KR101871804B1 - Cordyceps based upon a seahorse and method of cultivating thereof - Google Patents

Abstract

The present invention provides a method for cultivating Cordyceps sinensis and a method for cultivating it, which comprises culturing the hippocampus as a host and containing nutritional components of hippocampus. Accordingly, it is possible to provide a food containing a single component of horseradish cordyceps containing new nutritional ingredients which can not be expected in general caterpillar fungus, to activate sex hormone secretion, to provide anti-aging and antithrombogenic effect .

Description

[0002] Cordyceps based on a seahorse and method of cultivating < RTI ID = 0.0 >

The present invention relates to a method for cultivating Cordyceps and cultivated with hippocampus.

In winter, it is parasitic to insects, but in the summer it forms a mushroom in the body of an insect that becomes a host. The name of the mushroom is derived from the insect, which causes insects to die and the insects to die. , There is a kind that even if a bacterium invades an insect and kills an insect, it forms conidiospore which is an asexual reproductive organ on the surface of the host without forming a throat. Most of the caterpillar fungus forming the thyroid gland are Codiaceps, Cordyceps, Podonectria, Torrubiella, etc. belonging to the genus Cynoptera, , And it is widely distributed in Korea, China, and Japan.

The insects which are the hosts of these Cordyceps are parasitic to the spiders in addition to the lepidoptera (Cordyceps militaris), the cetacean (C. sobolifera), the loggerhead (C. sphecocephala), the other beetles have. For example, there are the beetles such as the beetle caterpillars, the beetle caterpillars, the beet caterpillars, the snowflake caterpillars, the parasitic caterpillars of the moth larvae, and the pupae.

Among them, the pharmacological effects have been acknowledged because it is collected only at the snowy alpine area of more than 4000 m above sea level, and thus artificial cultivation is difficult. And the bee caterpillars, beetle caterpillars and caterpillar caterpillars are unsuitable for use as edible because their host is a hateful insect. Therefore, artificial cultivation has been studied only for the pupae of the silkworms and the snowy flowers of the silkworms with silkworm or silkworm pupa.

When the mechanism of the formation of the mushroom is permanent, the mushroom forms a molecular spore or a concha spore and adheres to the joints and digestive parts of the living insects, so that the insects are killed by ingesting nutrients in the insect body In summer, fruiting bodies are formed by using the host insect nutrients.

Cordyceps is generally composed of moisture, carbohydrates, fats, ash and crude protein. The protein contains about 18 essential amino acids. In addition, Cordyceps sinensis contains special components such as polysaccharides and polysaccharides, and is known as a therapeutic agent for anemia, fatigue, hypertension, pulmonary tuberculosis and asthma.

In addition, it has been proved that cordyceps contains a component for enhancing immune function, a sexual stimulating substance and an anti-cancer component, and studies have been conducted actively, and the effect as an antidote for drug addicts is excellent, Therefore, it is very effective as a biological pesticide which can prevent environmental pollution while providing effective pest control.

Despite these excellent effects, it is very difficult to harvest the naturally occurring cucurbitaceae fruiting bodies, so artificial cultivation techniques for stably supplying the cucumber herbaceous fruiting bodies are required.

The known method of artificial cultivation of Cordyceps is based on the fact that caterpillar fungus forms fruiting bodies based on insects, and there is a method of producing fruiting bodies by inoculating larvae of Cordyceps sinensis and snowy flower caterpillar fungus to silkworm larva or silkworm pupa.

Korean Patent Laid-Open Publication No. 1998-023907 is known as a prior art. In this invention, a method of cultivating fruiting bodies by inoculating a fourth or fifth instar larva or a pupa with silkworms is disclosed.

Another prior art is Korean Patent Laid-Open Publication No. 1998-033558. The present invention discloses a method of cultivating a fruit body by inoculating a seedling of a Chinese cabbage flown with a seed culture medium so that the whole surface of the culture medium can be grown in a state in which the fruit body is completely covered with a culture medium supplemented with a pupa.

In addition, there is known a method for cultivating caterpillar fungus using a soybean fiber foil (Application No. 10-1999-025540), a method for producing caterpillar fungus based on a long beetle larva and its production method (Application No. 10-1999-026782).

On the other hand, hippocampus lives mainly in sub-tropical shallow water, which is not contaminated with fish meat and fish. The hippocampus has been prescribed as a special medicine for intractable diseases as well as energetic drugs in Chinese and other Chinese cultural countries. According to international trade data, at least 20 million dried seahorses were circulated globally only in 1995, presumably being used as traditional Chinese medicines and their derivatives.

However, a technique for cultivating Cordyceps sinensis with the hippocampus as a host has not yet been known.

Disclosure of Invention Technical Problem [8] The present invention has been conceived to solve the above-mentioned problems, and it is an object of the present invention to provide a method for cultivating Cordyceps and cultivated on the basis of hippocampus. More specifically, a method for cultivating a Chinese cabbage seedlings which can solve the problems of easiness of cultivation, productivity, and supply of raw materials in order to meet various needs of development of host animals for artificial cultivation of Chinese caterpillar fungus .

However, the objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned can be clearly understood by those skilled in the art from the following description.

The present invention relates to a method for culturing a Chinese cabbage seed culture in a seed culture medium; A medium forming step of forming a hippocampal medium based on seahorse; An inoculating step of inoculating the cultured Cordyceps spp. Into the hippocampal formation medium; And culturing the inoculated seedworm seeds in the hippocampal medium to form mycelia and fruiting bodies.

The hippocampus also provides a method for culturing hippocampal cordyceps, including cultured hippocampus.

The hippocampus also provides a method for culturing hippocampal cordyceps including a big-belly seahorse or a pot-bellied seahorse.

In addition, the above-mentioned medium-forming step provides a method for growing a hippocampus, which is a step of forming a hippocampal medium based on a hippocampus prepared by drying a hippocampus discharged with excrement at 30 to 60 DEG C for 5 to 10 hours.

In addition, the medium forming step may further include a desalting step of removing the salts of the hippocampus, wherein the desalting step is a step of mixing the brine having a salinity equivalent to that of the hippocampus and the dried hippocampus under an alcohol And cultivating the horseraceous caterpillar fungus.

Further, the desalting step further comprises a step of stationary processing for 15 to 20 hours after the mixing treatment, and a step of filtering the hippocampus to cultivate hippocampal cordyceps.

The culturing step further comprises a step of inoculating a liquid seed or solid seed of the Cordyceps sinensis into a seed culture medium in an aseptic condition.

Also, the inoculating step provides a method of culturing the hippocampal cordyceps, which is a step of sterilizing the hippocampus culture medium, cutting the outer skin of the hippocampus, and inoculating the cultured cordyceps spp. Into the hippocampus.

The step of culturing is carried out by culturing a mycelium at a temperature of 23 to 27 캜 and a humidity of 65 to 75%, irradiating light at a temperature of 15 to 20 캜 and a humidity of 80 to 95% And provides a cultivation method of the horseradish Cordyceps.

The present invention also provides hippocampal cordyceps containing the nutritional components of seahorse.

The nutritional ingredient may be selected from the group consisting of taurine, arginine, aspartic acid, alanine, glycine, proline and glutamic acid. The present invention provides a hippocampal cordyceps containing one or more amino acids.

The nutritional ingredients may also be selected from the group consisting of calcium, magnesium, potassium, sodium, iron, zinc, manganese, copper, and chromium. The present invention provides hippocampal cordyceps containing any one or more inorganic substances selected from the group consisting of

The present invention also provides a food comprising the hippocampal cordyceps as one component.

The present invention provides a hippocampal cordyceps containing new nutritional ingredients which can not be expected in a common caterpillar fungus, and provides a food containing the same as a single ingredient to activate sex hormone secretion, and to provide an anti-aging and antithrombogenic effect .

Also, the hippocampal cordyceps produced by the present invention can be used as a medicinal herb to treat diseases such as asthma, heart disease, and fracture.

Before describing the present invention in detail, it is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention, which is defined solely by the appended claims. shall. All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art unless otherwise stated.

Throughout this specification and claims, the word "comprise", "comprises", "comprising" means including a stated article, step or group of articles, and steps, , Step, or group of objects, or a group of steps.

On the contrary, the various embodiments of the present invention can be combined with any other embodiments as long as there is no clear counterpoint. Any feature that is specifically or advantageously indicated as being advantageous may be combined with any other feature or feature that is indicated as being preferred or advantageous. Hereinafter, embodiments of the present invention and effects thereof will be described with reference to the accompanying drawings.

The present invention provides a method for growing Cordyceps, based on a hippocampus, according to an embodiment of the present invention.

The method for culturing the hippocampal cordyer of the present invention comprises a culturing step (S1) for culturing the seedlings, a medium forming step (S2) for forming a hippocampus based on the hippocampus, an inoculation step for inoculating the seedlings cultured in the hippocampal medium (S3), and a growing step (S4) of cultivating the inoculated seedworm seeds to form a mycelium and a fruiting body.

The culturing step (S1) according to one embodiment of the present invention is a step of inoculating a liquid seed or solid seed of Cordyceps mushroom to a seed culture medium in an aseptic state, and the seedlings of Cordyceps sinensis may be cultured in a liquid culture using potato or soybean powder A common solid seed obtained by culturing in a solid medium such as liquid seed or brown rice can be used. The seed culture medium may be any medium such as a liquid medium, a solid medium, a natural medium, or a synthetic medium. It is needless to say that the seedling culture can be performed using a hippocampal medium prepared in the medium formation step S2 described later.

Paecilomyces japonica or Cordyceps militaris may be used as the Cordyceps mushroom.

More specifically, the prepared culture medium is sterilized, and then the Chinese cabbage fungus is inoculated. It is preferable that the sterilization is carried out through the second pasteurization at 100 to 120 ° C after the first pasteurization at 80 to 100 ° C. After the sterilized medium is cooled, the microorganisms are inoculated into the medium.

The step of forming a medium (S2) according to an embodiment of the present invention is a step of forming a culture medium based on the hippocampus, wherein the feeding of the food is stopped for at least 24 hours and the hippocampal slurry is drained and dried to form a hippocampal medium do.

A hippocampus can be used as a hippocampus, and it is preferable to use a big-belly seahorse or a pot-bellid seahorse. However, the present invention is not limited thereto, and any kind of hippocampus such as Greate seahorse (H. kelloggi), Spotted seahorse (H. kuda), Long horn seahorse (H. .

More specifically, the feeding of the hippocampus to be used as a medium is stopped for at least 24 hours, preferably 24 to 48 hours, to discharge the excrement. It is preferable that the hippocampus from which the excrement is discharged is dried at 30 to 60 DEG C for 5 to 10 hours. However, the drying temperature and the drying time can be adjusted depending on the state of hippocampus, the desired culture conditions, and the type of Cordyceps mushroom. The dried hippocampus may be used as a medium for use in a glass bottle, a plastic bottle, a bag, or the like, or may be used as a medium for itself.

The medium forming step S2 according to an embodiment of the present invention may further include a desalting step S201 for controlling the salinity of the hippocampus or a dehydrating step S202 for adjusting the moisture content of the hippocampus. In addition, it may include both the desalting step (S201) and the dehydrating step (S202), in which case desalting step (S201) is preferably performed first.

The salinity of the hippocampus is adjusted to a level suitable for the cultivation of Cordyceps mushroom through the step of desalting the hippocampus (S201) according to an embodiment of the present invention. In the desalting step (S201), a brine having a salinity equivalent to that of a hippocampus is mixed with a hippocampus under an alcohol, and the hippocampus is subjected to stationary for 15 to 20 hours, Is the step of obtaining the hippocampus.

The brine used in the desalting step (S201) includes a solution containing seawater or saline and containing salts such as calcium chloride. The salinity of the brine is equivalent to that of the hippocampus used as a medium.

Also, the desalting step (S201) is a step of precipitating salts by lowering the solubility of salts by mixing under an alcohol, and it is preferable to use ethyl alcohol, prethanol A among alcohols.

In the desalting step (S201), hippocampus is mixed with 80 to 120 parts by mass of 100 parts by mass of brine, and the mixture is subjected to a heat treatment under 500 to 1000 parts by mass of alcohol. The hippocampus, from which the salt has been removed, is separated by filtration and washed to obtain a salty hippocampus.

The water content of the hippocampus is regulated to a level suitable for the cultivation of Cordyceps, through the hippocampus dewatering step (S202) according to an embodiment of the present invention. It is preferable to dry the hippocampus by evaporating the water in the hippocampus at 20 to 40 캜 in a step of drying the hippocampus or the saline-adjusted hippocampus. If the water content is high, the fruiting body germinates well but the mycelium is not cultured well. If the water content is low, the mycelium is cultured well but the fruiting body does not germinate. Thus, there is a need for a process to adjust the water content of the hippocampus to an appropriate level. The drying temperature and drying time can be adjusted depending on the condition of the hippocampus, the desired culture conditions, the kind of the insects and the like to be inoculated. The hippocampus mixed medium can be constituted by hippocampus alone, that is, by constituting the medium with 100% hippocampus or further mixing medium components other than hippocampus.

The seeding step S3 according to an embodiment of the present invention is a step of seeding the cultured Cordyceps spp. On a hippocampus bearing hippocampal formed hippocampus, sterilizing the hippocampal prepared in the above step, And then inoculated with the cultivated Cordyceps spp. Cultured in the hippocampus. After cooling the sterilized hippocampal medium, liquid or solid seeds of Cordyceps sinensis are inoculated in an aseptic state at 20 ° C or lower.

As the Cordyceps specimens, general solid seeds cultured in a solid medium such as liquid seed or brown rice cultured in a liquid culture solution using potato or soybean powder can be used. In addition, as described above, according to the present invention, May be used.

According to an embodiment of the present invention, the step of growing (S4) includes culturing the inoculated seedworm seeds to form a mycelium and a fruiting body, and culturing the seedlings in a culture room under appropriate conditions. The culture conditions of the mycelia of Cordyceps mellifera are carried out at a specific incubation temperature and specific room humidity, and the mycelium spreads in the medium to complete the cultivation of the mycelium. Normally, when cultured at a temperature of 23 to 27 DEG C and a humidity of 65 to 75% for 8 to 10 days, the mycelium spreads in the culture medium to cultivate the mycelium.

Also, the step of growing (S4) according to an embodiment of the present invention includes a step of growing the mycelia of caterpillar fungus cultured in the above process under proper conditions. Growth conditions of Cordyceps sinensis are made at specific temperature and specific humidity. Normally, when a light of 100 to 200 lx is irradiated under the condition of a temperature of 15 to 20 ° C and a humidity of 80 to 95%, a fruiting body comes out, and when irradiated with light of 100 to 400 lx, . In the case of Millilitaris Cordyceps, 800-1000 lx of light is irradiated for about 3 days, resulting in a fruiting body. When irradiated with light of 100 to 400 lx on the firing body under the same conditions, a fruit body is grown.

If the fruit body grows more than 5cm, it will be harvested. In the case of the ginseng caterpillar fungus, it takes about 2 weeks from the time of harvest to the time of harvest, and about 4 weeks for the case of milletaris cordyceps. The total number of cultivation days is about 30 days. Harvest hatchlings and fruiting bodies harvested at the time of harvesting caterpillar harvesting through the growing stage.

The hippocampal cordyer cultivated by the cultivation method according to one embodiment of the present invention includes nutrients of seahorse used as a host. In addition, according to one embodiment of the present invention, the Cordyceps caterpillar fungus is cultivated on the basis of a bigbelly seahorse or a pot-bellid seahorse, and contains nutritional components of the Bigbell hippocampus or the potbellied hippocampus do. However, hippocampus can be included without limitation, such as Greate seahorse (H. kelloggi), Spotted seahorse (H. kuda), and Long horn seahorse (H.

The main nutritional elements included are taurine, arginine, aspartic acid, alanine, glycine, proline, glutamic acid, etc., which have high medicinal value. It contains 20 amino acids and is composed of calcium, magnesium, potassium, sodium, iron, zinc, manganese, copper, chromium ), And the like.

In another aspect of the present invention, there is provided a food comprising hippocampal cordyceps as one ingredient according to an embodiment of the present invention. The cordyceps of the present invention can be commercialized by freeze-drying according to a conventional method, or used as a water extract or alcohol. Cordyceps, including the nutritional components of the hippocampus, are used as therapeutic agents for sexual dysfunction, renal dysfunction and hypercholesterolemia. It also acts on the liver and kidneys to promote blood circulation, anti-thrombosis and anti-aging effects.

Example

Example 1

For the cultivation of Cordyceps, seedlings of Cordyceps militaris were cultured in liquid medium. The inoculum of the Cordyceps mushroom was obtained by cultivating the Cordyceps spp. At 22-30 DEG C for 5 days using PD broth.

The hippocampus used the Big Veli hippocampus cultivated in Jeju Island. The feed of the hippocampus was stopped for 36 hours to discharge the excrement, followed by drying at 40 ° C for 6 hours.

As a hippocampal medium, a medium having a composition of 100 g of dried hippocampus, 30 g of water and 2 g of sugar was formed. The medium was filled in a plastic container (about 850 ml) and autoclaved at 121 ° C and atmospheric pressure of 1.5 atm for 120 minutes.

Liquid culture, inoculated in a hippocampal medium cooled by pressurizing sterilized sterilized inoculum prepared in advance, cultured in a culture room at 25 ° C for about 7-10 days, and if the mycelium sufficiently spreads in the medium, the medium is taken out from the plastic container, It was transferred to a culture room with a temperature of 20 ° C, a humidity of 90% and an illumination of 500 Lux, and cultured for about 3 days. Seedlings started to form and harvested fully grown hippocampus through a growth period of about 15 days.

Example 2

After 36 hours, the feed of hippocampus was stopped and the feces was discharged. After drying at 30 ° C for 5 hours, 100 g of brine of 13% in salinity and 100 g of dried hippocampus were added to 800 g of prtanol A, mixed and left for 15 hours After the hippocampus was separated by filtration, the dried hippocampus was obtained by evaporating water at room temperature.

The hippocampal cordyceps were grown in the same manner as in Example 1 except that the desalted and dried hippocampus was used as a host.

Experimental Example

(1) Measurement of days of culture and days of growth

The mushroom was cultivated by the method of the above example, and harvested when the stem diameter of the fruiting body reached 7 cm. The number of days of culture and the number of days of growth were measured until harvesting and are shown in Table 1 below. The number of days of cultivation is the number of days required for mycelial growth from the inoculation to the bottom of the bottle. The number of days of growth is the number of days until the harvest date after the generation of fruiting bodies.

Days of culture Growing days Example 1 40 ~ 42 10 to 15 Example 2 32 to 35 7 to 13

Referring to the results shown in Table 1, it can be seen that when the desalinized hippocampus was used as a medium (Example 2), days of incubation and days of growth were shortened compared to the case of using dried hippocampus (Example 1) .

(2) General composition analysis

The general components were analyzed by Food Movement (2008), dry filtration method for crude oil, ether extraction method for crude fat, and Kjeldahl method for crude protein. The analysis of free sugar was performed by dissolving 1 ~ 5 g of the sample in 25 mL of water according to Food Code (2008), extracting with acetonitrile, diluting, filtering with 0.45 ㎛ membrane filter (Millipore, USA) USA).

The results of analysis of general components of hippocampal cordyceps cultivated according to the method of the above Example are shown in Table 2 below.

ingredient Example 1 Example 2 Unit Protein 52.8 64.1 % Saccharide 12.5 15.1 g / 100g Invert sugar 4.5 5.5 g / 100g

<Results of general composition analysis>

As shown in Table 2, in the case of Cordyceps mosaamed with the desalted and dried hippocampus of Example 2, 64.1% of the protein and 15.1 g / 100 g of the saccharides were cultured in the case of Cordyceps cultivated with the dried hippocampus of Example 1 (52.8% of protein, 12.5 g / 100 g of saccharides).

(3) Analysis of inorganic components

For inorganic analysis, 1 g of the sample is carbonized by taking the sample according to the Food Code (2008), and then heated at 550 ° C for several hours until a white whitish ash is obtained. This ash was sequentially decomposed using hydrochloric acid, diluted to a certain amount, filtered, and quantified by atomic absorption spectrometry using an ICP analyzer (ICP-OES, Varian).

The results of analysis of inorganic components of the hippocampal Cordyceps spp. Cultured according to the method of the above Example are shown in Table 3 below.

ingredient Example 1 Example 2 Ca 39.48 32.9 Mg 84.1 96.9 K 181.8 196.95 Na 100.92 109.33 Fe 23.28 25.22 Zn 7.56 8.19 Mn 1.82 1.68 Cu 1.8 1.8 Cr 1.4 1.5

Ca, Mg, K, Na, Fe, Zn, Mn, Cu and Cr were detected from the contents of inorganic components of Cordyceps sinensis. The content of Mg, K, Na, Fe, Zn and Cr was higher in the hippocampal cordyceps obtained according to the method of Example 2 than the hippocampal cordyceps obtained according to the method of Example 1.

(4) Analysis of amino acids

Amino acid analysis was analyzed by AccQTag amino acid analysis method (Waters). The analytical conditions were as follows: HPLC (Waters 2695, USA) and the column was detected with a fluorescence detector (EX: 250 nm, EM: 395 nm) using AccQTag (3.9 ㅧ 150 mm) The results are shown in Table 5 below.

Instrument HPLC (waters 2475)
Waters 2475 Multi-λ Fluorescence Detector Detector Grain: 1
Filter: 327 nm
Excitation: 297 nm
Emission: 395nm Pump Waters 2695 separations Module
AccQ · Tag column for hydrolyzate amino acid analysis Column 3.9 × 150 mm
Aqneous buffer, Water AccQ · Tag Eluente Mobile phase Acetonitrile
Water Flow rate 1.0 ml / min Injection volume 10 [mu] l

ingredient Example 1 Example 2 arginine 0.945 1.044 aspartic acid 1.222 1.664 field 0.851 1.090 glycine 0.522 0.596 taurine 0.543 0.692 proline 0.604 0.669 glutamic acid 1.575 1.606 serine 6.262 7.360 histidine 0.274 0.357 threonine 0.386 0.377 메티오 로 0.134 0.086 isoleucine 0.228 0.131 Total 9.710 10.566

As shown in Table 5, 12 kinds of amino acids including taurine were detected and identified. The total content of the amino acids of the hippocampal cordyceps obtained according to the method of Example was 9.710 g and 10.566 g per 100 g of the sample, respectively, and that taurine contained 0.712 g / 100 g and 0.878 g / 100 g.

The features, structures, effects, and the like illustrated in the above-described embodiments can be combined and modified in other embodiments by those skilled in the art to which the embodiments belong. Therefore, it should be understood that the present invention is not limited to these combinations and modifications.

Claims (13)

A culture step of cultivating the seedlings in a seed culture medium;
A medium forming step of forming a hippocampal medium based on seahorse;
An inoculating step of inoculating the cultured Cordyceps spp. Into the hippocampal formation medium; And
Culturing the inoculated Cordyceps specimens in the hippocampal medium to form a mycelium and a fruiting body;
Wherein the medium forming step further comprises a desalting step for removing salts of the hippocampus,
Wherein the desalting step comprises mixing the dried hippocampus with a brine having a salinity equivalent to that of the hippocampus under an alcohol.
The method according to claim 1,
Wherein said hippocampus is a cultivation method of hippocampal cordyceps including cultured hippocampus.
The method according to claim 1,
The hippocampus includes a big-belly seahorse or a pot-bellied seahorse.
The method according to claim 1,
Wherein the medium forming step is a step of forming a hippocampal medium based on a hippocampus prepared by drying the hippocampus from which excrement has been discharged at 30 to 60 DEG C for 5 to 10 hours.
delete delete delete delete delete delete delete delete A food comprising the hippocampal cordyceps prepared according to the method of claim 1 as one ingredient.