KR100826446B1 - Hericium erinaceus mycelium comprising rice bran and ginseng steamed red and it's cultivation method - Google Patents

Abstract

A production method of functional hyphae of Hericium erinaceum using rice bran or red ginseng as a substrate is provided to obtain effects such as the treatment of geriatric diseases, cancers and immunity system damage and the activation of brain function. A production method of functional hyphae of Hericium erinaceum using rice bran or red ginseng as a substrate comprises the steps of: (i) mixing 5% of glucose, 0.2% of a mixture obtained by mixing yeast extract and peptone in a ratio of 1:1, and 0.2% of a mixture obtained by mixing KH2PO4 and MgSO4 in a ratio of 1:1, to make a culture medium; (ii) culturing the spawn of Hericium erinaceum in the medium under conditions of a pH 4~6, a temperature of 20~30°C, a stirring velocity of 100~200rpm, a working volume of 50mL, and a hypha-inoculating ratio of 3~7%; (iii) firstly culturing the cultured spawn in a bio-reactor under conditions of a pH 5~6, a temperature of 25~30°C, a stirring velocity of 100~200rpm, and an airing velocity of 1~3vvm for 4~8days; and (iv) adding rice bran or red ginseng extract to a first culture liquid obtained from the step (iii), followed by secondly culturing.

Description

Method for producing functional mycelium of gingiva fungus with red rice bran or red ginseng as substrate method}

1 is a flow chart of a method for producing functional mycelium of roe deer fungus with substrate of rice bran or red ginseng according to one embodiment of the present invention.

The present invention relates to a method for producing a functional mycelium of roe deer fungus with a base of rice bran or red ginseng, and to a functional mycelium of roe deer fungus containing the active ingredient of rice bran or red ginseng produced using the same, and more specifically, By mass production of functional mycelium, it contains an active ingredient of rice bran or red ginseng, and it is a low-cost functional mycelium of roe deer fungus which has the effect of treating senile disease, cancer, immune system breakdown and brain function activity. The present invention relates to a method for producing a functional mycelium of roe deer fungus with a substrate of rice bran or red ginseng, which enables mass production, and a functional mycelium of roe deer fungus containing an active ingredient of rice bran or red ginseng produced using the same.

In general, Hepicum erinaceus is classified as basidiomycetes, Democratic mushrooms, Beard mushrooms, and Coral Sedimentary Mushrooms, and is one of the functional mushrooms of industrial interest due to its excellent functional efficacy.

Major pharmacologically active ingredients include NGF stimulators, nerve growth factor synthesis inhibitors, Hela cell proliferation inhibitors, immune function modulators, anticancer and dietary fiber (betaglucan). Especially, hericenone derived from fruiting body is known as the first substance that shows similar activity as adrenaline in natural products, and erinacine is also known to have the strongest activity.

It is well known that the sports drink Houtou, which became the driving force of the Chinese players during the 1990 Asian Games, was due to Hericium erinaceus.

In addition, the roe deer mushroom (Hericium erinaceus) is an edible mushroom widely distributed in the northern temperate hemisphere such as China, Japan, Korea, etc. This mushroom is a kind of white fungi parasitic on hardwoods such as oak and chestnut. Cultivation techniques were developed by Liu, which allowed artificial cultivation (Liu. CY 1981. Technique of cultivation of monkeyhead mushroom. Edible Fungi, No. 4, 33). In Japan, artificial cultivation methods have been established, such as cultivation of hazelnuts and fungal cultivation methods using polyethylene (PE) bottles, and are being shipped to the market all year round. In Korea, artificial cultivation has recently begun. H.K. 1998. Cultural characteristics of Hericium erinaceum. M.S. thesis, Korea Univ.]

The chemical composition of the roe deer mushroom is mainly composed of sugar and protein, and contains minerals but low fat content (Eisenhut, R., Fritz, D. and Tiefel, P. 1995. Investigations on nutritionally valuable constituents of Hericium erinaceus (Bull) .: Fr) Pers.Gartenbauwissenschaft, 60 (5), 212-218). There are many minerals in order of Na, P, Mg, etc., but ash composition is also affected by cultivation logs and fungi.

Taste components include free amino acids, free sugars (trehalose, manitol, arabinitol, glucose, etc.) and organic acids (malic acid, fumaric acid, citric acid, α-ketoglutaric acid, etc.). Nucleic acid components such as adenosine, guanoic acid and guanoadenic acid exist as other taste components, and these are known to give Jimmy by synergy with amino acids.

As vitamins, B1, B2 and niacin are present, but vitamins A and C are not detected. In addition, ergosterol (ergosterol), which is expected to play a role in preventing osteoporosis by changing to vitamin D2 by light and heat drying and helping to absorb metabolism of Ca, is commonly present.

However, studies to date have shown that in addition to these nutrients, snail mushrooms have pharmacologically active ingredients such as Hela cell proliferation inhibitors, nerve growth factor (NGF) -induced substances, immune function regulators (BRM effects), Anti-tumor polysaccharides, lectins, dietary fiber (β-glucan, chitin, hetro polysaccharides), and various biological activities have been reported by these [Mizuno, T. 1995. Yamabushitake, Hericium erinaceum : Bioactive substances and medicinal utilization. Food Reviews International, 11 (1), 173-175].

In China, it is famous as a herbal medicine. Hetero-polysaccharides of the fruiting beetle fruit body are known to have anticancer action (stomach cancer, food cancer, liver cancer, skin cancer, etc.) based on the regulation of immune function, and also the immune system of We have reported the effects of fortification on gastric, duodenal ulcer and gastric and esophageal cancer [Yang, QY, and Jong, SC 1989. Medical mushrooms in China. Mush. Sci., 12 (Part 1), 631-643], reported to promote anticancer and immune function. [Ahn, D.K. 1992. Medicinal fungi in Korea. Kor. J. Mycol., 20,154-165].

In particular, the central nervous system by phenol-related compounds derived from fruiting bodies (hericenone C, D, E, YA-8-c) exhibiting synthetic induction promoting activity of nerve growth factor (NGF) or erinacine A-I derived from mycelium It has been reported to improve the regeneration and Alzheimer's dementia, and to report its availability as a therapeutic agent.

The cultivation technology of roe deer mushroom with the above-mentioned efficacy was developed by Liu in 1981 to enable artificial cultivation of solid culture, but the production of fruiting bodies by solid culture requires control of light, temperature and humidity. Due to the low extraction yield, there was a problem that it is difficult to switch to the industrial production system due to high cost, low productivity, quality non-uniformity, quality change during distribution, and the like.

Since the first attempt by Humfeld to agaricus mushroom in the United States in 1948 to solve the above problems, H. eryaceus mushroom (Hericium erinaceus) has been applied to the liquid culture method applied to and reviewed about 60 kinds of edible mushrooms. Grigansky, Lomberh et al. Have been reported on the culture of liquids, but the results of these studies were made at the flask level. The yield of the mycelium was low at 2.7-9.2g / L, and the culture period was 10 days. Optimization is needed.

The present invention is to solve the above-mentioned problems, in particular, the present invention finds that the rice bran or red ginseng extract is effective in mass production of mycelia of roe deer fungus, and the extract of rice bran or red ginseng is added to the primary culture of The functional mycelium production method of the rotifera fungi cultured in the secondary culture and the functional mycelium of the roe deer fungus produced using the above method, such as high cost, low productivity and quality non-uniformity in the solid culture method, quality change in distribution, etc. By solving the problem, it is possible to mass-produce the functional mycelium of the high-quality locust fungus at low cost, as well as to treat senile disease, cancer, immune system breakdown and brain function activity by extracting rice bran or red ginseng used as a substrate. Roe deer beet based on rice bran or red ginseng to have the effect of To provide the functionality of the functional mycelium of mushroom mycelium production method and a Hericium erinaceus containing the effective components of the rice bran or red ginseng produced using this purpose.

Features of the present invention for achieving the above object, the step of preparing a primary culture using a mycelium of roe deer fungus, and adding the extract of rice bran or red ginseng to the primary culture solution prepared in the primary culture step After that, a method for producing a functional mycelium of roe deer fungus with a substrate of rice bran or red ginseng comprising a secondary culture step of secondary culture.

In addition, another feature of the present invention for achieving the above object is a functional mycelium of roe deer fungus containing an active ingredient of rice bran or red ginseng produced by the production method of the above features.

The primary culture step according to the characteristics of the present invention, glucose (glucose) 5%, yeast extract (yeast extract) and peptone (peptone) in a mixture of a mixture of 1: 1 and potassium phosphate anhydride (KH ₂ PO ₄ ) and 0.2% of a mixture of magnesium sulfate (MgSO ₄ ) 1: 1 mixed to form a medium to form a medium, the pH 4 ~ 6, the temperature 20 ~ 30 in the medium formed in the medium composition step ℃, agitation speed 100 ~ 200rpm, working volume 50mL, mycelial seeding incubation step of cultivating spawn at 3 ~ 10%, and the spawn culture in the spawn culture step in the bio-reactor (Bio-reactor) pH 5 ~ 6, temperature 20 ~ 30 ℃, agitation speed 100 ~ 200rpm, aeration rate 1 to 5vvm can be configured to include an embodiment comprising a culturing step for 4 to 8 days incubation.

In addition, the secondary culture step according to the characteristics of the present invention, after adding 1 ~ 7% of the hot water extract of rice bran or red ginseng to the main culture cultured in the main culture step, based on the dry amount, 100 ~ 200rpm, 20 ~ 30 An example can be comprised by the structure which further cultures for 2-3 days under conditions of ° C.

The functional mycelium of roe deer fungus containing the active ingredient of rice bran or red ginseng according to another aspect of the present invention is lyophilized, and then pulverized and powdered in a composition for treatment of senile disease, cancer, and immune system collapse. It is possible to configure the applied embodiment.

The objects, features and advantages of the present invention will be more readily understood by reference to the accompanying drawings and the following detailed description.

Hereinafter, with reference to the accompanying drawings will be described in detail the configuration and effect of the preferred embodiment of the present invention.

1 is a flow chart of a method for producing functional mycelium of roe deer fungus mushrooms based on rice bran or red ginseng according to an embodiment of the present invention, the production method of functional mycelium of roe deer fungus mushrooms based on rice bran or red ginseng of the present invention It is shown schematically. That is, the present invention comprises the steps of preparing a primary culture using a mycelium of roe deer fungus, and after adding the extract of rice bran or red ginseng to the primary culture prepared in the primary culture step, secondary culture As comprising a culturing step,

The primary culture step is prepared, including the medium composition step, spawn culture step, and the present culture step.

The medium composition step is to create an optimal medium for cultivating functional mycelia, glucose (5%), yeast extract (yeast extract) and peptone (peptone) in a 1: 1 mixture of 0.2% And, a medium is prepared by mixing 0.2% of a mixture of potassium phosphate anhydride (KH ₂ PO ₄ ) and magnesium sulfate (MgSO ₄ ) in a 1: 1 ratio,

The seed culture step is a step of culturing the seed in the medium prepared in the medium composition step, pH 4 ~ 6, temperature 20 ~ 30 ℃, stirring speed 100 ~ 200rpm, working volume 50mL, mycelia inoculation ratio 3 The seed is incubated at ~ 7%,

The culturing step is to produce functional mycelium by culturing the seed culture cultivated in the seed culture step, pH 5-6 in a bio-reactor, temperature 25-30 ° C., agitation speed 100-200 rpm, Incubate 4-8 days at aeration rate 1-5vvm.

The secondary culture step is to further culture by adding an extract, such as rice bran or red ginseng to the primary culture liquid prepared through the above culturing process, the hot water extract of rice bran or red ginseng to the main culture cultured in the main culture step After the addition of 1 to 7% on a dry basis, 100 to 200 rpm, 20 to 30 ℃ further incubated for 2 to 3 days, and centrifuged to produce a mycelium.

If the optimum medium composition of the medium composition step according to the embodiment, the comparison of the mycelial production amount according to the input time of the rice bran component, the optimum culture conditions during the spawn culture for the comparison experiment, etc. As follows.

Experimental Example 1

Factor mixture component (water, C-source, N-source, mineral) and 6-level amount (0, 1, 1/2, 1 / 3,1 / 4,1 / 6) Mixture experiment of the Simplex-centroid design using a mixture amount (MA, MA) was performed as shown in Table 1 below.

Table 1. Mixture test conditions of Simplex-centroid design using mixture amount (MA)

Based on the result of incubation for 9 days at the inoculation ratio of 5%, the temperature of 10 ~ 30 ℃, agitation speed of 100rpm in the medium having the optimum medium composition, the mixture ratio of the optimal medium composition was evaluated by 5% glucose (glucose), 0.2% mixture of yeast extract and peptone in 1: 1, 0.2% mixture of potassium phosphate anhydride (KH ₂ PO ₄ ) and magnesium sulfate (MgSO ₄ ) in 1: 1 Set.

In order to set the conditions of the optimum medium as described above, the culture broth was incubated for 9 days at 25-35 ° C., aeration rate 0.5-2.0vvm, and agitation speed 100-300 rpm for about 8 days, under the conditions shown in Table 2 below. After optimizing the culture conditions by performing liquid culture, the seed was incubated for about 4 to 8 days at pH 5 ~ 6, incubation temperature 25 ~ 30 ℃, stirring speed 100 ~ 200rpm.

Table 2. Variables and their steps to establish central elements of culture conditions

The primary liquid main culture was carried out under the optimum medium and culture conditions, and 5 days after the mycelial growth was sufficiently performed, the rice bran component was added to carry out the two-step culture to increase the production of the mycelia.

The amount of the rice bran extract added to the above is 1 to 7% (preferably 3 to 6%) on the basis of the dry amount, and the two-step culture by standing or shaking culture at 20 to 50 ℃ (preferably 20 to 30 ℃) It is preferable to carry out. Table 3 shows the results of the growth of the mycelia by the input time of the rice bran extract during the whole 8 days of liquid culture.

Table 3. Comparison of Mycelial Production

division General liquid culture  Rice bran added common liquid (stage 1 culture) The present invention (Bulb 2 stage culture) Incubation period 7-8 days 7-8 days 7-8 days Mycelial Production 4.25-10.77 g / L 1.5g / L 16.65-17.89 g / L Remarks Lab scale Lab scale 20, 75 and 300L Pilot Validation

As can be seen in Table 3 above, when starting from day 0 of culture, the growth of mycelium itself tended to be inhibited as in the results of mycelial culture experiments using garlic, but when the rice bran extract was added after 1 day or more, In the early stage, the growth of mycelium seemed to be somewhat decreased, but as the input time was delayed, the production amount increased rapidly, and after 4-5 days of incubation, the mycelium growth was much higher than that of the conventional liquid culture method.

In detail, the mycelial production was only about 1.5 g / L when the initial cultivation of rice bran or red ginseng extract was added with this method, but 16.65 ~ 17.89 in 20, 75, and 300 L fermenters after 4-5 days of primary culture. Up to g / L can be obtained, which is more than 160-300% improvement over 4.25-10.77 g / L of conventional liquid culture.

In addition to the functional mycelium production method of the present invention of the rice bran or red ginseng as a substrate, and the functional mycelium of the roe deer fungus containing the active ingredient of the rice bran or red ginseng produced by the present invention can be variously modified, Unless departing from the scope of the invention, all modified embodiments should be construed as being included in the scope of the invention.

According to the present invention, by solving the problems of high cost, low productivity and quality non-uniformity, quality change during distribution in the solid culture method, it is possible to mass-produce functional mycelium of roe deer fungus of high quality at low cost In addition, by using the rice bran or red ginseng extract used as a substrate can be obtained such as the effects of the treatment of senile diseases, cancer, immune system breakdown and brain function activity.

Claims (5)

delete In the method for producing a functional mycelium by liquid culture of roe deer mushroom, 5% glucose, 0.2% mixture of yeast extract and peptone in a 1: 1 ratio, potassium phosphate anhydride (KH ₂ PO ₄ ) and magnesium sulfate (MgSO ₄ ) 1: A medium composition step of forming a medium by mixing 0.2% of the mixture mixed with 1; A seed culture step of culturing the spawn with a pH of 4 to 6, a temperature of 20 to 30 ° C., a stirring speed of 100 to 200 rpm, a working volume of 50 mL, and a mycelium inoculation ratio of 3 to 7% in the medium prepared in the medium composition step. , The primary culture step of incubating the seed culture cultured in the seed culture step in a bio-reactor at pH 5-6, temperature 25-30 ° C., agitation speed 100-200 rpm, aeration rate 1-3 vem for 4-8 days. Wow, Method for producing a functional mycelium of roe deer fungus with a substrate of rice bran or red ginseng, characterized in that the step of adding the extract of rice bran or red ginseng to the primary culture prepared in the culture step and then secondary culture. In the method for producing a functional mycelium by liquid culture of roe deer mushroom, Preparing a culture solution using the mycelia of the roe deer fungus; After adding 1 to 7% of the hot water extract of rice bran or red ginseng to the culture based on the dry amount, the rice bran or red ginseng as a substrate, which is further cultured for 2 to 3 days under conditions of 100 to 200 rpm and 20 to 30 ° C. Method for producing functional mycelia of a Roe deer fungus. Functional mycelium of roe deer fungus containing an active ingredient of rice bran or red ginseng produced by the production method of any one of claims 2 to 3. delete

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