KR100723068B1 - Method for culturing flammulina velutipes including ginseng saponin - Google Patents

Abstract

The present invention relates to a cultivation method of the top mushroom containing ginseng components. The cultivation method of the fungi mushrooms production step of culturing by injecting the strain of the enoki mushroom in a natural medium to which ginseng saponin component is administered; The first medium for producing a first medium containing a solution in which the ginseng stock solution is diluted 1000-fold, soybean meal, white sugar, potassium phosphate, magnesium sulfate, and inoculated into the first medium the spawn produced by the prokaryotic preparation step A liquid inoculation source manufacturing step comprising a culturing step of culturing the liquid spawn after culturing; A second medium manufacturing step of preparing a second medium containing 500 times diluted ginseng stock solution, sawdust, corncob, rice bran, wheat bran, cottonseed foil, and waste fossil; A liquid spawn culture step comprising an inoculation step comprising an inoculation step of inoculating the liquid hyphae prepared by the liquid inoculum manufacturing step on the infected second medium; The step of scraping bacteria to remove the aged bacteria on the surface of the finished seed culture and spray the solution diluted 500 times the ginseng stock solution; It includes; the organic phase step of regenerating the mycelia on the surface of the medium removed by the bacteria scraping step.

 According to the cultivation method of the above-mentioned mushrooms, the ginseng extract is added to each step from the spawning, cultivation, inoculation and growth of the top mushroom, so that the ginseng component which performs useful functions to the human body may be included in the top mushroom. You can do that. It also provides the advantage that the process of incorporating ginseng into the enoki mushroom can be carried out very efficiently and reliably.

Description

Method for culturing flammulina velutipes including ginseng saponin}

1 is a test report of the ginseng component analysis for the sample of the top mushroom cultivated by the method of cultivating the top mushroom according to an embodiment of the present invention.

The present invention relates to a cultivation method of the top mushroom containing ginseng components.

Flammulina velutipes belongs to Dongdam ego, pine and pine (white porcelain). Among them, it is a type of white decaying fungus that rots timber. It is a mushroom that is used by oxidizing and decomposing cellulose and lignin, which are the main constituents of wood, through activating, and changing them into soluble substances such as sugars.

Wild top mushrooms that occur in dead wood, such as hackberry and persimmon, grow and give elasticity by gathering several pieces. The stem is thin, about 0.3 ~ 0.4cm, the length is 3 ~ 5cm relatively long, and the shade is flattened as it grows up to the size of 3 ~ 5cm in diameter, and the surface is sticky and glossy when dried. The color of the shade is light brown, and the bottom is dark brown and triangulation is occurring. Unlike shiitake mushrooms or oyster mushrooms, the stems are tubular, hollow in the middle, easy to break and clump, but the flesh is soft and elastic, and the shade is viscous and sticky.

Enoki mushroom is a mushroom with excellent palatability to people because it contains various essential amino acids with anticancer effect.

Enoki mushrooms have been grown in Korea since the late 1980s as a mechanized bottle cultivation system in a facility that can artificially control environmental conditions such as temperature, humidity, and light. In Korea, the market is increasing due to the unique color and taste of the top mushroom, but it is known to be more difficult to grow than other mushrooms because it requires the top mushroom growing facility, equipment, and skill.

The cultivation process of the top mushroom is cultivated through the process of cultivation, fermentation, suppression and fruiting part growth by inoculating the seedlings after sterilizing the medium in polypropylene bottles based on sawdust and rice bran.

Most of the spawns currently in use are inoculated with the strains inoculated with the medium in the form of sawdust and rice bran as sawdust spawns. Sawdust spawn causes late mycelial sticking due to spawning inoculation into sterilized bottle media and the media site. Especially, sawdust spawn uses cultivated strains for more than 3 weeks. I'm concerned.

The liquid spawn using the liquid medium has the advantage of not only controlling the spawn amount to some extent but also greatly shortening the culture period of the spawn. Currently, research on systematic culture techniques, substrates, and inoculation methods for liquid spawns has not been conducted.

On the other hand, ginseng has a number of active ingredients, including saponins. Saponin is an active ingredient of ginseng and is used as an herbal medicine such as cardiac medicine or diuretic, and is used as an index component of quality control of ginseng products. According to the recent analysis, ginseng contains not only saponin but also polyacetylene and acid polysaccharide which have antitumor activity and anti-tumor activity, as well as essential oil, plant sterol, tenol, protein, peptide, alkaloid, amino acid, nucleic acid, It is known to contain vitamins, lignin, minerals and the like.

The present invention was created in order to contain the active ingredient of ginseng in the top mushroom as described above, by adding the ginseng stock solution at each stage from the spawning, cultivation, inoculation and growth of the top mushroom, contains the ginseng component very efficiently and reliably It is an object of the present invention to provide a method for cultivating enoki mushrooms.

Method of cultivating the mushrooms according to the present invention for achieving the above object

Prokaryotic preparation step of culturing by injecting the strain of the enoki mushroom in natural medium to which ginseng saponin component is administered;

The first medium for producing a first medium containing a solution in which the ginseng stock solution is diluted 1000-fold, soybean meal, white sugar, potassium phosphate, magnesium sulfate, and inoculated into the first medium the spawn produced by the prokaryotic preparation step A liquid inoculation source manufacturing step comprising a culturing step of culturing the liquid seed after culturing;

A second medium manufacturing step of preparing a second medium containing 500 times diluted ginseng stock solution, sawdust, corncob, rice bran, wheat bran, cottonseed foil, and waste fossil; A liquid spawn culturing step comprising an inoculation step of inoculating a liquid spawn prepared by the liquid inoculation step on the encapsulated second medium;

The step of scraping bacteria to remove the aged bacteria on the surface of the finished seed culture and spray the solution diluted 500 times the ginseng stock solution;

It is characterized in that it comprises a; the organic phase step of regenerating the mycelium on the surface of the medium removed by the bacteria scraping step.

Hereinafter, a preferred embodiment of the cultivation method of the top mushroom containing the ginseng component according to the present invention will be described in detail.

Probiotic production stage (First stage)

The prokaryotic preparation step is to cultivate the strain of the enoki mushroom, a natural medium manufacturing step for producing a natural medium, and a strain using a tissue separation step and a natural medium to obtain a strain by transplanting some tissue from the fruiting body of the top mushroom It includes a culture step of culturing. This will be described in more detail as follows.

(1) Natural medium production stage

In order to manufacture natural medium mixed with nutrients and other special substances required for the culture to cultivate the tissues of the fungus strains, administration of ginseng saponin to agar medium containing potato extract It was prepared by. This natural medium was prepared by mixing 45 parts by weight of agar and 0.1 parts by weight of ginseng saponin component when 200 g of potato was added to 1 liter of distilled water and 100 parts by weight of the extract was extracted from hot water. Here, the natural medium may further include rice bran and ink extract.

(2) tissue separation stage

A strain of the enoki mushroom is obtained by transplanting a part of the tissue from the fruiting body. The strain used in the present invention was used by receiving a strain from the Agricultural Science and Technology Institute.

(3) culture step

In this culture step, the medium in which the strain was transplanted as described above was put in a thermostat at 20 to 22 ° C. and grown about 1 to 2 cm, and then grown in new medium.

In more detail, the strain obtained from the fruiting body was transplanted to the natural medium, the natural medium was grown to about 2cm while maintaining the temperature at about 20.5 ℃ using a thermostat, and transplanted into a new natural medium to grow. After transplanted into the new natural medium and propagated for 3 days, it was used as a strain of the first medium to be described later.

Strains not used here were stored in a cool dark place at about 4 ° C., removed and used whenever necessary, and used, and the aged strains were discarded.

Liquid Inoculation  Manufacturing stage (second stage)

This liquid inoculum preparation step is for producing a liquid seed bacterium, the first medium production step, the inoculation step for transplanting and culturing the strain prepared in the probiotic production step on the first medium and the culture for inoculating the inoculated strain With steps.

(1) first medium production step

The first medium is for liquid culture of the strain, and the material of the first medium includes potato extract, agar, ginseng stock solution, soybean meal, white sugar, potassium phosphate, and magnesium sulfate.

Such a first medium may be prepared by the following method.

When 100 parts by weight of the potato extract was extracted from hot water, 45 parts by weight of agar and 150 ml of ginseng stock solution were diluted 1000 times using groundwater. 2 parts by weight of ginseng solution, 5 parts by weight of soybean meal, 5 parts by weight of sugar, and phosphoric acid A mixed solution is prepared by mixing 0.03 parts by weight of potassium and 0.05 parts by weight of magnesium sulfate.

The mixture is stirred and then sealed to sterilize. Sterilization of the mixed solution is firstly sterilized at 100 ° C. for 80 minutes using a sterilizer and secondly at 121 ° C. for 40 minutes. When the 1st and 2nd sterilization is completed, cool down to an appropriate temperature (15 ℃) for inoculation.

(2) inoculation step

The cooling medium vessel was moved to the experimental culture chamber, and the strain in which the culture was completed in the first step was inoculated using an Erlenmeyer flask. That is, 50 ml of the strain was inoculated into the flask containing 400 ml of the first medium.

(3) culture step

This culturing step is to cultivate the strain of the inoculated top mushroom, the top mushroom is aerobic bacteria to cultivate the hyphae during the cultivation of liquid spawn, incubated while supplying air to prevent this. At this time, the temperature of the culture chamber is preferably about 14 to 15 ℃, the culture period took about 26 days.

To explain this in more detail, the Erlenmeyer flask cultured with mycelium was precultured at 130 RPM for 25 days at 25 ° C. using a shaker incubator. The main culture was inoculated with 200 ml of the preculture solution in a 5-liter three-neck flask (3neck fla or Duran, Germany) containing 3 liters of cultured mycelium cultured as above. Was carried out. Aeration at the time of incubation was carried out using two vents (24 mm in diameter and 34 mm in length) in a three neck flask.

On the other hand, the liquid spawn that has been cultured is used after performing a contamination test. The contamination test is a method of inoculating a part of the cultured culture solution in potato agar medium and incubating at 25 to 40 ° C. to test for bacterial contamination. It was.

Liquid spawn  Incubation stage (third stage)

The liquid seed culture step is a step for cultivating the top mushroom by inoculating the liquid seed culture incubated as described above.

(1) second medium production step

The second medium is for culturing the liquid spawn, and is prepared with the ginseng stock solution, sawdust, corn cob, rice bran, wheat bran, cottonseed foil, and waste fossil of the second medium.

To prepare the second medium, the mixture was prepared by stirring 15 parts by weight of corncob, 10 parts by weight of wheat bran, 5 parts by weight of rice bran, 5.5 parts by weight of cottonseed foil, and 5 parts by weight of waste fossil, based on 100 parts by weight of sawdust. In this mixture, 1500 ml of ginseng stock solution was diluted 500 times in groundwater to prepare a final 63% moisture in the medium.

(2) Entry stage

The initiation step is to put the second medium prepared as described above into a polypropylene bottle for cultivation, and the initiation is made as soon as possible in order to prevent the medium from being deteriorated due to delay.

(3) sterilization stage

In order to sterilize the second medium after completion of the disease, the bottle containing the second medium was first sterilized at 100 ° C. for 80 minutes using a steam autoclave, and sterilized at 121 ° C. for 40 minutes for aseptic conditions.

(4) cooling stage

The second medium sterilized in the stomach was cooled to a temperature (15 ° C.) capable of inoculating the seed mushrooms of the top mushroom.

(5) Inoculation step

The step of inoculating the mycelia to the liquid mycelium (liquid spawn) which is the culture medium in the liquid inoculum cultivation step in the second medium cooled above.

(6) culture step
After completion of the inoculation, the second medium, that is, the bottles containing the second medium, are transported and loaded into a culture chamber at 14 ° C. and incubated for about 25 to 26 days while controlling humidity and ventilation. After the incubation, the seedlings are screened for preparation of the germs and ready for the bacterium scraping step described below.

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At this time, the enoki mushroom is white, so any other color or turbidity of spawn disease is considered as a germ and removed.

Scratching (Fourth step)

In this step, the culture is carried out to remove the aged bacteria from the pigmented seed. At this time, a solution of 200 ml of ginseng stock solution diluted 500 times on one surface of the bacterium scraping was sprayed 2 to 3 times so that the surface of the bacterium scraping was wetted and moved to a foot chamber.

Organic hair  Step (5th step)

This step is to regenerate the mycelium on the surface of the fresh medium removed by the bacteria scraping in the fourth step to form the base of the young fruiting body of the mushrooms.

For the fungi of the fungus, the temperature was maintained at a temperature of 13 to 14 ° C., a relative humidity of 90% or more, a CO ₂ concentration of 1000 ppm or less, and a roughness of 100 to 200 lux. Unlike the environmental conditions at the time of culture, this step is to induce a stimulus to change the reproductive growth by changing the environment, such as low temperature, humidity, light irradiation.

7 to 8 days after the foot weaving, a grain-shaped lump is formed on the surface of the medium, and a lump of spirum continues to rise, and at the same time, the tip of the epoch is colored with brown to form a small bead. On the 10th day young fruiting bodies are formed. From this time, the fruiting step of fruiting body described later was performed.

Growth stage (sixth stage)

In this growth stage, the temperature was maintained at 4 to 5 ° C, the ginseng stock solution was diluted 1000-fold and humidified until the growth stage. However, it is not humidified from the mid-growth until the harvest stage.

Harvest stage (seventh stage)

When the top mushroom is grown up to about 10 to 12cm, harvested and shipped in various specifications.

Ginseng Components Analysis

As a result of analyzing the content of the mushrooms, which were cultivated by the cultivation method as described above by the Yeongunggwangju Environmental Life Research Institute, the ginseng component was found to contain 2.16 mg / g (Fig. 1 test). See test report).

The matters described in this specification and the drawings are presented by way of example to describe preferred embodiments of the present invention, and the scope of protection of the present invention should not be construed as being limited to these matters.

According to the cultivation method of the enoki mushroom according to the present invention, since the ginseng stock solution is added at each stage of the seed collection, cultivation, inoculation and growth of the enoki mushroom, a significant amount of the ginseng component which performs useful functions to the human body is included in the enoki mushroom. You can do that. In addition, according to the present invention provides an advantage that can be performed very efficiently and reliably the process of containing ginseng in the top mushroom. In addition, ginseng saponin removes the odor of mushrooms, which improves spawn vitality.

Claims (2)

Prokaryotic preparation step of culturing by injecting the strain of the enoki mushroom in natural medium to which ginseng saponin component is administered; The first medium for producing a first medium containing a solution in which the ginseng stock solution is diluted 1000-fold, soybean meal, white sugar, potassium phosphate, magnesium sulfate, and inoculated into the first medium the spawn produced by the prokaryotic preparation step A liquid inoculation source manufacturing step comprising a culturing step of culturing the liquid spawn after culturing; A second medium manufacturing step of preparing a second medium containing 500 times diluted ginseng stock solution, sawdust, corncob, rice bran, wheat bran, cottonseed foil, and waste fossil; A liquid spawn culture step comprising an inoculation step comprising an inoculation step of inoculating the seed spawned with the liquid spawn prepared by the liquid inoculum preparation step; The step of scraping bacteria to remove the aged bacteria on the surface of the finished seed culture and spray the solution diluted 500 times the ginseng stock solution; A reorganization step of regenerating mycelia on the medium surface removed by the bacterium scraping step; The method of cultivating the enoki mushroom containing the ginseng component, characterized in that it further comprises a growth step of growing while supplying a solution diluted in the ginseng stock solution 1000 times the fruiting body formed in the organic phase step. 200 g of potato in 1 liter of distilled water to make 100% by weight of the extract extracted from hot water, agar-producing step of culturing the fungi by adding strains of the enoki mushroom to natural medium prepared by mixing 45 parts by weight of agar and 0.1 parts by weight of ginseng saponin components Wow; When 100 parts by weight of the potato extract was extracted from hot water, 45 parts by weight of agar and 150 ml of ginseng stock solution were diluted 1000 times using groundwater. 2 parts by weight of ginseng solution, 5 parts by weight of soybean meal, 5 parts by weight of sugar, and phosphoric acid After sealing the mixed solution of 0.03 parts by weight of potassium sulfate and 0.05 parts by weight of magnesium sulfate, the first sterilization was carried out at 100 ° C. for 80 minutes, and the second sterilization was carried out at 121 ° C. for 40 minutes to prepare the first medium and the progenitor production step. A liquid inoculum manufacturing step comprising a culture step of inoculating the spawn made by the inoculation into the first medium to form a liquid spawn; A second medium manufacturing step of preparing a second medium containing 500 times diluted ginseng stock solution, sawdust, corncob, rice bran, wheat bran, cottonseed foil, and waste fossil; A liquid spawn culture step comprising an inoculation step comprising an inoculation step of inoculating the seed spawned with the liquid spawn prepared by the liquid inoculum preparation step; The step of scraping bacteria to remove the aged bacteria on the surface of the finished seed culture and spray the solution diluted 500 times the ginseng stock solution; A reorganization step of regenerating mycelia on the medium surface removed by the bacterium scraping step; The method of cultivating the enoki mushroom containing the ginseng component, characterized in that it further comprises a growth step of growing while supplying a solution diluted in the ginseng stock solution 1000 times the fruiting body formed in the organic phase step.

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