KR20010087699A - The cultivation method of Cordyceps sinensis et al. having a high concentration of gamma-aminobutyric acid - Google Patents

Abstract

PURPOSE: Provided is a cultivation method of vegetable worms with high gamma-aminobutyric acid content which treats by keto acid in germinating grains or beans and treats in an anaerobic way after germination so that it produces vegetable worms with high gamma-aminobutyric acid content. CONSTITUTION: The cultivation method of the vegetable worms with high gamma-aminobutyric acid content comprises the steps of: (i) germinating grains or beans as a culture source to obtain a germinated body; (ii) vaccinating culture liquid of the vegetable worms to a culture wherein the germinated body is mixed and sterilized; (iii) culturing at 25deg.C and 70(RH) in relative humidity for 2-3 weeks; (iv) inducing to germinate when spawn is rich; and then (v) generating a fruit body to finish the culture.

Description

Cultivation method of Cordyceps sinensis with high content of gamma-aminobutyric acid {The cultivation method of Cordyceps sinensis et al. having a high concentration of gamma-aminobutyric acid}

The present invention relates to a method of cultivating Cordyceps sinensis with a high content of gamma (γ) -aminobutyric acid, and more specifically, as a medium source, grains or beans such as rice, glutinous rice, brown rice, barley, wheat (rye), and soybeans. Cultivating Cordyceps sinensis using only germinated bodies obtained by germination or using edible insects such as germinated bodies and pupa, or a mixture of cereals or beans such as rice, glutinous rice, brown rice, barley, wheat (rye), and beans It relates to a cultivation method of Cordyceps sinensis with high content of γ-aminobutyric acid.

The word Cordyceps sinensis originated in China and is meant to live in the insect's body in the winter, absorbing nutrients, killing the insects, and making mushrooms in the dead insect's body in the summer. Cordyceps sinensis, along with ginseng and antler, has long been known as one of China's three rare medicines.

Cordyceps sinensis contains about 7 Chungzoic acid, Cordyceps sinensis helps qi and lubricates the lungs, stops bleeding, removes sputum, and is known to be effective in treating anemia and old age weakness. It has been favored as a leap of Gangjeong River. Herbal life in the Chinese record is said to be used to protect the lungs, benefit the kidneys, sputum sputum or treat tuberculosis cough.In the Medicinal Dictionary, it is sweet and mild, helps kidney function, strengthens the lungs, strengthens tonic, energizes, calms, It is recorded to be effective for anemia and the like. Recent studies have shown excellent efficacy in anti-cancer, immune enhancement, anti-fatigue, and anti-aging.

In addition, Cordyceps sinensis is known to be rich in polysaccharides and vitamin B12. Polysaccharides play an important role in defending the body from diseases. They increase immunity, protect the heart and liver, inhibit cancer, and are involved in anti-aging. It is known that it is good for pernicious anemia and is involved in brain activity, strengthens mental concentration and memory, and protects nerve activity or liver.

By pharmacological action of Cordyceps sinensis, the antibacterial action of the bacterium Mycobacterium tuberculosis, pneumococcal bacillus, low hemorrhagic sepsis bacillus, gypsum pylori, anthrax, nasal bacillus; Central nervous system actions such as gangjeong tonic, efficient sleep induction, suppression of central nervous system excitement, anticonvulsant action, body temperature lowering action; Respiratory system effects such as cold, chronic cough, asthma, tuberculosis effect, bronchial dilation, sputum removal, respiratory stability; Cardiovascular effects such as treatment of pulmonary dysfunction, slowing heart rate, lowering blood pressure, lowering cholesterol, endurance of oxygen deficiency, antiplatelet collection; Immunity enhancing effects such as natural healing and inflammation inhibition; Anticancer activity; Anti-exposure action; Chronic antifatigue; Drug addiction detoxification; It is known to be effective in treating jaundice.

Until now, most of the Cordyceps sinensis has been collected in a natural state around the Tibet Mountains, which are alpine highlands, but since the fruiting bodies are small and difficult to collect, they are difficult to popularize because they are expensive.

In recent years, artificial cultivation methods of various Cordyceps sinensis have been developed and commercially available. However, even though artificial cultivation is limited to insect larvae, larvae, and adults as host parasites, there is a limit to popularization because they are sold at high prices. There is no advantage, but also due to the use of insects as a host source, but also abomination is a fact that there is a limit to popularization despite the useful effect.

According to the contents of the domestic patent of the research on the cultivation of Cordyceps sinensis, the live silkworms were inoculated with the strains of Cordyceps sinensis as parasitic bacteria and parasitic on these cells, and when the silkworms become pupa after the cocoon during the metamorphosis process, The strain kills the pupa and predominates as an endogenous nucleus in the cell. At this time, the cocoon is removed (or melted) and fertilized (cultivation method of a sheep farming company under the Nassau Insect Research Institute), after the chrysalis are soaked in red ginseng gourd (shell), sterilized and inoculated, and grains are added to the chrysalis. It is known that it is possible to cultivate in a mixed medium, to place insects such as silkworms and pupa on top of grains, to sterilize them, and to grow them using a slug or centipede as a host source.

On the other hand, cultivation of Cordyceps sinensis can be cultivated only by pupa, grain or legume as a medium source, but it is known that the accumulation of certain useful substances is more accumulated when using additives. For example, in cultivating Cordyceps sinensis, a polysaccharide called codycepin, which is a useful ingredient, accumulates due to the red ginseng gourd (peel) that has been dried and then boiled and cultivated in the red ginseng gourd (peel). have.

However, little research has been done on methods using additives in growing Cordyceps sinensis, and as the present invention, chitosan is added to the germ bodies of cereals and legumes, and their germination as a medium source other than pupa. There is no mention of cultivation of Cordyceps sinensis which increases the content of γ-aminobutyric acid by anaerobic treatment after germination.

Recently, γ-aminobutyric acid,life GABA) is gaining attention as a source of new functional foods as well as supplements for health foods, and γ-aminobutyric acid is accumulated in large amounts during germination of rice embryos to be used as a food material or to produce γ-aminobutyrate (GABA). Method is known, skim food material in which a large amount of γ-aminobutyrate (GABA) is accumulated in the germination process is used, and the rough development process of GABA is as follows.

During the study of the biologically active components of rice embryos, a very effective ingredient in the treatment of rice was found in rice embryos and named as orizinine (vitamin B1). Pectin was extracted to find pectase (enzyme), and colorless crystals of bioactive activity were separated from rice bran and rice embryo oil and named as oranol. The pharmacological action of oranol was hepatic brain function control, UV absorption, antioxidant Actions, peripheral blood vessel dilation, and the like have been confirmed, and it is recognized as a therapeutic agent for symptoms of autonomic ataxia and cosmetic raw materials. From this point, GABA became known.

Also, Konoda of Japan's Agricultural Research and Development Bureau of Japan, and Nakagawa of Duck Liberation Co., Ltd. found that rice embryos are rich in valuable nutrients and bioactive components. Developed as a source of functional foods to increase physiological activity such as blood pressure drop by accumulating γ-aminobutyric acid (GABA) in large amounts during germination of rice, and adding water to rice bran and embryo containing rice bran It was found that the decarboxylase automatically initiated the reaction and rapidly produced large amounts of GABA.

Brown rice is divided into rind, seedling, stratum corneum and stratum corneum in the skin, seedling, stratum corneum (between shell and starch layer), endosperm starch layer, embryo and its tissue structure, and the thickness of the stratum corneum is 45μm. Embryos are around 3, and the embryos mixed in the river are separated by a sieve and separated by screening method using the difference in specific gravity. However, in the actual treatment process, the tissues are damaged, the lipids in the embryo are hydrolyzed by the action of endogenous enzymes, and the triglycerides are reduced, and the free fatty acid content is 10-15. Becomes Factors that deteriorate (badly) the taste and taste of embryos are known n-valaldehyde, n-gaproaldehyde, etc. produced by the automatic oxidation of free fatty acids and fatty acids. It is necessary to remove the lipids.

It acts on rice embryos, which are said to be fully nutritional foods and are used mostly as food ingredients, to solve the problem of improving food taste and residual pesticides, and by accumulating GABA in large amounts in the germination process, it enhances physiological activity such as lowering blood pressure. It was developed as a new food material. Compared to other food ingredients, the GABA content of the pre-processed rice embryos was 25mg-50mg / 100g compared to other food ingredients, but the GABA content of rice embryos accumulated in large amounts during the germination of GABA increased to 350-400mg / 100g. .

The physiological activity of GABA is reported to have lowering blood pressure, activating kidney function, improving liver function, preventing obesity, promoting alcohol metabolism, and removing odor. GABA as a dietary supplement is vitamin B. It can be used as a group, minerals, plant fiber, etc., and it is possible to use GABA accumulated in a large amount in the germination process of rice germ in food.

Modern people are gradually increasing their ability to prevent geriatric diseases in response to the reality of aging and the aging of adult diseases due to chronic exercise insufficiency, extreme stress, difficulty in eating, obesity caused by predation, and symptoms of adult diseases even in the low age group. More than 3 billion people around the world, such as rice and barley, grains and beans, are the food of life.

GABA obtained from brown rice germination extract is a functional food material having a physiologically active function for preventing adult diseases, and is considered to be used as a nutritional supplement containing vitamins and minerals. For example, there is a method for use as a beverage such as embryo tea, a method for mixing powdered foods with powder, or a form of granule tablets, for use as a separator.

However, techniques for growing Cordyceps sinensis using GABA have not been studied or developed.

On the other hand, chitin is contained in shells of shellfish such as shrimps and crabs, exoskeletons of arthropods such as beetles' shells, organs of mollusks, cell walls of fungi such as fungi, yeasts and mushrooms. Chitin is a straight-chain monoamino polysaccharide with (1-4) -bound N-acetyl-β-D-glucosamine, and chitosan is a derivative in which chitin is N-deacetylated.

Since chitin forms the shell of a living organism, in 1823, the French scientist Ohzizii named it chitin, which means envelope in Greek. Chitin has a digestibility of 24 to 38 in rabbits and 75 to 98 in chickens. While visible It is not decomposed by human digestive enzymes and has some of the same properties as dietary fiber, and has been studied for its antitumor, hemostatic, immunostimulating, cholesterol-lowering, and bifidus growth.

Chitin activates the enzymes involved in blood coagulation to promote blood coagulation, hemostasis, and immune enhancing effects. Immune-related macrophages recognize the invasion of pathogens in the body and play a major role in the production of antibodies to the pathogens, and chitin acts to enhance the activity of these macrophages.

Just as dietary fiber has an anticancer effect, chitin also has an anticancer effect and is called an animal dietary fiber.

However, no chitin or chitosan induced by N-deacetylation of chitin was used for cultivation in Cordyceps sinensis.

Anaerobic treatment was performed by Yoon et al. (Korean Society of CroScience, 1998, Vol. 43 (1), 19-22), where the anaerobic treatment of the ground tissues of barley seedlings grown for one week after germination resulted in GABA. The experiment was applied according to the literature that the content was increased in large quantities.

Accordingly, it is an object of the present invention to provide a method for cultivating Cordyceps sinensis with high content of γ-aminobutyric acid, which can be used as a dietary supplement or a functional food, and can be used as a raw material thereof.

Another object of the present invention to provide a dietary supplement or functional food using Cordyceps sinensis or Cordyceps sinensis culture grown by the method of the above object.

In order to achieve the above object as well as another object that can be easily expressed in the present invention, only the germinated body obtained by germinating grains or beans such as rice, glutinous rice, brown rice, barley, wheat (rye), soybean as a medium source Cultivated cordyceps and cordyceps cultures using edible insects such as germinated bodies and pupa, or a mixture of cereals or beans such as rice, glutinous rice, brown rice, barley, wheat (rye), and beans By preparing a dietary supplement or a functional food using was able to obtain a dietary supplement or a functional food that can be obtained not only the effect of the useful components of Cordyceps sinensis, but also the effect of γ-aminobutyric acid.

That is, after the cultivation of Cordyceps sinensis, Cordyceps mycelium, fruiting body or medium, Cordyceps sinensis culture, freeze-dried or hot-air dried and milled to crush the soybeans, confectionery, confectionery, confectionery, tea, retort foods, alcoholic beverages, noodles and powdered soup thereof By blending into processed foods such as dairy products, dairy products, beverages, extracts, gums, and candy products, the effect of γ-aminobutyric acid is expected while the effect of cordyceps is expected without compromising the original taste of each product. It is expected to have a useful effect of obtaining functional foods with no difference in taste, aroma and color.

1 is a schematic process diagram of the method of the present invention,

2 is a process diagram schematically illustrating an analysis process of γ-aminobutyric acid,

3 is a standard chromatogram of amino acids of Cordyceps sinensis grown according to the present invention,

Figure 4 is a chromatogram analysis of the entire Cordyceps sinensis culture grown in accordance with the present invention using a mixture of pupa and beans as a medium,

Figure 5 is a chromatogram of the medium after using a mixture of rice and beans as a medium and cultured Cordyceps sinensis according to the present invention.

The present invention is described in more detail as follows.

Cultivation method of Cordyceps sinensis with high content of γ-aminobutyric acid according to the present invention uses only germinated bodies obtained by germinating grains or legumes such as rice, glutinous rice, brown rice, barley, wheat (rye), soybean as a medium source, or Edible insects such as germinated bodies and chrysalis or mixed grains or legumes such as rice, glutinous rice, brown rice, barley, wheat (rye) and soybeans, but germinated with chitosan and anaerobic processing of grains or legumes as anaerobic treatment Exposure to N ₂ gas or CO ₂ gas in a 25 ° C. wet incubator for 12-20 hours promotes the deactivation of the decarboxylase and, if necessary, to dry the pupa or its flour or common grains, beans or its pretreated soybeans. After mixing the ground, sterilized and inoculated liquid culture solution of Cordyceps sinensis into the cooled medium, incubated for 2 to 3 weeks at a temperature of 25 ℃ and relative humidity 70 (RH) When the growth is completed, and then the growth is complete, freeze-dried for 40 to 50 hours in a freeze dryer or dried for 3 to 4 days at a temperature of about 60 ℃ with a hot air dryer and powdered with a grinding machine to supplement the health supplement or functional Manufactured with food.

GABA (Gaba) content in the Cordyceps sinensis culture according to the present invention was significantly higher than that of a conventional silkworm and pupa only, GABA (Gaba) is originally present in a small amount in the flora and fauna, but chitosan during germination of cereals and legumes, etc. After the addition or germination, it was confirmed that GABA (GABA) content is increased by the analytic treatment of decarboxylase by anaerobic treatment, thereby completing the present invention. In the present invention, it was also found at which point in time the germination of cereals or legumes accumulates the most GABA.

The cultivation of Cordyceps sinensis used in the present invention used a method commonly used in the technical field to which the present invention belongs, and using chitosan to add grains and beans in germination, and after anaerobic treatment after germination, mixed a certain amount of pupa As well as the fruiting body and mycelium of Cordyceps sinensis obtained using the medium source, the medium component is included.

The medium source used for the cultivation of Cordyceps sinensis is the pupal: germinating grains (brown rice, barley, rye and soybean) based on the total weight on dry basis: chitosan = 0 to 1: 1 to 2: 0.05 to 0.2 (V / V and W) was studied using a source of medium, and in the mixed medium using germination of grains rather than pupa alone, although the incubation period was a little longer, there was no significant problem in the production and growth of fruiting bodies. The enhanced their culture and processing them can be used as a source of health supplements.

Cordyceps sinensis can be obtained by a method commonly used in the art to which the present invention belongs.

However, in the present invention, it was most effective to cultivate Cordyceps sinensis by mixing the pupa with grains and beans through a pretreatment process by the following method. Cordyceps sinensis uses only edible insects such as chrysalis, or grains and legumes such as rice, glutinous rice, brown rice, barley and wheat, or mixtures of germinated bodies obtained by germinating grains and legumes. Cordyceps mycelium and fruiting bodies cultured as a medium can be used as raw materials for the preparation of processed foods. Chitosan was treated at the time of grain and legumes, and anaerobic treatment at the time of footing. As an anaerobic treatment, N ₂ gas or CO ₂ gas on the incubator was used.

In other words, N ₂ gas in a wet incubator with a temperature of 25 ° C. as an anaerobic treatment of grains and soybeans germinating with grains and beans and chitosan and germinated grains such as brown rice, waxy brown rice, barley and wheat (rye). B) Exposure to CO ₂ gas for 12 to 20 hours to promote the activity of decarboxylase, mix pupa powder with it, sterilize it, sterilize and cool it, and then inoculate the Cordyceps liquor medium into the inoculation medium as an inoculum. Then, incubate for 2 to 3 weeks at a temperature of 25 ° C. and a relative humidity of 70 (RH) and induce fermentation when hyphae spread. The germination process was carried out by maintaining the relative humidity of 95 (RH) at the temperature of 18-20 ℃ and the illuminance of the light source at 400 ~ 1000 Lux, and to control the concentration of CO ₂ gas in the air. As much as possible, entry and exit were inevitable, and inevitably, CO ₂ generators were installed to control the concentration of O ₂ / CO ₂ to induce elongation of fruiting bodies. This is generally due to the fact that a moderate concentration of O ₂ / CO ₂ maintains good mushroom quality rather than the presence of excessive O ₂ in mushroom cultivation and that the excess O ₂ conditions, ie CO ₂ concentration of 1,000 μl / l. In this case, the elongation of the fruiting body stops, the end of the fruiting body is branched, a large amount of spores are generated, and in a small amount of O ₂ condition, that is, when the concentration of CO ₂ is 9,000 µl / l, the end of the fruiting body is not branched. It was applied with reference to a research paper (Yamanaka et al., Mycoscience 39 : 43-48, 1998) in a strain of Isaria japonoca which was good and had less spores. Then, the lid closed during the cultivation was opened or the medium source was taken out, and the basket was placed with a wet sponge, and the culture medium was placed upside down to maintain the condition of the feet and to maintain a relative humidity of about 95 (RH) during the generation of the fruiting body. When the growth is complete, the culture is completed and harvested, and then lyophilized in a freeze dryer for 40 to 50 hours or dried in a hot air dryer at 60 ° C. for 3 to 4 days until moisture disappears and powdered with a grinding machine. It was used as a raw material in the manufacture of foodstuffs.

On the other hand, by the fungus strain is no additives, as well as coordination sepseu sinen sheath (Cordyceps sinensis), coordination sepseu Millar other less (Cordyceps militaris), coordination sepseu Scarab coke (Cordyceps scarabaecola), coordination sepseu press Tansu (Cordyceps nutans), coordination sepseu seupeko Seppala (Cordyceps sphecocephala), Cody sepseu tree Sentry (Cordyceps tricentri), Passage to my access tenwi Fez (Paecilomyceps tenuipes), payito Cody sepseu non chyuki Austrian Fora (Phytocordyceps ninchukiospora) all kinds of well-known, such as the use of Cordyceps sinensis fungus It may be.

The present invention is to improve the content of γ-aminobutyric acid (GABA) in the Cordyceps sinensis and its culture, the polysaccharide known as 'Codcepin' so far, as well as those components not known so far, than the conventional media source As the GABA content is increased in the medium source of the invention, it is possible to develop a new food material with higher added value and enhanced bioactive function.

As a food material rich in γ-aminobutyric acid, it is possible to develop products such as powders and extracts of GABA's new functional material contained in the culture of Cordyceps sinensis. Tea, retort foods, alcoholic beverages, noodles, dairy products, beverages, extracts, gums, candy, processed foods such as can be used as an additive source.

The following examples illustrate the invention in more detail, but do not limit the scope of the invention.

In the following examples, unless otherwise specified, the strain used was Paecilomyceps japonica (aka Snow Cordyceps sinensis) and Cordyceps sinensis was cultivated by the following method.

As a medium source, germinate using pupa, rice, brown rice, barley, wheat (rye), soybean, etc., and dilute the mixed solution in which 5 chitosan is dissolved in 5 lactic acid so that chitosan is well dissolved in water. As a control of chitosan treatment, 5-lactic acid was diluted 500-fold and inoculated into liquid culture after sterilization. After culturing for 4 to 5 days while shaking in a Erlenmeyer flask with medium as a preculture, expand it to about 10 L in a 12-18 L container, or finely grind mycelia with a homogenizer and inoculate it into an expansion incubator. Was incubated for 4 to 6 days at a culture temperature of 23 to 26 ° C. while filtering and aeration injection. The inoculation was a sterile spray spray, and the inoculation solution per bottle was inoculated by about 8 ml. The amount of medium used in this example was not limited to about 20 to 25g as a solid, but cultivated Cordyceps sinensis according to the process shown in FIG. 1, but the pupa: germinating grains (brown rice, barley, rye, etc.) and legumes: The ratio of chitosan = 0-1: 1-2: 0.05-0.2 (V / V, W) was adjusted, and the water content of the medium was adjusted so that the average water content of the medium after the sterilization was about 65. In the case of sterilization by treating the ratio of solids and water in a ratio of 1: 0.8 to 1.2 without the soaking of grains according to the conventional method, the growth of mushroom fruit bodies is so slow that the growth of mushroom fruit bodies is very slow. As a control of the water, at 4 ℃ put the water in the process of soaking, and treatment as in the same manner as the experimental treatment by the process of the present invention used by using the same process by maintaining the appropriate pores to improve the overall growth and increase the media source per pathogen There was an advantage.

After the inoculation of Paecilomyceps japonica , the culture process was maintained until the incubation of the hyphae was completed at a temperature of 25 ° C., and the germination process was performed at a temperature of 18 to 20 ° C. and a light intensity of 400 to 1000 lux (Lux). ) were line to maintain the relative humidity on the order of a 95 (RH), a contact capable of opening and closing were refrain to control the concentration of the air in the CO ₂ gas when the feet, grown in fact absolutely necessary to install a CO ₂ generator O ₂ The concentration of / CO ₂ was induced to elongate the fruiting body. Then, the lid closed during the cultivation was opened or the medium source was taken out, and a wet sponge was placed on the basket and the culture medium was placed upside down to maintain the condition of the feet and to maintain relative humidity at 95 (RH) when the fruiting body was generated. When the growth is completed, the culture is completed and harvested, freeze-dried for 40 to 50 hours in a freeze dryer or dried at 60 ° C. for 3 to 4 days in a hot air dryer until it is dried and powdered with a crusher. Treated as in 2 and used for analysis.

Examples 2 to 9 may be individual or sequential, but the final application of the method is to combine the same contents as in Example 9 into one unit of experiment. The samples obtained in Examples 1 to 9 were analyzed in the same manner as in Example 10, which were analyzed in Table 1 together with some schematics [x; x = n recovery] and these together. Table 1 calculates and describes the GABA (GABA) content based on the untreated bunde apparatus of Example 2.

Example 1

PS, PS + peptone, PS + peptone + wort (seed oil extract) were prepared as a solution, and cultured at 25 ° C. for 7 days. After sterilization of the culture medium, the solution was divided into two groups, control and experiment. Changes in inoculation were compared for those not inoculated with Cordyceps sinensis. After completion of the culture was extracted with boiling water and centrifuged at 1000g for 20 minutes at room temperature to remove the mycelium and the residue, and compared to each medium before and after the culture under the same conditions.

Example 2

The overall criterion for GABA content analysis was based on cultures obtained using only conventional chrysalis.

As a control, the frozen pupa medium was used as it was, and as a control, 5 chitosan was diluted to 1000X, 500X, and 250X in the frozen pupa medium source, and the control was treated with 500X of 5 lactic acid. After treatment with high pressure to infiltrate the treatment agent, the filtrate was discarded to adjust the humidity of the medium source and then sterilized by inoculation.

Example 3

As a control, 5 chitosan was diluted to 1000X, 500X, 250X in a rice medium source and a test medium in a rice medium source, and treated with 5X lactic acid 500X as a control and maintained for 5 days. As a control of the soaked water was maintained at 4 ℃ for 5 days, the filtrate was discarded by adjusting the humidity of the medium source and then inoculated by sterilization.

Example 4

The pupa powder for the following brown rice medium source was divided into the addition group (20) and the no addition group (0), and 5 chitosan was diluted to 1000X, 500X, 250X at the time of germination of brown rice medium source and the experimental group as a control and 5 as a control. Treatment with lactic acid 500X was maintained for 1, 2, 4, 6 days. As a control of soaking, water was maintained at 4 ° C. for 5 days, and the filtrate was discarded to adjust the humidity of the medium source and then sterilized and inoculated.

Example 5

The pupa powder for the following barley medium source was divided into the addition group (20) and the no addition group (0), and the 5 chitosan was diluted to 1000X, 500X, 250X at the time of germination of barley medium and the experimental group as a control and 5 as a control. Treatment with lactic acid 500X was maintained for 1, 2 or 3 days. As a control of soaking, water was kept at 4 ° C. for 3 days, and the filtrate was discarded to adjust the humidity of the medium source and then sterilized and inoculated.

Example 6

The pupa powder for the following soybean culture source was divided into the addition group (20) and the no addition group (0), and 5 chitosan was diluted to 1000X, 500X, 250X at the time of germination of soybean with the soybean source and the experimental group as a control. Treated with lactic acid 500X and maintained for 1, 2, 3 days or pretreated as above, dried and used after grinding. As a control of soaking, water was kept at 4 ° C. for 3 days, and the filtrate was discarded to adjust the humidity of the medium source and then sterilized and inoculated.

Example 7

Chrysalis powder for the mixed medium source of brown rice and soybean was divided into addition group (20) and no addition group (0), and 5 chitosan in the mixed germination of brown rice and soybean as control medium and experimental group Was diluted to 1000X, 500X, 250X and treated with 500X 5 lactic acid as a control to maintain for 1, 2, 4, 6 days for brown rice for 1, 2, 3 days for soybean. As a control of soaked, water was maintained at 4 ° C. for 5 days and 3 days, and the filtrate was discarded to adjust the humidity of the media source and then sterilized.

Example 8

The pupa powder for the mixed barley and soybean medium source was divided into the addition group (20) and the no addition group (0), and 5 chitosan in the mixed germination of barley and soybean as the control medium and barley as the control. Was diluted to 1000X, 500X, 250X and treated with 500X lactic acid 500X for 1, 2, 4, 6 days for barley, 1, 2, 3 days for soybeans and discarded the filtrate. After controlling the humidity, inoculated by sterilization by high temperature and high pressure treatment.

Example 9

The pupa powder for the mixed medium source of brown rice, barley and soybeans was divided into the addition group (20) and no addition group (0), and the mixed medium source of brown rice, barley and soybean as a control and the brown rice, barley and soybean as a control group Dilute 5-chitosan to 1000X, 500X, 250X at the time of mixed germination and treat with 500X 5-lactic acid as a control, for 1, 2, 4, 6 days for brown rice and barley, and 1, 2, 3 days for soybean. Germination treatment was performed. Anaerobic treatment was carried out for 12 to 20 hours in an incubator of CO ₂ (or N ₂ ) gas for the reaction of decarboxylase, and the control was kept in water at 4 ° C. for 5 days and 3 days, respectively. It was inoculated by sterilization with the experiment.

Example 10

Samples of the culture obtained by culturing in Examples 1 to 9 were lyophilized and analyzed. For analysis, the sample was ground, extracted with organic matter, freeze-dried, dissolved in a small amount of water, and analyzed for GABA (Gaba) by the automatic amino acid analysis system. Although it was divided into a culture part, a spore part, and a total culture part, the Cordyceps sinensis can be edible as the total culture part, and thus the analysis results are summarized only in the total culture part.

Standard Chromatogram of Amino Acids of Cordyceps Sinensis Cultivated According to the Present Invention RT (minutes) Peak name Area (mV ² sec area Height (mV) Height Concentration One 8.433 Asp 235.420 2.682 22.156 2.872 50.00 2 9.100 AMQ 85.433 0.973 4.320 0.560 0.00 3 10.200 Ser 566.879 6.458 37.103 4.809 50.00 4 11.800 Glu 278.984 3.178 21.453 2.781 50.00 5 12.383 His 427.528 4.870 32.907 4.265 50.00 6 13.950 NH ₃ 321.735 3.665 13.664 1.771 0.00 7 15.967 Arg 376.504 4.289 26.336 3.414 50.00 8 16.650 Thr 359.288 4.093 23.980 3.108 50.00 9 17.650 48.263 0.550 2.723 0.353 0.00 10 18.200 Ala 322.350 3.672 21.439 2.779 50.00 11 21.333 GABA 408.305 4.651 27.070 3.509 50.00 12 21.900 Pro 178.898 2.038 14.484 1.877 50.00 13 26.150 Tyr 458.872 5.227 45.278 5.869 50.00 14 27.117 Val 709.866 8.087 74.873 9.705 50.00 15 27.700 Met 557.669 6.353 62.094 8.049 50.00 16 30.217 Lys 352.715 4.018 39.725 5.149 50.00 17 30.550 Ilu 909.440 10.360 90.082 11.677 50.00 18 31.017 Leu 962.927 10.970 90.101 11.679 50.00 19 32.000 Phe 1216.948 13.864 121.682 15.773 50.00

Chromatogram analysis of the entire Cordyceps sinensis culture grown according to the present invention using a mixture of pupa and soybean as a medium RT (minutes) Peak name Area (mV ² sec area Height (mV) Height Concentration One 8.350 Asp 1060.087 3.233 94.569 3.794 220.15 2 9.267 AMQ 176.506 0.538 9.739 0.391 0.00 3 10.067 Ser 3758.919 11.464 236.748 9.498 328.81 4 10.867 19.960 0.061 1.917 0.077 0.00 5 11.417 5.596 0.017 0.782 0.031 0.00 6 11.733 Glu 823.661 2.512 57.757 2.317 146.19 7 12.300 His 250.178 0.763 17.419 0.699 28.70 8 12.850 25.277 0.077 1.733 0.070 0.00 9 13.117 17.300 0.053 1.335 0.054 0.00 10 13.917 NH ₃ 4571.771 13.943 156.076 6.626 0.00 11 15.183 42.696 0.130 2.580 0.103 0.00 12 15.633 156.999 0.479 17.247 0.692 0.00 13 15.900 Arg 3364.269 10.261 225.356 9.041 439.97 14 16.600 Thr 933.916 2.848 59.292 2.379 128.56 15 18.167 Ala 1639.202 4.999 86.744 3.480 248.13 16 19.933 1006.567 3.070 45.664 1.832 0.00 17 20.633 0.880 0.003 0.205 0.008 0.00 18 21.367 GABA 245.786 0.750 17.026 0.683 29.45 19 21.933 Pro 3248.996 9.909 260.008 10.431 878.18 20 22.583 49.884 0.152 4.108 0.165 0.00 21 23.107 5.405 0.016 0.560 0.022 0.00 22 23.400 4.797 0.015 0.551 0.022 0.00 23 23.750 39.866 0.122 4.146 0.166 0.00 24 23.950 29.256 0.089 3.622 0.145 0.00 25 24.250 788.755 2.406 91.651 3.677 0.00 26 24.667 2.103 0.006 0.446 0.018 0.00 27 24.883 139.592 0.426 18.377 0.737 0.00 28 25.400 38.190 0.116 4.060 0.163 0.00 29 25.583 26.605 0.081 3.405 0.137 0.00 30 25.767 39.737 0.121 5.879 0.236 0.00 31 25.883 64.931 0.198 6.967 0.279 0.00 32 26.217 Tyr 842.854 2.571 92.180 3.698 91.29 33 26.583 39.859 0.122 5.496 0.220 0.00 34 26.683 23.353 0.071 3.192 0.128 0.00 35 27.000 333.036 1.016 52.023 2.087 0.00 36 27.183 Val 1898.156 5.789 174.954 7.019 132.88 37 28.000 Met 21.484 0.066 2.985 0.120 1.88 38 28.233 92.511 0.282 10.901 0.437 0.00 39 28.667 3.724 0.011 0.801 0.032 0.00 40 28.850 27.006 0.082 3.748 0.150 0.00 41 29.283 34.151 0.104 5.257 0.211 0.00 42 29.450 1168.170 3.563 127.445 5.113 0.00 43 29.850 67.437 0.206 6.115 0.245 0.00 44 30.250 Lys 806.581 2.460 87.707 3.519 110.06 45 30.600 Ilu 1096.425 3.344 104.263 4.183 60.00 46 31.067 Leu 1998.481 6.095 192.821 7.736 104.43 47 31.700 1.936 0.006 0.308 0.012 0.00 48 32.033 Phe 1605.459 14.896 166.245 6.670 66.30 49 32.867 149.725 0.457 20.156 0.809 0.00

Chromatogram analysis of the medium after using a mixture of rice and beans as a medium and incubating Cordyceps sinensis according to the present invention RT (minutes) Peak name Area (mV ² sec area Height (mV) Height Concentration One 8.350 Asp 684.658 2.516 60.856 2.892 141.53 2 9.267 AMQ 247.667 0.910 13.778 0.655 0.00 3 10.083 Ser 2735.723 10.052 164.771 7.830 237.62 4 10.883 18.189 0.067 1.490 0.071 0.00 5 11.417 7.813 0.029 0.896 0.043 0.00 6 11.767 Glu 767.647 2.821 52.607 2.500 135.90 7 12.317 His 250.264 0.920 16.484 0.783 28.47 8 12.900 13.447 0.049 1.509 0.072 0.00 9 12.933 26.610 0.098 1.493 0.071 0.00 10 13.950 NH ₃ 3864.258 14.199 136.366 6.480 0.00 11 15.217 54.991 0.202 4.053 0.193 0.00 12 15.967 Arg 2766.591 10.166 176.187 8.372 359.97 13 16.650 Thr 852.522 3.133 52.841 2.511 117.25 14 17.900 382.515 1.406 23.227 1.104 0.00 15 18.217 Ala 1118.383 4.109 61.753 2.934 167.71 16 19.450 15.967 0.059 1.149 0.055 0.00 17 20.017 459.143 1.687 18.516 0.880 0.00 18 20.700 11.904 0.044 1.208 0.049 0.00 19 21.400 GABA 438.114 1.610 29.175 1.386 52.01 20 21.950 Pro 1881.686 6.914 148.397 7.052 493.53 21 22.600 83.785 0.308 5.543 0.263 0.00 22 23.033 10.574 0.039 0.913 0.043 0.00 23 23.400 0.768 0.003 0.162 0.008 0.00 24 23.767 36.206 0.133 4.215 0.200 0.00 25 23.950 84.739 0.311 8.729 0.415 0.00 26 24.250 921.213 3.385 104.461 4.964 0.00 27 24.683 16.379 0.060 2.282 0.108 0.00 28 24.883 180.817 0.664 22.411 1.065 0.00 29 25.383 48.926 0.180 4.845 0.230 0.00 30 25.733 137.363 0.505 8.578 0.408 0.00 31 26.200 Tyr 756.933 2.781 82.619 3.926 80.74 32 26.567 94.927 0.349 9.251 0.440 0.00 33 26.983 560.449 2.059 81.828 3.888 0.00 34 27.167 Val 1445.635 5.312 156.927 7.457 99.65 35 27.350 466.114 1.713 48.394 2.300 0.00 36 27.983 Met 119.896 0.441 14.010 0.666 10.33 37 28.200 84.741 0.311 10.428 0.496 0.00 38 28.417 23.248 0.085 2.811 0.134 0.00 39 28.650 2.959 0.011 0.615 0.029 0.00 40 28.833 16.980 0.062 2.494 0.119 0.00 41 29.250 47.751 0.175 6.590 0.313 0.00 42 29.450 1115.253 4.098 120.108 5.707 0.00 43 29.867 55.881 0.205 4.950 0.235 0.00 44 30.250 Lys 504.206 1.853 54.984 2.613 68.96 45 30.583 Ilu 791.589 2.909 74.575 3.544 42.50 46 31.050 Leu 1425.007 5.236 133.832 6.360 72.64 47 32.033 Phe 1439.634 5.290 152.293 7.237 57.67 48 32.850 144.956 0.533 18.988 0.902 0.00

Although it is a result of some of the results in this example compared to the pupa only control, the following results were obtained even in the mixed medium without germination. That is, the ratio of GABA content (x nmol / g FW = fresh weight)} in the total culture of Cordyceps sinensis analyzed using various media was mixed with pupa and soybeans with respect to the control medium containing only pupa medium. In the experimental section of the medium, 1.3 times, 1.6 times in the brown rice and soybean mixing section, and 2.3 times in the mixing section of rice and soybeans, it was confirmed that only the fruiting bodies cultured in the mixed medium containing rice were analyzed. There was more than 6 times improvement effect compared to the control of fruiting body produced in pupa. Therefore, it is expected that a much larger amount than cereals and cereals such as brown rice, barley, wheat (wheat), and the like may be further improved in the germination process as in the present invention.

According to the present invention as described above, using only the germination obtained by germinating grains or legumes such as rice, glutinous rice, brown rice, barley, wheat (rye), soybean as a medium source or edible such as germination and pupa Insect or rice, glutinous rice, brown rice, barley, wheat (rye), beans, etc., mixed with cereals or legumes, but when the germination of grains and legumes treated with chitosan, anaerobic treatment after germination It is possible to obtain a Cordyceps sinensis or Cordyceps sinensis culture having a high content of γ-aminobutyric acid to be used as a dietary supplement or as a functional food.

Claims (9)

After germination of the germ bodies obtained by germinating grains or legumes as a medium source, inoculate the liquid culture solution of Cordyceps sinensis into the cooled medium, and incubate for 2-3 weeks at a temperature of 25 ° C. and a relative humidity of 70 (RH). A method of cultivating Cordyceps sinensis with high content of γ-aminobutyric acid, which induces fertility when infestation, generates fruiting bodies, and completes culture. The method of claim 1, A method of cultivating Cordyceps sinensis with high content of γ-aminobutyric acid, characterized by treating chitosan during germination of cereals or beans. The method of claim 1, A method of cultivating Cordyceps sinensis with high content of γ-aminobutyric acid, characterized by performing anaerobic treatment after germination of cereals or legumes. The method of claim 3, wherein Anaerobic treatment is a method of cultivating Cordyceps sinensis with high content of γ-aminobutyric acid, characterized in that the method is exposed to wet N ₂ gas or CO ₂ gas for 12 to 20 hours. The method of claim 1, Cultivation method of Cordyceps sinensis with high content of γ-aminobutyric acid, characterized by mixing edible insects or general grains or legumes in the medium. The method of claim 1, Cordyceps sinensis is a method of cultivating cordyceps with high content of γ-aminobutyric acid, characterized in that the cordyceps mycelium. The method of claim 1, Cordyceps sinensis is a method of cultivating cordyceps with high content of γ-aminobutyric acid, characterized by the cordyceps fruiting body. The method of claim 1, Cordyceps sinensis is a method of cultivating Cordyceps sinensis with high content of γ-aminobutyric acid, characterized in that it contains a Cordyceps mycelium and a medium component. The method of claim 1, Cordyceps sinensis is a method of cultivating Cordyceps sinensis with high content of γ-aminobutyric acid, characterized by containing Cordyceps mycelium, fruiting body and medium components.