of the sesame bioconversion product of the present invention in intestinal intestinal permeation of Salmonella using Caco-2 cell line.
11 is a graph showing the effect of the sesame bioconversion of the present invention on the death of INS-1 cell line induced by alloxan and production of reactive oxygen species (ROS).
Fig. 12 (A) is a graph showing the liver damage protection effect of the sesame bioconversion product of the present invention in a type 1 diabetes mouse induced by alloxan.
Fig. 12 (B) is a photograph of the effect of protecting the liver damage of the sesame biotransformation of the present invention in a type 1 diabetes mouse induced by alloxan.
Fig. 13 is a photograph of the protective effect of the sesame biotransformation of the present invention on pancreatic damage in a type 1 diabetes mouse induced by alloxan.
14 is a graph showing the effect of the sesame biotransformation of the present invention in a type 2 diabetic mouse.
Fig. 15 (A) is a photograph showing the protective effect of the liver tissue of the sesame biotransformation of the present invention in a type 2 diabetic mouse. Fig.
Fig. 15 (B) is a photograph of the protective effect of pancreatic tissue of the sesame biotransformation of the present invention in a type 2 diabetic mouse.
16 is a graph showing an inhibitory effect of the sesame bioconversion of the present invention on alcohol-induced hepatocyte death.
FIG. 17 is a graph showing the inhibitory effect of the sesame bioconversion of the present invention on the generation of ROS in alcohol-induced hepatocyte death. FIG.
18 is a graph showing an effect of inhibiting liver weight increase due to the administration of the sesame bioconversion of the present invention in an alcohol-induced liver injury mouse.
FIG. 19 is a photograph showing the protective effect of liver damage caused by the administration of the sesame biotransformation of the present invention in an alcohol-induced liver damaged mouse.
20 shows a production process chart and a manufacturing process for obtaining a polysaccharide fraction from the sesame bioconversion product of the present invention.
FIG. 21 shows chromatogram results of HPLC analysis of polysaccharide fractions of the sesame bioconverted product according to the present invention.
22 shows the results of evaluating the macrophage activation ability of the polysaccharide fractions of the sesame biotransformation of the present invention.
FIG. 23 shows the results of investigation of cytokine secretion patterns of the sesame bioconversion product and the sesame bioconversion polysaccharide fraction of the present invention.
24 shows the result of inhibition of the secretion amount of cytokine of the sesame bioconversion and sesame bioconversion polysaccharide fractions of the present invention at the time of TLR4 inhibitor treatment.

In the present invention, sesame (Sesamum indicum ), comprising the steps of: (a) preparing a liquid medium; (B) a step (b) of adding a fibrinolytic enzyme to the culture after the step (a), and then adding a fibrinolytic enzyme to the culture after the step (c); A sesame bioconversion product prepared from a process comprising the steps of:

According to the experiment of the present invention as described below, it was confirmed that the sesame bioconversion obtained from the process of the present invention can increase the immunity against microbial infection through activation of macrophages. In addition, it has been confirmed that diabetes can be improved, hepatic damage or damage of the pancreas to Langerhans islet can be prevented, and the production of triglyceride is inhibited, thereby improving or preventing (or treating) hyperlipemia. In addition, it has been confirmed that the effect of protecting the liver such as alcoholic fatty liver or alcoholic fatty liver disease expressed by alcohol or preventing (or treating) liver disease can be exerted.

Meanwhile, in the present invention, a step (a) of adding distilled water to the sesame biological conversion product obtained through the step (c) to extract a water-soluble substance; Removing the residue through centrifugation after extracting the water-soluble substance, and separating the extracted supernatant (B); Adding EtOH to the extracted supernatant, mixing and maintaining (c); Separating the resulting precipitate after said holding (d); Adding distilled water to the separated precipitate to dissolve the precipitate, and then removing undissolved material by centrifugation or filtration; And performing dialysis against the sediment lysis solution after the centrifugation or filtration. The polysaccharide fraction obtained by the process further comprises:

In the present invention, sesame bioconversion and polysaccharide fractions isolated therefrom are recognized in a pattern recognition receptor, which is a specific receptor of immune cells, to confirm that signal transduction is mediated after the polysaccharide fraction is obtained from the sesame bioconversion.

As a result of the following experiments, it was confirmed that the TNF-α, IL-6, IL-10 and IFN-β were expressed in the sesame bioconversion and the polysaccharide fractions thereof. The activity was confirmed to be due to the polysaccharide fraction isolated therefrom.

Meanwhile, in the following experiment of the present invention, it was confirmed that the polysaccharide fraction isolated from the sesame biotransformant was similar to that of LPS (lipopolysaccharide) and the cytokine secretion pattern. Thus, it was confirmed that the polysaccharide fraction of the present invention is an antioxidant Which is an antagonist.

Meanwhile, TAK-242, which is a specific inhibitor for TLR4, was used to confirm whether the sesame bioconversion product and its polysaccharide fraction of the present invention were specifically recognized and mediated by TLR4. As a result of the following experiment, it was confirmed that the reactions represented by the treatment of the sesame bioconvert or its polysaccharide fraction were all inhibited by the TLR4-specific inhibitor TAK242. From this, TLR4 is a receptor for the sesame bioconversion and its polysaccharide fraction I could confirm.

From these results, it was confirmed that the sesame bioconversion product and its polysaccharide fraction produced by the bioconversion process pursued in the present invention have TLR4 agonist activity.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[Example 1: Preparation of Sesame Bioconversion]

 (1) Pretreatment process (foreign body and mold washing process)

To produce a sesame bioconversion, sesame ( Sesamum indicum ) was cleaned to remove contaminants such as foreign matter and fungi.

First, high-pressure air guns were used to remove foreign matter and dust from the surface of sesame seeds. Secondly, the sieve was sieved to remove other foreign matter. Thirdly, water or ethanol was treated to remove contaminants such as mold spores.

The sesame which had been subjected to the third washing process was used as a raw material after performing the microbial contamination test.

 (2) Enzymatic treatment with hydrolytic enzymes for culture

After the sesame seeds were removed from the soil and fungal contamination, the enzymatic treatment was carried out using various hydrolytic enzymes. The hydrolytic enzymes were tested using enzymes of amylase family and cellulase family which can decompose starch and cellulose of sesame seeds.

Each enzyme was treated with 0.3% (w / v), reacted for 1 hour at the temperature showing optimal activity, and sterilized to prepare a liquid medium.

 (3) Fermentation and enzyme treatment Bioconversion process

Sesame seedlings were inoculated on a liquid culture medium and cultured at 30 ° C. At the time when the residual carbon source was depleted, the cells were cultured for 8 days in a liquid phase process in which a concentrated liquid medium was added. It is possible to hydrolyse sesame seeds by cultivating ovarian mycelium by producing fibrotic enzymes. After cultivation, an enzyme treatment process, which is a secondary bioconversion process, was further added to the fermented sesame seeds produced by the fermentation process.

The above-mentioned fermentation and enzymatic treatment bioconversion process is intended to effectively produce an immunopotentiating substance derived from a cell wall from a fermentation product, a cultured fermentation product, together with sesame, which is a fermentation medium for a culture fermentation. , Which is an effective method for extracting and efficiently extracting an immunologically active substance.

Accordingly, the present inventors have found that a cell wall lytic enzyme complex (cellulase, hemi-cellulase, pectinase, and beta-glucanase, which is a cell wall hydrolase capable of degrading a cell wall, The enzyme / substrate reaction was optimized by shaking at 250 rpm at 50 to 60 ° C using an enzyme conjugate of β-glucanase (see FIG. 1). BRIEF DESCRIPTION OF THE DRAWINGS FIG.

 (4) Post-treatment process

The sesame bioconversion product produced by the secondary biotransformation process (enzyme reaction) was subjected to 'enzyme inactivation and sterilization' process at 90 ° C for 1 hour, followed by drying and pulverization, and used in the following experimental examples 2). FIG. 2 is a process chart of a post-treatment and recovery operation of a sesame bioconversion product.

[ Experimental Example 1: Measurement of in vitro NO production ability in macrophages ( macrophages ) by sesame and sesame biotransformation ]

In order to confirm the macrophage activation effect of sesame and sesame bioconversion, the NO production ability in macrophage (macrophage), RAW264.7 cell line was examined.

100 μl of a sesame biotransformant and a serially diluted sesame and sesame biotransformant and a standard were dispensed into a 96-well plate, and a 'Griess reagent' (① N- (1-naphtyl) ethlene diamine dihydrochloride: 0.5 g / 500 ml ② Sulfanilamide: 5 g / 85% H ₃ PO ₄ 29.5 ml / 470.5 ml) was added and reacted for 1 minute. Then, the immunoreactivity of the sample was measured by measuring the abundance of macrophage NO by measuring the absorbance at 540 nm (or 550 nm) using an ELISA leader.

The results showed that the sesame bioconversion showed NO production ability of macrophages at a concentration lower than 1 / 1,000 ~ 1 / 10,000 compared with sesame, and the NO production ability of macrophages by sesame bioconductors was remarkably improved, (Fig. 3). FIG. 3 is a graph showing the NO production ability and the concentration-dependent increasing NO production of macrophages remarkably improved by the sesame bioconversion.

[ Experimental Example 2: Analysis of cytokine secretion ability of sesame and sesame biotransformants in macrophages ]

In order to confirm the effect of sesame and the sesame bioconversion of the present invention on the innate immune response activation activity, cytokine secretion ability in macrophages (macrophages), RAW264.7 cell lines was examined.

Sesame and the sesame bioconversion of the present invention were treated at a concentration of 1, 10, and 100 / / ml, respectively. After 16 hours of sample treatment, the supernatant of the culture was taken and TNF-α, IL-1β, IL-4, IL-5, (Interleukin-6), IL-10 (Interlukine-10), IL-12p70 (Interlukine-12p70), IFN- Respectively.

As a result of the experiment, it was confirmed that TNF-α, IL-6, IL-10 and IFN-β were expressed when the sesame bioconversion was treated (FIG. 4 is a graph showing the cytokine expression potential of sesame and sesame biotransformants.

TNF-α is secreted from macrophages and mononuclear cells during phagocytosis, and is known to have an antiviral effect against various viruses such as influenza virus along with 'inducible nitrogen monoxide (NO)'.

IL-6 is a major mediator in antigen-specific immune responses, inflammatory responses, and acute responses, and is a representative cytokine that plays a key role in the in vivo defense system.

IFN-β is a type I interferon with IFN-α, and has antiviral and antiviral effects. It is a typical cytokine that induces Th1 immune response by contributing to Th1 cell differentiation, inducing Th1 immune response, while Th2 And Th17 immune response.

The culture supernatant was taken every 6 hours, 12 hours, and 18 hours after the sample treatment time, and the cytokine secretion was examined. The experiment was repeated three times.

Therefore, the sesame bioconversion product of the present invention was found to be capable of specifically activating the Th1 immune response, and at the same time, could suppress the exaggerated Th2 immune response. It is also expected that it may be effective in diseases where it is necessary to improve the Th1 immune response or that the Th2 immune response is exaggerated.

[ Experimental Example 3: Assessment of Inhibitory Effect of Sesame Bioconversion on Salmonella In Vitro in Macrophages ]

In this experiment, we investigated the effect of sesame bioconversion on the phagocytosis and inhibition of intracellular microbial survival by inducing macrophage activation in an in vitro experimental system using macrophages.

 (1) Antimicrobial activity measurement of sesame bioconversion product

In order to evaluate the ability of sesame bioconversion to inhibit Salmonella infection, we tried to determine whether sesame bioconversion has direct antimicrobial activity.

Salmonella typhimurium ATCC14028 strain was used as the indicator strain and nutrient broth (NBGP), which is a general salmonella culture medium, and phenol red and glucose for fermentation, was used as an indicator strain. The growth of salmonella and color change due to sugar fermentation were measured.

When the salmonella was cultured, the sesame bioconversion was treated at 1, 10, and 100 ㎍ / mL, respectively, to induce the survival inhibition of Salmonella, and the absorbance was measured at 0, 2, 4, Respectively.

As a result of the experiment, it was confirmed that the sesame bioconversion does not show direct antibacterial activity (FIG. 5). 5 is a graph showing the antimicrobial activity of salmonella in the sesame bioconversion product.

 (2) Measuring Macrophage Phagocytosis of Sesame Bioconversion

The RAW 264.7 cells were treated with sesame bioconverters at a concentration of 1, 10, and 100 ㎍ / mL for 4 hours, followed by Salmonella typhimurium ATCC14028 was infected for 30 minutes and 60 minutes.

As a result of the experiment, it was confirmed that the increase of the spermatogenesis caused by the treatment of the sesame biotransformation was increased to 4.24 times at the time of infection with Salmonella 30 minutes, and it increased up to 9.7 times at 60 minutes of Salmonella infection (FIG. 6). FIG. 6 is a graph showing the effect of the sesame bioconversion per concentration on the salmonellosis of macrophages.

 (3) Measurement of inhibition of salmonella proliferation of sesame bioconversion

The RAW 264.7 macrophage cell line was treated with sesame bioconducts at a concentration of 1, 10, and 100 μg / mL for 4 hours, followed by salmonella typhimurium ATCC14028) was infected for 2 hours, 4 hours, and 8 hours, and then an increase in Salmonella was observed.

As a result of the experiment, the increase of intracellular salmonella occurred for 2 hours, and then decreased. In the group treated with sesame biotransformation of 100 / / mL and infected with Salmonella, it was confirmed that salmonella present in cells significantly decreased after a maximum of 8 hours (FIG. 7). 7 is a graph showing the effect of the sesame bioconversion per concentration on the inhibition of salmonellosis in macrophages.

[ Experimental Example 4: Evaluation of modulating efficacy against mouse mortality caused by Salmonella infection ]

Through the above experiments, it was confirmed that the sesame bioconverted product induces macrophage activation in macrophage in vitro using macrophages to increase phagocytosis and suppress the survival of intracellular microorganisms after phagocytosis, ' Salmonella typhimurium ATCC14028)' strains were used to test the in vivo antimicrobial activity of sesame biotransformants.

In order to determine if the sesame bioconversion could reduce the mortality rate of mice due to Salmonella infection, 10 ⁵ CFU, the lethal concentration of Salmonella typhimurium ATCC14028 strain, was injected intraperitoneally to induce the death of mice by infection.

As a result of the experiment, when all mice were killed at 7 days after the infection, when the sesame bioconversion was administered at a dose of 10 mg / kg, the mouse survived up to 19 days, (Fig. 8). FIG. 8 is a graph showing the delayed effect of the sesame biotransformation on mortality caused by Salmonella infection in mice.

Experimental Example 5: Sesame bioconversion of water, in vivo live in Salmonella infection mice infected with Monel called inhibitory ability evaluation;

As a result of the above experiment, it was confirmed that the administration of the sesame bioconverting agent reduced the mortality rate of the mice due to the Salmonella infection at the lethal concentration. Thus, in this Example, the in vivo antimicrobial activity and the sesame bioconversion Induced activation of the immune response.

The sesame biotransformation was fed at a dose of 10 mg / kg for 2 weeks, and then ' Salmonella typhimurium ATCC14028 'strain was intraperitoneally injected at a concentration of ¹ × 10 ⁴ CFU to induce intraperitoneal Salmonella infection.

Two days after infection, the salmonella in the abdominal cavity of the mice was recovered using PBS, and the number of Salmonella surviving in the abdominal cavity was measured by spreading on an LB plate.

As a result of the measurement, it was confirmed that surviving salmonella was significantly reduced in the group treated with sesame bioconversion compared to the control group (Fig. 9). 9 is a graph showing a result of suppressing the salmonella in the abdominal cavity of the sesame bioconversion product.

[ Experimental Example 6: Induction of lymphocyte proliferation and evaluation of interferon-gamma production activation in Salmonella-infected mice ]

After the intraperitoneal salmonellar experiment, the spleen of the mouse was excised and splenic lymphocytes were isolated. The isolated lymphocytes were stimulated with concanavalin A, which is a T cell proliferation inducing substance, and LPS, which is a B cell proliferation inducing substance, to determine how cell proliferation is induced. The results are shown in Table 1 below.

Sample Lymphocyte proliferation (fold-increase) IFN-y (pg / mL) vehicle 1.38 + - 0.12 287.204 ± 13.586 10 mg / kg sesame bioconversion 1.79 + 0.13 341.094 + 24.265

Experimental results showed that mouse spleen lymphocytes were more increased in mice treated with sesame bioconversion than in the control group. In addition, it was confirmed that the expression level of interferon-gamma (IFN-y), a representative Th1 cytokine, was increased.

Test Example 7: Evaluation Th1 cytokine (cytokine) activation efficacy in mice infected with Salmonella;

In order to confirm whether the sesame bioconversion induces the activation of the Th1 immune response, the amount of Th1 cytokine expressed in the spleen of the mouse infected with salmonella at the above-mentioned concentration of saliva was measured to determine the Th1 activation effect Respectively. Four cytokines, IL-1β, IL-2, IL-6 and IL-12, were measured in the Th1 cytokine using ELISA.

The effect of the sesame bioconversion was compared between the mice not infected with Salmonella and the mice infected with Salmonella. The results are shown in Table 2 below.

sample cytokines (pg / mL) IL-1? IL-2 IL-6 IL-12 salmonella (-) vehicle 171 ± 12 41.7 ± 3.7 57.3 ± 3.2 240 ± 13 10 mg / kg
Sesame Bioconversion 184 ± 11 41.9 ± 2.9 58.3 ± 4.7 247 ± 13 salmonella (+) vehicle 239 ± 13 55.7 ± 3.8 73.4 ± 5.4 284 ± 13 10 mg / kg
Sesame Bioconversion 254 ± 20 61.5 ± 4.5 76.8 ± 6.0 324 ± 31

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As a result, when the sesame bioconversion was administered to normal mice, there was no significant difference in the amount of cytokine expression except that the expression level of IL-1β was slightly increased. On the other hand, when infected with Salmonella, the amount of cytokines such as IL-1β, IL-2, IL-6 and IL-12 is increased by Salmonella infection and the sesame bioconversion In the case of simultaneous administration, it was confirmed that more cytokine production was induced. In particular, IL-12, which is a representative Th1 cytokine, showed the most remarkable increase in the expression level.

These results show that the administration of the sesame bioconversion does not induce unnecessary activation of the immune response in a normal organism, and induces the activation of the Th1 immune response upon microbial infection, thereby effectively controlling the infected microorganism in vivo.

Test Example 8: Th2 cytokine (cytokine) expression efficacy evaluation in Salmonella infected mice;

As a result of evaluating the cytotoxic activity of Th1 cytokine, it was confirmed that the administration of sesame bioconversion activates the expression of Th1 cytokine, thereby activating Th2 immune response, which is an antagonistic reaction of Th1 immunoreactivity To investigate the expression level of Th2 cytokine,

Three cytokines, IL-4, IL-5 and IL-10, which are representative Th2 cytokines, were detected. Similar to Th1 cytokines, they were examined in both normal and Salmonella-infected mice not infected with Salmonella. The results are shown in Table 3 below.

sample cytokines (pg / mL) IL-4 IL-5 IL-10 salmonella (-) vehicle 52.1 ± 3.4 294 ± 17 44.7 ± 2.3 10 mg / kg
Sesame Bioconversion 51.0 ± 3.9 292 ± 20 44.1 ± 3.2 salmonella (+) vehicle 56.8 ± 4.3 289 ± 17 48.7 ± 3.3 10 mg / kg
Sesame Bioconversion 57.3 ± 3.5 290 ± 21 48.9 ± 3.5

As a result of the experiment, it was confirmed that administration of the sesame bioconversion does not increase the production of Th2 cytokine in both the normal mouse not infected with Salmonella and the infected mouse

These results show that the administration of the sesame bioconversion does not induce activation of unnecessary immune responses in normal organisms and does not induce activation of Th2 immune response upon microbial infection.

Experimental Example 9: Sesame bioconversion of water, in the intestinal cells Evaluation of inhibition of penetration of Salmonella in vitro ]

In this experiment, we evaluated the effect of sesame bioconversion on the intestinal cell adhesion and penetration inhibition of Salmonella using Caco-2 cell line. Since most of the Salmonella infections occur through intestinal infections, we wanted to see if the sesame bioconversion could inhibit salmonella intestinal cell adhesion and invasion. Who was evaluated in colon cancer derived cell line, Caco-2 cell line and in vitro experimental system using the Salmonella typhimurium SL1344 strain.

Caco-2 cell line was treated with 100 ㎍ / mL sesame bioconversion for 4 hours or with Salmonella typhimurium SL1344, and the number of salmonella infiltrated into cells was measured.

As a result of the experiment, it showed 11.4% inhibition of salmonella penetration in the case of pretreatment for 4 hours, and 54.5% inhibition of salmonella penetration in simultaneous treatment. Since the sesame bioconversion was confirmed to have no antimicrobial activity in the above experiment, it was judged that it acted directly on the Caco-2 cell line and inhibited microbial penetration (FIG. 10). 10 is a graph showing inhibitory activity of intestinal intracellular penetration of sesame bioconversion using Caco-2 cell line.

[ Experimental Example 10: Effect of salmonella biotransformation in vivo inhibition of Salmonella penetration]

In this experimental example, the ability to inhibit Salmonella permeation in vivo of sesame bioconversion was investigated.

 (1) Identification of salmonella excreted in feces

As a result of the experiment using the cell line, it was confirmed that the sesame bioconversion inhibits salmonella intestinal cell penetration, so that the salmonella infestation inhibitory effect was evaluated in a salmonella food poisoning mouse model.

Salmonella typhimurium SL1344 strain was orally administered with 10 ⁸ CFU of the sesame bioconversion at a dose of 10 mg / kg for 2 weeks, respectively, to induce Salmonella infection. The feces were sampled 24 hours and 48 hours after infection, and the viable cell counts of Salmonella in feces were measured. The results are shown in Table 4 below.

Sample Salmonella in feces (x10 ⁴ CFU / g) 1 day 2 day normal mice 0.00 ± 0.00 0.00 ± 0.00 SL1344 infection only 338 ± 18 420 ± 31 SL1344 infection +
10 mg / kg sesame bioconversion 609 ± 37 667 ± 58

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As a result of the experiment, it was confirmed that a large amount of salmonella did not penetrate into the intestinal tract and excreted into the feces when the sesame bioconversion was administered.

 (2) Identification of Salmonella infiltrated into tissues

From the results of Table 4, it was confirmed that when the sesame biotransformant was administered, Salmonella entered into the organism could not permeate into the tissues and was discharged to the outside of the body. Therefore, the fecal release experiment mice were administered to the cecum and intestinal membrane lymph nodes, spleen, liver And the number of Salmonella infiltrated into the tissues was measured.

① Cecum and intestinal membrane lymph nodes

After the infection of salmonella, salmonella in the intestinal membrane lymph node, the cecum, which is the primary infiltrating tissue, and the lymphatic tissue connected with it, were measured to confirm the tissue infiltration inhibition effect of sesame bioconversion. The results are shown in Table 5 below.

Sample Salmonella in tissue Cecum (x10 ⁵ CFU / g) Mesenteric lymph node (x10 ³ CFU / organ) normal mice 0.00 ± 0.00 0.00 ± 0.00 SL1344 infection only 489 ± 32 109 ± 8 SL1344 infection +
10 mg / kg sesame bioconversion 249 ± 21 37.7 ± 2.3

The results showed that the sesame bioconversion inhibited the penetration of Salmonella into the cecum, and the sesame bioconversion showed a suppressive effect of 49.1%. The sesame bioconversion was found to inhibit Salmonella in 65.4% of intestinal membrane lymph nodes, and it was found that the inhibition rate in the intestinal membrane lymph nodes was greater than in the cecum.

These results suggest that the immune cells in the intestinal membrane lymph node were activated by the sesame biotransformant and could be effectively removed even if Salmonella was infiltrated.

② spleen and liver

The amount of Salmonella in the spleen and liver which can penetrate into tissues and move through body fluids was measured.

Sample Salmonella in tissue Spleen (x10 ² CFU / organ) Liver (x10 ² CFU / organ) normal mice 0.00 ± 0.00 0.00 ± 0.00 SL1344 infection only 129 ± 11 77.4 ± 5.6 SL1344 infection +
10 mg / kg sesame bioconversion 26.8 ± 2.3 23.7 ± 2.0

The results showed that 79.2% of salmonella in the spleen was inhibited by the sesame bioconversion. Also, sesame bioconversion against Salmonella present in the liver showed a inhibition rate of 69.4%. Both the spleen and liver showed a higher inhibition rate than the cecum, and these results were also attributed to the activation of various immune cells.

[ Experimental Example 11: Preparation of Sesame Bioconversion in Beta Cells In vitro inhibition of type 1 diabetes evaluation]

In order to confirm that the sesame bioconversion has an inhibitory activity against type 1 diabetes, the activity of the sesame bioconversion was evaluated in an in vitro system using the INS-1 cell line, a mouse-derived pancreatic beta cell line.

 (1) Effect on beta-cell cytotoxicity and reactive oxygen production

Pancreatic beta cells and beta-cell specific reactive oxygen species (ROS) through INS-1 cell line through 'Glucose transporter 2 (GLUT2)', and treatment with alloxan, And that the sesame bioconversion can be treated to inhibit cell death.

The INS-1 cell line was dispensed into a 96-well plate at a concentration of ⁵ × 10 ⁵ cells / well, treated with 10 mM alloxan and 1, 10, 100 μg / mL sesame bioconversion And cultured for 48 hours. After incubation, cell viability was measured by MTT assay.

As a result of confirming the ability of the sesame bioconversion inhibitor to inhibit beta cell death and inhibit ROS formation, it was found to inhibit cell death and ROS production by alloxan in a concentration dependent manner. At a concentration of 100 ㎍ / mL, 39.7% inhibition of cell death and 60.3% inhibition of ROS formation (Fig. 11). 11 is a graph showing the effect of sesame bioconversion on the death of INS-1 cell line induced by alloxan and production of reactive oxygen species (ROS).

 (2) Effect on beta -cellular NO production and insulin secretion

Increased ROS and cell death in the beta cell, INS-1 cell line by treatment with alloxan increase the amount of nitric oxide (NO) produced and secreted by the cells and decrease the secretion of insulin do.

In this experiment, the amount of nitric oxide (NO) secreted by the INS-1 cell line and the amount of insulin secreted by the INS-1 cell line decreased as the amount of intracellular ROS increased and the cell death rate increased by the simultaneous treatment of the sesame biotransformation by alloxan treatment The amount was also measured. The results are shown in Table 7 below.

Sample Nitric oxide
(arbitrary fluorescent unit) Insulin release (ng / mL) vehicle 19.784 ± 1.253 1.809 ± 0.057 10 mM Alloxan only 82.479 ± 5.261 0.821 + 0.034 10 mM Alloxan +
Sesame
Bioconversion 1 / / mL 70.736 ± 6.034 0.911 + 0.078 10 [mu] g / mL 48.309 ± 2.242 1.154 + 0.105 100 / / mL 25.384 ± 1.931 1.351 + - 0.107

Experimental results showed that the secretion of NO increased by the treatment of alloxan was inhibited by the concurrent treatment of sesame bioconversion in a concentration dependent manner and that the secretion of insulin suppressed by treatment with alloxan was reduced by sesame biotransformation Water, and it was confirmed that the sesame bioconversion inhibited NO production and secretion by 69.2% at the maximum concentration of 100 ㎍ / mL.

In the case of insulin, it was confirmed that the secretion amount was recovered up to 64.6% at the maximum concentration of 100 ㎍ / mL.

[ Experimental Example 12: Effect of sesame biotransformation on type 1 diabetes induced mice in vivo Type 1 diabetes inhibiting ability evaluation]

As a result of confirming the antidiabetic effect of the sesame bioconversion on the type 1 diabetes in the in vitro experimental system, the antidiabetic effect of the sesame bioconversion in the mouse model of type 1 diabetes was examined in this experimental example .

(1) The effect of the type 1 diabetic mouse model on blood glucose, serum insulin, and hepatic glycogen level

The diabetic rats were fed diets containing 10 mg / kg of sesame bioconversion for two weeks followed by 100 mg / kg of alloxan administered intraperitoneally. After one week of diabetes induction, blood glucose level, blood insulin concentration, and hepatic glycogen concentration were measured as an indicator of diabetes, and the results are shown in Table 8 below.

Sample Blood glucose (mg / dL) Serum insulin
(ng / mg protein) Glycogen
(mg / g liver wt.) vehicle 127.0 ± 8.1 59.204 + - 4.283 3.041 + - 0.213 Alloxan only (100 mg / kg) 308.4 ± 15.7 9.407 ± 0.821 2.427 ± 0.154 Alloxan +
10 mg / kg sesame bioconversion 210.5 ± 10.6 39.845 +/- 2.450 2.883 + - 0.207

As a result, the blood glucose level was increased to 308.4 mg / dL in the control group in which the diabetes was induced, but it was confirmed that the blood sugar level was 210 mg / dL and the increase in the blood glucose level was suppressed in the mouse treated with sesame bioconversion.

In addition, blood insulin concentration was decreased to 15.9% compared with that of normal mice when diabetic induction was performed, and it was confirmed that 67.3% of sesame bioconversion was recovered. The glycogen level in the liver, which is decreased due to the decrease of blood insulin, was also recovered to 94.8% when the sesame bioconversion was administered, while the diabetic control group was reduced to 79.8%.

 (2) Evaluation of the ability to regulate glucose metabolism enzymes

To investigate the effect of sesame bioconversion on the activities of glucose-6-phosphatase (G6pase), phosphoenolpyruvate carboxykinase (PEPCK) and glucokinase (GCK) .

When the insulin concentration in the blood is reduced by diabetes, G6pase and PEPCK are activated to promote glucose synthesis, and inhibition of glycogen synthesis is inhibited by inhibition of GCK. Therefore, it was confirmed that the activity of these three enzymes can be controlled as the insulin secretion is restored when the sesame bioconversion is administered. The results are shown in Table 9 below.

Sample enzyme activity (nmol / min / mg protein) G6pase PEPCK GCK Vehicle 71.422 + - 4.231 17.540 ± 1.161 10.840 ± 0.893 Alloxan only (100 mg / kg) 172.559 ± 12.549 33.426 + 2.076 5.224 + 0.335 Alloxan +
10 mg / kg sesame bioconversion 93.043 + - 5.637 23.471 ± 1.052 9.274 ± 0.569

As a result, the activity of G6pase was 2.4 times higher than that of the normal mouse in the case of diabetic induction. In the case of PEPCK, the activity was increased about 1.9 times in the diabetic mouse, but it was found to be about 1.3 times in the case of the sesame bioconversion. In the case of GCK, the activity was decreased to 48.2% of normal mice when diabetic induced, but it was confirmed that the activity increased to 85.6% of normal mice when the sesame bioconversion was administered.

 (3) Protection effect of liver tissue and pancreatic tissue

GOT / GPT levels were measured and tissue staining was performed to determine if liver damage induced by type 1 diabetes could be inhibited by sesame bioconversion.

In the case of diabetic induction, GOT was increased 1.8 times and GPT was increased 3.1 times, but when the sesame bioconversion was administered, the increase was 1.3 times and 1.5 times, respectively, to effectively suppress the liver damage caused by diabetes (Fig. 12 (A)). Fig. 12 (A) is a graph showing the liver damage protection effect of a sesame bioconversion in a type 1 diabetes mouse induced by alloxan.

In addition, liver tissue was extracted, and paraffin blocks were stained with 'hematoxylene' and 'eosin Y' to examine the extent of damage to the tissues. As a result, it was shown that when alloxan induced diabetes, , It was confirmed that the degree of liver damage was alleviated upon administration of the sesame bioconversion (Fig. 12 (B)). Fig. 12 (B) is a photograph of the liver damage protection effect of the sesame bioconversion in a type 1 diabetes mouse induced by alloxan.

As shown in FIG. 13, when the pancreas tissue was stained in the same manner, the size of the Langerhans islet in the pancreatic tissue of the mouse induced diabetes was reduced due to the breakdown of the? -Cell and the result was inadequate, In mice treated with the diets, it was found that the tissue damage of the pancreatic islets was alleviated. FIG. 13 is a photograph showing the protective effect of the sesame biotransformation on pancreatic damage.

[ Experimental Example 13: Effect of sesame biotransformation on type 2 diabetes induced mice in vivo Type 2 diabetes inhibiting ability evaluation]

In this experiment, we aimed to evaluate the antidiabetic effect of sesame bioconversion in a type 2 diabetic mouse model induced by obesity induced by high fat diet.

(1) The effect of sesame bioconversion on the weight, liver weight, and white fat weight changes in the type 2 diabetic mouse model

The high fat diet for diabetes induction was manufactured by Dyet Company and the composition of the diet is shown in Table 10 below.

component normal diets (g / kg diet) high-fat diets (g / kg diet) casein 200.0 200.0 DL-methionine 3.0 3.0 corn starch 150.0 150.0 sucrose 500.0 150.0 cellulose 50.0 50.0 corn oil 50.0 salt mix # 200000 ^a 35.0 35.0 vitamin mix # 310025 ^b 10.0 10.0 choline bitartrate 2.0 2.0 beef tallow 400.0

(* ^A Dyets cat No. 200000, ^b Dyets cat.

On the other hand, the sesame bioconversion was dietary supplemented with 10 mg / kg of high - fat diets and fed for 7 weeks to induce type 2 diabetes. After 7 weeks, mouse weights, liver weights, and white fat weights were measured, and the results are shown in Table 11 below.

Sample initial weights (g) final weights (g) liver weights (g) white adipose tissue weights (g) feed intakes (g / day) normal control 23.15 ± 1.75 31.16 + - 2.24 1.79 ± 0.12 0.41 + 0.03 3.44 ± 0.22 HFD-control 23.54 + 1.43 35.58 ± 2.18 2.63 ± 0.16 1.77 + - 0.12 3.01 ± 0.19 HFD-10 mg / kg
Sesame Bioconversion 24.03 + - 2.02 33.12 ± 2.10 2.02 ± 0.08 0.89 + 0.07 3.06 ± 0.23

As a result, weight gain of 12.04 g in the type II diabetic mouse was observed in normal mice compared with 8.01 g in the weight gain at 7 weeks.

When the high fat diet was administered to the mice with the sesame bioconversion, the body weight gain was 9.09 g, indicating a decrease in body weight gain compared to the control group. Liver weight and white fat weight were also inhibited compared to diabetic control group. However, since the difference in the average food intake was not observed, it was confirmed that this effect was not due to unhealthy food.

 (2) Measurement of effect on blood glucose and insulin in serum

The effect of the sesame bioconversion on the blood glucose level and blood insulin concentration in the type 2 diabetic mouse model was measured and the results are shown in Table 12 below.

Sample Blood glucose (mg / dL) Serum insulin (ng / mL) normal control 113.5 ± 9.7 0.805 ± 0.053 HFD-control 216.7 ± 10.8 0.412 + 0.027 HFD-10 mg / kg sesame bioconversion 143.8 ± 10.5 0.556 + 0.041

As a result of blood glucose measurement, blood glucose level was increased about 1.9 times in the high fat diet mouse, but the blood glucose level was inhibited about 1.3 times in the high fat diet when the sesame bioconversion was administered to the mouse.

In the case of insulin concentration in blood, it was confirmed that the high-fat diet decreased to 51.1% of the normal level in the mouse and the type 2 diabetes was induced, and the high fat diet recovered to 69.1% .

(3) Effect on glucose tolerance

Oral glucose tolerance test was performed to measure blood glucose control disorder by type 2 diabetes. Seven-week high-fat diet-fed mice were fasted for 16 hours, and glucose was orally administered. Blood was collected from mouse tail vein at intervals of 30 minutes and blood glucose level was measured.

As a result, in the case of normal mice, the blood glucose level increased until 30 minutes after the administration of glucose, and then decreased after 120 minutes. However, in the case of mice in which type 2 diabetes was induced, It was confirmed that the descent effect was greatly reduced.

It was confirmed that the high-fat diet reduced the overall blood glucose increase when the sesame biotransformation was administered to the mice, and it was confirmed that the blood glucose increase inhibition and control ability was excellent (Fig. 14). 14 is a graph showing the effect of a sesame bioconversion in a type 2 diabetic mouse.

(4) Lipid analysis

Analysis of serum lipids increased by high fat diet showed that the amount of HDL decreased and the amount of triglyceride, cholesterol and LDL increased in type 2 diabetic mice. The high-fat diet regenerated the reduced amount of HDL, and increased the amount of triglyceride, cholesterol and LDL when the sesame biotransformation was administered to mice. This is shown in Table 13 below.

Sample triglyceride (mg / dL) total cholesterol (mg / dL) HDL (mg / dL) LDL (mg / dL) normal control 112 ± 9 141 ± 8 84.2 ± 5.6 28.3 ± 1.7 HFD-control 163 ± 12 169 ± 12 61.9 ± 6.0 35.7 ± 2.3 HFD-10 mg / kg
Sesame Bioconversion 129 ± 9 151 ± 10 72.6 ± 6.8 30.7 ± 2.2

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(5) Assessment of metabolic enzyme-regulating ability

The activity of G6pase, PEPCK, and GCK was measured as the sesame bioconversion recovers insulin secretion reduced by type 2 diabetes. The results are shown in Table 14 below.

Sample enzyme activity (nmol / min / mg protein) G6pase PEPCK GCK Vehicle 62.419 + 5.056 22.908 ± 1.087 11.524 ± 0.993 HFD control 183.442 ± 10.115 41.438 ± 3.240 5.113 + 0.427 HFD-10 mg / kg
Sesame Bioconversion 85.037 + - 4.426 28.834 + 2.090 9.306 ± 0.718

The results showed that sesame bioconversion inhibited the activity of G6pase and PEPCK, and regulated the activity of GCK, as did type 1 diabetes. From these results, it can be concluded that the sesame bioconverting product has an excellent effect of controlling the enzyme activity.

 (6) Inflammatory cytokine analysis

Although the induction mechanism of type 2 diabetes due to obesity is not yet known, inflammatory cytokines secreted from adipose tissue have been reported to play an important role. Therefore, we examined the effect of sesame bioconversion on the expression of cytokines in adipose tissue and the cytokines secreted into blood. The results are shown in Table 15 below.

Sample serum (pg / mL) adipose tissue (pg / mL) TNF-a IL-1? IL-6 TNF-a IL-1? IL-6 normal control 2.87 ± 0.13 13.2 ± 1.1 16.9 ± 1.3 23.0 ± 1.1 58.3 ± 4.2 77.3 ± 5.8 HFD-control 15.2 ± 1.2 109 ± 8 58.9 ± 4.0 74.9 ± 5.6 175 ± 9 212 ± 16 HFD-10 mg / kg
Sesame Bioconversion 7.83 + - 0.47 45.3 ± 3.5 36.1 ± 2.6 38.1 ± 2.5 113 ± 6 132 ± 10

As a result of the experiment, it was confirmed that both type 2 diabetes induced increase in adipose tissue and serum TNF-α, IL-1β, and IL-6, and the expression of all three cytokines during sesame bioconversion Lt; / RTI >

In the case of IL-1β, which is known to play the most important role in the insulin resistance of the beta cells, both the serum and the adipose tissue showed a suppressive effect when the sesame bioconversion was administered.

 (7) Protection effect of liver tissue and pancreatic tissue

When liver and pancreatic tissues were extracted and stained in mice induced type 2 diabetes, tissue damage and fat accumulation were observed in the liver during diabetic induction, and Langerhans' islet was observed in the pancreas. On the other hand, when the sesame biotransformation was administered, such liver tissue damage and fat accumulation were alleviated, and the pancreatic Langerhans is also protected (FIG. 15 (A) and FIG. 15 (B)). Fig. 15 (A) is a photograph showing the liver tissue protective effect of the sesame biotransformation in the type 2 diabetic mouse, Fig. 15 (B) is a photograph showing the pancreatic tissue protective effect .

[Experimental Example 14: Evaluation of in vitro liver protective function of sesame bioconduct in hepatocytes ]

In this experiment, HepG2 cell line, a human-derived liver cancer cell line, was used to examine the alcohol-induced hepatotoxicity of sesame biological transformants.

 (1) Cytotoxicity measurement using HepG2 cell line

HepG2 cells were seeded in a 96-well plate at a density of 1 × 10 ⁵ cells / well, treated with sesame biotransformers at a concentration of 1, 10, 100 μg / mL and 200 mM of ethanol to kill the hepatocytes Respectively. After 24 hours of culture, cell viability was measured by MTT assay.

As a result of the experiment, the survival rate of the sesame biotransformant was increased in a concentration dependent manner, and the survival rate was 80.1% at the concentration of 100 ㎍ / mL (FIG. 16). 16 is a graph showing an inhibitory effect of sesame bioconversion on alcohol-induced hepatocyte death.

 (2) ROS scavenging activity induced by ethanol treatment

We confirmed that the generation of ROS, one of the mechanisms of cell death induced by ethanol treatment of hepatocytes, can be inhibited by the administration of sesame bioconversion. Sesame bioconversion was carried out for 30 minutes at 1, 10, and 100 ㎍ / mL, respectively, followed by treatment with 200 mM ethanol for 30 minutes to induce the development of ROS. Then, DCFH-DA, a fluorescent marker of ROS, was treated and labeled, and the fluorescence generated by ROS was measured.

As a result, the sesame bioconversion showed a concentration-dependent inhibition of ROS production, and it was confirmed that ROS inhibition ability was 37.3% at a concentration of 100 ㎍ / mL (FIG. 17). FIG. 17 is a graph showing the inhibitory effect of sesame bioconversion on the generation of ROS in alcohol-induced hepatocytes.

[ Experimental Example 15: Alcohol-Induced Liver Damage Induced Mice, Sesame Bioconversion Evaluation of in vivo liver protection]

In this experiment, we investigated the effect of sesame bioconversion on the inhibition of alcoholic fatty liver disease in an in vivo experimental system.

(1) Changes in liver weight in alcoholic fatty liver mouse models induced by alcohol administration

The ethanol-induced liver mouse model using the 'Liber-Decarli' liquid basic diet was used in the in vivo experimental system. The composition of the 'Liber-Decarli' liquid basic diet is shown in Table 16 below, and the same calories as the ethanol contained in the normal diets (AIN-93G) and the ethanol diets fed with the normal diet were substituted with maltodextrin Control diet (control diet).

component control diets (g / kg diet) ethanol diets (g / kg diet) casein (100 mesh) 41.4 41.4 L-cysteine 0.5 0.5 DL-methionine 0.3 0.3 corn oil 8.5 8.5 olive oil 28.4 28.4 safflower oil 2.7 2.7 dextrin maltose 115.2 25.6 cellulose 10.0 10 choline bitartrate 0.53 0.53 xanthan gum 3.0 3.0 salt mix ^a 8.75 8.75 vitamin mix ^b 2.5 2.5 ethanol 0 48

(* ^A salt mix (g / kg mix): calcium phosphate, dibasic, 500; sodium chloride, 74; potassium citrate, monohydrate, 220; potassium sulfate, 52 magnesium oxide, 24 manganous sulfate, , 4.95; zinc carbonate, 1.6; cupric carbonate, 0.3; potassium iodate, 0.01; sodium selenite, 0.01; chromium potassium sulfate, 0.55; sodium fluoride, 0.06; sucrose, finely powdered, 117.92./ b vitamin mix (g / kg mix ), vitamin A12 (0.1%), vitamin A acetate (500,000 IU / ml), thiamin HCl, 0.6, riboflavin, 0.6, pyridoxine HCl, 0.7, niacin, 3.0, calcium pantothenate, 1.6, folic acid, 0.2, biotin, g), 4.8, vitamin D3 (400,000 IU / g), 24 menadione sodium bisulfite, 0.08, p-aminobenzoic acid, 5, inositol, 10, dextrose, 939.)

Sesame bioconverts were added to ethanol diet at a dose of 10 mg / kg for a total of 8 weeks to induce fatty liver and hepatitis by ethanol.

After 8 weeks, the weight and liver weight of the mice were measured. As a result, it was confirmed that the liver weight / body weight ratio was increased by 13.0% compared to that of the normal mice due to inflammation and fatty liver in the ethanol diet group. However, when the sesame bioconversion was administered, it was confirmed that it increased by 1.1%, and thus it was judged to inhibit the inflammatory reaction and fatty liver formation in liver tissue (FIG. 18). 18 is a graph showing the effect of inhibiting liver weight increase due to administration of a sesame bioconversion in an alcohol-induced liver damaged mouse.

 (2) Measurement of serum biochemical indicators

After isolating the mouse serum, biochemical markers present in the serum were measured to confirm liver damage. The results are shown in Table 17 below.

Sample Serum index (U / L) GOT GPT γ-GTP billirubin normal control 63.7 ± 5.2 22.1 + 1.7 5.56 + - 0.47 0.49 + 0.03 control diet 58.9 ± 4.9 26.7 ± 1.5 5.53 + - 0.39 0.51 + 0.04 Ethanol diet-control 75.5 ± 5.9 45.1 ± 3.6 6.38 + - 0.42 0.81 ± 0.05 Ethanol diet-10 mg / kg
Sesame Bioconversion 60.3 ± 4.2 28.1 ± 1.4 5.71 + - 0.43 0.60 + 0.04

Experimental results showed that all of the four biochemical markers were increased in ethanol - induced liver injury, but they were found to be close to the normal values when the sesame bioconversion was administered together.

 (3) Changes in antioxidants in liver tissues

Reduced GSH shows cytoprotective effect against oxidative stress, but it is known that when alcohol induced liver toxicity, glutathione is oxidized to GSSG and does not control liver toxicity. Therefore, the amount and type of glutathione (GSH), a representative antioxidant in liver tissue, were measured. The results are shown in Table 18 below.

Sample Total GSH
([Mu] g / mg tissue) Reduced GSH
([Mu] g / mg tissue) GSSG / GSH ratio
([Mu] g / mg tissue) normal control 4.27 ± 0.28 3.75 ± 0.27 0.16 ± 0.01 control diet 4.95 + - 0.45 4.30 0.38 0.17 ± 0.01 Ethanol diet-control 3.82 ± 0.29 3.01 ± 0.21 0.22 0.02 Ethanol diet-10 mg / kg
Sesame Bioconversion 4.41 0.25 4.08 ± 0.33 0.17 ± 0.01

In the alcohol-fed mice, both the total amount of GSH and the amount of reduced GSH were decreased, and the ratio of GSSG / GSH was 1.38 times higher than the normal value. On the other hand, when treated with sesame bioconversion, the total GSH amount and the reduced GSH amount were increased, and the GSSG / GSH ratio was found to be decreased to the normal value.

 (4) Protection effect of liver tissue

The mice were sacrificed and liver was extracted and stained with 'hematoxylene' and 'eosin Y'.

Experimental results showed that normal mice and control mice had no hepatic lesions and no fat accumulation, whereas hepatocyte damage, hemorrhage, and fat accumulation occurred in ethanol - fed mice. However, when the sesame bioconversion was administered, liver damage caused by ethanol was strongly inhibited, and fat accumulation was also found to be very low. Thus, it was confirmed that the liver protection effect of the sesame bioconversion was excellent (Fig. 19) . FIG. 19 is a photograph showing the protective effect of liver damage caused by administration of a sesame bioconversion in an alcohol-induced liver damaged mouse.

[Example 2: Isolation and purification of immunological active ingredient (polysaccharide)] [

(1) Separation and purification process

The production process for obtaining polysaccharide fractions from sesame bioconverts is shown in Fig. To the pulverized sesame bioconversion (Example 1) about 20 times of the third distilled water was added to extract the water-soluble substance. After extracting the water soluble substance, the residue was removed by centrifugation and the extract supernatant was separated. To the extract supernatant was added EtOH in an amount corresponding to four times that of the supernatant, mixed well, and stored at 4 ° C overnight.

The resulting precipitate was separated by centrifugation, and the supernatant was removed. The precipitate was dissolved by adding tertiary distilled water, and the undissolved material was removed by centrifugation or filtration. Dialysis was carried out using a dialysis tube for the filtered precipitate solution. The third distilled water was changed every 2 hours, and this was repeated 10 times or more. After completion of the dialysis, it was pulverized through lyophilization.

(2) Analysis of polysaccharide fractions

The instrument used for the analysis of the polysaccharide fractions was Shimadzu LC 010Avp (Shimadzu Co., Japan), and the analysis conditions were as follows.

device Condition Instrument SHIMADZU_HPLC, SEDEX 75_ELSD Column Ultrahydrogel (TM) 1000 (7.8 x 300 mm) Waters, WAT011535 Column oven 45 ° C Flow rate 0.6 ml / min Solvent HPLC grade water Elution Isocratic, HPLC water 100% Run time 40 min Injection 20 μl Sample extraction 100 mg of the sample was shaken with 10 ml of HPLC water at 250 rpm for 1 hour,
Analysis after centrifugation

Polysaccharide fractions were prepared by separation purification using sesame bioconducts of the first batch produced in a 50 L fermenter. Analysis of the polysaccharide fractions of sesame bioconverted by HPLC showed that the fractions of immunologically active polysaccharides were well removed from all chromatogram peaks. Table 20 below shows the recovery rates of the polysaccharide fractions.

Recovery of polysaccharide fractions (%) Sesame Bioconversion 50L 1st batch 12.16

On the other hand, a sample was collected for each polysaccharide fraction production process of the sesame bioconversion, and the chromatogram pattern change according to the process was analyzed. As a comparative substance, Mesima Capsule 550, which is an immunoactive polysaccharide derived from Mycelia mycelium and used for chemotherapy, was used.

The supernatant from which the insoluble material of the sesame biotransformation was removed was analyzed by gel filtration HPLC. As shown in FIG. 21, the peak of the low molecular weight substance was observed at 18.7 minutes and the peak of the polysaccharide fraction of the polymer was detected at 8.3 minutes .

As a result of measuring the macrophage NO production ability of the eluate of the peak of the low molecular substance and the eluate of the polysaccharide fraction peak of the polymer, only macrophage NO production ability was observed in the polysaccharide fraction peak of the polymer.

After the EtOH precipitation process and the dialysis process, the low molecular weight material was removed and the peak of the low molecular weight material was decreased. Also, it was confirmed that no significant change was observed after freeze drying.

[Experimental Example 16: Assessment of macrophage activation ability by measurement of macrophage NO production]

Polysaccharide fractions were prepared using the first batch of sesame bioconversion produced in a 50 L fermenter and the ability of macrophage NO production was measured to evaluate macrophage activation.

As a result of the measurement, it was confirmed that the NO production ability of the polysaccharide fraction prepared from the sesame biotransformant of the first batch of 50 L was significantly improved as compared with the sesame biotransformant. It was confirmed that the polysaccharide fraction prepared from the sesame biotransformant produced in the first batch had an immunostimulatory activity MEC ₁₀₀ of about 1/4 占 퐂 / ml (FIG. 22).

[ Experimental Example 17: Analysis of cytokine secretion pattern in macrophages and analysis of receptor solubility ]

Recently, research results on various physiological activities such as macrophage activation, NK cell stimulating activity and stimulation of cytokine production by polysaccharide showing immunological activity have been continuously announced.

In addition, the specific structure of the polysaccharide exhibiting immunological activity is known to be a pattern recognition receptor (PRR), which is a specific receptor expressing on the cell surface, such as macrophages, and it is known that it mediates signal transduction.

Based on these results, we tried to confirm whether the polysaccharide fractions of sesame bioconverts produced by the separation and purification process established above were recognized in pattern recognition receptors, which are specific receptors of immune cells, and mediated signal transduction. First, a specific receptor was deduced from the comparison of cytokine secretion patterns in macrophages. In order to confirm this, an experiment was carried out to examine whether signaling is blocked by treating an inhibitor against the specific receptor expressed .

The TLR4 (Toll-like receptor4) receptor was identified as a specific receptor for the sesame bioconversion. TLR4, a TLR4 ligand, and TLR4 agonist, The immunoreactivity of sesame bioconductors was investigated by comparison with MPLA, a proven immunomodulator.

First, the secretory pattern of cytokines in macrophages was examined in order to confirm the immunological activity of polysaccharide fractions of sesame bioconvert. As a control, LPS (Lipopolysaccharide) was treated at concentrations of 0.01 ㎍ / mL, 0.1 ㎍ / mL and 1 ㎍ / mL, respectively. The sesame bioconduct was treated with 0.08 ㎍ / mL, 0.8 ㎍ / mL and 8 ㎍ / mL While the polysaccharide fractions of the sesame bioconversion were tested at the concentrations of 0.01 ㎍ / mL, 0.1 ㎍ / mL, and 1 ㎍ / mL, which are the same as those of LPS. The concentration of the sesame bioconversion was determined based on the recovery rate of the polysaccharide fractions from the sesame bioconversion, which was about 12% through repeated experiments.

(2) Evaluation of TNF-α secretion amount

Sixteen hours after the sample treatment, the supernatants of the macrophage culture were taken and the secretion and concentration of TNF-α (Tumor necrosis factor-α) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In the case of sesame bioconversion, it was confirmed that TNF-α was secreted from the treatment concentration of 0.08 μg / mL. As the treatment concentration increased, the secretion amount of TNF-α also increased. In the sesame bioconverted polysaccharide fraction, the secretion of TNF-α increased from 0.01 μg / mL to 0.1 μg / mL and 1 μg / mL. And the level of secretion was similar to that of TNF-α in the sesame bioconversion at the concentration determined based on the recovery rate. In addition, it was confirmed that the TNF-α secretion amount was similarly increased with increasing concentration in both sesame bioconversion and sesame bioconversion polysaccharide fractions.

It was confirmed that polysaccharide component was recognized as a specific receptor of macrophage among several substances contained in sesame bioconversion, and thus it was confirmed that TNF-α is secreted by mediating signal transduction.

In addition, when LPS was secreted at a concentration of 0.01 ㎍ / mL, the amount of TNF-α secreted by the polysaccharide fraction of sesame bioconverted polysaccharide was higher than that of LPS at the concentration of 0.1 ㎍ / mL and 1 ㎍ / mL TNF-a < / RTI >

(3) Evaluation of IL-1β secretion amount

16 hours after the sample treatment, the supernatants of the macrophage culture were taken and the secretion and concentration of IL-1β (Interlukine-1β) were analyzed by ELISA (enzyme linked immuno solvent assay).

As a result, the LPS, sesame bioconversion, and sesame bioconversion polysaccharide fractions exhibited a remarkably low value or no value at all, regardless of their concentrations (not shown).

(4) Evaluation of IL-4 secretion amount

At 16 hours after the sample treatment, the supernatant of macrophage culture was taken and the secretion and concentration of IL-4 (Interlukine-4) were analyzed by ELISA (enzyme linked immuno solvent assay).

As a result, the LPS, sesame bioconversion, and sesame bioconversion polysaccharide fractions exhibited a remarkably low value or no value at all, regardless of their concentrations (not shown).

(5) Evaluation of IL-5 secretion amount

16 hours after the sample treatment, the supernatant of macrophage culture was taken and the secretion and concentration of IL-5 (Interlukine-5) were analyzed by ELISA (enzyme linked immuno solvent assay).

As a result, the LPS, sesame bioconversion, and sesame bioconversion polysaccharide fractions exhibited a remarkably low value or no value at all, regardless of their concentrations (not shown).

(6) Evaluation of IL-6 secretion amount

Sixteen hours after the sample treatment, the culture supernatant of macrophages was collected and the secretion and concentration of IL-6 (Interlukine-6) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In the case of sesame bioconversion, IL-6 secretion started from the treatment concentration of 0.8 ㎍ / mL, and the secretion of IL-6 was increased at 8 ㎍ / mL.

In the sesame bioconverted polysaccharide fraction, the amount of IL-6 secretion increased as the concentration of IL-6 started to increase from 0.01 ㎍ / mL. In addition, it was confirmed that the IL-6 secretion amount showed a similar amount as the concentration of sesame bioconversion and sesame bioconversion polysaccharide fraction increased.

It was confirmed that among the various substances contained in the sesame bioconversion, the polysaccharide component was recognized as a specific receptor of macrophages, and thus, it was confirmed to be effective for mediating signal transduction and secretion of IL-6.

When the IL-6 secretion level of the sesame bioconverted polysaccharide fraction was compared with that of LPS, the level of IL-6 secretion by LPS was slightly lower at the concentrations of 0.1 ㎍ / mL and 1 ㎍ / mL.

(7) Evaluation of IL-10 secretion amount

16 hours after the sample treatment, the supernatant of macrophage culture was taken and the secretion and concentration of IL-10 (Interlukine-10) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In sesame bioconversion, the amount of IL-10 secretion was measured at 8 ㎍ / mL. In the sesame bioconversion polysaccharide fraction, the secretion amount of IL-10 was measured at 1 ㎍ / mL.

It was confirmed that IL-10 secretion levels of sesame bioconversion and sesame bioconversion polysaccharide fractions were similarly measured at high concentrations. It was confirmed that polysaccharide component of various substances contained in sesame bioconversion is recognized as a specific receptor of macrophages, and thus, it is an efficacy that mediates signal transduction and secretes IL-10.

When IL-10 secretion level of LPS was compared, IL-10 secretion increased from 0.01 ㎍ / mL to 0.1 ㎍ / mL and 1 ㎍ / mL, and IL-10 secretion increased . Compared with the sesame bioconversion polysaccharide fraction, the amount of IL-10 secretion in the polysaccharide fraction of LPS and sesame bioconvert at 1 ㎍ / mL was similar to that of the polysaccharide fraction.

(8) Evaluation of secretion of IL-12p70

16 hours after the sample treatment, the supernatant of the macrophage culture was taken and the secretion and concentration of IL-12p70 (Interlukine-12p70) were analyzed by ELISA (enzyme linked immuno solvent assay).

As a result, the LPS, sesame bioconversion, and sesame bioconversion polysaccharide fractions exhibited a remarkably low value or no value at all, regardless of their concentrations (not shown).

(9) Evaluation of IFN-beta secretion amount

At 16 hours after the sample treatment, the supernatant of the macrophage culture was taken and the secretion and concentration of IFN-β (Interferon-β) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In the sesame bioconversion, IFN-β was secreted from 0.08 ㎍ / mL at the treatment concentration, and the secretion of IFN-β was also increased with increasing treatment concentration. In sesame bioconversion polysaccharide fractions, the secretion of IFN-β increased from 0.01 μg / mL to 0.1 μg / mL and 1 μg / mL, indicating that the secretion of IFN-β increased And the amount of secretion was similar to the amount of IFN-β secreted from the sesame bioconversion at the concentration set based on the recovery rate. In addition, it was confirmed that the IFN-β secretion level of the sesame bioconversion and sesame bioconversion polysaccharide fractions increased similarly as the concentration increased.

It was confirmed that the polysaccharide component was recognized as a specific receptor of macrophages among the various substances contained in the sesame bioconversion, and thus it was confirmed that it is effective to secrete IFN-β by mediating signal transduction. When the amount of IFN-beta secreted by LPS was compared, it was confirmed that the amount of secretion increased in a concentration-dependent manner. The LPS was slightly higher than the sesame bioconversion polysaccharide fraction.

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(10) Analysis of results

TNF-α, IL-6, IL-10 and IFN-β were expressed in the sesame bioconversion. As a result, the immunological activity of the sesame biotransformant was confirmed by the polysaccharide fraction .

In comparison with LPS, the cytokine secretion patterns of TNF-α, IL-6 and IFN-β in LPS were similar to those of sesame bioconvert polysaccharide fractions and IL-1β, IL-4 and IL-5 , And IL-12p70 were not secreted together.

LPS is well known as TLR4 (Toll-lokreceptor4), and it can be interpreted as a polysaccharide fraction of sesame biotransformers recognized by TLR4 and mediating signals.

On the other hand, TNF-α is secreted from macrophages and mononuclear cells during phagocytosis and is known to have an antiviral effect against various viruses such as influenza virus along with inducible nitric oxide (NO). In addition, IL-6 is a major mediator in antigen-specific immune responses, inflammatory responses, and acute responses, and is a representative cytokine that plays a key role in the in vivo defense system. IFN-β is a type I interferon in combination with IFN-α and has antiviral and antiviral effects. In particular, it induces Th1 immune response as a typical cytokine which induces Th1 immune response by contributing to Th1 cell differentiation, while Th2 and Th17 It is a cytokine with an immunoregulatory function that suppresses the immune response.

In conclusion, it was confirmed that the production of interferon-β was induced in the activated macrophages, and it was judged that the Th1 immune response could be specifically activated. All of these immunological activities were confirmed by the polysaccharide fraction of the polymer.

[Experimental Example 18: Confirmation of Receptor]

(1) Outline of experiment

Many of the immunomaterials developed so far have not been identified with receptors, and only some beta glucan products are known to bind to receptors called dectin-1. However, even beta-glucan products are known to bind to Dextin-1 in the case of insoluble materials, while water-soluble materials do not bind to Dextin-1.

If the receptor is known, the biomarker can be determined, and on the basis of this, there is an advantage that the biomarker can be used in the clinical test for the development of pharmaceuticals and veterinary pharmaceuticals.

Based on the results of previous experiments, it was concluded that TLR4 is a receptor for the sesame bioconversion and sesame bioconversion polysaccharide fractions, and that sesame bioconversion and sesame bioconversion polysaccharide fractions are specific to TLR4 and mediate the signal We used TAK-242, a specific inhibitor for TLR4, to confirm the results.

(2) Evaluation of TNF-α secretion amount

Sixteen hours after the sample treatment, the supernatants of the macrophage culture were taken and the secretion and concentration of TNF-α (Tumor necrosis factor-α) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In the case of LPS, it was confirmed that when the TLR4 inhibitor TAK-242 was treated at a concentration of 0.01 μg / ml and 0.1 μg / ml, the secretion amount of TNF-α was completely blocked. At a high concentration of 1 μg / ml And it was confirmed that the decrease greatly greatly. In the case of the sesame bioconversion, it was confirmed that the TNF-α secretion level was completely blocked when the TLR4 inhibitor TAK-242 was treated at a concentration of 0.08 μg / ml, and a high concentration of 0.8 μg / ml and 8 μg / ml It can be confirmed that the decrease is remarkably greatly reduced. In the case of the sesame bioconversion polysaccharide fraction, it was also confirmed that when TAK-242 was treated at a concentration of 0.01 μg / ml and 0.1 μg / ml, the secretion amount of TNF-α was completely blocked, and 1 μg / ml It was confirmed that the concentration at the high concentration was greatly reduced greatly.

Based on the results that the secretion of TNF-α was completely blocked or greatly reduced during treatment with the TLR4 inhibitor TAK-242, polysaccharides of sesame biotransformants and sesame bioconductors were specifically bound to TLR4 and recognized as TNF -α secretion signal was mediated.

(3) Evaluation of IL-6 secretion amount

Sixteen hours after the sample treatment, the culture supernatant of macrophages was collected and the secretion and concentration of IL-6 (Interlukine-6) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In the case of LPS, the TLR4 inhibitor TAK-242 was completely blocked at all three concentrations, confirming that IL-6 secretion was completely blocked. In the case of the sesame bioconversion, it was also confirmed that when the TLR4 inhibitor TAK-242 was treated at all three concentrations, the secretion of IL-6 was completely blocked. In the case of sesame bioconverted polysaccharide fractions, it was confirmed that when TAK-242 was treated at all three concentrations, secretion of IL-6 was completely blocked as well.

The TLR4 inhibitor, TAK-242, completely blocked the secretion of IL-6, and the polysaccharide of the sesame bioconversion and sesame bioconversion was specifically bound to TLR4 like IL-4, and IL-6 secretion signal Respectively.

(4) Evaluation of IL-10 secretion amount

16 hours after the sample treatment, the supernatant of macrophage culture was taken and the secretion and concentration of IL-10 (Interlukine-10) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In the case of LPS, when the TLR4 inhibitor TAK-242 was treated at all three concentrations, the secretion of IL-10 was completely blocked. In the case of sesame bioconversion, it was also confirmed that when the TLR4 inhibitor TAK-242 was treated, the secretion of IL-10 was completely blocked. In the case of the sesame bioconversion polysaccharide fraction, it was also confirmed that when the TLR4 inhibitor TAK-242 was treated, the secretion of IL-10 was completely blocked as well.

The TLR4 inhibitor, TAK-242, completely blocked the secretion of IL-10, and the polysaccharide of the sesame bioconversion and sesame bioconversion was specifically bound to TLR4 like IL-10, Respectively.

(5) Evaluation of IFN-beta secretion amount

At 16 hours after the sample treatment, the supernatant of the macrophage culture was taken and the secretion and concentration of IFN-β (Interferon-β) were analyzed by ELISA (enzyme linked immuno solvent assay).

The results of the analysis are shown in FIG. In the case of LPS, the secretion of IFN-β was completely blocked by treatment with the TLR4 inhibitor TAK-242 at all three concentrations. In the case of sesame bioconversion, the secretion of IFN-β was completely blocked by treatment with the TLR4 inhibitor TAK-242 at all three concentrations. In the sesame bioconversion polysaccharide fraction, When the TLR4 inhibitor TAK-242 was treated, it was confirmed that the secretion amount of IFN-β was completely blocked.

On the basis of the result that the secretion of IFN-β was completely blocked during the treatment with the TLR4 inhibitor TAK-242, the polysaccharide of the sesame bioconversion and the sesame bioconversion was specifically bound to TLR4 similarly to LPS, .

(6) Analysis of results

The macrophage immunoactivity of the sesame bioconversion was confirmed by the polysaccharide fraction. The responses induced by macrophage activation were all suppressed by the TLR4-specific inhibitor TAK242, confirming that TLR4 is a receptor for sesame bioconversion, The sesame bioconversion product and the sesame bioconversion polysaccharide fraction produced by the bioconversion process pursued in the present invention have activity of the TLR4 agonist.

Claims (6)

(A) adding at least one selected from the group consisting of amylase and cellulase to sesame seeds, reacting them, and sterilizing them to prepare a liquid medium;
(B) after the step (a), inoculating the mushroom mycelium into the liquid medium and culturing to produce a culture;
(C) after the step (b), adding the fibrolytic enzyme to the culture and reacting; Which comprises a sesame bioconversion product produced from a process comprising the steps of:
Wherein the sesame bioconversion product is a polysaccharide.
The method according to claim 1,
The sesame bioconversion product,
Wherein said composition inhibits the production of triglyceride and low-density cholesterol (LDL-cholesterol) and increases the production of high-density cholesterol (HDL-cholesterol).
The method according to claim 1,
The polysaccharide,
(A) adding distilled water to the sesame bioconversion obtained through step (c) to extract a water-soluble substance;
Removing the residue through centrifugation after extracting the water-soluble substance, and separating the extracted supernatant (B);
Adding EtOH to the extracted supernatant, mixing and maintaining (c);
Separating the resulting precipitate after said holding (d);
Adding distilled water to the separated precipitate to dissolve the precipitate, and then removing undissolved material by centrifugation or filtration; And
(Vi) dialyzing the sediment lysis solution after the centrifugation or the filtration step. [Claim 11] The composition for improving hyperlipemia according to claim 1,
(A) adding at least one selected from the group consisting of amylase and cellulase to sesame seeds, reacting them, and sterilizing them to prepare a liquid medium;
(B) after the step (a), inoculating the mushroom mycelium into the liquid medium and culturing to produce a culture;
(C) after the step (b), adding the fibrolytic enzyme to the culture and reacting; Which comprises a sesame bioconversion product produced from a process comprising the steps of:
Wherein the sesame bioconversion product is a polysaccharide.
5. The method of claim 4,
The sesame bioconversion product,
(HDL-cholesterol) by inhibiting the production of triglyceride, triglyceride, and low-density cholesterol (LDL-cholesterol), and increasing the production of high-density cholesterol (HDL-cholesterol).
5. The method of claim 4,
The polysaccharide,
(A) adding distilled water to the sesame bioconversion obtained through step (c) to extract a water-soluble substance;
Removing the residue through centrifugation after extracting the water-soluble substance, and separating the extracted supernatant (B);
Adding EtOH to the extracted supernatant, mixing and maintaining (c);
Separating the resulting precipitate after said holding (d);
Adding distilled water to the separated precipitate to dissolve the precipitate, and then removing undissolved material by centrifugation or filtration; And
And (ii) dialyzing the sediment lysis solution subjected to centrifugation or filtration. The pharmaceutical composition for preventing or treating hyperlipidemia according to claim 1,

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