KR20020048848A - A grain culture composition of inonotus obliquus, a method of preparing the antitumor and immuno-activity polysaccharide from it and their use - Google Patents

Abstract

PURPOSE: Provided are a grain a culture composition of obliquus and a method for preparing antitumor and immuno-activity polysaccharide therefrom used as active ingredients for functional foods and health food supplements. CONSTITUTION: The grain culture composition contains mycelium of Inonotus obliquus Inonotus which is prepared by: mixing rice including brown rice, glutinous brown rice, black rice, glutinous rice or regular rice, and grain including barely, wheat, corn or job's tear; preparing a medium using the mixture raw or optionally roasted; inoculating Inonotus obliquus; and culturing it at 25-30 deg.C under dark condition for 15-30 days. Polysaccharide is prepared by the steps of: drying and pulverizing the composition; placing the pulverized composition into a tea bag then dipping the tea bag in water of 30-100 deg.C for 5-10 minutes to elute active ingredients therefrom; vacuum-concentrating the eluate; and spray-drying the concentrate to obtain mycelium-containing extract.

Description

Grain culture composition of Inotus olivius and method for isolating and producing anticancer immunoactive polysaccharides from the same, and uses thereof USE}

The present invention relates to a grain culture composition of Inonotus oblivus, a method for separating and producing anticancer immunoactive polysaccharides therefrom, and uses thereof.

Modern people are exposed to various pollutants and carcinogens, and the incidence of cancer is increasing every year. There are also genetic factors in the carcinogenesis, but the immune function of the body is lowered and cancer occurs when the treatment is very difficult, and the survival rate is low, so prevention is the best.

Various edible and medicinal mushrooms are currently being used as an attempt to prevent cancer. Mushrooms are known to have various physiological functions, for example, anticancer activity, immune enhancing effect, blood pressure and hypoglycemic effect, antioxidant effect, and the like. The use form of mushrooms is taken as it is or used as fruiting body and mycelium extract. have.

The present invention is directed to an Inotus oblivus mushroom, which is unfamiliar in Korea, and relates to artificial culture of the mushroom and a method for producing a substance having an anti-cancer immunoactive effect in vivo, and the use of the active substance. will be.

Inonotus obliquus (Pers) pilot is a perennial basidiomycete fungus belonging to the genus Hynoenochaetaceae, Inonotus , which in its natural state is more than 45 degrees north latitude of Siberia and Hokkaido. It is distributed in and is a mushroom that grows on stems and stumps of birch, alder and rowan. It is a kind of white fungus that has a diaphragm and does not have a clamp connection for nuclear movement. When grown in nature, it becomes a black fungal nucleus mass and adds a trunk such as birch. The fungus tissue has the characteristics of KOH and Xantochronic reactions.

Inonotus oblivus is still an unrecorded species in Korea, and nine mushrooms belonging to the same genus have been reported. In the case of I. dryadeus (Pers.:Fr.) Murr of the same genus, it is known to secrete a large amount of antibacterial activity against bacteria on the surface of the lampshade. In Korea, there is an evergreen mushroom ( I. nodulosus (Fr.) Pilat.) Recorded in brain cancer (1613) as encephalopathy, and its effectiveness and safety have already been proven for several centuries. .

Inonotus obliquus has been used as a tea and a drink in Russia since ancient times, and is currently used to treat all cancers related to the stomach, and is used as a treatment for AIDS, O-157, and liver cancer in Japan.

It is also known as Chaga or Chaga, Cabanoanatake, Bot Hump Mushroom, White Birch Mushroom, Black Birch Mushroom, Fuscosporia obliqua , and has a strong anti-cancer effect and strong immune activity. (41st Annual Banquet Research Presentation of the Japanese Society of Pharmacognosy). It is also known to exhibit anti-AIDS activity by acting on both cells and viruses (7th Japan AIDS Society).

Betaglucan and heteroglucan contained in the polysaccharide of Innoteus obliquus mushroom are recognized to have strong antioxidant and cellular activity effects (research on anticancer polysaccharides, Japanese Pharmacognosy No. 27). The water-soluble and water-insoluble polysaccharides of the polysaccharides extracted from the fungal nucleus and mycelium of this mushroom were investigated to have antitumor activity and hypoglycemic activity.

However, Inonotus oblivus is difficult to obtain as a species that has not yet been reported in Korea, and because it is rare in the world and is widely distributed in nature, its production is extremely limited. It is traded at a high price, and mycelium is not easily separated and purely cultivated.

Therefore, the present inventors have successfully identified artificial strains having excellent anti-cancer immunity and microbiological properties from natural Inonotus obliquus collected in Russia, and successfully cultivated artificial cultures. Isolation of a superior anti-cancer immunoactive substance that is nearly equivalent to Notus obliquus has led to the present invention.

Existing patents related to Innotus obliquus have been used as a natural extract of this mushroom as an additive in natural complexes with immune and tonic effects (Russian Federation, RU2125460C1), and vodka production with plant materials It may be used as a kind of fragrance additive in China (Russian Federation RU2141517C1). It is also used as a kind of toothpaste with bad breath and anti-inflammatory effects (Russian Federation RU2131725C1).

Previous studies have reported that extracts from natural mushrooms with distilled water inhibit the proliferation of human uterine cancer cells on in vitro (Rzymowska, 1997). Also, extracts of natural mushrooms are effective in gastric ulcers on in vitro. Has been reported (Pashinsky et al., 1998).

One of the objects of the present invention is to artificially incubate Innoteus obliquus.

To this end, in the present invention, a strain having excellent anticancer immunological activity and microbiological characteristics was isolated and identified from the natural Inonotus obliquus collected in Russia. Inonotus obliquus strain used in the present invention is a strain of Accession No. KCTC 0894BP deposited on November 28, 2000 at the Genetic Research Center of Biotechnology. In the present invention, the strain culture medium of the strain may be rice such as brown rice, brown rice glutinous rice, black rice, glutinous rice, non-glutinous rice, or edible grains such as barley, wheat, corn, soybean, yulmu, sorghum, or two or more of them. The mixture is used at a predetermined ratio, but is inoculated in a medium prepared as it is or roasted, and cultured for 15 to 30 days at 25 to 30 ° C. and dark conditions, thereby obtaining a grain culture composition including the Inotus olivius mycelium.

Another object of the present invention is to isolate and purify mushroom-derived polysaccharides having anticancer immunoactivating activity from the Inototus obliquus strain.

In vivo anticancer immune activity of Inotus oculus mushroom extract is well known and its bioactive component is known as glucan of polysaccharides. However, purified pure glucan is not absorbed in oral administration and shows its effect only by injection. Thus, in the case of oral administration, purified glucan is not used and is used as a glycoprotein with a peptide bound to glucan.

Therefore, the present invention extracts a complexed polysaccharide form of Crude from Innoteus obliquus so as to exhibit anti-cancer immune activity even by oral administration. Extraction method is dried and pulverized grain culture composition containing Inonotus obliquus mycelium, and the powder is placed in a tea bag and immersed in hot water of 5 to 40 times the volume of 30 to 100 ℃ for 5 to 10 minutes After eluting, the eluate was concentrated in vacuo, and the concentrate was spray-dried to obtain a grain cultured mycelium extract containing polysaccharide.

It is a further object of the present invention to provide a grain culture composition comprising Inonotus obliquus mycelium and the use of a complex polysaccharide isolated therefrom, in particular as a dietary supplement and a functional food.

In addition, the present invention compares the anticancer immune activity of the fruiting body of the mushroom obtained from the artificial culture and the polysaccharide extracted from the fruiting body collected in nature, the mycelium produced by the liquid culture produced in and out of the mycelium, the polysaccharides, grain medium and solid from wood The anticancer immune activity of the cultured mushroom products is compared. Through these comparative experiments, the present invention provides basic bioactivity data for the industrial formulation of Innoteus obulicus mushroom.

Mushroom products in the present invention includes the mycelium and fruiting body forming the body of the mushroom or produced by the mushroom strain, polysaccharide in the mycelium in the case of liquid culture and the polysaccharide secreted out of the mycelium.

In the present invention, the polysaccharide refers to a complex polysaccharide including components other than alcohol-insoluble polysaccharides and water-soluble polysaccharides, intracellular polysaccharides, extracellular polysaccharides, and sugars extracted from the mushroom product of Innoteus obliquus. ), For example, it is used as a concept that includes all of the protein polysaccharide having anticancer activity.

1 is a block diagram showing a process of extracting anti-cancer immuno-active polysaccharides by grain culture of Innoteus Olivius.

In the present invention, isolates from the natural state of Inonotus obliquus collected in Russia were isolated and identified strains excellent in anti-cancer immunity and microbiological characteristics were used in artificial culture. Inonotus obliquus strain used in the present invention was deposited on November 28, 2000, accession number KCTC 0894BP in the Center for Genetic Resources of the Biotechnology Research Institute.

Innotus Aubricius Inonotus obliquus Artificial Culture of Mushroom Strains

In the present invention, artificial culture of Inonotus obliquus uses a grain culture method.

Grain culture composition

Artificial culture using edible grains is carried out to obtain mushroom products of Inotus olivus strains to be used as functional foods. For grains, rice such as brown rice, brown rice glutinous rice, black rice, glutinous rice, non-glutinous rice, or barley, wheat, corn, soybeans, yulmu, or sorghum, or used after roasting, these grains are used alone or in two or more kinds. Use a mixture at a certain ratio. The grains are immersed in cold water for 12 to 48 hours to control moisture, and then sterilized by enclosing them in a polypropylene culture bottle. After sterilization, when the medium is at room temperature, inoculate about 10% of the Inotus oculus cultivar and incubate at 25 to 30 ° C for 15 to 30 days under dark conditions. After the incubation is completed, the culture bottle is removed and hot-air dried at 50 to 70 ° C., thereby obtaining a grain culture composition comprising the Inotus oculus mycelium of the present invention. The composition is pulverized with a hammer mill or the like to make a mesh size of about 1.4 to 2 mm or a powder.

Extraction of Anticancer Immune Active Substances

Extraction of anticancer immunoactive substance from mycelium composition obtained from grain culture

The process of extracting an anticancer immunoactive substance from the culture composition of Inonotus obulius obtained by the grain culture may include: (a) drying and pulverizing a culture composition comprising the inonotus obliqueus mycelium; (b) immersing the ground product in a tea bag and immersing the active ingredient for 5 to 10 minutes in a warm water of 5 to 40 times the volume of 30 to 100 ° C .; (c) vacuum concentration of the eluate; (d) spray drying the concentrate to obtain a mycelium extract containing the polysaccharide.

Figure 1 shows a block diagram of this process, in this process, the dried grains containing the cultured mycelia, that is, the pulverized product of the culture composition is put in a tea bag 5 to 40 times the hot water of 5 to 40 times the volume of 30 to 100 ℃ After dipping for 10 minutes, the mixture was eluted sufficiently, and the obtained eluate was concentrated in vacuo and spray dried to obtain a mycelium extract containing polysaccharides.

Usage

The grain culture composition comprising the Inonotus obliqueus mycelium obtained by the above method and the extract prepared separately therefrom, that is, the complex polysaccharide of anti-cancer immune activity, are obtained in the form of a functional food and anti-cancer, immune-enhancing, etc. It can be used for health supplements and the like.

When formulated for the use of dietary supplements, it may have various forms such as powders, tablets, capsules, granules, liquids, pills, dispersants, syrups, and also in the case of functional foods may have various forms as described below. Can be.

The active polysaccharides of Inonotus obliquus obtained in the present invention are effective over a wide range of dosages. For example, the adult daily dosage of the active substance is usually in the range of 200 mg to 5,000 mg, preferably 1,000 mg to 3,000 mg in several divided doses. However, since the amount of the active substance to be administered may vary depending on the purpose of use, the scope of the present invention is not limited by the above administration range.

In addition, when used as an additive of a functional food or a favorite food complex polysaccharide of the present invention, may be preferably added at 0.1% to 40% of the total weight of the food, more preferably 0.05% to 20% have. However, very small amounts of up to 0.1% may be added depending on the food or the intended use.

When formulating the grain culture composition of Innoteus obulicus obtained in the present invention and the polysaccharide separated therefrom into a dietary supplement or a functional food in the same manner as described above, sweeteners in a range that does not affect the active ingredient, Auxiliaries such as colorants, flavors, emulsifiers, suspending agents, preservatives and the like may be added.

For example, in the case of preparing a polysaccharide, it is used by adding a grain culture composition or an extract (complex polysaccharide) obtained from the conventional coffee, black tea, green tea, barley tea, honey tea, round tea, or the like, or Grain culture composition or the extract can be used as it is in a paper bag. At this time, the usual additives used in foods such as flavoring agents can be added.

In addition, the complex culture of the grain culture composition of the present invention or the extract isolated therefrom can be used as a substitute for soups, porridges, cooking additives, sun foods, alcoholic beverages such as health drinks, flavored beverages, carbonated drinks, juices, and candy. When added to chewing gum, chocolate, ice cream, yogurt, etc. to make functional foods, it is possible to prevent cancer from occurring and enhance immune function naturally even by ingestion of ordinary favorite foods.

In addition, when the grain culture composition of the present invention or the extract isolated from it is added to various liquors such as soju, sake, etc., it is possible to make a functional alcoholic beverage having the effects of anticancer, immune enhancement, etc. while utilizing the unique flavor and aroma of mushrooms. have.

In addition, the active polysaccharide extracted in the present invention, can also be used for the purpose of the prevention of bad breath, anti-inflammatory, etc. that is the use of the known Inotus Oculus.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the examples are only for illustrating the present invention and the scope of the present invention is not limited by the following examples.

Example 1

Innotus Oculus KTCT0894BP Artificial Liquid Culture of Mushroom Strains

For the liquid culture of the Inotus oblivus strain, a medium containing 50 g of glucose per liter of medium, 10 g of peptone, 10 g of yeast extract, 0.5 g of potassium diphosphate, 0.5 g of magnesium sulfate, and 0.4 g of calcium chloride was used. In addition, the mass production medium is 20 g of soluble starch per liter of medium, 40 g of glucose, 10 g of yeast extract, 5 g of soybean meal, 0.5 g of potassium diphosphate, 0.5 g of magnesium sulfate, 0.4 g of calcium chloride and trace elements of iron sulfate, zinc sulfate, copper sulfate. A medium containing a back was used. All media were used after adjusting the pH to 6 after sterilization.

After the sterilization of the 250ml shake culture Erlenmeyer flask containing 50ml of the spawn medium was sterilized at 121 ° C. for 20 minutes, the inotus olivus KTCT0894BP mycelium on the malt extract medium was inoculated and shaken at 28 ° C. for 3 days. To the primary seed culture solution. The cultured primary seed culture solution was homogenized at 5,000 rpm for 30 seconds, and then inoculated in a jar fermenter containing 2 liters of sterilized seed medium under the same conditions as above, and incubated at 28 ° C. to prepare a secondary seed culture solution.

Subsequently, a 500L fermentation vessel containing 300L of production medium was sterilized under the same conditions as above, followed by inoculation with a secondary seed culture solution and incubated for 5 days to form a third seed culture solution. The cultured tertiary seed culture broth was inoculated in a sterile 3,000 liter fermentation vessel containing 3,000 liters of production medium and cultured for 5 days to produce a large amount of mycelium. A total weight of 28 g / L was obtained. .

Example 2

Culture of fruiting bodies of Innoteus obliquus KTCT0894BP

Innotus olivus KTCT0894BP mycelium cultured in malt extract medium was removed from the medium added with rice bran to oak sawdust for the production of seedlings to obtain the fruiting body of the strain of Inotus Olivius KTCT0894BP for functional food and medical use. Inoculation was incubated under dark conditions at 28 ° C. for 22 days.

Logs harvested between November and February were naturally dried in the shade and cut into pieces of 10 to 15 cm in diameter and 20 to 25 cm in length. The cut logs were put in a polypropylene bag and then inoculated with sterilization at 121 ° C. for 15 hours, and then cooled to room temperature. The mycelial culture was carried out in the incubation room until the white mushroom mycelium adhered to the whole wood and the color of the mycelium on the surface of the wood began to turn black for 32 days at the optimum temperature of 28 ℃.

To induce the fruiting bodies of the mushrooms, the cultured logs were removed from the polypropylene bags and buried in the soil of well-drained soil ganoderma lucidum growers so that the upper layer of the logs protruded to the ground, and the mycelium used as inoculum was removed and sand was removed. Thinly spread and then watered. After 27 days, the mushrooms begin to form on the surface of the wood, providing sufficient ventilation and inducing the formation of fruiting bodies. The adherent bacterium developed into a cylindrical form, and 12 months later, 32 g of fruiting body per 1 kg of wood was obtained.

Example 3

Grain cultivation of Inonotus oblivus KTCT0894BP mushroom strain using edible grains

As a method for obtaining a mushroom product of the Inototus Aubricus KTCT0894BP strain to be used as a functional food application, artificial culture using edible grains was performed. The types of grains used were brown rice, brown rice glutinous rice, black rice, glutinous rice, non-glutinous rice, barley, wheat, corn, soybeans, yulmu, sorghum, etc., or used after roasting. Used. The grains were immersed in cold water for 24 hours to control moisture, and weighed 600 g per bottle, and sealed in a polypropylene culture bottle and sterilized for 40 minutes. After sterilization, the medium was inoculated with 10% of the culture medium of Example 1 when the medium was brought to room temperature, and cultured at 28 ° C. for 20 days under dark conditions. After the incubation was completed, the culture bottle was removed, hot air dried, and then ground into a hammer mill (Icasa, Germany) to make powder.

Example 4

Extraction of Anticancer Immune Active Substances

1) 100 g of the mycelium obtained in Example 1 was mixed with a suitable amount of the culture filtrate, homogenized for 1 hour, hot water extracted at 95 ° C. for 6 hours, and centrifuged at 6000 rpm for 30 minutes to remove the mycelium as a precipitate and hot water as a non-precipitate. An extract was obtained. It was concentrated to vacuum 100ml to make a final concentration of 60% ethanol was added and left at 4 ℃ for 24 hours. The precipitate was obtained by centrifugation at 6,000 rpm for 30 minutes, and the precipitate was dissolved in 100 ml of water heated at 70 ° C., and then dialyzed with dialysis membrane to separate polysaccharides having a molecular weight of 3,000 or more, and then lyophilized at -70 ° C. Extracellular polysaccharide and 4.3 g of polysaccharide having components other than sugar were obtained. In addition, 100 g of the mycelia obtained in Example 1 were mixed with the culture filtrate in the same manner as described above, homogenized, and extracted by hot water, followed by centrifugation to obtain a hot water extract, which is a non-precipitate, concentrated, dialyzed, and spray dried to obtain 3.5 g of the extract. Got it. A total of 7.8 g of the complex polysaccharide extract (hereinafter referred to as "extract A") was obtained by combining the extract of 4.3 g and 3.5 g of the polysaccharide obtained above.

2) The fruiting bodies of the natural Inonotus obliquus mushrooms and the fruiting bodies produced in Example 2 were dried at room temperature until there was no change in weight, and then each was ground with a hammer mill to obtain a powder. 100 g of each powder was suspended in distilled water of 10-fold volume, hot water extracted at 95 ° C. for 6 hours, homogenized, and centrifuged at 6,000 rpm for 30 minutes to remove mushroom mycelium, which was a non-precipitated hot water extract. This was concentrated to a vacuum to make 100ml, ethanol was added to the final concentration of 60% and left at 4 ℃ for 24 hours. The precipitate was obtained by centrifugation at 6,000 rpm for 30 minutes, and the precipitate was dissolved in 100 ml of water heated at 70 ° C., and then dialyzed at molecular weight, and then lyophilized at −70 ° C. to 3.2 g of polysaccharide (natural acid), respectively. 3.7 g (artificial culture) were obtained. In addition, 10 times the volume of water was added to the fruiting body obtained through centrifugation, extracted with hot water at 70 ° C., homogenized, and then centrifuged to concentrate the non-precipitated material, followed by dialysis and spray drying, respectively, 4g (natural acid) and 3.2g ( Artificial water) to obtain a hydrothermal extract. A total of 7.2g of natural fruiting bodies (hereinafter referred to as "extract B") and a total of 6.9g of artificial fruiting bodies (hereinafter referred to as "extract C") were obtained.

3) Extraction of the anticancer immunoactive substance from the mycelium of the Inonotus oblivus mushroom cultured in the grain medium in Example 3 was carried out by the following method. 100 g of dried and crushed grains containing the cultured mycelium were immersed in a tea bag and then immersed in hot water of 20 ° C. and 20-fold volume for 6 minutes to elute well. 5 g (hereinafter referred to as "extract D") were obtained.

Example 5

Acute Toxicity Experiment

Extracts A, C, and D of Innoteus obliquus KTCT0894BP obtained in Example 4 were administered intraperitoneally (5) and orally (5) to 10 ICR mice each 4 weeks old at a concentration of 10,000 mg / kg. Observed daily. As a result, no abnormality was found in all 10 animals.

Example 6

Tumor growth inhibition rate investigation experiment

The following anticancer test was carried out using the Inonotus oblivus KTCT0894BP mushroom extract obtained in Example 4. The control and treatment groups were divided into 6 groups of 5 weeks old ICR male mice, and about 1 × 10 ⁶ plural tumors of Sarcoma 180 / mouse were implanted subcutaneously in the lateral abdomen of all mice. From the day after transplantation, the control group was intraperitoneally administered once a day for 10 days, and the treated group was dissolved in physiological saline after dissolving the complex polysaccharide extract obtained in Example 4 at a ratio of 10, 30 and 100 mg / kg, respectively. Sterilized with a bacterial filtration membrane and administered once a day for 10 days. After 5 weeks, the solid cancer was extracted and compared with the average weight of the treated group and the average weight of the control group. The results are shown in Table 1.

Inhibitus Oculus KTCT0894BP Extract Inhibits Tumor Growth Dose (mg / kg) 10 30 100 Natural Inonotus Oculus Mushroom Extract (B) 76% 89% 100% Extract of Inonotus oblivus mycelium product through liquid culture (A) 66% 75% 99% Wood-cultivated Inototus Oculus fruiting body extract (C) 72% 88% 100% Extract of Inonotus oblivus mycelium cultured in brown rice medium (D) 24% 40% 68%

Example 7

Tumor growth inhibition rate investigation experiment

Using the same sample as in Example 6, the same Sarcoma 180 solid tumor was transplanted to 5-week-old ICR male mice, and 500 mg / Kg, 1,000 mg / kg, orally, were administered orally once a day from the day after transplantation. After 5 weeks, the solid tumor was removed and the average weight was compared with the control. The obtained results are shown in Table 2.

Inhibitus Oculus KTCT0894BP Extract Inhibits Tumor Growth Dose (mg / kg) 500 1000 Natural Inonotus Oculus Mushroom Extract (B) 85% 100% Extract of Inonotus oblivus mycelium product through liquid culture (A) 75% 97% Wood-cultivated Inototus Oculus fruiting body extract (C) 83% 100% Extract of Inonotus oblivus mycelium cultured in brown rice medium (D) 42% 63%

Example 8

Immunopotentiation Investigation Experiment

After treating mouse leukemia cells L-1210 with 0.013% glutaraldehyde-phosphate buffer solution, the cells were again treated with concanavalin A to prepare GA-conA-L-1210 cells. These cells were injected intraperitoneally with 1 × 10 ⁶ mice / mouse into mouse BDF ₁ males.

After the first injection, a second injection was performed after one week. The next day, 500 mg / kg orally of the multi-polysaccharide of Innoteus obliquus KTCT0894BP obtained in Example 4 was administered to each group of mice. On day 6 after drug administration, 1 × 10 ² live mice / mouse of L-1210 were transplanted intraperitoneally in all mice, and immunoreactivity was measured by the average days of survival of mice. The results are shown in Table 3.

Immune Enhancing Activity of Innotetus Olivius KTCT0894BP Extract sample Average survival days Life rate Control 13th - Natural Inonotus Oculus Mushroom Extract (B) 19th 46.2% Extract of Inonotus oblivus mycelium product through liquid culture (A) 18 days 38.5% Wood-cultivated Inototus Oculus fruiting body extract (C) 19th 46.2% Extract of Inonotus oblivus mycelium cultured in brown rice medium (D) 14 days 7.7%

Example 9

Immunopotentiation Investigation Experiment

The vaccine of L-1210 was administered intraperitoneally to BFD male mice similarly to Example 8. The administration day was twice and administered over one week. The cell number was 1 × 10 ⁶ cells / mouse. Inototus Oculus KTCT0894BP mushroom extract obtained in Example 4 was prepared and administered intraperitoneally by 100 mg / Kg. Six days after drug administration, 1 × 10 ² live cells / mouse of L-1210 were implanted intraperitoneally in all mice. The average survival days at this time are shown in Table 4.

Immune Enhancing Activity of Innotetus Olivius KTCT0894BP Extract sample Average survival days Life rate Control 13th - Natural Inonotus Oculus Mushroom Extract (B) 16th 23.1% Extract of Inonotus oblivus mycelium product through liquid culture (A) 15.5 days 19.2% Wood-cultivated Inototus Oculus fruiting body extract (C) 16th 23.1% Extract of Inonotus oblivus mycelium cultured in brown rice medium (D) 14 days 7.7%

In the present invention, artificial culture of Innoteus obliqueus in a grain medium provides a grain polysaccharide composition comprising an innotetus obliqueus mycelium and an extract isolated therefrom. As can be seen from the above examples, the complex polysaccharide of the present invention has an anticancer immune activity with almost the same effect as that of the extract of native Inototus Olivius. Inonotus oculus grain culture composition of the present invention and the complex polysaccharide extract thereof is an active ingredient having anti-cancer immune activity can be used for the use of health supplements and functional foods.

Claims (7)

Inino on rice containing brown rice, brown rice glutinous rice, black rice, glutinous rice, non-glutinous rice, or edible grains such as barley, wheat, corn, soybeans, yulmu, or sorghum, or by mixing two or more kinds A grain culture composition comprising Inotus oculus mycelium obtained by inoculating seed spawn of Tooth Oculus and cultured at 25 to 30 ° C. for 15 to 30 days under dark conditions. Using the grain culture composition of claim 1, comprising: (a) drying and grinding the composition; (b) immersing the ground product in a tea bag and immersing the active ingredient for 5 to 10 minutes in a warm water of 5 to 40 times the volume of 30 to 100 ° C .; (c) vacuum concentration of the eluate; (d) spray drying the concentrate to obtain a mycelium extract containing the polysaccharide. A dietary supplement and functional food comprising a grain culture composition comprising the inonotus obliquus mycelium of claim 1. The food product according to claim 3, wherein the food is a functional food having one of a variety of teas, soups, porridges, cooking additives, foods, yogurt, health drinks, gum, sweets, bread, chocolate, and ice cream. Health supplement and functional food comprising a complex polysaccharide prepared by the method of claim 2. 6. The food product according to claim 5, wherein the food is a functional food having one of a variety of teas, soups, porridges, cooking additives, foods, yogurt, health drinks, gum, confectionery, bread, chocolate, and ice cream. The food according to claim 5, wherein the food is a health supplement having one of powders, tablets, capsules, granules, liquids, pills, dispersants, and syrups.