KR102053612B1 - Method for artificial cultivation of white grub Cordyceps using fermented sawdust medium - Google Patents

Abstract

The present invention relates to a method for artificial cultivation of white grub Cordyceps using a fermented sawdust medium, wherein the fermentation sawdust medium of the present invention can greatly increase a yield and productivity of Cordyceps, thereby being able to be utilized as a culture medium for artificial cultivation of Cordyceps and being expected to be applied to functional foods or pharmaceuticals by using cultivated white grub Cordyceps.

Description

Method for artificial cultivation of white grub Cordyceps using fermented sawdust medium}

The present invention relates to an artificial cultivation method of slugs Cordyceps sinensis using fermented sawdust medium.

Cordyceps sinensis is a small mushroom belonging to the Ascomycetes Clavicipitales, Clavicipitaceae. It is named after an insect in the winter and sprouting in the summer. The mechanism by which cordyceps is formed describes the mechanism by which cordyceps fungi form conidia or follicle spores, which are attached to the joints and digestive organs of living insects, ingest the nutrients in the insect body, form mycelia and eventually kill the insects. As winter passes and summer increases in temperature and humidity, mushrooms emerge from the body of insects buried in the ground and sprout out of the ground. Cordyceps sinensis means a combination of dead insects filled with mycelium and mushrooms from their bodies. Cordyceps sinensis is divided into three parts: head, sack, and host. The head is a organ containing spores used for breeding, and the long part supporting the head is sack. The head and the sack are called fruiting bodies or mushrooms, and the end of the sack sprouts from the body of the pupa. The head is shaped like a ball, a pointed cylinder at both ends, and a spatula, and has several asymptomatic angles on the surface or under the epidermis.

Cordyceps sinensis is generally composed of water, fat, crude protein, carbohydrate and ash, and protein contains about 18 kinds of amino acids essential for human body. In addition, Cordyceps sinensis contains special ingredients such as Choong-Cho Polysaccharide and Cho-Cho-Acid, which is known as a treatment for anemia, fatigue, hypertension, pulmonary tuberculosis and asthma, and because of its parasitic properties, insects can provide effective pest control while preventing environmental pollution. It is also very useful as a biopesticide. Cordyceps sinensis has been handed down in China since ancient times as 'Emperor's Buddha's Elixir', and it is also recorded in the herbaceous tree as 'the obvious effect is to strengthen weakness and strengthen immunity'. It is known to produce mannitol for medicinal use as well as other useful substances. The fruiting body also contains a component called cordycepin, known as an isomer of quinic acid, which acts as a toxin against host insects and has antibacterial, antiviral and anticancer effects. It is known to have excellent effects on various diseases. In spite of such excellent effect, it is very difficult to collect Cordyceps fruiting body grown in a natural state, so artificial cultivation technology for stably supplying Cordyceps fruiting body is required.

Meanwhile, Korean Patent No. 1871804 discloses 'Cordyceps sinensis cultivated using hippocampus and its preparation method', and Korean Patent Registration No. 0876820 discloses 'Cordyceps sinus cultivation method using Gasiogapi extract'. There is no description on the artificial cultivation method of slugs Cordyceps sinensis using fermented sawdust medium.

The present invention is derived from the above requirements, the present inventors mixed raw sawdust, rice bran, sugar and EM (effective microorgarnism) and fermented by controlling the moisture content to 60-65% fermented sawdust medium for cordyceps production Was prepared, and the result of incubation of the fermented sawdust medium and slugs were treated with cordyceps fungi, and it was confirmed that the growth of cordyceps was significantly increased at the same time compared to the conditions in which only the slugs were treated with fungus cordyceps fungi. In addition, when the rice bran medium was used instead of the fermented sawdust medium, the fruiting body of Cordyceps sinensis was not formed, so that the medium of the slugs of Cordyceps sinensis was confirmed that the fermentation sawdust medium was optimal, thereby completing the present invention.

In order to solve the above problems, the present invention (1) the step of culturing the Cordyceps fungus in a solid medium to separate the spores; (2) preparing a cordyceps fungi solution by culturing the separated spores of step (1) in a liquid medium; And (3) inoculating and incubating the Cordyceps fungus microbial solution prepared in step (2) in a medium and sterilized slugs to induce the Cordyceps sinensis fruiting body.

The present invention also provides slugs Cordyceps sinensis cultivated by the above method.

The slugs Cordyceps sinensis artificial cultivation method of the present invention can produce a high-quality Cordyceps sinensis in a short time, compared to the conventional bulk cordyceps production method, will be useful as an artificial cultivation technology for the stable supply of cordyceps.

1 is a photograph of fermented sawdust medium used for scavenging Cordyceps sinensis cultivation.
Figure 2 is a photograph of isolated single spores by culturing the collected Cordyceps fungi in a solid medium.
3 is a photograph of slugs used only for slugs of Cordyceps cultivation.
Figure 4 is a photograph of the slugs Cordyceps under cultivation using a fermented sawdust medium.
5 is a photograph of slugs Cordyceps sinensis cultivated using a fermented sawdust medium.
FIG. 6 is a photograph comparing growth of slugs Cordyceps sinensis grown under conditions in which the Streptococcus microorganisms were cultured in both the medium and the slugs (experimental group) and in which the Streptococcus fungi were treated only in the slugs (cultivation group).

In order to achieve the object of the present invention, the present invention

(1) culturing cordyceps fungi in a solid medium to separate monospores;

(2) preparing a cordyceps fungi solution by culturing the separated spores of step (1) in a liquid medium; And

(3) provides a method of artificial cultivation of slugs Cordyceps sinensis, comprising the step of inoculating the Cordyceps fungus solution prepared in step (2) in the medium and sterilized slugs and inducing the Cordyceps Sinensis fruiting body.

In the artificial cultivation method of slugs Cordyceps sinensis according to an embodiment of the present invention, the spores of the step (1) is a solid of the natural Cordyceps fungi collected from the valley PDA (potato dextrose agar) or SDA (sabouraud dextrose agar) It may be isolated by culturing in the medium, but is not limited thereto. Cordyceps sinensis of the present invention means Cordyceps fungi (fruiting body).

In the artificial cultivation method of slugs Cordyceps sinensis according to an embodiment of the present invention, the Cordyceps fungi of step (2) is 3 ~ 5 in the liquid medium of the yeast malt broth (YMB) separated the spores of step (1) It may be obtained by culturing for one day, but is not limited thereto.

In the artificial cultivation method of slugs Cordyceps sinensis according to an embodiment of the present invention, the medium of step (3) may be a fermented sawdust medium, the fermented sawdust medium specifically, raw sawdust, rice bran, sugar and EM (effective microorganisms) may be prepared by mixing and fermenting for 55 to 65 days by controlling the moisture content to 55 to 70% and sterilizing for 10 to 30 minutes at 115 to 130 ℃, more specifically, raw sawdust, rice bran, Sugar and EM are mixed at a weight ratio of 38-42: 2-4: 1-3: 0.5-1.5, fermented for 60 days by adjusting the moisture content to 60-65%, and sterilized for 15-20 minutes at 121 ° C. The fermented sawdust medium may be prepared, but is not limited thereto. The raw sawdust according to the present invention is a by-product generated in the wood processing process such as cutting, grinding, sanding, etc., there is no particular limitation.

In addition, in the artificial cultivation method of slugs Cordyceps sinensis according to an embodiment of the present invention, the slugs of step (3) is the chafer slugs of the dedicated stage soaked in boiling water for 2-7 minutes in boiling water 10 ~ 10 ~ 115 ~ 130 ℃ It may be sterilized for 30 minutes, preferably after rubbing the scarab slugs through a dedicated step 3 to 5 minutes in water of 90 ~ 110 ℃ and then sterilized for 15 to 20 minutes at 121 ℃, but is not limited thereto. Do not.

As the host of the slugs of the present invention Cordyceps sinensis, any host that can be used for cultivation Cordyceps in the prior art, such as slugs or silkworm pupae, may be applied, preferably, but not limited to slugs.

In general, slugs are larvae that occur in metamorphic insects, and scarab slugs that have been successfully bred in farms include Allomyrina dichotoma , white spotted radish ( Protaetia brevitarsis seulensis ) and spotted radish ( Protaetia orientalis submarmorea ). There are slugs, and the slugs used in the present invention are white spotted flower larvae, but are not limited thereto.

In artificial breeding the slugs of the scarabs, the sawdust ingredients supplied to the slugs feed remain in the intestines of the slugs, and during the sterilization process of the host, the intestinal moisture and the sawdust components are leaked to contaminate the cordyceps during the cultivation of cordyceps. Not only does it generate a unique smell, it can also prevent mycelia from sticking to the inside of the carcass during cultivation of Cordyceps sinensis, or it may not be able to grow smoothly. Therefore, if the main stage is the beetle larvae slugs before the transformation from larva to pupa, the slugs stop the growth after releasing impurities in the body can solve problems such as contamination, sluggish growth, odor generation.

In addition, in the artificial cultivation method of slugs Cordyceps sinensis according to an embodiment of the present invention, the induction of cordyceps fruiting body of step (3), put the slugs on the fermented sawdust medium and fermented Sawdust fermented with Cordyceps fungi in a sterile box with a syringe 13 to 17 ml of the medium is inoculated, 0.3 to 2.5 ml of the slug is inoculated after temperature of 23 to 25 ℃, humidity less than 70%, ventilation may be cultured for 8 to 17 days at 7 to 8 times a day, preferably Preferably, put the slugs on the fermented sawdust medium and inoculate 15 ml of Cordyceps fungi liquid into the fermented sawdust medium with a syringe in a sterile box, inoculate 1 to 2 ml in slugs and then incubate temperature 24 ° C., humidity less than 70%, and ventilate for 1 day. It may be cultured for 10 to 15 days at 4-6 times conditions, but is not limited thereto.

When the fruiting body starts to grow by the above method, it can be harvested if it grows more than 1 cm after culturing 40 to 60 days by irradiating light of 500 lux or less (16 hours per day) at 18 to 21 ° C., but is not limited thereto.

Specifically, the slugs Cordyceps sinensis cultivation method according to an embodiment of the present invention,

(1) separating the single spores by culturing the Cordyceps fungus in a PDA (potato dextrose agar) or SDA (sabouraud dextrose agar) solid medium;

(2) preparing the cordyceps fungi by culturing the isolated monospores of step (1) in a YMB (yeast malt broth) liquid medium for 3 to 5 days; And

(3) the cordyceps fungi prepared in step (2) above, raw sawdust, rice bran, sugar and EM are mixed and fermented for 55 to 65 days by adjusting the moisture content to 55 to 70% and then 10 to 30 at 115 to 130 ° C. 14-16 ㎖ in fermented sawdust medium prepared by sterilization for 1 minute, inoculated 1 ~ 2 ㎖ in sterilized slugs and then incubated for 10-15 days at temperature 24 ℃, humidity less than 70%, ventilation 4 to 6 times a day By including the step of inducing Cordyceps fruiting body, but is not limited thereto.

The present invention also provides slugs Cordyceps sinensis cultivated by the above method. Slugs cultivated by the above cultivation method can be applied to any type of cordyceps that can be artificially cultivated in the prior art, Cordyceps militaris, Cordyceps takaomontana (Cordyceps takaomontana), Cordyceps Cordyceps scarabaeicola, Cordyceps sphecocephala, Cordyceps tricentri, Cordyceps sinensis, Cordyceps nutans, and papillocaice (Paecilomyces japonica), Paecilomyces tenuipes (Paecilomyces tenuipes), but is not limited thereto.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Preparation Example 1 Preparation of Fermented Sawdust Medium

Raw sawdust, rice bran, sugar and effective microorganisms (EM) are put in a sack at a rate of 40 kg: 3 kg: 2 kg: 1ℓ, adjusted to 60 ~ 65% of moisture content, and fermented for 60 days in a space with low heat loss. (FIG. 1). The EM was used by mixing Lactobacillus casei , yeast ( Saccharomyces cerevisiae ) and Bacillus subtilis .

Example 1. Isolation of Single Spores and Culture of Cordyceps Bacillus

From the fresh fruiting bodies of Cordyceps sinensis mushrooms collected from Jeonsi-si Valley, Seongpoi were isolated from PDA (potato dextrose agar) or SDA (sabouraud dextrose agar) solid medium (FIG. 2), and the separated spores were YMB (yeast malt broth). Incubated for 3 to 5 days in the test was used.

Example 2 Preparation of Host for Slugweed Cordyceps Sinensis Cultivation

Slugs of the dedicated stage, which is the stage before the transformation from larva to pupa, were soaked in boiling water (temperature 90-110 ° C.) for 3 to 5 minutes to allow the protein components in the larva to coagulate and then washed with cold water and refrigerated.

Example 3 Growth of Slugs Cordyceps Sinensis

Slugs were placed in a culture vessel containing fermented sawdust medium and sterilized at 121 ° C. for 20-30 minutes and cooled at 25 ° C. In a sterile box, Cordyceps sinensis solution was treated with 15 ml of fermented sawdust medium and 1 to 2 ml of slugs, and then incubated at 24 ° C. for 10 to 15 days. 40 ~ 60 days of incubation under the condition that the light (16 hours a day) of 500 lux or less at ~ 21 ℃.

The final cultivated slugs Cordyceps sinensis is shown in Figure 5, the result of comparing the method of the present invention (methods of Cordyceps fungi on both medium and slugs) and the conventional method (stomach fungus treatment only on slugs), the method of the present invention is a conventional method Compared to the same time, it was confirmed that the growth of Cordyceps sinensis significantly (Fig. 6). Therefore, it is considered that the Cordyceps sinensis cultivation method of the present invention can provide a beneficial effect of obtaining a good quality Cordyceps sinensis in a shorter time than the conventional cultivation method.

Comparative example. Growth of Cordyceps Cordyceps sinensis Using Rice Bran Media Instead of Fermented Sawdust Badger

In the same manner as in Example 3, except that the rice bran was used instead of fermented sawdust medium, slugs were grown Cordyceps sinensis. As a result of the use of rice bran medium, fungus only appeared in Cordyceps sinensis, and no fruiting body was formed.

Claims (6)

(1) culturing cordyceps fungi in a solid medium to separate monospores;
(2) preparing a cordyceps fungi solution by culturing the separated spores of step (1) in a liquid medium; And
(3) the cordyceps fungi prepared in step (2) were mixed with raw sawdust, rice bran, sugar and EM (effective microorganisms) and sterilized with fermented sawdust medium fermented for 55-65 days by adjusting the moisture content to 60-65%. As an artificial cultivation method of slugs Cordyceps sinensis, comprising the step of inoculating each of the slugs and incubating at 23-25 ° C. for 10-15 days to induce Cordyceps sinensis fruiting body,
EM is an artificial cultivation method of slugs Cordyceps sinensis, characterized in that the mixture of lactic acid bacteria, yeast and Bacillus subtilis. delete delete delete The method of claim 1, wherein the slugs are slugs slugs go through the dedicated step artificial slugs cultivation method, characterized in that the cordyceps. delete

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