TWI461206B - Method for producing ferment of glycine max (l.), ferment and uses thereof - Google Patents

Description

Method for producing black bean fermented product, black bean fermented product and use thereof

The present invention relates to a nutritional supplement. In detail, the present invention is based on Coprinus beans (Glycine max (L.)) of (Coprinus sp.) Was fermented as nutritional supplements and its application.

Nowadays, the competition in the industrial and commercial circles is fierce. All kinds of pressures make people's emotions tense. The body is also in a state of fatigue and even overwork. It causes mental inconsistency, irritability and metabolism to decline. If it is not properly relieved, it is easy to cause physical and mental trauma. In addition, In order to improve the quality of life and enhance competitiveness, various nutritional supplements have become an indispensable supplement in people's lives.

Most of the refreshing beverages come from caffeine and chemically synthesized vitamins. Although it can refresh people quickly, the effect fades quickly. After the fade, the mental condition of the drinker is worse than before drinking. It is not a proper nutrition. Supplements.

Chicken essence has now been shown to aid in the elimination of urea nitrogen and lactic acid in the blood. Because urea nitrogen is a product of protein metabolism in the human body, when the blood urea nitrogen content is increased, it is easy to cause fatigue and burnout; on the other hand, lactic acid is an important cause of muscle soreness after exercise, if it can accelerate the lactic acid in the blood. Metabolism can reduce muscle soreness after exercise and accelerate physical recovery. Most of the production of chicken essence is directly extracted from the concentrate of chicken by high temperature and high pressure. The main health care related ingredients are the total branched amino acids containing leucine, isoleucine and valine, which are important components of protein. . Because chicken essence is rich in amino acid, it can keep the drinker's mental state for a long time. Even if the effect fades, the spirit and physical strength of the drinker will not drop sharply immediately, so it is often used as the most postoperative rehabilitation for patients. Good tonic.

However, vegetarians are not able to enjoy the benefits of chicken essence, so it is necessary to develop nutritional supplements that contain nutrients similar to chicken essence and can be used by vegetarians.

The invention provides a nutritional supplement which is edible and has the effect and nutrient composition similar to chicken essence, so that the vegetarian can also get good health care.

The nutritional supplement also has the effect of inhibiting the production of nitric oxide and can prevent and/or treat inflammation.

The present invention provides a method for producing a fermented product of Coprinus sp., which comprises fermenting a sterilized medium of black beans with Coprinus comatus.

The present invention also provides a fermented product of Coprinus comatus, which is obtained by the aforementioned method.

The present invention further provides a nutritional supplement comprising the aforementioned Black Bean Coprinus comatus ferment.

The present invention further provides a use of a fermented product of Coprinus comatus, which is used for the preparation of a medicament for preventing and/or treating inflammation.

The invention is described in detail in the following sections. Other features, objects, and advantages of the invention are apparent from the embodiments of the invention and the appended claims.

The invention utilizes the black bean fermented product as the main component of the nutritional supplement, and the protein content of the black bean is as high as 36 to 40, which is equivalent to twice the meat content, three times the egg, and twelve times the milk. The eighteen kinds of amino acids of black beans are rich in content, especially the eight amino acids required by the human body are higher than the advanced protein standards stipulated by the US Food and Drug Administration. Compared with soybeans which are also beans, black beans contain higher protein content, and black beans are more effective in reducing arterial atherosclerosis and preventing vascular embolism than soybeans. In addition, black beans contain many anti-oxidant and anti-aging ingredients, especially the content of isoflavones and anthocyanins are significantly higher than soybeans, so its antioxidant capacity is stronger than soybeans.

The black bean according to the present invention can be changed by the fermentation of Coprinus comatus, and the composition metabolized by fermentation is different from that before fermentation, and has different curative effects.

Unless otherwise defined herein, the scientific terms used in connection with the present invention shall have the meaning commonly understood by one of ordinary skill. The meaning and scope of the terms should be clear; however, if there is any potential ambiguity, the definitions provided herein take precedence over any dictionary or foreign definition.

Unless the context requires otherwise, singular terms shall include the plural and plural terms shall also include the singular.

Unless otherwise indicated, it is to be understood that the following terms as used in this context have the following meanings: The term "prevention" as used herein refers to delaying the onset of a condition or reducing the onset of a condition in a susceptible individual.

The term "treating" as used herein means reducing and/or improving the symptoms of a rickety individual.

The term "individual" as used herein refers to an animal, particularly a mammal. According to a preferred aspect of the invention, "individual" refers to a human.

The term "therapeutically effective amount" as used herein means an amount of an active ingredient, either alone or in combination with other treatments/drugs, which is sufficient to demonstrate therapeutic efficacy.

The term "carrier" or "pharmaceutically acceptable carrier" as used herein refers to diluents, excipients, acceptors or the like which are well known to those of ordinary skill in the art of making pharmaceutical compositions. .

Preparation

A method for producing a fermented product of Coprinus comatus according to the present invention, which comprises fermenting a sterilized medium of black beans with Coprinus comatus.

The term "black bean" as used herein refers to the root, leaf, stem, seed or any combination or whole plant of a black bean plant, preferably a seed. The seed preferably comprises a seed coat, an embryo and a cotyledon, which may be used throughout the seed or in any part or in combination. The black beans according to the present invention are preferably samples that have not been processed or processed by processes such as drying, peeling, slicing or grinding.

The variety according to the present invention is not particularly limited. In a preferred embodiment of the present invention, it is a Hengchun (Huangren) black bean, a Hengchun black bead, a yellow kernel black bean, a green kernel black bean, a Danbo bean, Black lentils or black peas; more preferably, they are Hengchun black peas. In a preferred embodiment of the invention, the scientific name is Glycine max (L.).

The term "sterilization" as used herein refers to a measure of killing or removing microorganisms and bacterial spores on an object. Sterilization methods commonly used in the art include a pressure sterilization method, an ultraviolet irradiation sterilization method, a filtration sterilization method, and a chemical sterilization method; preferably, a pressure sterilization method. The pressure sterilization method is generally carried out in a sealed high-pressure steam pot. When the vapor pressure in the pot is raised to 1.05 kg/cm 2 , the steam temperature can reach 121 ° C. At this high temperature, it is preferably about 15 minutes or more. Sterilization can be accomplished for about 15 to 120 minutes, more preferably about 15 to 60 minutes.

In a preferred embodiment of the invention, the sterilized black bean is from about 5 to about 20% (v/w); preferably from about 8 to about 17% (v/w), more preferably about The ratio of 10 to about 15% (v/w) is mixed with the culture medium, and the concentration of the strain is about 6 g biomass g/L to about 12 g biomass g/L; preferably about 7.5 g biomass g/ L to about 10.5 g biomass g/L; more preferably about 8.5 g biomass g/L to about 10.5 g biomass g/L.

According to the method of the present invention, the species of Coprinus comatus used is any commercially available Trichoderma sinensis, and in a preferred embodiment of the present invention, the scientific name is Coprinus sp., more preferably obtained. Coprinus comatus BCRC 35671, 36031, 36055, 36074, 36075, 36076, 36077, 36078, 36079, 36099, 36100, 36101, 36102, 36103, 36104, 36302, 36317, 36390, 36391, 36478, 36479, 36616, 36517, 36597, 36649, 36658 and 36918 strains; more preferably BCRC 36658.

In a specific embodiment of the present invention, the medium may further comprise other components. Preferably, the medium further comprises a flavoring agent to impart a special flavor to the fermented product, which is more convenient for consumption.

In a preferred embodiment of the present invention, the flavoring agent is brown sugar, wheat, black sesame, buckwheat, oat, hazelnut, brown rice, buckwheat, millet, sorghum, pumpkin, walnut or corn; more preferably brown sugar , wheat or black sesame seeds. The flavoring agents may be used singly or in combination, and the content of each component may be adjusted according to the flavor of a person having ordinary knowledge in the technical field of the invention, wherein wheat can enhance the sweet taste; black sesame can enhance the flavor; brown sugar increases the brown sugar. Sweet.

In a preferred embodiment of the present invention, the medium comprises: from about 80 to about 250 parts by weight of black beans; from about 10 to about 30 parts by weight of brown sugar; from about 10 to about 30 parts by weight of wheat; and about 5 to About 25 parts by weight of black sesame seeds.

Those of ordinary skill in the art to which the present invention pertains can select the temperature conditions for fermentation between about 20 ° C and about 30 ° C according to their knowledge. In a preferred embodiment of the invention, the fermentation is incubated at about 25 °C. On the other hand, those having ordinary skill in the art to which the present invention pertains may select other conditions or fermentation forms of fermentation, such as liquid fermentation or solid state fermentation, according to their own knowledge; preferably, the fermentation is solid state fermentation.

In a preferred embodiment of the invention, the method further comprises a drying step after fermentation. Those of ordinary skill in the art to which the present invention pertains may determine the conditions under which the drying is carried out. In a preferred embodiment of the invention, the drying step is carried out at a temperature of from about 50 to about 70 ° C and a pressure of from about -0.4 to about 0.9 kg/cm ² .

In order to be convenient for consumption, the method according to the invention comprises a step of obtaining a fermentation broth. The step of obtaining the fermentation broth may be determined by a person having ordinary knowledge in the technical field of the present invention. In the specific example of the present invention, it comprises hot water or contains methanol, ethanol, propylene glycol, isopropanol or The fermented medium is extracted with a solution of ethyl acetate; preferably hot water or ethanol, and the water-soluble portion thereof is taken. In a preferred embodiment of the invention, the methanol, ethanol, propylene glycol, isopropanol or ethyl acetate liquid is an aqueous solution having a concentration of from about 10 to about 100 v/v%; preferably about 50 to About 90 v/v%; more preferably from about 50 to about 80 v/v%. In a more preferred embodiment of the invention, the solution is from about 50 to about 70 v/v% solution.

According to the method of the present invention, the weight ratio of the fermentation medium to the extraction solution is from about 100 w/v% to about 2000 w/v%, preferably from about 200 w/v% to about 1000 w/v%, more The best is about 400 to about 700 w/v%.

According to the method of the present invention, the step of obtaining the fermentation broth can be carried out by any conventional method such as boiling, soaking, autoclaving, ultrasonic extraction, stirring, shaking or a combination thereof. The extraction time is from about 10 minutes to about 100 minutes; preferably from about 10 minutes to about 50 minutes; more preferably from about 15 minutes to about 30 minutes. The extraction temperature is from about 70 ° C to about 121 ° C, preferably from about 95 ° C to about 121 ° C.

In one embodiment of the invention, the method further comprises the step of removing solids from the fermentation broth, and those of ordinary skill in the art may select suitable removal conditions, which may be by any conventional method. The removal, for example via gauze filtration, centrifugation or a combination thereof; is preferably removed via filtration or precipitation filtration.

In a specific embodiment of the invention, the method further comprises the step of sterilizing the product obtained by fermentation.

In a preferred embodiment of the present invention, the method further comprises adding the culture medium to water, heating and extracting, filtering the medium, and further precipitating the aqueous extract, filtering, and then filtering the obtained filtrate. Sterilize.

Preferably, the method according to the present invention further comprises concentrating the fermentation broth, which can be concentrated by any conventional method, such as freeze drying or evaporation (J. Pharmacol. Sci., 99:294-300, 2005 and J. Chromatography, 932: 91-95, 2001) to obtain a concentrated mint fermentation extract, and optionally freeze-dried by adding an excipient such as methyl cellulose.

Fermentation

The present invention provides a fermented chicken of the black bean, which is obtained by the aforementioned method.

In a specific embodiment of the present invention, the result of the high pressure liquid chromatography (HPLC) analysis of the fermented product is shown in Fig. 1, and the chromatography column used is Inersil ODS-2, 5 μm, ( GL Sciences, ToKyo, Japan), the column has a length of 250 mm and an inner diameter of 4.6 mm. The chromatograph used is Hitachi. PDA L2455, the extract (A) is 0.1% phosphoric acid solution, the extract (B) is 100% acetonitrile, the extraction procedure is 0 minutes to 60 minutes: 90% extract (A)/10% extract (B), 60 minutes to 71 minutes: 0% of the extract (A) / 100% of the extract (B) and 71 minutes to 80 minutes: 90% of the extract (A) / 10% of the extract (B Wherein the residence time is about 7.3, about 7.8, about 9.2, about 13.0, about 13.4, about 16.0, about 21.0, about 21.6, about 25.0, about 25.4, about 37.0, about 39.1, about 40.0, about 43.2, about 53.2 There are absorption peaks at about 56.8, about 60.9 and about 67.2 minutes.

The black beans according to the present invention have different compositions before and after fermentation, and the results of high pressure liquid chromatography analysis of the black beans unfermented are shown in Fig. 2, wherein the residence time is about 7.3, about 7.8, about 9.2. There are absorption peaks of about 11.8, about 13.0, about 13.4, about 16.0, about 21.0, about 21.6, about 25.0, about 25.4, about 37.0, about 39.1, about 40.0, about 67.2 minutes.

Comparing the black beans before and after fermentation, the absorption peak content of the retention time of about 21.0, about 21.6, about 25.0, and about 25.4 minutes was increased, and the new residence time was about 43.2, about 53.2, about 56.8, about 60.9. The absorption peak component of the minute shows that the fermentation does change the composition of the medium.

In a preferred embodiment of the present invention, the fermented product has a total branched chain amino acid (leucine, isoleucine, valine) content of 0.83 g/68 mL, and is compared with a commercially available health food. The certified standard total branched chain amino acid needs to be greater than or equal to 0.32 g/68 mL, which is 1.6 times higher.

combination

The present invention provides a fermented product of Coprinus comatus comprising a therapeutically effective amount of the black bean of the present invention. The fermented product of the present invention can be used as a food composition, a pharmaceutical composition or a nutritional supplement.

The compositions of the invention may be administered to an individual by any suitable route, such as oral administration. Suitable formulations of the compositions of the present invention include, but are not limited to, lozenges, blister packs, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs. If desired, the composition can be sterilized or mixed with any pharmaceutically acceptable carrier, such as a stabilizer, a wetting agent, and the like.

The compositions of the present invention may be admixed by any conventional method with conventional pharmaceutically acceptable excipients. Thus, the compositions of the invention to be used orally may comprise, for example, one or more color formers, sweeteners, flavor enhancers, and/or preservatives.

The compositions of the present invention can be used in combination with one or more other drugs currently in use to treat diseases associated with excessive production of nitric oxide.

use

The present invention provides a use of a fermented product of Coprinus comatus according to the present invention for preventing and/or treating a disease associated with excessive production of nitric oxide.

In a preferred embodiment of the invention, diseases associated with excessive production of nitric oxide, such as inflammation, septic shock, hemorrhagic shock, traumatic shock, anaphylactic shock, toxic shock syndrome, hypovolemic shock, Absence of blood, brain, cytokines, cytokine overexpression, ulceration, inflammatory bowel disease (such as ulcerative colitis or Crohn's disease), diabetes, metabolic syndrome, diabetic nephropathy, diabetes Neuropathy and diabetic ulcer, arthritis, rheumatoid arthritis, asthma, Alzheimer's disease, Parkinson's disease, multiple sclerosis, hardening, allograft rejection, encephalomyelitis, meningitis, pancreatitis , peritonitis, vasculitis, lymphocytic choriomeningitis, glomerulonephritis, uveitis, ileitis, inflammation (such as liver inflammation, kidney inflammation, etc.), burns, infections (including bacteria, viruses, fungi and parasitic Sexual infections), influenza, respiratory tract inflammation associated with upper respiratory tract infections, blood dialysis, chronic fatigue syndrome, stroke, cancer (eg breast cancer, melanoma, Body tumor, leukemia, myeloma, lymphoma, etc.), cardiopulmonary bypass, hematopoietic/reperfusion injury, thrombotic or hemorrhagic stroke, subarachnoid hemorrhage, cerebral vasospasm, myocardial infarction, shock, gastritis, ulcerative colon Inflammation, adult respiratory distress syndrome, cachexia, myocarditis, autoimmune disorder, eczema, psoriasis, heart failure, heart disease, arteriosclerosis, myocardial infarction, acute coronary syndrome, restenosis after vascular surgery, hypertension and vasculitis , dermatitis, atopic dermatitis, acne, urticaria, systemic lupus erythematosus, AIDS, AIDS dementia, chronic neurodegenerative diseases, chronic pain, abnormal erection, cystic fibrosis, amyotrophic lateral sclerosis, ALS and muscle Atrophy, dystonia, myasthenia gravis (MG), schizophrenia, depression, premenstrual syndrome, anxiety, addiction (eg medication, alcohol and nicotine dependence), morphine-induced tolerance and withdrawal symptoms, Bipolar disorder, post-traumatic stress disorder, anorexia nervosa, ADHD and autism spectrum disorders, migraine, inflammatory pain and neuralgia, Hunting Disease, epilepsy, neurodegenerative disorders, gastrointestinal motility disorders, obesity, overfeeding, solid tumors (such as neuroblastoma), malaria, blood cancer, spinal cord fibrosis, lung injury, transplantation versus host disease, head injury, CNS Trauma, hepatitis, renal failure, pancreatitis, liver disease (such as chronic hepatitis C), drug-induced lung injury (such as Paraquat), macular degeneration, diabetic retinopathy, glaucoma, retinitis, hearing loss, tinnitus, asthma, chronic Obstructive pulmonary disease, acute respiratory distress syndrome, cystic fibrosis, osteoporosis, osteoarthritis, other degenerative conditions of hard bone and cartilage, acute or chronic graft failure or rejection and transplant-versus-host disease, and cancer therapy (including Radiation therapy or chemotherapy) related mucositis, severe burns, insufficient platelet aggregation, imbalance of homeostatic processes, tissue damage, etc.

Preferably, the disease associated with excessive production of nitric oxide according to the present invention is inflamed.

Those of ordinary skill in the art to which the present invention pertains can select the appropriate route of administration and dosage for treatment without difficulty. According to the invention, the preferred route is oral administration. The dosage will depend on the nature of the disease and its condition, the age and health of the individual, the route of administration and pretreatment. Those of ordinary skill in the art to which the invention pertains will appreciate that the dosage will vary with the age, weight, health, and other relevant factors of the individual.

The following non-limiting examples are intended to assist those of ordinary skill in the art to practice the invention. The examples are not to be construed as limiting the invention in any way. Modifications and variations of the embodiments discussed herein may be made without departing from the spirit and scope of the invention, and still fall within the scope of the invention.

Instance

Example 1. Black bean of Coprinus comatus fermentation

Take 155 grams of Hengchun black peas, 20 grams of brown sugar, 20 grams of wheat and 5 grams of black sesame. After mixing evenly, add water. The ratio of the medium ingredients to the water is 150 w/v%, and then at 121 °C. Incubate for at least 30 minutes to sterilize into medium.

Then, solid inoculation was carried out, and Coprinus comatus BCRC 36658 was inoculated into the above-mentioned medium, and the ratio of the medium to the culture medium of Coprinus comatus was 10% (v/w), and the concentration of the strain was about 10 g of biomass g/L. The mixture was prepared and stirred well, and cultured at 25 ° C for 30 days.

The fermented material is dried after the completion of the fermentation, and the step is mainly carried out at a temperature of about 50 ° C and a pressure of about 0.5 kg / cm ² to obtain a fermented product of the black bean.

Example 2: High-pressure liquid chromatography analysis of the fermentation of the black bean Map

1 g of the fermented product obtained in Example 1 was weighed and added to 10 ml of methanol solution, and ultrasonically extracted for 1 hour, centrifuged, filtered, and subjected to high pressure liquid chromatography analysis, and the column used was Inersil ODS. -2,5 μm, (GL Sciences, ToKyo, Japan), the length of the column is 250 mm, the inner diameter is 4.6 mm, and the chromatograph used is Hitachi PDA L2455, the extract (A) is 0.1% phosphoric acid solution, the extract (B) is 100% acetonitrile, the extraction procedure is 0 minutes to 60 minutes: 90% extract (A)/10% extract (B), 60 minutes to 71 minutes: 0% of the extract (A) / 100% of the extract (B) and 71 minutes to 80 minutes: 90% of the extract (A) / 10% of the extract (B ).

The results are shown in Figure 1, wherein the residence time is about 7.3, about 7.8, about 9.2, about 13.0, about 13.4, about 16.0, about 21.0, about 21.6, about 25.0, about 25.4, about 37.0, about 39.1, about 40.0, There are absorption peaks of about 43.2, about 53.2, about 56.8, about 60.9, and about 67.2 minutes.

Further, the unfermented medium was analyzed according to the above conditions, and the results are shown in Fig. 2, wherein the residence time was about 7.3, about 7.8, about 9.2, about 11.8, about 13.0, about 13.4, about 16.0, about 21.0, about 21.6. There are absorption peaks of about 25.0, about 25.4, about 37.0, about 39.1, about 40.0, about 67.2 minutes.

Comparing the black beans before and after fermentation, the absorption peak content of the retention time of about 21.0, about 21.6, about 25.0, and about 25.4 minutes was increased, and the new residence time was about 43.2, about 53.2, about 56.8, about 60.9. The absorption peak component of the minute shows that the fermentation does change the composition of the medium.

Example 3: Analysis of amino acid content of fermented food of Coprinus comatus

The fermented product obtained in Example 1 was analyzed for its total branched amino acid (leucine, isoleucine, valine) content, and the content was determined by SGS, and the test result was 0.83 g/68 mL. The standard total branched chain amino acid to be certified against commercially available health foods is greater than or equal to 0.32 g/68 mL, which is 1.6 times higher.

Example 4, black bean, chicken leg mushroom fermentation broth

168 g of the fermented product obtained in Example 1 was boiled with 840 g of water (cold water was charged, and the water was rolled for 30 minutes), then cooled, and then the medium was filtered and removed, and the precipitate was refrigerated for 4 hours, and then filtered again, and water was added. The weight was added back to 750 grams and sterilized at 121 °C for 25 minutes.

Example 5: Analysis of the inhibition of nitric oxide production by the fermentation of Coprinus comatus

Inhibition of nitric oxide free radical (NO) production is one of the main methods for evaluating anti-inflammatory activity. The principle is to use lipopolysaccharide (LPS) to stimulate mouse macrophage RAW264.7, and to induce inflammatory reaction by inducible one. Inducible nitric oxide synthase (iNOS), iNOS produces a large amount of free radicals. The extract is used to remove the NO radicals to assess whether the components have anti-inflammatory activity.

The nitrite concentration was determined by using the Griess reagent [0.1% N-(1-naphthyl) ethylenediamine) and 1% Sulfanilamide in 5% phosphoric acid] and The nitrate reaction, the color change produced, the absorbance at 548 nm, and the absorbance-concentration standard curve, the content of NO ₂ in the tissue culture solution can be obtained. Since the half-life of NO is very short, it is quickly oxidized to nitrite and further oxidized to nitrate. Therefore, the amount of nitrite can be measured in a short time by using the Griess reagent to indirectly indicate the release amount of NO (Paul et Al., 1994, Anal Biochem. 1982 Oct;126(1): 131-8).

Weigh 1 g of the fermented material obtained in Example 1 and add 10 mL of 95% ethanol or H ₂ O, mix well, and then bathe at 70 ° C for 1 hour, then centrifuge at 10,000 rpm, 4 ° C for 10 minutes, and take the supernatant. . It is then dried at 45 ° C until it is completely dry, taking about 48 hours. And weighed dry weight.

Then, using ultrasonic vibration, the dry matter was reconstituted with 10% dimethyl sulfoxide (DMSO) to a concentration of 10 mg/mL. Each sample was diluted with water, and each sample was subjected to three concentrations of 10, 5, and 1 mg/mL, and filtered in a sterile operation table with a 0.45 μm filter to test its IC ₅₀ , and the IC ₅₀ was an inflammatory reaction. The concentration required for the sample at a inhibition rate of 50%, that is, the lower the IC ₅₀ value, the better.

The test was carried out using the RAW264.7 cell line, and 2 × 10 ⁵ cells/100 μL were seeded into a 96-well plate so that the number of cells per well was 2 × 10 ⁵ . And incubated at 37 ° C for 24 hours. The culture solution was removed, and the culture solution and the sample were added as described above, and the total test volume was 100 μL. It was further cultured at 37 ° C for 24 hours.

For the detection of the content of nitric oxide, first prepare the standard and background value for use, standard sample, background value and sample each take 50 μL to the new 96-well plate, add the reagents, and use enzyme immunoassay (Enzyme- Linked immunoassay (abbreviated as ELISA) reads the OD ₅₄₀ value, wherein the standard curve prepared by the standard is shown in Figure 3, with a slope of 0.0063, an intercept of 0.0719, and a linearity of 0.9993.

The results are shown in Table 1 below.

The medium before fermentation was used as a control group, and the preparation method was as described above, and the results are shown in Table 2.

As is apparent from the results of Tables 1 and 2, the black bean fermented product according to the present invention has an effect of suppressing the production of nitric oxide, and can prevent and/or treat a disease associated with nitric oxide production, particularly inflammation. On the other hand, the effect of unfermented black beans is poor.

Fig. 1 shows a high pressure liquid chromatogram of a black bean fermented product according to the present invention.

Figure 2 shows a high pressure liquid chromatogram of black bean medium before fermentation.

Figure 3 shows a standard curve of the standard for testing the amount of nitric oxide produced.

(no component symbol description)

Claims (11)

A method for producing a fermented product of Coprinus sp. of Glycine max (L.), which comprises fermenting a sterilized medium of black beans with Coprinus comatus, wherein the Coprinus comatus is obtained from the food industry of the consortium. Institute, and the registration number is BCRC36658. The method of claim 1, wherein the medium further comprises a flavoring agent. The method according to claim 2, wherein the flavoring agent is selected from the group consisting of brown sugar, wheat, black sesame, buckwheat, oat, hazelnut, brown rice, buckwheat, millet, sorghum, pumpkin, walnut, and corn. The method according to claim 2, wherein the medium comprises: about 80 to 250 parts by weight of black beans; about 10 to 30 parts by weight of brown sugar; about 10 to 30 parts by weight of wheat; and about 5 to 25 parts by weight of black sesame. The method of claim 1, wherein the fermentation is a solid state fermentation. According to the method of claim 1, it further comprises a step of obtaining a fermentation broth. The method of claim 6, wherein the step of obtaining the fermentation broth comprises extracting the fermented medium with hot water or a solution containing methanol, ethanol, propylene glycol, isopropanol or ethyl acetate, and taking a water-soluble portion thereof. A black bean sauerkraut fermented product obtained by the method according to any one of claims 1 to 7. The fermented product according to claim 8, wherein the fermented product is analyzed by High Pressure Liquid Chromatography (HPLC) using a chromatography column of Inersil ODS-2, 5 μm, (GL Sciences, ToKyo, Japan), the length of the column is 250mm, the inner diameter is 4.6mm, the chromatograph used is Hitachi® PDA L2455, the extract (A) is 0.1% phosphoric acid solution, and the extract (B) is 100. % acetonitrile, the extraction procedure is from 0 minutes to 60 minutes: 90% of the extract (A) / 10% of the extract (B), 60 minutes to 71 minutes: 0% of the extract (A) / 100% Liquid (B) and 71 minutes to 80 minutes: 90% of the extract (A)/10% of the extract (B), wherein the residence time is about 7.3, about 7.8, about 9.2, about 13.0, about 13.4, about 16.0. There are absorption peaks of about 21.0, about 21.6, about 25.0, about 25.4, about 37.0, about 39.1, about 40.0, about 43.2, about 53.2, about 56.8, about 60.9, and about 67.2 minutes. A nutritional supplement comprising the black bean squash fermented product according to claim 8. A use according to claim 8 for the fermented product of the black bean of Coprinus comatus, which is used for the preparation of a medicament for preventing and/or treating inflammation.