KR20040032001A - A method of vegetable worms culture using a honeybee - Google Patents

Abstract

PURPOSE: Provided is a method for culturing Cordyceps sinensis using a medium composed of honeybee larvae and containing abundant amino acids and various kinds of trace nutrients, thereby producing Cordyceps sinensis which is excellent in nutrients and efficacies. CONSTITUTION: A method for culturing Cordyceps sinensis using a honeybee comprises the steps of: preparing a culture medium using honeybee larvae; sterilizing the medium and inoculating Cordyceps sinensis thereinto; culturing the medium to give mycelium; and growing the mycelium to a fruit body of Cordyceps sinensis.

Description

How to grow cordyceps based on honeybees {A METHOD OF VEGETABLE WORMS CULTURE USING A HONEYBEE}

The present invention relates to a method for cultivating Cordyceps sinensis, and more particularly, to a method of artificially cultivating Cordyceps sinensis by directly inoculating Cordyceps perilla liquid seedlings in an infiltrating manner on a host of Cordyceps sinensis.

Cordyceps sinensis is a small mushroom belonging to the genus Cordyceps sinensis, which is a parasitic insect in winter and turns into a mushroom in summer. Cordyceps sinensis is a kind of medicinal mushroom produced by insects infected with cordyceps, a kind of filamentous fungus, known to grow native to China, Nepal, Japan and Korea. Cordyceps fungus forms follicular spores or conidia and attaches to soft parts of living insects such as respiratory organs, digestive organs, joints, etc., and when conditions such as temperature and humidity are formed, germinate and invade the insect body into mycelial state. After growth and growth, the mycelium in the body forms fruiting bodies on the epidermis of host insects. Fruiting body forms a shape of 1.5 ~ 10cm size, club-shaped, cylindrical, etc. consisting of the diseased part and the distal end of the tip, and the color varies in various ways such as white, black, red, and brown.

Cordyceps sinensis has not only been used as an herbal medicine since ancient China, but recently it has been found to be effective as an anti-cancer and antidote to drug addiction, and research is being actively conducted to scientifically determine the efficacy of cordyceps. According to recent studies, Cordyceps sinensis has been shown to be effective for kidney disease (Wak-dong, 'Lyoning magazine', February 2, 1995), to support the function of the spleen, and to have an excellent effect on hair growth ('The journal of Dermatology', 1986). , Japan), reported to be effective in lowering blood sugar levels.

Cordyceps sinensis is usually based on insects, except that it is parasitic to a small number of fungi and seeds, so the classification of cordyceps also includes the classification of the host insect. Insect necks, known as hosts of Cordyceps sinensis, include many types of insects, such as dragonfly, larva, lepidoptera, fly, lumber, cicada, stink wood, locust, and arachnid.

Commonly known cordyceps is Cordyceops Militaris, Snowfly cordyceps (Paedilomyces Japonica), Cordyceps sinensis Berk. Sacc., C. sobolifera, Bee bee cordyceps (C. shpecocephala) There is this.

Cordyceops Militaris is a complex of fruiting bodies and chrysalis that are parasitic in the chrysalis of Lepidoptera. Militaris Cordyceps sinensis has a host specificity based on pupa and prefers a stable ecosystem without external disturbances. It is distributed in many countries around the world, and artificial cultivation is possible using silkworm pupae (100%).

Snowfly Cordyceps (Paedilomyces Japonica), also known as silkworm cordyceps, is easy to manage and has a short growing period, so it is easy to mass produce most of the currently marketed cordyceps. Snow fungus Cordyceps sinensis is an incomplete fungus that forms conidia and is not significantly affected by the habitat environment and occurs on the basis of various insects.

Cordyceps sinensis Berk. Sacc., Also known as Cordyceps sinensis or Choongcho, is a complex of fruiting bodies and larvae that occurs around the body of a bat moth larva.

These Cordyceps sinensis have excellent effect, but it is very difficult to collect Cordyceps fruit body grown in nature, and many artificial cultivation technologies have been developed to stably supply Cordyceps fruit body.

On the other hand, bees are insects that collect honey, pollen and propolis from nature and produce royal jelly by themselves and provide useful natural foods for humans. In China, Japan and Europe, bee larvae are actively used for food. have. In particular, honeybee larvae contain about 50% of the amino acids in the water except moisture, and are rich in micronutrients such as vitamins, minerals, and serine. It is also known to be effective in the activation of sexual function, such as promotion, increased sperm productivity, and increased egg activity.

Bees and larvae of bees are used for medicinal purposes in our country by using honeycombs and larvae that have been naturally collected in the country.In foreign countries, large quantities of tablets and capsule products using canned bees and lyophilized powders of bee larvae are produced and distributed. have.

Bees have also been identified as Cordyceps Sphecosephala, Cordyceps Oxycephala, as one of the main hosts of natural Cordyceps, together with the above mentioned insect species.

As described above, there are many insects such as bat moth, chrysalis, scarab, stink bug, bee, etc., but until now, the insect cultivation method of cordyceps is known to be mainly of Cordyceps militaris and Paecilomyces. In most cases, it is about inoculating silkworm larvae or silkworm pupae to produce fruiting bodies.

For example, Korean Laid-Open Patent Publication No. 1998-00013 discloses a method of culturing pupa Cordyceps spp. After cultivating silkworm chrysalis in water, boiling and adjusting the moisture content to 60-65%. 1998-023907 discloses a method for culturing fruiting bodies by inoculating Cordyceps fungi to silkworm larvae or chrysalis of the silkworm, and Korean Patent Laid-Open No. 1998-33558 uses a medium in which pupa is added to a grain medium. It describes a method for culturing fruiting bodies by inoculating the liquid seedlings of Cordyceps sinensis, and Korean Patent Laid-Open Publication No. 2001-16739 discloses fruiting bodies in silkworm larvae or chrysalis after inoculation of the dryworm larva or chrysalis with snowflake verminus or pupa On the artificial cultivation method of Cordyceps sinensis which facilitates mass production by adjusting cultivation conditions in forming process Korean Laid-Open Patent Publication No. 2001-4812 discloses Cordyceps sinensis by inoculating snow cordyceps fungi into a liquid medium with a high content of soybean fiber foil remaining after the production of tofu and soy milk, instead of a gauze or chrysalis used as a host of Cordyceps sinensis. It describes a method of artificial culture.

However, all of the above-described prior art are using silkworm larvae or silkworm chrysalis as the host, and when using silkworm larvae or chrysalis, the hygiene problem of host insects is serious. Because of the situation, quality, reliability of the machining process, and the like are problematic.

In addition, the development of a variety of host insects are required to produce a variety of cordyceps close to natural cordyceps in terms of ingredients and effects.

Accordingly, the present inventors can develop a new cordyceps with excellent components by combining the impact characteristics of bees with the properties of cordyceps, in particular, bees are about 100 nationally among the various insects that can be the host of cordyceps. The present invention has been completed in view of the fact that a large number of dogs are bred in large numbers and artificial spawning and planned spawning can be performed smoothly and continuously as needed.

Therefore, an object of the present invention is to provide a method of cultivating Cordyceps sinensis, which is based on honey bees with excellent nutritional properties and easy artificial cultivation.

In addition, an object of the present invention is to provide a method for cultivating Cordyceps sinensis suitable for the growth stage of bee larvae.

In addition, an object of the present invention is to provide an inoculation method that can significantly increase the inoculation success rate, and shorten the culture period, and a method of cultivating Cordyceps sinensis accordingly.

In order to achieve the above object, the present invention comprises the steps of preparing a bee medium, inoculating Cordyceps liquid spawn on the prepared bee medium, culturing the inoculated culture medium into Cordyceps mycelium and growing the cultured mycelium into Cordyceps fruiting body In the cordyceps cultivation method consisting of steps, in the step of preparing the bee medium, the larvae or adults of the bee is collected and classified to prepare a single medium or a mixed medium mixed with cereal components such as brown rice, corn, wheat and sterilized Drying to maintain the proper water content, inoculating the Cordyceps fungi seedlings infiltrated into the prepared liquid spawn and stirring to inoculate the seedlings of Cordyceps fungi, and in the mycelial culture step, the culture medium inoculated with spawn seedlings in a sealed state The Cordyceps mycelia were incubated at a culture temperature of 22 to 25 ° C. In the somatic growth step, the Cordyceps sinensis mycelium is grown to 15.20 ° C. and 80-95% in humidity to grow Cordyceps herb fruit.

Hereinafter, the present invention will be described in more detail.

(1) Bee stage preparation

Cordyceps sinensis cultivation method according to the present invention does not use silkworm pupa medium as in the prior art, it uses bee medium. Bees include worker bees, charcoal bees, queen bees and their larvae.

For use in the bee medium of the present invention, (i) mature larvae such as worker bee larvae immediately before the age of 17-18 days old, charcoal bee larvae that are just 20-22 days old, and (ii) immature younger than 17 days old. Immature larvae such as worker bee larvae, immature charcoal bee larvae before 20 days of age, immature queen bee larvae within 6 to 8 days of age, and (i) adult bee larvae.

The reason for using the above-mentioned honey bee is as follows.

The classification of bee medium is related to the growth stage and allegory period according to the age of bee larvae. After 3 days of laying, the bees hatch into chrysalis, and after 6 days of feeding, the bees grow into chrysalis in the labyrinth-scattered egg room, and allegorize themselves to release themselves. The allegory period is usually from 24 to 26 days for charcoal bees, 20 to 21 days for worker bees, and 16 days for queen bees.

The honeybee has the highest amino acid content in the pupa, but it grows into a stratified layer as the amino acid is transformed into protein just before the allegory, and the emergence of amino acids accelerates as the wings rise and the epidermal layer becomes harder when it is about to emerge. Larvae 2-3 days before the period have the highest content of amino acids.

Therefore, worker bees can be used as a medium containing the most nutrients of worker bees are 17 to 18 days larvae, charcoal bees 20 to 22 days larvae.

However, in the case of queen bees, artificial cultivation of larvae is very difficult, and the amount thereof is extremely small, and thus the larvae of queen bees are extracted through the collection process of royal jelly. The collecting process of Royal Jelly is 3 days after spawning, and insects are fed to the queen bee nesting room, where artificially produced hatched larvae are fed, and worker bees supply Royal Jelly for 6 days. Royal jelly of the highest quantity and high quality is supplied between days and 3 days), and from the 5th to 6th days, gas is ejected as a result of metabolism in the larvae. The larvae (6 to 8 days old) are picked and stored separately. Queen bee larvae are known to have almost the same nutritional properties as royal jelly.

Therefore, in the present invention, the queen bee larva 6 days to 8 days after spawning with the highest nutritional content is used as the host of the cordyceps.

Mature larvae and adults are prepared in a single medium and sterilized for 60 to 90 minutes at a temperature of 120 ~ 125 ℃, 1.5 atm in a bottled state. After sterilization, it is transferred to a cooling room that is maintained aseptic state, and the water content of the medium is maintained at about 50 to 55% through a drying process that ventilates sterile air while cooling at 20 to 25 ° C. for 20 to 30 minutes.

On the other hand, the immature larvae are mixed with grains such as brown rice, corn, wheat at a ratio of about 50: 50, and sterilized under the same conditions as described above. The mixed medium absorbs grains of less than 10% moisture during the sterilization process to maintain the water content of larvae and grains of about 50-55% during sterilization.

Looking at the nutritional components of the charcoal beetle larva used in the present invention is shown in Table 1.

Nutritional components of charcoal larvae (per 100g) ingredient content ingredient content amino acid 42.25% Total Vitamin C 488mg Vitamin A 20 IU sign 451 mg Vitamin D 8,250 IU calcium 83.1mg Vitamin B1 0.71mg iron 7.72mg Vitamin B2 1.83mg zinc 68.3 ppm Pantothenic Acid 2.37mg Seren 0.07ppm

In addition, looking at the type of the amino acid of Table 1 in detail as shown in Table 2.

Types of Charcoal Larva Amino Acids amino acid ratio Arginine 2.23% Lee Sin 2.91% Histidine 1.20% Phenylalanine 1.77% Tyrosine 2.73% Leucine 3.65% Isoleucine 2.16% Methionine 0.90% Valine 2.56% Alanine 2.54% Glycine 2.49% Proline 2.55% Glutamic acid 5.63% Serine 2.08% Threonine 1.77% Aspartic acid 4.14% Tryptophan 0.54% Cystine 0.40% Sum 42.25%

As shown in the table, bees have higher amino acid content than silkworms, and it can be seen that they are rich in various vitamins.

(2) Inoculation of Bacillus subtilis liquid spawn

Pour the liquid spawn into the jar containing the bee medium prepared for each growth stage as described above and inoculate the spawn by stirring. The medium having a water content of 50 to 55% absorbs the water of the liquid spawn in the state of infiltrating the liquid spawn, thereby maintaining a water content of 60 to 65% suitable for mycelial culture and rapidly absorbing the spawn.

In the prior art, the liquid spawn is mostly sprayed using a nebulizer or an air compressor. However, in this case, the liquid spawn is difficult to inoculate evenly on the surface of the medium due to the characteristics of the honeybee having a relatively small volume per individual. As a result, infection or decay of microorganisms other than spawn causes extremely low inoculation rate and yield.

In addition, the infiltration method applied in the present invention compared to the conventional spray method is inoculated in a state in which the water content of the medium is adjusted to about 50% before inoculation to rapidly inoculate the water and the seed of the liquid spawn using the osmotic pressure inside the medium. By penetrating into the interior of the mycelia can be further promoted.

According to the experiment carried out by the present inventors, the inoculation by the spray method was not inoculated at all or the yield remained up to 50% or less, but the inoculation method by the infiltration method showed 90-100% inoculation success rate.

The liquid spawn is usually used by the Korea Agricultural Microbiological Conservation Center or another institution that preserves the spawn, or collected from the wild. Since the fruiting body does not come out in the cultivation process when it is passaged several times, for the fruiting body development, the Militaris cordyceps fungi (Cordyceps militaris) is transplanted two or three times, and the bacteria are used again in the wild. Must be used separately.

(3) mycelium culture step

The culture medium inoculated with Cordyceps spp. Is incubated with Cordyceps spp. Mycelia in the culture chamber. The culture conditions are usually the culture temperature of 22 ~ 25 ℃ of Cordyceps mycelium mycelium, and the humidity does not need to be artificially adjusted. Generally, the incubation temperature of Cordyceps sinensis is 25 ~ 27 ℃, but if the medium is infiltrated with liquid spawn, the heat of culture is generated and it becomes the inhibitor of normal mycelial culture. . After 8 to 10 days, the spawn is spread evenly in the medium to complete the culture of the mycelium, and if cultured continuously for 30 days, it is cultured as a mycelium that can be used as a product or raw material. At this time, the culture vessel in which the medium and the liquid spawn are introduced should be kept tightly sealed to prevent decay through the introduction or infection of foreign substances or the cultivation of the defective culture.

(4) Cordyceps fruit fruit growth stage

Cordyceps mycelium cultured for 8 to 10 days in the process is grown as Cordyceps fruiting body. The conditions for growing into fruiting bodies are usually suitable for the growth conditions of Cordyceps sinensis, temperature 15-20 ° C, humidity 80-95%, preferably about 90%.

Under these growth conditions, the fruiting body comes out after three days of irradiation of 1000 lx light, and the fruiting body grows when 100-400 lx light is irradiated under the same conditions. Militaris cordyceps takes about 3 to 5 weeks from harvesting to harvesting. Cordyceps sinensis is a combination of mushroom and host, so harvest host and fruiting body together. Thus, fruiting bodies do not fall from the host bees at harvest.

The harvested Cordyceps sinensis is lyophilized according to a conventional method and commercialized, or used as a water extract or liquor.

EXAMPLE

The following examples are provided to aid the understanding of the present invention, and the present invention is not limited to the following examples.

Example 1

18-day-old worker bee larvae are sterilized at 120 ° C. and 1.5 atm for 60 minutes. While maintaining the sterilization state, the sterile air is ventilated while cooling at 20-25 ° C. for 25-30 minutes to make the water content of the medium 50-55%. In the culture vessel containing the medium prepared as described above, Militaris cordyceps fungus (Cordyceps militaris: Korea Agricultural Microorganism Conservation Center strain No. KACC50171) is poured into a little more than the amount of medium and stirred. Incubate for 10 days at the temperature of 22-25 ℃ while keeping the culture vessel sealed. When the cultured mycelium is kept at a temperature of 15 to 20 ° C. and a humidity of 90% for 3 days, 1000lx of light is irradiated for 3 days, and when the fruiting body is grown, the fruiting body is grown by irradiating 100 to 400lx of light under the same conditions. Three to five weeks after harvesting, Cordyceps sinensis fruit is harvested.

Example 2

15-day-old immature worker bees larvae are mixed with brown rice at a ratio of 50:50 and then sterilized at 121 ° C. and 1.5 atm for 70 minutes. While maintaining the sterilization state, the sterile air is ventilated while cooling at 20-25 ° C. for 25-30 minutes to make the water content of the medium 50-55%. Into the culture vessel containing the medium prepared in this way, Pleistococcus aeruginosa (Paecilomyces japonica: Korea Agricultural Microorganism Conservation Center strain No. KACC50170) is poured into a little more than the amount of medium and stirred. Incubate for 10 days at the temperature of 22-25 ℃ while keeping the culture vessel sealed. When the cultured mycelium is kept at a temperature of 15 to 20 ° C. and a humidity of 90% for 3 days, 1000lx of light is irradiated for 3 days, and when the fruiting body is grown, the fruiting body is grown by irradiating 100 to 400lx of light under the same conditions. Three to five weeks after harvesting, Cordyceps sinensis fruit is harvested.

Example 3

8-day-old immature queen bee larvae are mixed with brown rice in a 50:50 ratio and sterilized for 70 minutes at 121 ° C and 1.5 atm. While maintaining the sterilization state, the sterile air is ventilated while cooling at 20-25 ° C. for 25-30 minutes to make the water content of the medium 50-55%. Into the culture vessel containing the medium prepared in this way, Pleistococcus aeruginosa (Paecilomyces japonica: Korea Agricultural Microorganism Conservation Center strain No. KACC50170) is poured into a little more than the amount of medium and stirred. Incubate for 10 days at the temperature of 22-25 ℃ while keeping the culture vessel sealed. When the cultured mycelium is kept at a temperature of 15 to 20 ° C. and a humidity of 90% for 3 days, 1000lx of light is irradiated for 3 days, and when the fruiting body is grown, the fruiting body is grown by irradiating 100 to 400lx of light under the same conditions. Three to five weeks after harvesting, Cordyceps sinensis fruit is harvested.

Cordyceps sinensis cultivation method according to the present invention is cultivated by cultivating Cordyceps fungi in a bee medium rich in amino acids and various nutrients, so the nutrition and efficacy is excellent, stable mass production is possible through the continuous and smooth supply of host insects.

The present invention can also maximize the yield and minimize the waste of raw materials by varying the composition of the medium, such as a single medium or mixed medium, depending on the growth stage of the bees to be the host.

In addition, by directly inoculating the liquid spawn in the inoculation medium by infiltration method, there is an advantage of further inoculating and inoculating the spawn evenly in all the medium to further promote the culture of the mycelia.

Claims (8)

Preparing a Cordyceps sinensis culture medium; Sterilizing the Cordyceps sinensis medium and inoculating the Cordyceps fungi seedlings; In the method of cultivating Cordyceps sinensis comprising the step of culturing the inoculated culture medium with Cordyceps mycelium in the culture chamber and the step of growing the culture medium cultured mycelium into Cordyceps sinensis fruiting body, The cordyceps herb culture medium preparation step sterilize the bees for 60 to 90 minutes at 120 to 125 ℃, 1.5 atm and then cooled to 20 to 30 minutes at 20 to 25 ℃ to cultivate the water content to 50 to 55% Preparing a medium; Inoculating the Cordyceps sinensis liquid spawn on the prepared culture medium; Culturing the mycelium for 8 to 10 days at a temperature of 22-25 ° C. inoculated with the Cordyceps fungus; And Cordyceps cultivation method based on the bees, characterized in that it comprises the step of growing the cultured mycelia into fruiting bodies. The method of claim 1, wherein the bees are selected from the group consisting of mature larvae, immature larvae, and worker bee larvae. The method of claim 2, wherein the immature bee larvae are mixed with grain and used as a host. The method of claim 2, wherein the mature larva is a bee larva of 17 to 18 days of age or a charcoal bee larva of 20 to 22 days of age. According to claim 2, wherein the immature larvae are selected from the group consisting of worker bees larvae 17 days old, charcoal bee larvae 20 days old, and queen bee larvae 6 to 8 days old. One Cordyceps cultivation method. The method of claim 1, wherein the seed spawn inoculation step is infiltrating the liquid spawn in the culture medium and inoculated by stirring directly inoculate the Cordyceps sinensis, characterized in that the inoculation. The method of claim 1, wherein the mycelium culture step is Cordyceps cultivation method based on bees, characterized in that in a thorough sealed state. A cordyceps based on a bee, characterized by inoculating a liquid seed in a caterpillar herb host of a single medium consisting of mature bee larvae or a mixed medium in which immature bee larvae and grains are mixed and cultured.