KR101572638B1 - Paecilomyces hepiali CBG-CS-1 strain isolated form Cordyceps sinensis and method for culturing thereof - Google Patents

Abstract

The present invention relates to a process for the production of pacilomyces herpesviruses The present invention relates to a food composition comprising hepalin CBG-CS-1, a culture method of the above strain, a mycelium cultured by the above culture method, an extract of said mycelium, an extract of said mycelium or mycelium as an active ingredient, Paecilomyces The present invention relates to a culture medium for the culture of CBG-CS-1 strain and to a method for producing an extract using the cultured mycelium of the above- mentioned Lactobacillus cereus CBG-CS-1, Production and continuous raw material production and supply, it can be used industrially.

Description

[0002] The present invention relates to a method for producing Saccharomyces cerevisiae CBG-CS-1 isolated from Chinese caterpillar fungus, and a method for culturing Saccharomyces cerevisiae CBG-

The present invention relates to a strain of Fabolomyces hesiphilus CBG-CS-1 isolated from Cordyceps, and a method for culturing the strain. More particularly, the present invention relates to a strain of Paecilomyces hesiphilus isolated from Cordyceps militaris, The present invention relates to a food composition comprising hepalin CBG-CS-1, a culture method of the above strain, a mycelium cultured by the above culture method, an extract of said mycelium, an extract of said mycelium or mycelium as an active ingredient, Paecilomyces hepiali CBG-CS-1 strain, and a method for producing an extract using the cultured mycelium of the above- mentioned Fabolomyces hesiphil CBG-CS-1.

Cordyceps are also called caterpillar fungus in that they absorb insects' nutrients in the insect's body during the winter, kill the insects, and then pass through the body of insects in the summer to form grass-like fruiting bodies. Cordyceps has been traditionally used by some noble people along with Korean ginseng as a material of medicinal herbs from ancient times in China. Originally Cordyceps is Cordyceps ( Cordyceps ) from the larva of the moth and Hepialidae sinensis ) extracts. Today, however, insects as well as mushrooms from spiders and fungi are collectively called caterpillar fungus. Cordyceps is effective for hepatitis, diabetes, tuberculosis, etc. It is known that it has various effects such as improvement of blood circulation, effect as a tonic agent, and strengthening of immune function.

As the strains of the Cordyceps sinensis, the strains of the pupae ( Cordyceps millitaris , Paecilomyces japonica ), Scarab Cordyceps scarabaeicola , Cordyceps mutans ), cicadas Cordyceps sobolifera ), Bat moth larva Cordyceps sinensis ) are known. In particular, the moth larvae Cordyceps sinensis ) has been known for some time as its most effective drug.

The Cordyceps sinensis ) grows in the high mountains of more than 3,000 meters above sea level in the Tibet Mountains, and in Sichuan, Yunnan, Guizhou, Gansu, Qinghai, and Xizang in China, but it grows only in the highlands so it is not only very difficult to collect, There are no examples of successful artificial cultivation. Cord moth caterpillars Cordyceps sinensis ) is infested with mosquitoes and silent spores or hyphae in the moth larvae in winter and mycelium grows in the body of moth larvae in winter and the larvae gradually die. In summer, Cordyceps sinensis develops, and fruiting body grows at the head of the host and forms a baseball stick shape after penetrating the host. The total length of Cordyceps is about 10cm, circular shape with a diameter of about 0.5cm, which is initially light green and turns dark brown. The head of the fruiting body is slightly bulged and has a columnar shape and an elliptical shape. One nucleus contains many thin and long ones, and one sac contains eight cytoplasms.

Pupae Cordyceps militaris and Paecilomyces japonica have been successfully cultivated and have entered the stage of mass production. However, Cordyceps ( Cordyceps), which is known to have the greatest efficacy, Sinensis ) has not succeeded in artificial cultivation to date, so it depends only on natural acid. It is very difficult to collect it. Cordyceps sinensis ) in Lhasa area of Tibet city of Sichuan province is currently very expensive to trade at 35 million won / kg. In this way, because of its high price, fakes that artificially created a shape similar to that of Bat moth larvae are widely distributed. This is because the problem of the separation technique of the seed and the culture technique of the separated seed have not been developed. Therefore, only two techniques need to be developed to enable mass production of artificial insects, and it is possible to supply raw materials and standardize products for commercialization.

On the other hand, Korean Patent Registration No. 0415893 discloses a method of isolating Corysepssinensis bacteria, and Korean Patent No. 0459869 discloses a method of culturing Corysepssinensis in a large amount by using silkworm or silkworm pupa as a host Discloses a method for culturing Codishessinensis using silkworm or silkworm culture medium. However, the present invention relates to a method for culturing Facillomyces hepifolia CBG-CS-1 isolated from Cordyceps, There is no description.

The present invention has been made in view of the above-mentioned needs. In the present invention, strains were isolated, identified and cultivated from the tiger moth caterpillars of Tibetan wild moth, and the strains of the strains were identified. The strains were cultured, The present inventors completed the present invention by confirming that the extract of the artificial cultured mycelium has no significant difference from the active ingredient of the wild moth larvae Cordyceps sinensis extract of the present invention.

In order to solve the above-mentioned problem, the present invention relates to a method for the production of pacilomyces herpesvirus hepiali ) CBG-CS-1 strain.

The present invention also relates to a method for producing a yeast extract, which comprises inoculating the strain into a medium containing 3-4% (w / v) dextrose, 0.5-2% (w / v) yeast extract and 0.5-4% (w / v) , And culturing the cells at a temperature of 25 to 40 캜 for 5 to 6 days, wherein the method comprises culturing the strain of Fabylomyces heffilii CBG-CS-1.

The present invention also relates to a method for the production of a pharmaceutical composition which is cultured by the above method and wherein the main ingredient is selected from the group consisting of cordycepic polysaccharide, cordycepic acid, cordycepin and adenosine Or more of the mycelium of B. subtilis CBG-CS-1.

In addition, the present invention provides an extract of cultured mycelia of the above-mentioned Fabolomyces hesiphil CBG-CS-1.

In addition, the present invention provides a food composition comprising the cultured mycelium of the above-mentioned Lactobacillus bisporus CBG-CS-1 or an extract thereof as an active ingredient.

In addition, the present invention relates to a pharmaceutical composition comprising as an active ingredient a proteinaceous agent comprising 3 to 4% (w / v) dextrose, 0.5 to 2% (w / v) yeast extract and 0.5 to 4% (w / v) Paecilomyces hepiali CBG-CS-1 culture medium.

In addition, the present invention is (a) My process hepi Ali (Paecilomyces Indeed the L hepiali CBG-CS-1 strain for 5 to 6 days;

(b) concentration under reduced pressure at 58 to 62 ° C for 10 to 14 hours until the liquid culture medium of step (a) reaches 8 to 12% of the total volume;

(c) mixing 20 to 30% of the initial culture volume of 94 to 96% (v / v) ethanol with the reduced-pressure concentrate of step (b);

(d) extracting the mixture of step (c) at 85 to 95 ° C for 4 to 8 hours; And

(e) concentrating the extract of step (d) at a reduced pressure of 3 to 7% of the initial culture volume. The present invention also provides a method for producing a culture mycelial extract of Facillomyces hepifolia CBG-CS-1.

In the present invention, a large amount of the strain isolated from Tibetan wild mosquito larvae and the extracts from the cultured mycelium were compared with the extracts derived from wild mosquito larvae Cordyceps sinensis extracts, and the major components were found to be the same level. Since the instability of the raw mosquito larvae and the irregularity of the ingredient content are the greatest disadvantages, the strains isolated from the Tibetan wild moth larvae of the present invention can be used to produce raw materials of standardized standards and to continuously produce and supply raw materials It can be very useful for industrial applications.

FIG. 1 is a Tibetan wild Cordyceps bat moth larvae (Cordyceps sinensis ).
FIG. 2 is a photograph (A) of a mycelium cultured in a liquid phase and a result (B) in a solid culture dish after culturing a strain isolated from Tibetan wild mosquito larvae.
FIG. 3 is a graph showing the amount of change in cell weight over time in a large-scale 4-ton-scale culture of a strain isolated from Tibetan wild mosquito larvae.
FIG. 4 is a graph comparing the content of major components in a mycelial body artificially cultivated with a strain isolated from Tibet natural wild mosquito larvae and Tibet natural wild mosquito larvae, and (A) cordycepic polysaccharide and Cordycepic acid, cordycepin, and adenosine in (C), (C), and (C), respectively. Natural cordyceps; Unidentified Cordyceps, Fermented mycelia; Artificial culture mycelium.
FIG. 5 is a graph showing changes in the contents of major components with time in the culture of a strain isolated from Tibetan wild mosquito larvae, Cordyceps militaris, (cordycepic acid), (B) cordycepin and (C) adenosine.

In order to accomplish the object of the present invention, the present invention relates to a method for the treatment of Paecilomyces herpesvirus hepiali ) CBG-CS-1 strain.

The strains of the present invention, Facillomyces hesiphilus CBG-CS-1, were obtained from Cordyceps As a result of analysis of the 5.8S ribosomal DNA sequence, 99% or more identity with the Facillomyces herpeticus strain which can be isolated from Tibetan wild-branded Cordyceps were confirmed, and the strain was identified as a microorganism of the Korean Biotechnology Research Institute Deposited at the Resource Center on March 21, 2014 (Accession No .: KCTC 18275P).

Tibetan wild fungus of the invention is N-Sys hepi alruseu rhinorrhea (Hepialus biruensis), hepi alruseu dingyen sheath (Hepialus dingyeensis ), Hepialus namensis ), Hepialus namlinensis , Hepialus pui ), Hepialus zingazeensis ) or Hepialus ( Hepialus yadongensis ), and the like, but the present invention is not limited thereto.

The present invention also relates to the use of the above-mentioned < RTI ID = 0.0 > Paecilomyces & Hepiali The CBG-CS-1 strain was inoculated on a medium containing 3-4% (w / v) dextrose, 0.5-2% (w / v) yeast extract and 0.5-4% (w / v) And culturing the cells at a temperature of 25 to 40 ° C for 5 to 6 days. The method of culturing the strain of Fabilomyces hesieri CBG-CS-1 is also provided.

In the culture method of the present invention, the culture medium preferably contains 4% (w / v) dextrose, 1% (w / v) yeast extract, Peptone may be, but is not limited to, 1% (w / v).

In the method of culturing the strain of the present invention, the culture temperature is preferably 28 to 37 ° C, and the number of days of culture may be 5 to 6 days, but the present invention is not limited thereto.

The present invention also relates to a process for the production of a pharmaceutical composition which is cultured by the above method and wherein the main ingredient is selected from the group consisting of cordycepic polysaccharide, cordycepic acid, cordycepin and adenosine Characterized in that it comprises at least one < RTI ID = 0.0 > Paecilomyces < / RTI > hepiali ) CBG-CS-1.

In the cultured mycelium of facilomyces herpes complex CBG-CS-1 of the present invention, the content of the main component is 200-300 mg of a codispic polysaccharide per gram of cultured mycelia, 50-100 mg of a codisipic acid, 10 to 20 占 퐂, and adenosine 1200 to 1,500 占 퐂.

The present invention also provides an extract of the cultured mycelium.

The extract of the present invention may include, but is not limited to, one or more of the main components selected from the group consisting of a codisiperic polysaccharide, a codisipic acid, a codisepine, and adenosine.

The present invention also relates to the use of the above-mentioned < RTI ID = 0.0 > Paecilomyces & hepiali ) CBG-CS-1 or an extract thereof as an active ingredient.

The food composition of the present invention is preferably cultured in a medium containing at least one major component selected from the group consisting of a codedispic polysaccharide, a codedipic acid, a cordycepin, and adenosine. Which is a food composition containing an extract of mycelium as an active ingredient.

The food of the present invention may contain a functional food. In the functional food of the present invention, in addition to the above-mentioned active ingredients, various auxiliary ingredients may be added as necessary. In the case of the functional food of the present invention, vitamins such as vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B12, folic acid, vitamin C, vitamin D3 and vitamin E, Iron, magnesium, potassium, zinc, etc., or lactic acid bacteria.

In addition, among the functional foods of the present invention, health drinks may contain various flavors or natural carbohydrates as additional components such as ordinary beverages. Examples of the flavoring agent include natural sweetening agents such as tau martin and stevia extract, and synthetic sweetening agents such as saccharine and aspartame. Examples of natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol.

In addition, the present invention relates to a pharmaceutical composition comprising as an active ingredient a proteinaceous agent comprising 3 to 4% (w / v) dextrose, 0.5 to 2% (w / v) yeast extract and 0.5 to 4% (w / v) Paecilomyces hepiali CBG-CS-1 culture medium. The culture medium composition of the present invention may preferably include, but is not limited to, 4% dextrose (w / v), yeast extract 1% (w / v) and peptone 1% (w / v). In the culture medium composition of the present invention, the Fabolomyces helieri strain may be, but is not limited to, a strain of Paecilomyces hepiali CBG-CS-1.

In addition,

(a) contacting the above Paecilomyces < RTI ID = 0.0 > hepiali CBG-CS-1 strain for 5 to 6 days;

(b) concentration under reduced pressure at 58 to 62 ° C for 10 to 14 hours until the liquid culture medium of step (a) reaches 8 to 12% of the total volume;

(c) mixing 20 to 30% of the initial culture volume of 94 to 96% (v / v) ethanol with the reduced-pressure concentrate of step (b);

(d) extracting the mixture of step (c) at 85 to 95 ° C for 4 to 8 hours; And

(e) concentrating the extract of step (d) at a reduced pressure of 3 to 7% of the initial culture volume. The present invention also provides a method for producing a culture mycelial extract of Facillomyces hepifolia CBG-CS-1.

The method for producing the mycelial extract of cultured Mycosis herpirii CBG-CS-1 is preferably

(a) Paecilomyces < RTI ID = 0.0 > Hepiali The CBG-CS-1 strain was inoculated on a medium containing 3-4% (w / v) dextrose, 0.5-2% (w / v) yeast extract and 0.5-4% (w / v) Followed by liquid culture at a temperature of 25 to 40 DEG C for 5 to 6 days,

(b) concentration under reduced pressure at 60 占 폚 for 12 hours until the liquid culture medium of step (a) becomes 10% of the total volume;

(c) mixing 95% (v / v) ethanol of 25% of the initial culture volume with the reduced pressure concentrate of step (b);

(d) extracting the mixed solution of step (c) at 90 DEG C for 6 hours; And

(e) concentrating the extract of step (d) under reduced pressure to 5% of the initial culture volume, but it is not limited thereto.

In addition, in the method for producing the mycelial extract of cultured Mytilus CBG-CS-1 of the present invention, the cultured mycelial extract of the above-mentioned Fabolomyces hesiphil CBG-CS-1 is a cordycepic polysaccharide but are not limited to, one or more major components selected from the group consisting of polysaccharide, cordycepic acid, cordycepin, and adenosine.

The present invention also provides cultured mycelial extracts of Fabilomyces hesieri CBG-CS-1 produced by the above method.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  1. Isolation, culture and identification of the strain of the present invention

Tibetan wild Cordyceps bat moth larvae (Cordyceps sinensis, washed in FIG. 1) to sterile distilled water, and washed for 1 minute in an ultrasonic cleaner filled with a 95% ethanol 5 times to remove contaminants attached to the surface fungus. Immersed in 0.1% mercuric chloride (HgCl ₂ ) for about 5 minutes, and then subjected to shaking water treatment with sterilized water to prepare a sample.

In order to obtain spores, the fruiting body portion of the sample was cut into thin pieces and cultured in a PDA medium (300 g of potato pieces, 20 g of glucose, 15 g of agar, and 1,000 ml of distilled water) as a fungal solid medium for 5 days at 25 ° C. 2 (B)) were analyzed for the identification of 5.8S ribosomal DNA sequences for identification and identification of Paecilomyces herpesviruses hepiali ) was confirmed. The strain was deposited on March 21, 2014 at the Microbiological Resource Center of the Korea Research Institute of Bioscience and Biotechnology with Paecilomyces hepiali ) CBG-CS-1 (KCTC18275P).

Example  2. Large-scale liquid culture conditions

In order to maximize the cell yield according to the culture conditions of the isolated strains, the media components, culture time and temperature conditions were compared.

2-1. The amount of mycelium according to the medium composition

The medium was prepared by cultivating a culture medium for fungal growth culture, and cultured at 28 ° C. The mycelia were harvested according to the culture days for each culture medium, and the weight was measured after drying. Table 1 shows the composition of each medium, and Table 2 shows the result of measurement of the amount of cells according to the number of days of culture for each type of medium.

Dextrose (%, w / v) Yeast extract (%, w / v) Peptone (%, w / v) Badge 1 2 2 2 Badge 2 2 One 2 Badge 3 3 2 One Badge 4 3 One 3 Badge 5 4 One One Badge 6 2 3 3

division Number of cells per culture days (g / ℓ) 2 days 3 days 4 days 5 days 6 days 7 days Badge 1 0.0 0.1 0.4 0.6 0.6 0.5 Badge 2 0.0 0.1 0.3 0.5 0.5 0.5 Badge 3 0.0 0.1 0.5 0.8 0.8 0.7 Badge 4 0.0 0.2 0.5 0.8 0.8 0.7 Badge 5 0.0 0.2 0.6 1.0 1.0 0.9 Badge 6 0.0 0.1 0.2 0.4 0.6 0.5

The medium with the best cell mass was the medium 5 (dextrose 4% (w / v), yeast extract 1% (w / v), peptone 1% (w / v)), Showed the maximum amount of cells when cultured for 5 to 6 days (Fig. 3).

2-2. Incubation temperature

The medium was assayed by measuring the amount of the cells while culturing at 18, 28 and 37 ° C, respectively, using Medium 5, which produced the best amount of cell mass.

division Number of cells per culture days (g / ℓ) 2 days 3 days 4 days 5 days 6 days 7 days 18 ℃ 0.0 0.1 0.4 0.6 0.6 0.5 28 ℃ 0.0 0.2 0.6 1.0 1.0 0.9 37 ℃ 0.0 0.2 0.6 1.0 0.9 0.8

As a result of the measurement of the amount of microbial cells by the incubation temperature, it was confirmed that the microbial strain showed the maximum value at 28 ° C and 37 ° C for 5 days after the culture.

Example  3. Analysis of major components

The content of cordycepic polysaccharide, cordycepic acid, cordycepin and adenosine in the mycelial body after artificially cultivated and cultivated in the dry powdered state of the Tibetan wild moth larvae Respectively.

In order to maximize the utilization of the active ingredient from the cultured mycelium, the mycelial body was cultured for 5 days on a 4-ton scale, and then concentrated under reduced pressure at 60 ° C for 12 hours until it became about 400ℓ. Then, 100 L of 95% ethanol was mixed and subjected to an extraction reaction at 90 ° C. for 6 hours, and then the mixture was concentrated under reduced pressure to about 200 L. The samples for the analysis were dried and measured immediately. The raw materials for the manufacture of the product were lyophilized and powdered after mixing the excipients.

Codisepic polysaccharide content analysis was analyzed using the phenol-sulfuric acid method. Glucose was dissolved in water to make a standard solution of 0.1 mg / ml. Add 0.1 ml of phenol-sulfuric acid solution (80% of sulfuric acid solution) to each well of a 10 ml test tube, pour water up to 2 ml, add 0.6 ml of phenol-sulfuric acid solution 100 ml of phenol was added to dissolve the phenol, and the mixture was homogeneously mixed). Then, the mixture was heated in a thermostatic chamber at 100 ° C for 1 hour, and then transferred to an ice bath and cooled for 15 minutes. The absorbance of each standard solution was measured at a wavelength of 625 nm using a spectrophotometer, and then a standard curve was prepared by setting the measured absorbance to the ordinate and the concentration to the abscissa. For the preparation of the experimental solution, 0.06 g of the sample powder was placed in a round bottom flask, and 100 ml of 80% ethanol was extracted by reflux extraction for 1 hour. After the filtration, the remaining residue was washed three times with 10 ml of 80% ethanol. 100 ml of distilled water was poured into the beaker and the filtrate was immediately filtered after 1 hour of heating at 90 ° C. The residue was washed three times with 10 ml of warm water and the liquid was combined with the filtered liquid. And diluted to the scale. 2 ml of the test solution was placed in a 10 ml test tube, 6 ml of phenol-sulfuric acid was added to measure the absorbance, and the weight (mg) of the glucose contained in the test solution was read using a standard curve. Respectively. As a result, it was confirmed that the content of the corysepic polysaccharide of cultured mycelium was 440% higher than that of wild-type Cordyceps (Fig. 4A).

About 0.5 g of the cultured mycelium was extracted with 5 ml of 30% methanol (v / v) for 30 minutes for centrifugation. 1 ml of the supernatant was taken, 5 ml of p-nitrobenzoly chloride (PNBC) was added, and the derivatization reaction was carried out at 50 ° C for 90 minutes. To stop the reaction, 5 to 6 drops of methanol were added, concentrated, and re-dissolved in 5 ml of chloroform. A redissolved sample was passed through a Sep-pak cartridge activated with n-hexane and then a 10% ethyl acetate / n-hexane mixture was poured off and then 25 ml 10% ethyl acetate / n-hexane mixture. The eluate was redissolved in acetonitrile after concentration in nitrogen, each sample was diluted to a certain concentration, and the supernatant was used as the sample solution. 10 [mu] l of this was analyzed by high performance liquid chromatography (HPLC). The results are shown in Fig. It was confirmed that the content of the cultivated mycelium of cordispic acid was higher by about 10% than that of wild-type Cordyceps (Fig. 4A).

Approximately 1 g of cultured mycelium was extracted for 10 minutes and 30 ml of methanol was extracted for centrifugation. The sample was diluted to a certain concentration and the supernatant was used as the sample solution. 15 μl of the sample was analyzed by high performance liquid chromatography (HPLC). As a result, the content of cordisepin and adenosine in cultured mycelium was found to be almost similar to that of wild-type Cordyceps (Fig. 4B and C).

Example  4. Analysis of main ingredient content according to culture date

According to the optimized culture conditions of Example 2, strains isolated from Tibetan wild mosquito larvae were artificially cultured and analyzed for changes in the contents of the codispic polysaccharide, codisepic acid, cordisepin and adenosine in the cultured mycelium . The analysis method is as described above.

As a result, the content of the codicepic polysaccharides, the codedipic acid, the codedispin and the adenosine started to increase from the third day after the culture and reached the highest level on the fifth day after the culture (FIG. 5).

Korea Biotechnology Research Institute KCTC18275P 20140321

Claims (12)

A cordycepic polysaccharide high-production Paecilomyces hepiali ( Pseudomonas spp. ) Isolated from Tibetan wild mosquito larvae Cordyceps sinensis, and the content of codispic polysaccharides per g of cultured mycelium is 200 to 300 mg. ) CBG-CS-1 strain (KCTC 18275P). [3] The method of claim 1, wherein the T. naturally occurring Chinese caterpillar fungus is selected from the group consisting of Hepialus biruensis , Hepialus dingyeensis , Hepialus namensis , Hepialus namlinensis , A cordycepic polysaccharide high production filamentous herpes virus CBG ( HepG2 ), which is hosted by Hepialus pui , Hepialus zingazeensis or Hepialus yadongensis , -CS-1 strain. The strain of claim 1 is inoculated into a medium containing 3-4% (w / v) dextrose, 0.5-2% (w / v) yeast extract and 0.5-4% (w / v) And culturing the cells at a temperature of 40 占 폚 for 5 to 6 days. The method of culturing the strain of Cordycepic polysaccharide-producing Fabolomyces hesieri CBG-CS-1. A cordycepic polysaccharide high-producing Paecilomyces hepiali CBG-CS, which is cultured by the method of claim 3, wherein the major component is a cordycepic polysaccharide. -1. delete An extract of the cultured mycelium of cordycepic polysaccharide high production Paecilomyces hepiali CBG-CS-1 of claim 4 . A method for the production of cordycepic polysaccharide high production Paecilomyces hepiali CBG-CS-1 cultured mycelium of claim 4 or high production of cordycepic polysaccharide of claim 6 A food composition comprising an extract of cultured mycelium of Paecilomyces hepiali CBG-CS-1 as an active ingredient. delete (a) culturing the Cordycepic polysaccharide high producing Paecilomyces hepiali CBG-CS-1 strain of claim 1 for 5 to 6 days;
(b) concentration under reduced pressure at 58 to 62 ° C for 10 to 14 hours until the liquid culture medium of step (a) reaches 8 to 12% of the total volume;
(c) mixing 20 to 30% of the initial culture volume of 94 to 96% (v / v) ethanol with the reduced-pressure concentrate of step (b);
(d) extracting the mixture of step (c) at 85 to 95 ° C for 4 to 8 hours; And
(e) concentrating the extract of step (d) at a reduced concentration of 3 to 7% of the initial culture volume to produce a cordycepic polysaccharide high-production filamentous herpes complex CBG-CS-1 A method for producing a cultured mycelium extract. 10. The method according to claim 9, wherein the culture of step (a) comprises culturing the cordycepic polysaccharide high production strain Paecilomyces hepiali CBG-CS-1 of claim 1 in an amount of 3-4% (w / v) dextrose, 0.5-2% (w / v) yeast extract and 0.5-4% (w / v) peptone, Wherein the cultured mycelial extract of cordycepic polysaccharide high-yielding filamentomyces hesieri CBG-CS-1 is cultured in a culture medium. delete A cultured mycelium extract of cordycepic polysaccharide high-production filamentomyces hesieri CBG-CS-1 produced by the method of claim 9.

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