KR101124783B1 - Medium composition for Phellinus linteus culture - Google Patents

Abstract

The present invention relates to a medium mushroom mycelium culture medium composition with a higher content of beta glucan, and more particularly, to increase the content of beta glucan by culturing the mycelium mushroom mycelium in an herbal medicine extract.

Situation Mushroom, Medium Composition, Beta Glucan

Description

Medium composition for Phellinus linteus culture

The present invention relates to a medium composition for cultured mushroom mycelium.

Mushrooms are microorganisms belonging to basidiomycetes in the classification system of organisms, and have been used for food for a long time and have been introduced as folk traditional medicine for various diseases. There are about 15,000 mushrooms growing naturally on earth, but most of them can be used for food, except those that are toxic. These mushrooms are reported to contain a large amount of polysaccharide glucan and its derivatives, polysaccharides such as glucan is known to have an anticancer action.

Beta-glucan is a type of polysaccharide that activates the immune function of human normal cellular tissues, inhibits the proliferation and recurrence of cancer cells, and promotes the production of interleukin and interferon that activate immune cells. Active beta glucan enters the body where cancer cells are located and produces cytokines (Cytokine) to support the activity of immune cells, T cells and B cells, thereby activating the immune function of cellular tissues.

In addition, mushrooms are mostly used for food, so it is known to have fewer side effects, safer in terms of toxicity, strengthen human immune function, and have excellent anti-cancer effects. In particular, the situation mushroom is known to be excellent as an immune enhancer and anticancer agent compared to other mushrooms, but it is a perennial and does not reproduce well in nature. In addition, there is a problem of resource depletion and ecosystem destruction, so it is very difficult to obtain fruiting bodies. Situation mushrooms are widely distributed throughout the world, but the fruiting bodies are not well formed in nature.

Conventional situation mushroom artificial cultivation method is a variety of wood by-products, such as sawdust, a mixture of grains, there is a method of cultivating fruiting body by using herbal medicine by-products. However, the conventional method for artificially cultivating the situation mushroom fruiting body takes a long period of 3-6 months of cultivation is difficult to mass production and does not meet the demand. On the other hand, the cultivation of the situation mushroom mycelium has the advantage that not only shortens the time required for fruiting body formation, but also obtains a large amount of polysaccharides in a short time. However, the conventional situation mushroom mycelium culture method is difficult to mass production due to the difficult culture conditions. As such, although the situation mushroom is effective as an anticancer agent and an immune enhancer, its supply is limited and mass production and rapid production are not easy, and thus are not widely used.

As a prior art related to artificial mushroom culture, Korean Patent Registration No. 51056 (Registration date 1992.04.24) discloses a method of cultivating a mushroom mushroom mycelium by using a mixture of mulberry and mulberry sawdust rice bran as a medium. Mulberry or the old tree, which is the main raw material of the medium, is difficult to obtain, so it is difficult to manufacture a large amount of culture medium, and the culture period is 50 to 60 days, which takes a long time.

In addition, Korean Patent Registration No. 204984 (Registration Date 1999.03.31) discloses a method of using cereals as a culture medium component of Felinus linteus mycelium, but this method also requires 60 days of incubation time, which is not economical. have.

Thus, the present inventors have confirmed that the content of beta glucan in the culture product when the cultured mycelium mushroom mycelium using the herbal extracts to complete the present invention.

Therefore, the main object of the present invention is to provide a medium composition for culturing mushroom mycelium to increase the content of beta glucan in the extract of mushroom mushroom mycelium.

In order to achieve the above object, the present invention is composed of ginseng, Astragalus, larvae (mugwort), Angelica, Sangbaekpi (Mulberry bark), and medicinal herb extract 5-20 wt%, potato starch 0.2-1 wt%, dextrose It provides a medium mushroom culture mycelium culture medium composition comprising 1-5% by weight of woods and 75-90% by weight of distilled water. In addition, it is characterized in that it comprises 5-20% by weight of the herbal medicine extract consisting of ginseng, Astragalus, larvae (mugwort), Angelica, sangbaekpi (Mulberry bark) and Ogapi extract, 1-5% by weight of malt extract and 75-90% by weight of distilled water. It provides a medium composition for culturing mushroom mycelium.

The present invention relates to a medium composition for culturing situation mushroom mycelium having a high content of beta glutan.

The first aspect of the present invention is 5-20% by weight of Chinese herbal medicine extract consisting of ginseng, Astragalus, larvae (mugwort), Angelica, Morus bark (Mulberry bark) and Ogapi extract, 0.2-1% by weight potato starch, dextrose 1- It provides a culture medium composition for the situation mushroom mycelium culture comprising 5% by weight and 75-90% by weight of distilled water.

The second embodiment of the present invention is 5-20% by weight of the herbal medicine extract consisting of ginseng, Astragalus, Aerobaia, Mugwort, Morus bark (Mulberry bark) and Ogapi extract, 1-5% by weight of malt extract and 75-90% by distilled water. It provides a culture medium composition for the situation mushroom mycelium culture comprising a%.

More specifically, the herbal medicine extract ginseng 200g, Astragalus 200g, Aegye (mugwort) 200g, Angelica 200g, Morus bark (mulberry bark) 100g and 100g of ogapi in 2L purified water and then extracted by boiling water at 110 ℃ for 2 hours It features.

Hereinafter, the present invention will be described in more detail with reference to Examples.

However, the following examples are merely to illustrate the present invention, the contents of the present invention

It is not limited to an example.

< Example  1> Herbal Medicine Extract Potato Dextrose  Situation Mushroom Mycelium Culture in Medium

The herbal extracts were prepared by adding 200 g of ginseng, 200 g of Astragalus, 200 g of Aegye (mugwort), 200 g of Angelica vulgaris, 100 g of Morus bark (Mulberry bark), and 100 g of Ogapi in 2 L of purified water and then hot water at 110 ° C for 2 hours.

10% herbal extract, 0.4% potato starch and 2% dextrose were added to 1 L of distilled water to prepare a liquid medium and sterilized for 15 minutes in an autoclave. Aseptic inoculation of the spawn mushroom mycelium in the sterile medium obtained above and incubated for 20 days at 30 ℃ to obtain a culture solution.

< Example  2> Herbal Medicine Extract Malt Extract  Situation Mushroom Mycelium Culture in Medium

The herbal extracts were prepared by adding 200 g of ginseng, 200 g of Astragalus, 200 g of Aegye (mugwort), 200 g of Angelica vulgaris, 100 g of Morus bark (Mulberry bark), and 100 g of Ogapi in 2 L of purified water and then hot water at 110 ° C for 2 hours.

10% herbal extract and 1% malt extract were added to 1 L of distilled water to prepare a liquid medium and sterilized for 15 minutes in an autoclave. Aseptic inoculation of the spawn mushroom mycelium in the sterile medium obtained above and incubated for 20 days at 30 ℃ to obtain a culture solution.

< Comparative example  1> Potato Dextrose  Situation Mushroom Mycelium Culture Using Medium

0.4% g of potato starch and 2% dextrose were added to 1 L of distilled water to prepare a liquid medium and sterilized for 15 minutes in an autoclave. Aseptic inoculation of the spawn mushroom mycelium in the sterile medium obtained above and incubated for 20 days at 30 ℃ to obtain a culture solution.

< Comparative example  2> Malt Extract  Situation Mushroom Mycelium Culture Using Medium

Malt extract 2% was added to 1 L of distilled water to prepare a liquid medium, and then sterilized for 15 minutes in an autoclave. Inoculated aseptic seed of the mushroom mushroom mycelium in the sterile medium obtained above and incubated for 20 days at 30 ℃ to obtain a culture solution.

< Example  3> Situation Mushroom Mycelia Extract Powder Preparation and Extract Powder Beta Glucan  Content measurement

The cultures prepared in Examples 1 and 2 and Comparative Examples 1 and 2 were collected at 5 day intervals, extracted at 100 ° C. for 3 hours, filtered, and then 3 times the amount of 95% alcohol (v / v) was added to the filtrate. To precipitate the polysaccharide. The precipitate was recovered by centrifugation and then dried under vacuum at 50 ° C. to prepare a situation mushroom mycelium extract powder.

Beta-glucan was measured by using a Megazyme β-glucan assay kit (Megazyme International Ireland Ltd., Ireland) to determine the sugar content other than total glucan and glucan, and then excluding alpha-glucan and glucan. Obtained by subtracting the value of sugar content.

To 100 mg of mycelial powder of 0.5 mm or less, 1.5 mL of hydrochloric acid (37%, v / v) was added, and then the contents were mixed well, followed by mixing at 30 ° C. for 60 minutes at 15 minutes intervals. In order to decompose and remove starch and protein, 10 mL of distilled water was added thereto and then left in boiling water (100 ° C.) for 2 hours.

The contents were then cooled to room temperature, adjusted to pH by addition of 2N KOH, and diluted with 200 mM sodium acetate buffer (pH 5.0). Diluted contents were centrifuged to obtain supernatant, and then, 100 μL of the supernatant was extracted with axole, 3-β-glucanase (exo1,3-β-Glucanase; 100 U / mL) + β-glucosidase (β-Glucosidase; 20). unit / mL) solution was added to each 100uL and left for 1 hour at 40 ℃, 3mL of glucose determination reagent was added and then left at 40 ℃ 20 minutes. Thereafter, absorbance was measured at 510 nm to obtain a total glucan and sugar content values other than glucan. In addition, sugar content values other than alpha-glucan and glucan were added to 2 mg KOH 2mL in 100mg of mycelium powder, and stirred for 20 minutes in an ice-filled water bath using a magnetic bar, followed by 8mL of 1.2M sodium acetate buffer (pH 3.8). Added.

In addition, 200uL of a solution of Amyloglucosidase (Amyloglucosidase; 1630 unit / mL) + Invertase (500 unit / mL) was added thereto, and then allowed to stand at 40 ° C for 30 minutes, followed by 200 nM sodium acetate buffer. ; pH 5.0). Then, 3 mL of glucose determination reagent was added and left at 40 ° C. for 20 minutes. The absorbance was then measured at 510 nm to obtain sugar content values other than alpha-glucan and glucan.

Beta-glucan content was calculated by the following equation.

Beta glucan = (total glucan + sugar other than glucan)-(alpha glucan + sugar other than glucan)

<Experimental Results>

The mycelium extract powder prepared above was measured and compared with the culture solution, and the results are shown in Table 1.

<Table 1> Yield of powdered mushroom mycelium extract (%)

Incubation days 5 days 10 days 15th 20 days Example 1 0.27 0.58 0.69 0.66 Example 2 0.29 0.41 0.61 0.62 Comparative Example 1 0.17 0.25 0.31 0.39 Comparative Example 2 0.12 0.21 0.34 0.35

As shown in Table 1, it was found that the highest number of situation mushroom mycelium extract powder in Example 1 cultured in potato dextrose medium to which the cultivation days were added on the 15th day. It was found that the extraction powder yields of Examples 1 and 2 to which the medicinal herb extract was added were significantly higher than Comparative Examples 1 and 2 to which the medicinal herb extract was not added.

In addition, Examples 1 and 2, and Comparative Example 2 can be seen that the yield of the situation mushroom mycelium extract powder does not increase after 15 days of incubation, it is economically most suitable to use the culture date 15 days and extract the situation mushroom mycelium The powder yield was also found to be the highest.

<Table 2> Beta-glucan content (%) in the situation mushroom mycelium extract powder

Incubation days 5 days 10 days 15th 20 days Example 1 31.8 37.5 49.7 51.1 Example 2 30.4 33.6 38.4 39.7 Comparative Example 1 25.9 26.4 29.2 33.7 Comparative Example 2 23.4 27.6 28.7 31.4

As shown in Table 2, the beta glucan content of Examples 1 and 2 to which the medicinal herb extract was added was higher than that of Comparative Examples 1 and 2 to which the medicinal herb extract was not added. In addition, it can be seen that the highest beta glucan content was significantly higher in Example 1 cultured in a potato dextrose medium to which the medicinal herb extract was added. In Examples 1 and 2, the beta glucan content increased 12.2% and 4.3%, respectively, between 10 and 15 days of cultivation, and the highest increase was found to be less than 2% in subsequent cultures. .

 Comparing Examples 1 and 2, it was found that adding the medicinal herb extract to potato dextrose was higher than 10% of the beta glucan. It was found that the beta glucan content of Examples 1 and 2 to which the medicinal herb extract was added was higher than Comparative Examples 1 and 2 to which the medicinal herb extract was not added.

 The conventional method of artificially cultivating the situation mushroom fruiting body takes a long culture period of 3 to 6 months, making mass production difficult and not meeting the demand. On the other hand, the present inventors can not only shorten the incubation time by culturing the situation mushroom mycelium in the herbal medicine extract, but also obtain a large amount of beta glucan in a short time. Therefore, the situation mushroom mycelium culture medium which can increase the content of beta glucan in the situation mushroom culture medium can be widely used for artificial culture of situation mushroom mycelium.

Claims (3)

Herbal medicine extract consisting of ginseng, Astragalus, larvae, Mugwort, Angelica bark, Mulberry bark and Ogapi extract 5-20% by weight, potato starch 0.2-1% by weight, dextrose 1-5% by weight and distilled water 75-90 Situation mushroom mycelium culture medium composition comprising a weight%. Herbal medicine extract consisting of ginseng, Astragalus, larvae (mugwort), Angelica, Sangbaekpi (Mulberry bark) and Ogapi extract 5-20% by weight, malt extract 1-5% by weight and distilled water 75-90% by weight Situation mushroom mycelia culture medium composition. The method of claim 1 or 2, wherein the herbal extract is ginseng 200g, Astragalus 200g, 200g of Aegye (mugwort) 200g, Angelica 200g, 100g of Morus bark (Mulberry bark) and 100g of ogapi in 2L purified water and then 2 at 110 ℃ Situation mushroom mycelium culture medium composition, characterized in that extracted by hot water bath.