KR101151111B1 - Device for culturing mycelium and method of culturing mycelium using the same - Google Patents

Abstract

The present invention relates to an apparatus for culturing Mycelial Cordyceps mushroom mycelium for the production of blood circulation improving agent and a method for culturing mycelial Cordyceps mushroom mycelium using the same. The present invention also provides a method for culturing Mycelial Cordyceps mushroom mycelium for the production of a blood circulation improving agent which is capable of achieving high productivity at low cost in a facility, and a method for culturing mycelial Cordyceps mushroom mycelium using the same.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for culturing Mycelial Cordyceps mushroom mycelium and a method for cultivating a Mycelial Cordyceps mushroom mycelium using the same,

The present invention relates to an apparatus for culturing mycelium of Cordyceps mellifera and a method for culturing mycelia of Cordyceps militaris using the same. The present invention relates to a method for culturing mycelia of Cordyceps mellitus, The present invention relates to a method for culturing Mycelial Cordyceps mushroom mycelium for the production of a blood circulation improving agent which is capable of high productivity at a low cost, and a method for culturing a high concentration mycelial of Cordyceps militaris using the same.

Cordyceps fungus belongs to the Cordyceps family (Cordyceps), which is widely distributed throughout the world. Cordyceps, which have been used medicinally in China, are predominantly C. sinensis, which is a host of larvae of moths. They live in the highlands of more than 3,000 meters above sea level in Tibet, It is a medicinal plant that does not have much collecting amount because only fruiting bodies occur in summer.

Cordyceps are mycelial organisms parasitic to the insect itself or larvae or larvae of insects. At first, the insects are alive, but mycelium grows and spreads gradually in the insect holders, forming sclerotia, and then the insects die while maintaining their appearance. When the conditions such as temperature and humidity are improved, the fungus forms in the body of the insect, and fruiting bodies are formed. When it matures, the ascospores or the spermatozoa are formed and the insect is parasitized again. Cordyceps is named as C. pneumophilus (C. militaris) and C. sphecocephala according to host. As a result of these clinical studies, it has been known that such a Chinese caterpillar fungus is known as a hepatocyte protective action, an immunity enhancing effect, a cancer inhibiting effect, a virus inhibiting ability, and is now being sold as a functional food. In particular, cordycepin, ophiocordin and the like among the functional materials have anticancer and antimicrobial properties, and there are many other functional substances which are unknown.

Generally, the production of Cordyceps is used as a host for a pupa or the like, and the grown fruiting bodies are recovered and then liquidated and commercialized, or powdered and sold as a ring. When cultivated using such a pupa as a host, the production is influenced by the actual situation of the paddy farmhouse and there is no accumulation of the material technology. However, it is impossible to cultivate high concentration because of the poor facilities, The problem often occurs. Because of this relationship, manufacturers are anxious about the quality and supply of raw materials. In order to solve this problem, it is necessary to cultivate a large amount of Chinese cabbage by using a liquid medium.

In recent years, studies on physiological activity against the Cordyceps, and researches on active substances have been actively conducted, and commercialization using the results of these studies has been made. However, due to the growth characteristics of the insects, it has a disadvantage that the growth is stable and the supply of the caterpillar fungus is not smooth.

On the other hand, there are about 800 fungi entering the insects known to date. Among them, about 300 kinds of fungi belonging to the genus Mytilus are among the representative bacteria known to form mushrooms. In Korea, 76 papers were identified. Among them, C. falciparum (C. sinensis) and C. militaris are the internationally recognized fungi. Domestically, these fungi are parasitized in the pupa, and grown fruiting bodies are sold as extracts or powder products. However, such a culture method has disadvantages of non-uniformity of production quality of the product and a long incubation period. In order to solve this problem, it is necessary to cultivate a large amount of Chinese cabbage by using a liquid medium. However, productivity and efficiency of conventional culture incubators are low, so it is urgent to develop a low cost high efficiency incubator.

An object of the present invention is to provide a culture apparatus for mycelium of Cordyceps mellifera mycelium for the production of a blood circulation improving agent which is capable of mass culturing mycelia of caterpillar fungus organisms on the basis of seaweeds and having a simple facility and having high productivity at a low cost even in a small- And to provide a method for culturing mycelia of Cordyceps militaris using the same.

In order to solve the above-mentioned problems, the present invention provides a method for producing a health food using the culture of a Mycelialis fungus hyphae mycelium,

According to the present invention, there are provided a culture tank in which a culture raw material is received in a sealed state; An air injector for injecting air required for culture into the culture tank through an air inlet tube; An air exhaust unit for exhausting the air exhausted from the culture tank through the air exhaust pipe during culturing; An inoculum for inoculating the culture source in the culture tank with the insect pest; And a heater for supplying heat to the culture tank; The present invention also provides a method for culturing a mycelia of Cordyceps mushroom mycelium.

Preferably, the air injection tube is provided with a horizontal discharge portion whose end is horizontally bent in the culture tank, and the horizontal discharge portion is located below the liquid layer of the culture raw material during the culture.

Preferably, the lateral discharge portion is provided with a plurality of air discharge holes.

Preferably, the air injector filters air through a filter to supply air to the air injection pipe.

Preferably, the ventilation rate of the air supplied into the culture tank through the air injector is 0.5 to 3 vvm.

Preferably, the culture tank is provided with a cover, and the cover is provided with an observation window.

Preferably, the air evacuator is filled with alcohol to purify the contaminated air used in the culture.

According to another aspect of the present invention, there is provided a method for producing a fermented beverage comprising the steps of: (a) mixing boiled potatoes and algae, removing the husks, pulverizing and kneading the mixture into a culture tank as a culture raw material; (b) injecting a growth promoter into the culture material and mixing and sealing the culture tank; (c) sterilizing the internal culture material by supplying heat to the culture tank; (d) cooling the culture tank and inoculating the culture source with the bacterium; And (e) stirring and culturing the culture material by injecting and discharging air into the culture tank. The present invention also provides a method of cultivating the organism mycelia.

Preferably, in the step (a), the seaweed is at least one of sea tangle and seaweed.

Preferably, in step (a), the potato as a culture source and the seaweed are introduced at a ratio of 2: 1.

Preferably, the growth promoter of step (b) is glucose.

Preferably, the air injected into the culture tank in the step (e) is discharged through a horizontally discharging portion of the air exhaust pipe located below the liquid layer of the culture material, and the horizontally extending discharging portion is bent at the distal end thereof in a horizontal direction.

Preferably, the lateral discharge portion is provided with a plurality of air discharge holes.

Preferably, the air injected through the air exhaust pipe is filtered and injected through a filter, and the polluted air exhausted from the culture tank is purified and discharged through an alcohol-filled air exhaust unit.

Preferably, the ventilation rate of the air supplied into the culture tank in the step (e) is 0.5 to 3 vvm.

In general, since insect is necessary for the growth characteristic of the Chinese cabbage, it is necessary to cultivate the seedlings, to grow the cells for a long period of time, to increase the possibility of contamination, and to provide a new and constant culture method. However, Thereby making it possible to uniformize the product and mass-produce the product.

In addition, according to the present invention, not only the final culture but also the liquid medium itself has commerciality and can be mass-produced. Thus, it is possible to earn a high income even in small and medium-sized facilities due to the reduction of raw material cost of caterpillar fungus.

In addition, there is also an effect of improving the agitation capacity and productivity by improving the ventilation structure so as to increase the efficiency of the political incubator suitable for the reality of the company operating the small and medium facilities.

In addition, the culturing method according to the present invention is different from that of the fruiting body culturing by culturing the mycelial mycelium, and the purity of the mycelial mycelium is 80% or more, which is excellent in its efficacy and can be commercialized without any processing, (cordycepin) content is high.

In addition, sterilization and cultivation can be performed through the same culture tank, so that a large amount of the Mycelialis mycorrhizae mycelia can be produced even in a culture room having a small area, and the contamination can be remarkably reduced during cultivation, which is efficient.

1 is a view for explaining an apparatus for culturing Mycelial Cordywacker mycelia according to an embodiment of the present invention.
FIG. 2 is a photograph showing the appearance of a Mycelial Cordywacker mycelium culture apparatus according to an embodiment of the present invention.
FIG. 3 is a photographic image for explaining the inside of a culture apparatus for mycelia of Cordyceps mellitus mycelium according to an embodiment of the present invention.
FIG. 4 is a photograph showing a cultured state of a mycelial organism mycelia culture apparatus according to an embodiment of the present invention.
FIG. 5 is a flow chart of a method for culturing mycelia of Cordycephalomyces mycelium according to an embodiment of the present invention. FIG.
6 is a graph for explaining experimental results on the cordycepin content of a culture according to the present invention.
Fig. 7 is a graph showing the cultivation curve of Cordyceps sinensis according to the present invention. Fig.
8 is a view for explaining the blood circulation efficiency of a culture according to the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 is a view for explaining an apparatus for culturing Mycelial Cordywacker mycelia according to an embodiment of the present invention.

FIG. 2 is a photograph of a real image for explaining an appearance of a cultivation apparatus for mycelia of Cordyceps mellifera mycelium according to an embodiment of the present invention, and FIG. 2 is a photograph of a real object for clearly confirming the configuration of FIG. FIG. 3 is a photograph of the inside of a culturing apparatus for mycelia of mycelia of mycelia according to an embodiment of the present invention, and FIG. 4 is a graph showing the state of culture of the mycelial culture of mycelia of mycelia It is a real photograph for.

1 to 4, the apparatus for culturing the organism mycelia according to the embodiment of the present invention comprises a culture tank 10 having a constant internal space for culturing and culturing the culture material, An air injector 20 for injecting the air required for culturing, an air exhaust 30 for discharging the air to be discharged in the culture tank 10, An inoculation port 40 for inoculating the culture tank 10, and a heater 50 capable of supplying heat to the culture tank 10.

First, the culture tank 10 has a predetermined inner receiving space open on the upper side, and the opened upper side is covered by the lid 11 so that the receiving space is sealed. The cover 11 is provided with an observation window 12 made of a transparent or semitransparent material so as to observe a culture state in the internal accommodation space.

At this time, the culture tank 10 is made of a stainless steel material having good corrosion resistance, and the internal accommodation space can be variously configured as 5 L and 200 L depending on the desired production amount of the installation facility. In the following embodiments, The description will be made with reference to the culture tank 10 having the accommodation space.

In this culture tank 10, a culture material for culturing the mycelia of Cordyceps sinensis is injected. Such culture materials include seaweeds. At this time, the algae can be used in combination with at least one of brown algae such as red sea bream, seaweed, kelp, rhubarb, and mackerel, green algae such as mugwort, And / or seaweed.

That is, the culture material is a mixture of boiled potatoes and algae from which peel is removed, and the mixture is pulverized into a dough (gel) state and put into the culture tank 10.

In the case of the culture tank 10 having a storage space of 200 liters, the input amount of the culture material is preferably 600 g of potato and 300 g of seaweed.

A lid 11 for sealing the culture tank 10 is provided with a first through hole 11a for air injection, a second through hole 11b for inoculation of the bactrian fungus and a third through hole 11c for air exhaust, .

The air injector 20 injects air into the culture tank 10 for culturing the culture material in the culture tank 10. The air injector 20 is composed of a compressed air generator, Air required for cultivation is injected into the tank 10. [

That is, the air supplied from the air injector 20 injects clean uncontaminated oxygen through the filter 21 into the culture tank 10 along the air injection pipe 22.

One end of the air injection pipe 22 is connected to the filter 21 to receive the filtered air and the other end passes through the first through hole 11a of the lid 11 to the inside of the culture tank 10 .

Here, in the present invention, the ventilation structure of the air injection tube 22 in the culture tank 10 is improved so that the distal end is bent in a transverse direction. That is, the vertical air inflow pipe 22 having passed through the first through hole 11a of the lid 11 is bent horizontally at the end horizontal discharge portion 22a. And this horizontal discharge portion 22a is positioned below the liquid layer height for agitation of the culture raw material. At this time, the horizontal discharge portion 22a may be made of a porous material to increase the dispersing force of air. That is, the horizontal discharge portion 22a has a plurality of air discharge holes.

Therefore, the air supplied from the air injector 20 is discharged through the horizontal discharge portion 22a in the culture tank 10. Since such a horizontal discharge portion 22a is located below the liquid layer height of the culture material, By appropriately stirring together, the growth of the fungi is facilitated and a high yield is obtained in the culture.

At this time, the ventilation rate of the air supplied into the culture tank 10 through the air injector 20 is preferably maintained at 0.5 to 3 vvm, preferably 2 vvm.

The air exhaust pipe 30 removes air used for culturing the culture material in the culture tank 10. The air exhaust pipe 31 of the air exhaust unit 30 has one end connected to the cover 11, Through the third through hole 11c of the culture tank 10 and the other end thereof is connected to the air exhaust unit 30 to discharge the air used for cultivation to the outside. In this exhaust pipe 31, a pressure gauge 32 is provided to measure the air pressure in the culture tank 10.

At this time, the air exhaust unit 30 through which the air used for cultivation is discharged is filled with alcohol so that the contaminated air used for the culture is not directly discharged to the atmosphere.

On the other hand, the inoculation opening 40 inoculates the culture material in the culture tank 10 with the microorganism Cordyceps mucilage. The seedling tube 40 is provided with a seeding tube 41 and the seedling tube 41 passes through the second through hole 11b of the lid 11 and the end of the seedling tube 41 is positioned in the culture tank 10. The inoculation pipe 41 is provided with a thermometer 42 to measure the temperature in the culture tank 10.

At this time, the Cordyceps mushroom bacterium is cultured in a flask beforehand, and the ready-made mushroom bacterium is inoculated into the liquid phase through the inoculum 40 (within about 1 second).

Meanwhile, the heater 50 is provided at a lower portion of the culture tank 10 to directly raise the temperature of the culture tank 10 to sterilize the culture tank 10.

A discharge tube (51) is connected to the bottom surface of the culture tank (10) to discharge the cultured medium.

Now, a method for culturing mycelial Cordyceps mushroom mycelium using the apparatus for cultivating mycorrhizal fungus mycelium having the above-described structure will be described in detail with reference to FIG.

The culture tank 10 is provided with an air injector 20 and an air injection tube 22 for air injection and also includes an air exhaust 30 for air exhaust and an air exhaust And a micro filter is installed in the air injection pipe 22 to inject clean oxygen that is not contaminated.

Hereinafter, as an example, the culture tank 10 is assumed to have a capacity of 200 liters.

On the other hand, as a step S20 of culturing the raw material, boiled potatoes and seaweeds which are removed from the shell are mixed and pulverized into a dough (gel) state and put into the culture tank 10.

At this time, the algae can be used in combination with at least one of brown algae such as red sea bream, seaweed, kelp, rhubarb, and mackerel, green algae such as mugwort, And / or seaweed.

In the case of the culture tank 10 having a storage space of 200 liters, the input amount of the culture material is preferably 600 g of potato and 300 g of seaweed.

Next, as a step of injecting the growth promoter (S30), a growth promoter is injected for continuous growth growth of the mycelium of mulberry leaves.

Here, the growth promoter is preferably glucose, and it is preferable that a dosage of 1 kg is applied to the culture tank 10 having an accommodation space of 200 liters of glucose. At this time, the growth promoter may further include inorganic materials and vitamins.

Next, as a mixing and sealing step (S40), the culture tank 10 into which the culture material and the growth promoter have been introduced is sufficiently mixed and the lid 11 of the culture tank 10 is closed as described above, Respectively.

At this time, the mixing of the culture material and the growth promoter may be performed through a separate external mixer.

Next, as a sterilization step (S50), heat is supplied to the culture tank (10) through the heater (50) to sterilize the inner culture material. Such sterilization is sufficiently sterilized at 121 ° C and 1.5 atm or more for 60 minutes.

Next, in step S60, the culture tank 10 is sterilized through the sterilization step, the culture tank 10 is cooled to 21 ° C or lower, and the culture tank 10 is cultured in a flask. (Approximately 1 second or less) in a liquid phase through the inoculation port 40. [0064]

Next, in the culturing step S70, the air filtered through the air injector 20 is injected into the culture tank 10 and the inside air is discharged through the air exhaust 30, 12) to check the condition of sclerotium daily and cultivate until the sclerotia is fully formed.

Here, this incubation period may take about 25 to 30 days.

The mycelium of the solid component can be separated from the medium after the completion of the subsequent culturing, and the powdery or ringy mycelium can be produced through the lyophilization process, or the liquid mycelium can be produced. In addition, not only the final culture but also the liquid medium itself has excellent commercial properties.

Now, the productivity and the functionality of the culture apparatus of the mycelial Cordyceps mellus mycelium according to the present invention and the method of culturing the mycelial Cordyceps mellus mycelium using the same will be described in detail.

In the following, DPPH (2.2-diphenyl-1-picryl hydrezyl) method is used to measure the antioxidative activity. 16 mg of DPPH (α, α'-diphenyl-β-picrylhydrazyl) is dissolved in 100 mL of methanol, 100 mL of distilled water is mixed therewith, and the solution is filtered on Whatman filter paper No. 2. The sample prepared in 5 mL of this filtrate was diluted 4 times with 3 rd distilled water so that the absorbance was between 0.1 and 2, mixed with 1 mL, and the absorbance was measured at 528 nm to measure the scavenging effect on DPPH radicals And shows the antioxidative effect by converting the difference in absorbance between the sample added and the non-added sample as a percentage. All results are expressed as average values of the results obtained by three repeated experiments. BHA (2 [3] -t-Butyl-4-hydroxy-anisole, Sigma Chemical Co., USA), known as a synthetic antioxidant, is prepared at a concentration of 0.02%.

In the productivity evaluation, the cells of the solid fraction were separated from the culture medium after completion of the culture, and then lyophilized and evaluated by weight.

Now, with reference to Table 1 (based on 5 L culture tank), regarding the productivity and functional evaluation, it is as follows.

The cells of the solid fraction were separated from the culture medium after completion of the culture and lyophilized to evaluate the weight. Table 1 below shows the yield of solid body cells. At aeration rate of 2 vvm, the yield was similar to that of other cordyceps cultures. Therefore, It can be seen that the cell proliferation is good under very aerobic conditions. In addition, 2 vvm and 5% glucose showed the highest antioxidant and hepatocellular toxicity.

yield(%) Anti-oxidant Cancer
Ventilation rate

0.5 40 1.0 60 1.5 95 2 98 70 69
glucose
Addition amount
100g 75 150g 84 200g 90 250g 100 70 69

Next, the cordycepin productivity evaluation will be described with reference to Table 2 below.

The cultured tuna cultures were subjected to ethanol extraction (0.041 g / 2 mL) and the amount of cordycepin was analyzed by HPLC (High Performance Liquid Chromatography). The HPLC analysis conditions are shown in Table 2.

Oven temp 40 ℃ Flow rate 0.8 mL / min Mobile phage Methanol: DW = 30: 70 Column Mightysil RP-18, 250-4.6 Detector UV-254 nm Injection volume 10 uL

FIG. 6 shows that the peak of cordycepin by HPLC conditions was confirmed by standard cordycephin (Sigma Co.), and the peak at 5.23 minutes was codistepin. From the standard curve of the concentration and peak area, it was found that the codistin content of the cultured product was 54 mg per 100 g of the cultured ginseng. However, the content of the present invention at an optimum condition was about 0.3 g / 100 g-cell as shown in FIG.

As a result, the yield was increased to a level similar to that of other cordyceps cultures when the aeration rate of the culture tank 10 was 2 vvm. Therefore, It can be seen that the cell proliferation is good under very aerobic conditions. Also, antioxidant and hepatocellular toxicity were found to be 70% and 69% in 2 vvm and 5% glucose, respectively. The content of cordycepin in the optimum conditions of the present invention was found to be about 0.3 g / 100 g-cell.

Meanwhile, FIG. 8 shows a blood photograph for explaining the effect of taking the organism mycelia of the present invention. As can be seen from the drawing, it can be seen that the blood is purified and the blood circulation is promoted as the time passed after the mycorrhizal fungus body mycobacterium is administered. Therefore, it is possible to produce a blood circulation improving agent which makes blood flow smooth through the mycelium of Cordyceps mellus.

As described above, an optimal embodiment has been disclosed in the drawings and specification. Although specific terms have been employed herein, they are used for purposes of illustration only and are not intended to limit the scope of the invention as defined in the claims or the claims. Therefore, those skilled in the art will appreciate that various modifications and equivalent embodiments are possible without departing from the scope of the present invention. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims.

Claims (14)

delete delete delete delete delete delete delete A culture tank in which a culture raw material is received in a sealed state;
An air injector for injecting air required for culture into the culture tank through an air inlet tube;
An air exhaust unit for exhausting the air exhausted from the culture tank through the air exhaust pipe during culturing;
An inoculum for inoculating the culture source in the culture tank with the insect pest; And
A heater for supplying heat to the culture tank; Lt; / RTI >
The air injection tube is provided with a horizontal discharge portion whose end is horizontally bent in a culture tank, the horizontal discharge portion is located below the liquid layer of the culture raw material during the culture,
Wherein the horizontal discharge portion has a plurality of air discharge holes,
The air injector filters air through a filter to supply air to the air injection pipe,
The culture tank is provided with a cover, an observation window is provided on the cover,
The air vent is filled with alcohol to purify the contaminated air used in the culture,
(a) mixing the boiled potatoes and the seaweeds with the husks removed, crushing them into a kneaded state, and putting them into a culture tank as a culture raw material;
(b) injecting a growth promoter into the culture material and mixing and sealing the culture tank;
(c) sterilizing the internal culture material by supplying heat to the culture tank;
(d) cooling the culture tank and inoculating the culture source with the bacterium; And
(e) stirring and culturing the culture material by injecting and discharging air into the culture tank; Lt; / RTI >
The growth promoter of step (b) comprises at least one or more of glucose, minerals, and vitamins,
Wherein the flow rate of the air supplied into the culture tank in step (e) is 0.5 to 3 vvm.
9. The method of claim 8,
In the step (a)
Wherein the seaweed is at least one of sea tangle and seaweed.
9. The method of claim 8,
In the step (a)
Wherein the potato as a raw material for culture and the algae are introduced at a ratio of 2: 1.
delete 9. The method of claim 8,
Wherein the air injected into the culture tank in the step (e) is discharged through a horizontally discharging portion of an air exhaust pipe located below a liquid layer of the culture material, and the horizontally extending discharging portion is bent at an end thereof in a horizontal direction.
9. The method of claim 8,
Wherein the air injected through the air exhaust pipe is filtered and injected through the filter, and the polluted air exhausted from the culture tank is purified and discharged through an air exhausting device filled with alcohol.
delete

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