KR101459541B1 - Culture medium composition for Paecilomyces tenuipes including purified hemolymph from silkworm larvae and culture medium for Paecilomyces tenuipes including the same - Google Patents

Abstract

The present invention relates to a medium composition for culturing Paecilomyces tenuipes including purified hemolymph derived from silkworm larvae and to a medium for culturing Paecilomyces tenuipes including the same. The medium composition for culturing Paecilomyces tenuipes of the present invention comprises purified hemolymph gained by inputting the residual silkworm tissue after removing silk gland from silkworm larvae to distilled water, thermal treating, and obtaining supernatant through the continuous centrifugation. According to the present invention, provided are a medium composition for culturing Paecilomyces tenuipes which has high generation rate and growth rate of a fruit body of Paecilomyces tenuipes by comprising purified hemolymph derived from silkworm larvae as an active ingredient; and a medium for culturing Paecilomyces tenuipes including the same.

Description

TECHNICAL FIELD [0001] The present invention relates to a culture medium for cultivating snowy flower Cordyceps sinensis comprising a silkworm larvae derived from silkworm, and a medium for culture of snowy flower Cordyceps sinensis including the silkworm larvae derived from silkworm larvae derived from silkworm,

The present invention relates to a culture medium for cultivating snowy flower Cordyceps sinensis containing refined hemolymph derived from silkworm, and a culture medium for snowy flower Cordyceps sinensis containing the same, and more particularly, to a culture medium for snowy flower Cordyceps, To a culture medium for cultivating this highly snowy Chinese caterpillar fungus, and to a culture medium for snowy Chinese caterpillar fungus containing the same.

Insect body fluid has long been used as an additive to media for insect cell culture. Especially silkworms ( Bombyx mori fluids prolong cell survival by inhibiting cell apoptosis by a variety of chemicals such as baculovirus, actinomycin D, camptothecin, staurosporine, It is known that there is an effect of making.

The silkworm is a bulky insect that has been raised for a long time and is an insect that is easy to obtain. These silkworms, unlike other crops, are very sensitive to cleanliness and pesticides at the time of cultivation, and thus are widely used as experimental insects for stability testing. In addition, the silkworm body fluids have been approved for stability in items such as endotoxins.

The silkworm fluid is a solution that nourishes silkworms, called the silkworm hemolymph. This hemolymph has nutritional components necessary for the growth of animal cells including insect cells.

Thus, for experimental cultivation of insect tissue cells, silkworm hemolymph has been used as a culture medium by itself, and thereafter synthetic media have been developed which are defined based on the chemical analysis of the silkworm components of the silkworm.

Related Prior Art Korean Patent Publication No. 10-1996-0029454 entitled " Culture medium for animal cell culture containing silkworm fluid and method for culturing animal cells using the same, " and Korean Patent Publication No. 10-2001-0012745 ' And the cultivation method of Millitaris caterpillar fungus was used.

It is an object of the present invention to provide a culture medium for snow flower budworm culture which has a high rate of occurrence and growth rate of fruiting body of caterpillar fungus including the silkworm larvae derived from silkworm as an active ingredient and a culture medium for snow flower budworm seeds containing the same.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

In order to accomplish the above object, the present invention provides a culture medium for cultivating snowcone cicadenus, comprising a silkworm extract obtained from a silkworm, removing the silkworm from the silkworm, placing the remaining silkworm tissue in distilled water, heat treating it and then continuously centrifuging to obtain a supernatant, .

It is characterized in that the purified hemolymph contains 70 kDa protein.

The silkworm period is characterized by a nap.

The silkworm-derived purified hemolymph is characterized in that it is contained in an amount of 0.001 to 10% by weight based on the total weight of the culture medium for cultivating snow flower budworm seeds.

The heat treatment is performed at 40 to 80 ° C for 10 to 120 minutes

The method for producing a culture medium for snow flower corm bornea comprising the silkworm-derived purified hemolymph of the present invention comprises a first step of preparing a culture medium for cultivating snow flower budwing herb selected from among the first to fifth aspects, A second step of preparing a culture medium by mixing agar and distilled water in a culture medium for preparing a snowclow-line insect cell culture, and a second step of sterilizing the medium solution prepared in the second step at 115 to 125 ° C for 10 to 20 minutes, And a third step of manufacturing the second electrode.

In the second step, the agar is contained in an amount of 0.001 to 10% by weight based on the total weight of the culture medium.

In the second step, the distilled water is contained in an amount of 80 to 99.998% by weight based on the total weight of the culture medium.

The medium for cultivating ginseng Chinese cabbage seeds containing the purified blood lymph derived from silkworm of the present invention is characterized by being formed in a solid form by being produced by the above-mentioned production method.

According to the present invention, there is provided a culture medium for cultivating snowy flower cochassar larvae comprising a purified hemolymph derived from silkworm as an active ingredient, and a medium for cultivating snowy flower cochassaride containing the same, wherein the incidence and probability of snowy flower cochlear fruiting body is high, It will be possible to mass-produce mushrooms of snow flora.

Fig. 1 is an analysis of the protein of the purified silkworm-derived hemolymph of the present invention before application of the Q column.
FIG. 2 is a view showing proteins recovered through the Q column of the protein analyzed in FIG.
Fig. 3 is a diagram showing the thermal stability of the silkworm-derived hemolymph. Fig.
4 is a graph showing the antioxidative activity of the silkworm-derived hemolymph purified in Example 1 of the present invention.
Fig. 5 is a graph showing the generation characteristics of fruiting bodies of the snowy larvae cultured in the media of Examples 2 and 3 and Comparative Example 1 of the present invention.
Fig. 6 is a graph showing the incidence rate of fruiting bodies of the snowy caterpillar fungus grown in the media of Examples 2 and 3 and Comparative Example 1 of the present invention.
FIG. 7 is a graph showing the growth rates of mycelia of snowy larvae cultivated in the media of Examples 4 to 6 and Comparative Example 1 of the present invention. FIG.
FIG. 8 is a graph showing the growth rates of the mycelia of snowflake orchid crocus cultured in the media of Examples 4 to 6 and Comparative Examples 1 and 2 of the present invention.

The terms, techniques, and the like described in this specification are used in the meaning commonly used in the technical field to which the present invention belongs, unless otherwise specified.

Cordyceps is a medicinal mushroom that penetrates into the body of the host insect and forms fruiting body through host nutrients. Therefore, the nutrient source of Cordyceps is generally a nutrient of living insects. As such, the Chinese cabbage contains animal protein as a nutritional ingredient. Therefore, various physiologically active substances derived therefrom include useful substances that can not be found in ordinary mushrooms, that is, useful ingredients that can be used for the treatment of various human diseases, There is a high added value.

However, such a high value-added cordyceps has not been widely used due to difficulty in artificial cultivation due to its natural habitat. Therefore, the inventors of the present invention have found that it is possible to easily artificially cultivate the snowy caterpillar fungus having the above-mentioned functions, and in particular, to increase the incidence and yield of the females of the snowy caterpillar fungus, As a result of several studies, it has been found that refined hemolymph obtained by specific purification process of the previously abolished silkworm blood lymphed was used as an active ingredient of the culture medium for the snowy Chinese caterpillar fungus.

Hereinafter, the purification method of silkworm-derived purified hemolymph as an active ingredient of the culture medium for snowy flower cochassa larvae of the present invention will be described in detail as follows.

1. First step; Acquired tissue containing silkworm tissue from remaining silk gland

First, the silkworm is freeze-dried to remove the false gland from the silkworm.

Normally silkworm blood lymph has been collected by hand. However, when the blood lym- phs are taken from the raw silkworms by hand, the amount of melanin is reduced due to the low yield and is not economical. In addition, the blood silk is destroyed by the silkworm in the extraction process (the phenomenon that the silkworm vomits the digestive juice), so that a lot of impurities are contained.

Accordingly, the present invention first lyophilizes the silkworm to obtain a large amount of silkworm blood without impurities. In this case, it is preferable to treat the silkworm with liquid nitrogen and then freeze-dry the silkworm using a freeze-dryer to obtain a silkworm purification product.

The freeze-drying is preferably carried out at -40-70 ° C for 12-80 hours, which allows obtaining the most optimal silkworm blood without impurities.

The silkworm period may be any kind, but it is preferable that silkworms in which the mulberry leaves of the digestive tissues are fully digested and the false glands are maximally developed are preferably used to form silkworm cocoon.

The silkworm is the 5th to 7th day of silkworms. When the mulberry leaves in the silkworms of the silkworm are fully digested and the silkworms are fully developed and become clear and transparent, the silkworms are ready to become pupae. It is most economical because it can block the contamination of impurities due to large amount of blood lymphatic purification and digestive juice.

The lysosomal silk was removed from the lyophilized silkworm silk to obtain a tissue containing silkworm tissue. At this time, any method could be applied to remove the silkworm, but using a ball mill (diameter of 1 to 5 cm) Removal of the false gland can be used for other purposes, making it economically feasible.

2. The second step; Obtain a refined hemolymph

The separated silkworm tissue is placed in distilled water, heat-treated, and then subjected to continuous centrifugation to obtain a supernatant to obtain a purified hemolymph.

At this time, it is preferable that the distilled water contains 10 to 20 times of distilled water based on the weight of the tissue containing the separated silkworm tissue.

The heat treatment is carried out at 40 to 80 ° C. for 10 to 120 minutes. When the heat treatment is carried out at the same time, the tissue containing silkworm tissue is dissolved in distilled water and the tyrosinase activity in hemolymph by heat treatment Can be removed. In other words, toxic quinone protein produced by tyrosinase is terminated by heat treatment.

When the centrifugation is carried out at 12,000 to 15,000 rpm for 0.5 to 1 minute (flow rate: 0.8 L / min), the protein loss is minimized to obtain purified hemolymph.

At this time, the used centrifugal separator is a continuous centrifugal separator, which separates the supernatant while centrifuging, and is much more economical than the conventional centrifugal separation method (centrifugal separation followed by filtration to separate the supernatant), and the yield is high .

The purified hemolymph thus obtained may be used as an active ingredient of a culture medium for snow flower budworm seed culture, and a protein useful for the growth rate of budworm budworm fruit body in the purified hemolymph may be isolated and applied to the active ingredient It is also used in some cases.

The obtained purified hemolymph may be used as an active ingredient by separating the 70 kDa protein that affects the growth rate of the fruit fly mushroom fruiting body through the column.

At this time, any column may be used as the column, but preferably a Q column is suitable for separating the 70 kDa protein.

The Q column is a kind of ion exchange chromatography, which adjusts the pH of a buffer made with 20 mM Tris as a column having a strong negative charge. In other words, only the proteins with positive charges existing in the sample are adsorbed on the Q column, and then the proteins are separated by the difference of the charge value of the adsorbed protein by flowing 20 mM Tris and 1 M NaCl elution buffer. .

It is also possible to use size exclusion chromatography in addition to the Q column. In other words, size exclusion chromatography is a method of separating substances by size based on the interaction with the filler depending on the size and shape of the solute molecules in the sample. Therefore, 30 kDa and 70 kDa proteins, which are the major components of heat- .

Therefore, the 30 kDa protein and the 70 kDa protein can be separated using the size exclusion chromatography.

The protein purified through the above purification method is a 70 kDa protein, which has higher antioxidative effect than the 30 kDa protein and shows a high inhibitory effect on ROS production.

Thus, it becomes possible to use it as an active ingredient of a culture medium for cultivation of Snow Flower Cordyceps.

The purified hemolymph thus produced is characterized in that the growth rate of fruiting bodies of the seedlings is significantly higher than that of the medium containing the unpurified hemolymphs. It is preferable that the amount of the purified hemolymph is 0.001 to 10 wt% By weight, more preferably 1.0 to 5.0% by weight, and the balance of the remaining amount may include an acceptable material in the culture medium for other snowy larvae.

When the purified hemolymph is contained in an amount of less than 0.001% by weight, the content of the purified hemolymph is less than that of the purified hemolymph, The effect is not obtained and the blending ratio with other additional substances is not appropriate, which may adversely affect the growth rate of the fruit budworm fruit body.

Another aspect of the present invention is to provide a medium for cultivating snowy flower Cordyceps sinensis, which enhances the incidence and growth rate of fruiting body of snowy larvae, comprising the purified hemolymph or the 70 kDa protein isolated from the purified hemolymph, The medium for insect growth is prepared by mixing agar and distilled water with the active ingredient to prepare a culture medium, and then preparing a culture medium for sterilization.

At this time, the agar may be contained in an amount of 0.001-10 wt%, more preferably 1.5-10 wt%, based on the total weight of the culture medium. If the agar content is less than 0.001% by weight, the content of the agar is less than 0.001% by weight and the fruit body of the seedlings is difficult to grow. If the agar content is more than 10% by weight, There is a possibility that growth may be interrupted.

The distilled water is contained in an amount of 80 to 99.998% by weight, more preferably 93.5 to 97.5% by weight, based on the total weight of the culture medium. When the distilled water is contained in an amount of less than 80% by weight, the content thereof is too small and the fruiting body of the seedlings is difficult to grow. When the distilled water is contained in an amount exceeding 99.998% by weight, it becomes too thin to lose its shape as a solid medium .

In order to minimize the contamination degree during growth of the fruiting body of the snowy flower cochinacea and to minimize the change in the composition of the active ingredient contained in the culture medium for culture of the fir of the snowy flower cochinacea, It should be done for ~ 20 minutes.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples, but the scope of the present invention is not limited thereto.

Example 1 Production of silkworm-derived purified hemolymph of the present invention

One kilogram of silkworms (5th and 7th day) of silkworms were freeze-dried at -50 ° C for 70hours after liquid nitrogen treatment, and the remaining silkworms were recovered using a ball mill (diameter 1-5cm).

10 times of distilled water was added based on the weight of the recovered dried silkworm, and the solution was heat-treated for 60 to 30 minutes to dissolve. Then, centrifugation was performed at 12,000 to 15,000 rpm for 1 minute using a continuous centrifuge (flow rate: After separation, the supernatant was collected to obtain a purified hemolymph.

Example 2 Production of Medium 1 for Snow Flower Cordyceps Root Culture Including Silkworm-derived Purified Hemolymph of the Present Invention

0.6 g of purified hemolymph (hemolymph 2.4%) obtained in Example 1 and 0.375 g of agar powder were mixed in 25 mL of tertiary distilled water, and the mixture was sterilized at 115 to 125 DEG C for 10 to 20 minutes, Medium 1 for culture of Snow Flower Cordyceps.

Example 3 Production of Medium 2 for Cultivating Snow Flower Cordyceps, Including Purified Hemolymph from Silkworms of the Present Invention

1.2 g (hemolymph 4.8%) of the purified hemolymph obtained in Example 1 and 0.375 g of agar powder were mixed in 25 ml of the third distilled water and the mixture was sterilized at 115 to 125 ° C for 10 to 20 minutes, Medium 2 for cultivation of Snow Flower Cordyceps.

Example 4 Production of Medium 3 for Culture of Snow Flower Cordyceps sinensis Containing Purified Hemolymph Derived from Silkworm Silk of the Present Invention

0.5 g of the purified hemolymph obtained in Example 1 (hemolymph 2%) and 0.375 g of agar powder were mixed in 25 mL of tertiary distilled water, and the mixture was sterilized at 115 to 125 DEG C for 10 to 20 minutes, And medium 3 for culture of Snow Flower Cordyceps.

Example 5 Production of Medium 4 for Culture of Snow Flower Cordyceps, Including Purified Hemolymph from Silkworm of the Present Invention

0.75 g (hemolymph 3%) of purified hemolymph obtained in Example 1 and 0.375 g of agar powder were mixed with 25 mL of tertiary distilled water, and the mixture was sterilized at 115 to 125 DEG C for 10 to 20 minutes, Medium 4 for cultivation of Snow Flower Cordyceps.

Example 6 Production of Medium 5 for Culture of Snow Flower Cordyceps sinensis Containing Purified Hemolymph Derived from Silkworm Silk of the Present Invention

The purified hemolymph of 1.0 g (hemolymph 4%) obtained in Example 1 and 0.375 g of agar powder were mixed in 25 mL of tertiary distilled water and the mixture was sterilized at 115 to 125 ° C for 10 to 20 minutes, And medium 5 for cultivation of Snow Flower Cordyceps.

COMPARATIVE EXAMPLE 1 Production of PDA (Potato Dextrose Agar) Medium for Snow Flower Cordyceps

0.375 g of PDA (Potato Dextrose Agar) powder (PDA 2.4%) was mixed with 25 ml of distilled water, and the mixture was sterilized at 115 to 125 ° C for 10 to 20 minutes to prepare a culture medium for snow flower budworm seed culture.

≪ Comparative Example 2 > Preparation of medium for cultivating snowy flower Cordyceps sinensis containing 5th-3rd day silkworm-derived hemolymph

A needle was injected from the stomach to separate the blood lymph. 0.6 g of the silkworm blood (2.4% of hemolymph) and 0.375 g of agar powder were mixed in 25 ml of distilled water, and the mixture was sterilized at 115 to 125 ° C for 10 to 20 minutes, .

<Experimental Example 1> Effective protein separation in purified hemolymph of the present invention

One kilogram of the silk was treated with liquid nitrogen and then lyophilized (at -50 ° C for 70 hours) to produce silkworm dried matter.

From the silkworm dried material, the common glands were removed by using a ball mill (diameter of 1 to 5 cm), and the remaining hemorrhoids were recovered and separated.

The separated hemolymph was pulverized, 10 times of distilled water was added based on the weight of the pulverized material, and heat treatment was performed at 60 캜 for 30 minutes to prepare hemolymph lysate.

The hemolymph lysate was centrifuged at 12,000 rpm for 30 minutes to obtain a supernatant.

The obtained supernatant was separated from the glycoprotein through anion exchange chromatography.

As shown in FIG. 1, it was confirmed that 30 kDa and 70 kDa proteins were composed as main components. As a result, 30 kDa protein was recovered by 5 mg, and about 70 kDa protein was recovered by 42 mg.

In other words, the recovery of the 70 kDa protein having excellent physical properties among the 30 kDa and 70 kDa proteins was found to be 66% and the recovery rate was about 8 times higher than that of the 30 kDa protein.

<Experimental Example 2> Analysis of components of silkworm-derived hemolymph

1. Test Method

After removing silkworm silk gland, purified silkworm blood lymphedema was lyophilized and analyzed for general components such as moisture, crude protein, crude fat, ash, and inorganic matter.

2. Experimental results

Item Example 1-5 days 7 days (% by weight) 5 days 3 days Silkworm powder (% by weight) moisture 1.09 0.75 Views min 9.51 11.76 Crude fat 8.61 4.84 Crude protein 55.74 61.13 Crude carbohydrate 25.05 21.52

As shown in Table 1, the crude protein content was 55.74% by weight, the crude carbohydrate content was 25.05% by weight, the ash content was 9.51% by weight and the crude fat was 8.61% by weight in the order of the general components of the silkworm- .

Experimental Example 3: Thermal stability test of silkworm-derived hemolymph

1. Experimental Method

The tissues containing the hemolymph obtained from the silkworms after removal of the silkworms were divided into the before and after the heat treatment and the stability test of hemolymph was performed. At this time, the heat treatment proceeded at 60 ° C and 121 ° C.

SDS-PAGE analysis was performed to confirm the presence of hemolymph by heat treatment.

2. Experimental results

As shown in FIG. 3, it was confirmed that the hemolymph was maintained in the heat treatment condition at 60 ° C. (30 minutes). On the other hand, it was confirmed that the hemolymph was completely decomposed under the heat treatment condition at 121 ° C. (20 minutes).

In other words, the toxin quinone protein produced by tyrosinase was removed under the heat treatment condition at 60 ° C (30 min) and the hemolymph remained intact. However, in the heat treatment condition at 121 ° C (20 min) It means disassembled. Therefore, in the present invention, as a purification condition suitable for separating hemolymph from silkworm, heat treatment is performed at 60 DEG C for 30 minutes.

<Experimental Example 4> Antioxidant activity analysis of purified silkworm hemolymph in the present invention

1. Experimental Method

DPPH assay was performed to measure antioxidant activity of silkworm derived hemolymph.

In other words, 40 μl of the purified hemolymph of Example 1 was added to 160 μl of 0.2 mM DPPH dissolved in methanol, blocked with light, and reacted at 30 ° C. for 30 minutes.

Absorbance was measured at 517 nm using a microplate reader for the sample which had been completed.

Vitamin C was used as a control.

The antioxidant capacity of the measured hemolymph was shown in Table 2 and FIG.

2. Experimental results

sample IC ₅₀ ([mu] g / ml) Vitamin C 0.1 ± 0.06 Silkworm-derived hemolymph (Example 1) 15.3 + 4.8

As shown in Table 2 and FIG. 4, the antioxidant power of Example 1 of the present invention was 15.3 ± 4.8 g / ml, whereas the antioxidative power of 15% vitamin C, which is commonly used as a cosmetic material exhibiting antioxidant activity, was 0.1 ± 0.06 Mu] g / ml. That is, the antioxidative effect of the purified hemolymph of Example 1 of the present invention was found to be similar to that of 15% vitamin C.

<Experimental Example 5> Measurement of incidence rate of fruiting body

1. Experimental Method

The strains used in the experiment were Paecilomyces Tenuipes , strain J2) was used as a domestic strain isolated from Jamsa bean material and product utilization laboratory of the National Academy of Agricultural Science. The culture of the strain was carried out by using PDA (Potato Dextrose Agar) medium at 20 ~ 25 ° C at 150 ~ 200 rpm for 5 days Lt; / RTI &gt;

The cultured snowy parotyping strain was homogenized using a homogenizer, and then inoculated into the media for snowy Chinese caterpillar fungus cultivation of Examples 4 and 6 and Comparative Examples 1 and 2.

At this time, the cultivation conditions of the culture medium for the snowy flower Cordyceps sinensis were incubated for 10 days at 20 ~ 30 ° C under dark condition, and then for 16 days under dark condition at 15 ~ 25 ° C.

2. Experimental results

As shown in FIG. 5, the incidence of fruiting bodies of the seedlings was higher in the fruiting bodies of the present invention than those of the fruiting bodies of the present invention.

In order to measure the incidence of fruiting body, the cultured fruiting bodies were separated and lyophilized to measure the total weight. The results are shown in Fig.

As shown in FIG. 6, the productivity of the fruit bodies isolated in Example 2 was 396.98%, and that of the fruit bodies isolated in Example 3 was 722.64%, compared with the weight of the fruit bodies isolated in Comparative Example 1.

<Experimental Example 6> Measurement of Growth Rate of Mycelia of Snowy Flower Cordyceps mellifera

1. Experimental Method

The strains used in the experiment were Paecilomyces Tenuipes , strain J2) was used as a domestic strain isolated from Jamsa bean material and product utilization laboratory of the National Institute of Agricultural Science and was cultivated at 20 ~ 30 ℃ at 150 ~ 200 rpm for 5 days using PDA (Potato Dextrose Agar) Lt; / RTI &gt;

The culture of the above-mentioned snowflake Cordyceps sinensis strain was homogenized using a homogenizer, and then inoculated into the medium for the snow-crocodile culture of Examples 4 to 6 and Comparative Example 1.

At this time, the cultivation conditions of the culture medium for the snowy flower Cordyceps sinensis were incubated for 16 days at 20 to 25 ° C under dark conditions.

2. Experimental results

The growth rate was measured by the area of the mycelium grown on the medium using an image analyzer, and the result is shown in FIG.

As shown in FIG. 7, in the condition of culturing for 16 days, the growth characteristics were as high as 304% in Example 4, 308% in Example 5, and 316% in Example 6, respectively, as compared with Comparative Example 1 which is a control group.

The standard error, respectively Comparative Example 1 is 63.0mm ^2, Example 4 is 66.3mm ^2, Example 5 was 205mm ^2, Example 6 was analyzed to 117mm ^2.

<Experimental Example 7> Measurement of the growth rate of mycelia of the snowy flowers of various plants according to the amount of purified hemolymph from silkworm

1. Experimental Method

The strains used in the experiment were Paecilomyces Tenuipes , strain J2) was used as a domestic strain isolated from Jamsa bean material and product utilization laboratory of the National Academy of Agricultural Science. The culture of the strain was carried out by using PDA (Potato Dextrose Agar) medium at 20 ~ 25 ° C at 150 ~ 200 rpm for 5 days Lt; / RTI &gt;

The culture of the above-mentioned snowflake Cordyceps sinensis strain was homogenized using a homogenizer, and then inoculated into the medium for the snow-crocodile culture of Examples 4 to 6 and Comparative Example 1.

At this time, the cultivation conditions of the culture medium for the snowy flower Cordyceps sinensis were incubated for 16 days at 20 to 25 ° C under dark conditions.

2. Experimental results

The growth rate was measured by the area of the mycelium grown on the medium using an image analyzer. The results are shown in FIG.

As shown in FIG. 8, the growth rates of Example 4, Comparative Example 2, and Comparative Example 2 were 204%, 227% and 218%, respectively, The growth rate of Example 4 was 12.9%, that of Example 5 was 21.6%, and that of Example 6 was 18.3.

As described above, the culture medium for culturing the snow-capped Cordyceps mushroom containing the silkworm-derived purified hemolymph as an active ingredient and the culture medium for the snow-capped Cordyceps sinensis containing the silkworm, It can be seen that this is possible.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. You can implement the examples. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.

Claims (10)

The silkworm was removed from the silkworm, the remaining silkworm tissue was placed in distilled water, heat-treated, and then subjected to continuous centrifugation to obtain a supernatant, which contained the purified hemolymph as an active ingredient.
A medium composition for cultivating snowy flower Cordyceps sinensis comprising a silkworm derived from silkworm.
The method according to claim 1,
Wherein said purified hemolymph comprises a 70 kDa protein.
A medium composition for cultivating snowy flower Cordyceps sinensis comprising a silkworm derived from silkworm.
The method according to claim 1,
Characterized in that the silkworm period is a sleep-wake,
A medium composition for cultivating snowy flower Cordyceps sinensis comprising a silkworm derived from silkworm.
The method according to claim 1,
Wherein the silkworm-derived purified hemolymph is contained in an amount of 0.001 to 10% by weight based on the total weight of the culture medium for cultivating snow flower budworm,
A medium composition for cultivating snowy flower Cordyceps sinensis comprising a silkworm derived from silkworm.
The method according to claim 1,
Wherein the heat treatment is performed at 40 to 80 DEG C for 10 to 120 minutes,
A medium composition for cultivating snowy flower Cordyceps sinensis comprising a silkworm derived from silkworm.
A first step of preparing a culture medium for cultivating snow flower budworm seed according to any one of claims 1 to 5;
A second step of preparing a medium solution by mixing agar and distilled water in a medium composition for cultivating snow flower budworm prepared in the first step,
And a third step of sterilizing the medium solution prepared in the second step at a temperature of 115 to 125 ° C for 10 to 20 minutes to prepare a medium for cultivating snowy flower cochinacea.
A method for producing a medium for cultivating snowy flower Cordyceps sinensis containing refined hemolymph derived from silkworm.
The method according to claim 6,
Wherein the agar is contained in an amount of 0.001 to 10 wt% based on the total weight of the culture medium.
A method for producing a medium for cultivating snowy flower Cordyceps sinensis containing refined hemolymph derived from silkworm.
The method according to claim 6,
Wherein the distilled water in the second step is contained in an amount of 80 to 99.998% by weight based on the total weight of the culture medium.
A method for producing a medium for cultivating snowy flower Cordyceps sinensis containing refined hemolymph derived from silkworm.
A method for producing a microorganism of the present invention, comprising the steps of:
A culture medium for snow-white Cordyceps mushroom containing a silkworm-derived purified hemolymph.
10. The method of claim 9,
The medium for cultivating snowy flower Cordyceps sinensis is a solid form,
A culture medium for snow-white Cordyceps mushroom containing a silkworm-derived purified hemolymph.

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