JPH1042691A - Artificial culture of cordyceps sinensis sacc. - Google Patents

Abstract

PROBLEM TO BE SOLVED: To efficiently and artificially culture Cordyceps sinensis Sacc. useful in the fields of medicines, health foods, cosmetics, etc., to produce the Cordyceps sinensis Sacc. in a large amount by using a culture medium containing the components of the chrysalises of silkworms as main components to utilize the chrysalises of the silk-removed silkworms or the chrysalises of defective cocoons. SOLUTION: This method for artificially culturing Cordyceps sinensis Sacc. comprises using a culture medium such as an extract solution which is obtained by boiling down the chrysalises of silkworms, such as the chrysalises of silk- removed silkworms, with water in a weight amount of 2-50 times those of the chrysalises at a boiling temperature for >=2hr and which contains the components of the chrysalises as main components. The culture medium preferably further contains one or more kinds of components selected from carbon sources, amino acids, minerals, and vitamins in a total amount of 2-8 pts.wt. per 100 pts.wt. of the extract of the chrysalises of the silkworms.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to 18 kinds of amino acids, such as calcium, chromium, nickel, iron, zinc, manganese, adenine, adenosine, uracil, eicosan alcohol, mushroom sugar, ergostan, alkaloid, vitamin B12 and the like. It mainly has effects on the central nervous system (sedative effect), immunity and immune regulation, effects on the cardiovascular system such as arteriosclerosis, effects on the respiratory system such as asthma, effects on energy metabolism, The present invention relates to a method for artificially cultivating Cordyceps sinensis, which is believed to have a nutrient tonic effect and other anticancer effects, and is effective in fields such as pharmaceuticals, health foods, and cosmetics.

[0002]

BACKGROUND OF THE INVENTION Cordyceps has been prized by some people as a secret drug transmitted from ancient China. In recent years, effects of cordyceps as a secret drug and component analysis have been performed, and its use as a carcinostatic has attracted particular attention. In addition, due to changes in lifestyle and heightened health consciousness, their use as health foods is also increasing. On the other hand, looking at the state of supply of cordyceps, many of them are still obtained by collecting those in nature. However, Cordyceps is difficult to collect and cannot be supplied in large quantities because it is not parasitic on plants like other mushrooms, but also on insects. Therefore, the following artificial culturing method has been proposed. First, as a first artificial culturing method, the hypha or the spores of Cordyceps sinensis are garlic and garlic.
The culture broth mixed with soy sauce and sugar is sterilized and subjected to liquid culture by a conventional method, or this liquid culture broth is used for cereals such as rice, barley, corn, etc. or adult insects of arthropods such as silkworms and cicadas.
A method of performing solid culture by adsorbing to pupae, larvae, and the like has been proposed (Japanese Patent Application Laid-Open No. 54-80486). As a second artificial culturing method, the mycelium of Cordyceps sinensis is composed mainly of one or several kinds of sugars, protein substances, vitamins, nucleic acids, etc., and cereals are added to these main ingredients to form a solid solid medium. After transplanting and culturing, the mycelium cultured in this three-dimensional solid medium is composed mainly of one or several kinds of saccharide protein substances, vitamins, nucleic acids, etc., and one or more kinds of amino acids are added thereto. A method has been proposed in which the cells are transplanted into a liquid medium having a pH of 4.0 to 7.0 and used as a substrate for culturing, and cells are formed on the surface of the liquid medium (JP-B-61-53033). The third artificial culture method is to directly infect and inoculate living insects with the mycelium of Cordyceps sinensis, or to add a one-third amount of insect tissue to a pure isolation medium using agar as a substrate. (Japanese Patent Application Laid-Open No. 107725/1987). In addition, when using a living insect, the silkworm immediately before forming a cocoon is used. As a fourth artificial culture method, a method has been proposed in which a cordyceps cordis present in the field is collected, hyphae and conidia are grown on an agar medium or a liquid medium, and final mass growth is performed by field culture ( JP-A-6-233627).

[0003]

As the inventor who proposed the third artificial culturing method has pointed out the second artificial culturing method proposed by himself, the second method is a continuous method. However, it cannot be said that this is a sufficient culture, and the problem of a decrease in pharmacological activity is also considered. The biggest cause is that components other than insects are used as a medium. Therefore, the same problem can be considered in the first method, and it is not preferable. On the other hand, in order to solve the above problem, the third method proposes to use a living insect itself as a medium composition as in nature. However, various germs are breeding on and in the body of the field insects, and the infection rate of Cordyceps is low, so that it is impossible to supply the same species stably. Also, in the fourth artificial culturing method, in order to perform field culture, the ratio of contamination with other bacteria cannot be said to be low, and continuous culturing should be performed stably as in the third method. Is not easy. In addition, there is also a problem that spores are scattered when field culture is performed, which may have a great effect on the natural world. On the other hand, the pupa after silkworm cocoon reeling is now washed and dried and used for animal feed, etc. Therefore, effective utilization of pupae after spinning is desired.

[0004] Therefore, an object of the present invention is to propose a method for artificially cultivating Cordyceps sinensis capable of stably cultivating Cordyceps sinensis in mass production. Specifically, an object of the present invention is to propose a method for artificially cultivating cordyceps, which effectively prevents the decrease in pharmacological activity while effectively utilizing the components of the insect itself. It is another object of the present invention to increase the survival rate of the silkworm pupa itself, prevent a decrease in pharmacological activity, and propose a method for artificially cultivating Cordyceps with a high infection rate. Another object of the present invention is to propose a method for artificially cultivating Cordyceps sinensis, which allows stable and large-scale growth and subculture with nutrients in a state closer to the natural world. A further object of the present invention is to effectively utilize silkworm pupae after cocoon reeling and defective cocoon pupae that cannot be used for reeling.

[0005]

Accordingly, the method for artificially cultivating cordyceps according to the present invention is characterized in that the cultivation is carried out using a medium mainly containing the composition of silkworm pupae. Claim 2
The method for artificially cultivating cordyceps according to the present invention is characterized in that, in the method for artificially cultivating cordyceps according to claim 1, as the pupa, a pupa after thread reeling is used. The method for artificially cultivating Cordyceps according to the third aspect of the present invention is the first aspect.
Alternatively, in the method for artificially cultivating Cordyceps sinensis according to claim 2, an extract obtained by extracting a composition component of silkworm pupa using water at high temperature and heating is used as the medium.
The method for artificially cultivating cordyceps according to the present invention according to claim 4 is the method for artificially cultivating cordyceps according to claim 3, wherein the extract uses 2 to 50 times the weight ratio of water to pupae, It is characterized by being extracted by boiling over at the boiling temperature for 2 hours or more. Further, the method for artificially cultivating a cordyceps according to the present invention according to claim 5 is the method for artificially cultivating a cordyceps according to claim 3, wherein the extract uses water at a weight ratio of 2 to 50 times the pupa, It is characterized by being extracted by boiling for 20 minutes or more at a temperature higher than the boiling temperature. The method for artificially cultivating cordyceps according to the present invention according to claim 6 is the method for artificially cultivating cordyceps according to claim 1 or 2, wherein the pupa is pulverized as the medium and water is added thereto. Is used. The method for artificially cultivating cordyceps according to the present invention according to claim 7 is the method for artificially cultivating cordyceps according to any one of claims 3 to 6, wherein
It is characterized by adding at least one of carbon sources, amino acids, minerals, and vitamins. The method for artificially cultivating cordyceps according to the present invention according to claim 8 is the method for artificially cultivating cordyceps according to claim 7, wherein the amount is 2 to 8 by weight with respect to 100 extracts of silkworm pupae. It is characterized by having done. Further, the method for artificially cultivating cordyceps according to the present invention according to claim 9 uses aseptic silkworm as the silkworm, takes out a pupa after the silkworm has formed a cocoon, and the aspergillus or spores formed in the fruit body of the cordyceps in the pupa. It is characterized by culturing by inoculating conidia. In a tenth aspect of the present invention, there is provided a method for artificially cultivating a Cordyceps sinensis according to the ninth aspect, wherein a primary pupa is used as the pupa. Claim 11
The method for artificially cultivating a cordyceps cordis according to the present invention is as follows: the cordyceps cordis is grown and cultured in a liquid medium mainly containing the composition of silkworm pupae, and thereafter, ascospores or conidiospores formed in the fruit body of the cordyceps. It is characterized in that subculture is performed by inoculating spores directly into aseptically reared silkworm pupae. According to a twelfth aspect of the present invention, there is provided the method for artificially cultivating a cordyceps as set forth in any one of the first to eleventh aspects, wherein the cordyceps cordyceps is used as the cordyceps.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION Cordyceps type (Cordyceps), which produces ascospores (complete generation), and naked conidiospores (incomplete generation), are produced as cordyceps plants to be cultured in the present invention. Any of the Isaria type (genus Isaria) may be used. In addition, it is necessary to obtain a seed fungus in advance. For this purpose, actually matured cordyceps is collected in the field and separated and cultured on an agar medium according to a conventional method.

First, an artificial culturing method using a liquid medium containing a silkworm pupa as a main component will be described below. First, the silkworm pupa used as the main component of the medium may be a live pupa cut from a cocoon and taken out, or a dried pupa dried at the cocoon stage. Further, a raw pupa or a dried pupa after the reeling can be used. The pupa after spinning contains 60% of crude protein and 9% of total nitrogen, as well as a large amount of nutrients necessary for the development of cordyceps, such as ash and glycogen.

[0008] As a method of extracting the constituent components of the pupa, there are a method of boiling and a method of extracting with a high-pressure steam sterilizer. When extracting below the boiling temperature, use the boiling method. When a high-pressure steam sterilizer is used, extraction can be performed at a temperature higher than 100 ° C., so that an effective ingredient can be extracted in a short time. The silkworm pupa extract thus extracted is appropriately diluted with water to obtain a culture solution. When extracting pupal components under high temperature, water that is 2 to 5 times the weight of pupa 1 is used. Furthermore, when used as a culture solution, this extract solution may be diluted with 5 to 10 times the weight of water. In the case where the dilution is not performed, the pupa 1 may be boiled with 10 to 50 times the weight of water. In addition,
The amount of water lost by evaporation is added.

The pupal extract extracted as described above contains:
Among carbon sources, amino acids, minerals, and vitamins, necessary components are added depending on the type of cordyceps, sterilized at 121 ° C. for 15 minutes using a high-pressure steam sterilizer, and used as a liquid medium after natural cooling. These additional components are
It is about 2 to 8% based on the pupa extract. Examples of the carbon source include glucose, mannose, maltose, pectose, sucrose, and starch. The mineral components include phosphorus, calcium, potassium, sodium, and magnesium. Next, the separated strain previously separated and cultured is planted and cultured in the liquid medium thus prepared. In addition, subculture can be performed by inoculating ascospores and conidia formed by artificial culture. At this time, the culture conditions were set at a temperature of 15
It is preferable to keep the temperature between 25 ° C and 25 ° C and between 75% and 95%. The present invention can prevent a decrease in pharmacological activity while effectively utilizing the components of the insect itself by culturing using a medium containing an extract component of silkworm pupa as a main component, Continuous culture can be performed stably.

[0010] As a method for extracting pupa composition components, a method of boiling and a method of extracting with a high-pressure steam sterilizer have been described. In these boiling methods, it is more preferable to grind the pupae in advance. In addition to these boiling methods, the dried pupa may be crushed and sterilized, and sterilized water may be added thereto. At this time, not only water but also a carbon source, amino acids, minerals,
It is preferable to add nutrients such as vitamins and the like as appropriate. As described above, the essence of the silkworm pupa extract is the main component, and further components, such as carbon sources, amino acids, minerals, and vitamins, which are more preferable for the cultivation of cordyceps, are added as needed. Stable culture can be performed stably. Further, the above method has been described with reference to a liquid medium, but an agar medium may be used.

Next, an artificial culturing method using sterile silkworms and using the pupa itself as a medium will be described below. Here, the sterile silkworm is a silkworm reared by the artificial feed aseptic breeding method, and the sterilized egg surface is sterilized, and the aseptically hatched silkworm is fed with steam-sterilized feed, and sterilized environment is used by using a sterile apparatus or the like. Refers to silkworms raised below. Here, aseptic means that it does not originally contain any microorganisms, including viruses.
More than 00 and less than 3000 do not cause disease ("A new anniversary sericulture in Japan based on artificial feed aseptic breeding method", Kyoto Institute of Technology, Faculty of Textile Science, vol. 16, separate volume, published on March 16, 1992, Fujiwara Matsubara Good book)
Germ-free silkworm here is not necessarily germ-free
free) It is not necessarily silkworm. That is, even in the artificial feed aseptic breeding method, even microorganisms in the body of the silkworm are not sufficiently controlled. Therefore, the term “germ-free silkworm” as used herein refers to one that excludes the main pathogens that cause silkworm disease. Regarding the method of aseptic breeding of artificial feed, see "Study on the introduction of aseptic breeding of artificial feed into sericulture systems", Kyoto Institute of Technology, Academic Report, Faculty of Textile Science, Vol. 15, separate volume, March 1991.
Published on March 15th, "Sterile breeding of silkworms at the age of 5 years using artificial feed containing okara as a main component", The Japanese Society of Silk Science, Vol. 60, No. 6,
And Japanese Patent Application No. 260927.
The pupa after the silkworms, which have been germ-free aseptically and formed a cocoon, are used as a medium. Primary pupae immediately after pupation are most preferred. Here, the primary pupa is a pupa in a state before the compound eye turns black. Since the pupae are usually covered with cocoons, the pupae are artificially removed from the cocoons. And to this pupa,
Ascospores or conidiospores formed on the fruiting bodies of Cordyceps sinensis cultured in the aforementioned liquid medium are directly inoculated. At this time, the culture temperature is set to 15 ° C. to 25 ° C., and the culture is continued under aseptic conditions. In the artificial culture method of the present invention, silkworms that have been cleaned and grown for a limited period, such as during the age of 5 years, may be used.

As described above, according to the present invention, the use of silkworms bred using an artificial feed in an aseptic apparatus eliminates the propagation of various bacteria, increases the infection rate of Cordyceps sinensis and the formation rate of fruiting bodies, and enables continuous culture. Can be performed stably. Furthermore, since the pupa after the formation of the cocoon is directly inoculated, the infection rate of the fungus to the silkworm is higher than in the larva stage, and the culture can be performed stably. In addition, the present invention can be cultured in a nutrient in the same state as in nature by growing and culturing Cordyceps sinensis in a liquid medium, and then performing subculture by directly inoculating pupae of aseptically reared silkworms. At the same time, stable supply of the medium is easy, and propagation of various bacteria is eliminated, and stable mass production becomes possible.

[0013]

Embodiments will be described below. Example 1 Using a liquid medium containing a pupa extract as a main component, an artificial cultivation of a flower of Isaria Japonica was performed. The pupa extract is obtained by removing the dried pupa dried in the cocoon stage from the cocoon,
Boiled for 2 hours at boiling temperature using twice the weight of water. Then, this extract was further diluted with water five times the weight to obtain a liquid medium. About 1/3 of this liquid medium was placed in a test tube, and an isolated strain of Hanasagitake was planted on the liquid medium. Hyphae began to form in the liquid medium two to three days after planting, a mycelial layer was formed in one week, and loci were formed two weeks later. After 45 to 60 days, a 4-8 cm fruiting body was formed and matured. The culture was performed at a temperature of 15 ° C. to 25 ° C. and a humidity of 75% to 95% under sunlight and natural light. Also,
1.5% glucose against pupae extract 100, peptone 0.2%, those of MgSO ₄ · 7H ₂ O was added 0.4% were artificially cultured as a liquid medium. The addition of these materials resulted in brighter fruit body and better growth than when they were not added. In addition, Konasanagitake (Is
aria Farinosa) was also artificially cultured using a pupa extract as a liquid medium in the same manner as in the above-mentioned artificial culture of Anopheles mushroom. White hyphae began to form 3 days after the planting of the Pleurotus terrestris, planting loci 11 days later, and matured 45 to 50 days later. The formed fruiting bodies were 2-3 cm thin and pale yellow. In addition, when artificial culture was performed using the pupal extract with a carbon source added as a liquid medium, there was no difference from the pupal extract with no addition during the formation of mycelia or loci and the maturation period. However, when the carbon source was added, a broom-like, light brown, and vivid fruit body was formed, which was slightly longer, at 3 to 4 cm, than that without the carbon source.

Example 2 The spores of Anopheles velutipes obtained above were collected, and the spores were suspended in a 5000-fold solution of Tween 40. The pupa immediately after pupation of the silkworm grown on a sterile artificial feed was taken out. The body surface was inoculated directly. The culture temperature was 15 to 25 ° C. Two to three days after the inoculation, the silkworms became hard, and hyphae were formed from the joint membrane and the stomata of the body surface, and the infection rate was 99.5% or more. After 12 to 13 days, a loci was formed, and after 30 to 50 days, a 5-7.5 cm fruiting body was formed and matured. In addition, a locus is formed in the same manner for Konasanagitake,
The fruiting body formed and matured. Table 1 shows the results of the component analysis of the thus obtained Hanasagi mushrooms.
As a comparative example, a natural Cordyceps cordis from Tibet, China (Cordyc
eps Sinensis). As the specimen, Hanasa-nagatake was dried at about 60 ° C. and pulverized. Natural caterpillars were purchased dried. In addition, selenium which is an analytical test item is said to have a cancer preventing effect, β-glucan has an effect of enhancing the body's immune function, and ergosterol is said to have an effect of enhancing immunity and suppressing cancer.

[0015]

[Table 1]

The analysis method in Table 1 is as follows.分析 Analysis method 1) Selenium 2,3-diaminonaphthalene-based on fluorometry. Nitric acid (20 ml) and perchloric acid (10 ml) were added to 1 g of the sample, and the mixture was heated and decomposed. Then, 10% hydrochloric acid was added, and the mixture was heated in boiling water for 30 minutes, so that the pH became 1 to 1.5. Was adjusted as follows. Thereafter, a 2,3-diaminonaphthalene solution was added, the mixture was warmed at 50 ° C., shaken and allowed to stand, and then the cyclohexane layer was measured with a fluorescence spectrophotometer.
The excitation wavelength is 378 nm and the fluorescence wavelength is 520 nm. 2) β-glucan Enzyme method. Add 0.08 M phosphate buffer and Termamyl to 1 g of sample, warm with boiling water for 30 minutes,
After adjusting the pH to 7.5, a protease solution was added thereto, and the mixture was allowed to stand at 60 ° C. for 30 minutes. After adjusting the pH to 4.3, an amyloglucosidase solution was added, and the resulting solution was quadrupled with 95% ethanol. Thereafter, the mixture is left at room temperature for 1 hour to precipitate β-glucan. Thereafter, treatments such as filtration, washing, hydrolysis, neutralization, constant volume, and filtration were performed, and the filtrate was quantified for glucose by a glucose oxidase method. β-glucan (%) = glucose (%) × 0.9 3) Ergosterol By high performance liquid chromatography. 0.5% sample was added with 1% sodium chloride solution, 1% pyrogallol-ethanol solution, 60% potassium hydroxide solution and potassium hydroxide, and saponified in 70 ° C. water for 30 minutes, and 1% sodium chloride solution was added. And perform extraction. Further, a mixed solution of ethyl acetate and hexane was added thereto, and the solvent was distilled off. As the column, Nova-Pak C18 was used, and 35
C. 1.0 ml / min, UV 282 nm.

Example 3 Next, the infection rate of Cordyceps sinensis using pupae of silkworm grown on a sterile artificial feed was tested as a comparative example using pupae of silkworm grown on mulberry leaves. As the fungus species of Cordyceps, Hanasagitake was used. The spores of pupae are Tween 40 5000
The pupa of the silkworm grown on aseptic artificial feed and the silkworm grown on mulberry leaf immediately after pupation were taken out and inoculated directly to the body surface of the pupa. The culture temperature was 15 to 25 ° C. The results are as shown in Table 2.

[0018]

[Table 2]

The infection rate of Cordyceps sinensis using a pupa of a silkworm reared with aseptic artificial feed was infected at a high rate close to 100%, and fruiting bodies were formed. On the other hand, when mulberry-grown silkworm pupae were used, the infection rate of various bacteria was as high as 85%. Most of those infected with these bacteria were septic due to bacteria, and the tissues were disintegrated, resulting in a rotted corpse with only the outer skin left. Therefore, the target Cordyceps fungi died before the infection. This is considered to be because bacteria adhering to mulberry leaves, silkworms and the like invaded the respiratory tract and trauma of the silkworm. Particularly in the late autumn silkworm season, the amount of bacteria attached to the mulberry leaf is large, and the amount of bacteria attached to the silkworm itself is also large. If conditions such as moisture and temperature are in place,
Bacteria multiply and cause silkworm disease. Even in the case of silkworm pupae grown on mulberry leaves, those infected with Cordyceps fungi formed fruiting bodies.

Example 4 Next, a comparative experiment will be described in which inoculation is carried out on the primary pupa of the silkworm grown on the sterile artificial feed and the mature silkworm grown on the sterile artificial feed. Inoculation was carried out in the same manner as in the above-mentioned example, using cereal mushrooms (Cordyceps Militaris) as the bacterial species. When inoculated into primary pupae, the infection rate is 100% and the fruiting body formation rate is 9%.
It was 9.5%. 12 days after inoculation, a loci is formed,
After 30 to 50 days, fruiting bodies formed and matured. The fruiting body was club-shaped, light vermilion orange and averaged about 7 cm. On the other hand, when inoculated into mature silkworms, the infection rate was 82% and the fruiting body formation rate was 74%. At 13 days after inoculation, the loci are formed, and after 50 to 60 days, the growth of the fruiting body is 0.5 to
It was only about 1.5 cm.

Example 5 An experiment was conducted on the culture temperature used for the liquid medium and the extraction temperature and the extraction time. Table 3 shows the results. In addition,
Extraction was performed using water at a weight ratio five times that of pupa 1, and more water was added by the amount of evaporation. Further, the pupa extract was further diluted with 8-fold water to obtain a liquid medium. About 1/3 of this liquid medium was placed in a test tube, and an isolated strain of Hanasagitake was planted on the liquid medium. The dry weight ratio is obtained by drying the fruiting bodies and mycelia formed in each sample, and expressing the weight of the fruiting body and the mycelium as 100, using the heaviest one (using an extract extracted at 120 ° C. for 60 minutes). It is.

[0022]

[Table 3]

According to Table 3, when the extraction temperature is 100 ° C., extraction is preferably performed for 2 hours or more, more preferably 5 hours or more. When the extraction temperature exceeds 100 ° C., good results are obtained at least 10 minutes or more, but more preferably 20 minutes or more.

Example 6 Next, an experiment was conducted on the culture temperature and the extraction time for a culture solution used as a liquid medium when pupae containing water after the spinning were used. Table 4 shows the results. The extraction was performed using water three times the weight ratio of pupa 1. In the same manner as in Example 5, water was further added by the amount of evaporation. Also,
The pupal extract was further diluted with water twice to obtain a liquid medium. About 1/3 of this liquid medium was placed in a test tube, and an isolated strain of Hanasagitake was planted on the liquid medium.

[0025]

[Table 4]

From Table 4, it can be seen that 1 hour to 4 hours at 100 ° C.
When the extraction time is 30 minutes to 60 minutes at 107 ° C and 20 minutes to 40 minutes at 120 ° C, the infection rate of Cordyceps sinensis is 100%, the fruiting body formation rate is 98% or more, and 45%.
After 4 days to 60 days, 4-8 cm fruiting bodies formed.

Example 7 A comparative experiment was conducted on a method for extracting pupal composition components when a liquid medium was used. Experiments were conducted on a method of adding water to the dried pupa and boiling it, a method of crushing the dried pupa and then adding water and boiling, and a method of adding water to the crushed dried pupa.
In the case of boiling, water was used in an amount of 5 times the weight of the dried pupa 1, and water was further added for the evaporation. The extract was further diluted with water eight times to obtain a liquid medium. Other culturing methods were performed in the same manner as in Example 1.
Regarding the method of adding water to the crushed dried pupa, first, the dried pupa is crushed, and the crushed pupa is subjected to a sterilization treatment at 121 ° C. for 15 minutes using a high-pressure steam sterilizer. Performed by adding 40 times sterile water.
Although the fruiting bodies were well formed by any of the above methods, the condition of the fruiting bodies with respect to the method of adding water to the pulverized dried pupae was the best. In addition, the method of pulverizing and boiling the dried pupa was better than the method of boiling the dried pupa as it was.

[0028]

According to the present invention, continuous cultivation of cordyceps can be performed stably and in mass production by using silkworm pupae as a medium. In particular, aseptic artificial feed breeding of silkworms allows high-density breeding, and it is possible to supply a certain amount of pupae on a daily basis regardless of the season. Moreover,
When the pupae of aseptic artificial feed-fed silkworms were used, the infection rate was high and constant, the fruiting body formation after infection was stable because no germs were observed, and the production efficiency of Cordyceps was extremely high. High and unrivaled in space / time collection rate. Therefore, it is inexpensive and can be mass-produced. For example, when field insects are used, they are produced only once or twice a year, so that the production is limited, and even if the production amount is compared with the field cultivation system, the production can be several tens of thousands or more per year. Furthermore, the produced Cordyceps can be dried or supplied as a dry powder as it is, which is efficient and economical as an industrial production method. Needless to say, a product with higher purity can be supplied by an operation such as extraction and purification of the active ingredient, if necessary. In the case of culturing in a liquid medium, it is also possible to extract an active ingredient from a residue obtained by recovering fruiting bodies and a culture solution.

In addition, the present invention can utilize silkworm pupae after reeling, so that not only can it be obtained inexpensively and in large quantities, but also it is possible to effectively reuse silkworm pupae that are currently causing contamination problems. it can. As described above, the present invention particularly produces ascospores or conidia as a mass growth source of Cordyceps sinensis in a liquid medium containing a pupa extract as a main component. By establishing a mass production system, Cordyceps can be produced industrially at low cost and in large quantities.

Continuation of front page (72) Inventor Fujiyoshi Matsubara 2-507 Nabeshima, Momoyamacho, Fushimi-ku, Kyoto-shi, Kyoto Momoyama Plaza 507

Claims (12)

[Claims] 1. A method for artificially cultivating Cordyceps sinensis, which comprises culturing using a medium mainly containing a composition component of silkworm pupa. 2. The method for artificially cultivating Cordyceps according to claim 1, wherein the pupa is a pupa after the reeling. 3. The method for artificially cultivating Cordyceps according to claim 1, wherein an extract obtained by extracting a composition component of silkworm pupa using water at high temperature and heating is used as the culture medium. . 4. The extract has a weight ratio of 2 to pupa.
The method for artificially cultivating cordyceps according to claim 3, characterized in that the cordyceps is extracted by boiling it at a boiling temperature for 2 hours or more using 50 times water. 5. The extract has a weight ratio of 2 to pupa.
The method for artificially cultivating Cordyceps sinensis according to claim 3, characterized in that the cordyceps is extracted by boiling it for 50 minutes or more at a temperature higher than the boiling temperature using 50 times the water. 6. The method for artificially cultivating Cordyceps according to claim 1 or 2, wherein a liquid medium obtained by crushing pupae and adding water thereto is used as the medium. 7. The artificial Cordyceps of any one of claims 3 to 6, wherein at least one of a carbon source, amino acids, minerals, and vitamins is added to the medium. Culture method. 8. The method for artificially cultivating Cordyceps sinensis according to claim 7, wherein the amount added is 2 to 8 weight ratio with respect to 100 extracts of silkworm pupae. 9. A germ-free silkworm is used as a silkworm, and after the silkworm has formed a cocoon, a pupa is taken out, and the pupa is cultured by inoculating an ascospore or a conidium formed in the body of Cordyceps sinensis. A method for artificially cultivating cordyceps. 10. The method for artificially cultivating Cordyceps sinensis according to claim 9, wherein a primary pupa is used as the pupa. 11. A silkworm, wherein cordyceps sinensis is multiplied and cultured in a liquid medium containing a composition component of the silkworm pupa as a main component, and thereafter, ascospores or conidia formed in the fruit body of the cordyceps are aseptically reared. A method for artificially cultivating Cordyceps sinensis, wherein subculture is performed by directly inoculating the pupa of the plant. 12. The method for artificially cultivating Cordyceps according to any one of claims 1 to 11, wherein a Cordyceps sinensis is used as the Cordyceps.