KR101479209B1 - Manufacturing Process of Lentinus edodes Culture - Google Patents

Abstract

The present invention relates to a method for producing a mushroom culture. Specifically, the present invention relates to a method for producing a mushroom culture, characterized in that the solid medium is introduced into a muzzle, the liquid medium is inoculated and cultured, and then the solid medium is deglossed and the mushroom is again bagged.

Description

{Manufacturing Process of Lentinus edodes Culture}

The present invention relates to a method for producing Lentinus edodes ) culture. Specifically, the present invention relates to a method for producing a mushroom culture, characterized in that the solid medium is introduced into a muzzle, the liquid medium is inoculated and cultured, and then the solid medium is deglossed and the mushroom is again bagged.

Shiitake mushroom is a mushroom of the wormwood mushroom thorny mildew, and it occurs in the dry tree of the hardwood such as an oak tree, a chestnut tree, a crescent tree twice in spring and autumn. The mushroom is 4 to 10 cm in diameter and initially hemispherical, but gradually flattened and flattened. The surface is fresh brown and covered with a scaly piece of blackish-brown, thin, fluffy fur. Sometimes it bursts to show white flesh. Artificial cultivation is carried out by wood, and it is distributed in Korea, Japan, China, and Taiwan.

The effectiveness of shiitake mushrooms has been studied extensively in China since shiitake mushrooms contain a substance called eridadine, which reduces cholesterol levels in the blood. In addition, since it also acts to lower blood pressure, it is suitable for prevention of hypertension and arteriosclerosis. Because eridademin melts when dry mushrooms are concerned about water, it is better to use the juice without throwing away. Vitamins B1 and B2 are also abundant. The richness of shiitake mushrooms is a component of nucleic acid-based seasoning as guanylate. The smell is due to the wrench onion. In addition, shiitake mushrooms contain a lot of ergosterol that has the effect of vitamin D, changes to vitamin D when the body receives ultraviolet light. On the other hand, it was found that vitamin B12, which was known not to exist in plants, was found in the altitude. About 600 years ago, in the Ming Dynasty of China, a person called Oshi (吳瑞) called the efficacy of the elevation "a feast of blood and blood". When we analyze this modernly, it is that the spore of shiitake mushroom has a component that governs the influenza and cancer belonging to the cancer nowadays. And when you add shiitake mushrooms to various vegetable dishes, it tastes better than meat. This is because it contains guanylic acid, the most potent taste ingredient.

On the other hand, it is known that when liquid mushroom is used in the culture of mushroom, the growth efficiency is high because it can spread evenly over the solid medium and utilize a wide area of the medium compared to the solid medium. However, when the liquid seedlings are inoculated in the cultivation of the shiitake mushroom, there is a problem that the proportion of contaminated by other bacteria is high before the hyphae completely spread over the solid medium, and the amount of produced shiitake is much smaller. Because of this, in the case of shiitake mushrooms, solid seeds with a yield of less than half and a much longer adaptation period are still used.

KR 2011-0037447 Agricultural company corporation Hana Biotech Co., Ltd. 2011.04.13. KR 1988-0005854 Kim Wonhong 1988.07.21).

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a method for cultivating mushroom It is an object of the present invention to provide a method for producing a culture.

In order to accomplish the above-mentioned object, the present invention provides a method for producing a mushroom culture product,

(A) 100 parts by weight of sawdust and 10 to 40 parts by weight of rice bran, and adding 150 to 250 parts by weight of water to prepare a solid medium,

(B) sterilizing the solid medium at a temperature of 100 to 135 캜 for 2 to 3 hours and cooling in a sterilization chamber,

(C) inoculating the solid medium with 5 to 15 parts by weight of the mushroom culture on the basis of 100 parts by weight of the sawdust in a sterilization chamber, incubating the mixture at 15 to 25 DEG C for 40 to 55 days, and

(D) removing the cultured cultures from the vials, re-introducing them into porous bags, culturing them at 15 to 25 DEG C for 45 to 60 days

Lt; / RTI >

The mushroom culture of step (C)

100 parts by weight of water,

1 to 5 parts by weight of a saccharide selected from the group consisting of glucose, fructose, galactose, maltose, lactose and mixtures thereof,

0.2 to 0.5 parts by weight of a powder selected from the group consisting of corn flour, corn, peptone and mixtures thereof,

Selected from the group consisting of K ₂ HPO ₄ , K ₃ PO ₄ , NaH ₂ PO ₄ , Na ₂ HPO ₄ , Na ₃ PO ₄ , KHCO ₃ , K ₂ CO ₃ , NaHCO ₃ , Na ₂ CO ₃ and mixtures thereof 0.01 To 0.1 part by weight,

0.03 to 0.1 part by weight of MgSO ₄ .7H ₂ O, and

NaNO ₃ , Ca (NO ₃ ) ₂ and mixtures thereof 0.02 to 0.2 parts by weight

And the mushroom seeds are cultured in a liquid medium containing the mushroom seeds.

In addition, the method for producing a mushroom culture of the present invention may further comprise 10 to 20 parts by weight of wheat bran per 100 parts by weight of sawdust in the solid medium of step (A).

In addition, the method of the present invention may further comprise 5 to 10 parts by weight of cotton seed hull per 100 parts by weight of sawdust in the solid medium of step (A).

The liquid medium of step (C) preferably further comprises 0.002 to 0.05 parts by weight of a defoamer or edible oil per 100 parts by weight of the water.

In addition, it is preferable that the mushroom culture of step (C) is subjected to an insecticidal test before inoculating the solid medium.

In addition, in the method for producing a mushroom culture of the present invention, it is preferable that the humidity at the time of putting the culture cultured in the step (D) in the porous bag back into the porous bag is 1 to 60%.

In addition, in the method for producing a mushroom culture of the present invention, it is preferable that the temperature at which the culture cultured in step (D) is put back into the porous bag is 10 to 20 ° C.

It is preferable that the light intensity of step (D) is in the range of 150 to 400 Lux.

In addition, it is preferable that the humidity at the time of light cultivation in step (D) is 50 to 70%.

The sawdust is preferably oak sawdust.

The method of producing the mushroom culture of the present invention has an advantage that the sterilization can proceed perfectly by including the step of entering into the muzzle bottle before the step of pouring the solid medium into the bag. This has the effect of shortening the work time required for the entrance and the insertion, and significantly reducing the contamination rate caused by the germs. This pollution-reducing effect is due to the complete sterilization, but since the mushroom hyphae have already been sufficiently grown in the mushroom, the mushroom is dominant in the initial stage of the solid culture and the growth of the mushroom can be sufficiently inhibited.

In addition, since the present invention provides the hyphal culture in the solid medium in such a state that the hypha is already sufficiently grown, the total cultivation days can be shortened in terms of farmers who intend to cultivate the mushroom by purchasing the solid medium of the present invention. Specifically, in the case of culturing by inoculation with conventional solid organisms, it took 3 to 4 months for the mycelium to completely spread throughout the solid medium, and the mushrooms could not be harvested during this period. However, in case of inoculating liquid organisms It takes only 1 to 2 weeks for the mycelium to spread completely throughout the solid medium, and productivity is improved remarkably.

The present invention also allows for commercial production of at least 11 to up to 17 times per solid medium, resulting in a significant increase in efficiency compared to commercial production of up to six times of the conventional solid organism inoculation method. In addition to the complete sterilization described above, securing the dominance status of the shiitake mushroom, efficiency of the liquid seed bacterium, and deoxidizing and mixing the primary admitted solid medium, sufficient oxygen can be supplied to the solid medium, and the growth of the shiitake mushroom It seems to be because of

Fig. 1 is a photograph showing a step of mixing a solid medium material.
Fig. 2 is a photograph of the stage of entry into which a solid medium is put into a vial bottle.
FIG. 3 is a photograph of a step of sterilizing and cooling a masonry bottle.
FIG. 4 is a photograph showing a step of inoculating a liquid seedling into a vial bottle.
FIG. 5 is a photograph showing a step of culturing in a vial.
FIG. 6 is a photograph showing a degassing step for taking out a solid medium from a masonry bottle.
Fig. 7 is a photograph showing a step of filling a solid medium into a bag.
8 is a photograph showing a step of culturing in an encapsulation.
Fig. 9 is a photograph of a culture medium that is completed and awaited for sale.

Hereinafter, preferred embodiments of the present invention will be described in detail. In the following description, numerous specific details, such as specific elements, are set forth in order to provide a thorough understanding of the present invention, and it is to be understood that the present invention may be practiced without these specific details, It will be obvious to those who have knowledge of. In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

The method for producing the shiitake culture of the present invention starts with mixing 100 parts by weight of sawdust and 10 to 40 parts by weight of rice bran and adding 150 to 250 parts by weight of water to prepare a solid medium. Here, the sawdust is particularly preferable for oak mushroom cultivation. It is further preferable that the solid medium further contains 10 to 20 parts by weight of wheat bran per 100 parts by weight of sawdust, and further may include 5 to 10 parts by weight of cotton seed hull.

The solid medium thus prepared should be sterilized before the shiitake mushroom is inoculated. In the present invention, the sterilization effect is maximized by sterilizing the solid medium at 100 to 135 ° C for 2 to 3 hours. The sterilized solid medium was cooled to room temperature in the sterilization chamber.

Separately, the solid medium of the present invention is characterized in that a liquid seed is inoculated instead of the conventional solid seed inoculation. Generally, it is known that liquid seedlings are more efficient than solid seedlings because they can penetrate evenly into the solid medium and utilize a wide area of the medium. However, inoculation of liquid seedlings during the cultivation of shiitake showed that the proportion of contaminated by other bacteria was high before the mycelium completely spread throughout the solid medium, and the amount of shiitake produced was much smaller. This is because the nutrients in the liquid medium cause other bacteria to grow first in the solid medium, resulting in loss of efficiency as the mushroom loses its dominant position and consequently grows in competition with other microorganisms.

However, according to the present invention, the solid medium is first put into a bottle before being packed in a bag, and is subjected to a complete sterilization process, thereby remarkably lowering the possibility of contamination of germs, and as a result, it is possible to grow only shiitake mushrooms alone.

Specifically, in the present invention, the method for producing a liquid seed bacterium firstly starts from the step of obtaining the bacterium. A plate medium containing a PDA (Potato dextrose agar) was placed in a Petri dish, and shiitake mushrooms having a diameter of 2 to 7 cm were cut to have an average edge length of 1 to 7 mm, Lt; / RTI > After the tissue-implanted Petri dishes were cultured at 20 to 25 DEG C for 10 to 15 days, the grown shiitake mycelia were cut with a plate medium to an average length of 5 to 10 mm, transferred to a new PDA plate medium and cultured , And this process is repeated one to two more times to obtain a mushroom mold.

1 to 5 parts by weight of saccharide selected from the group consisting of glucose, fructose, galactose, maltose, lactose and a mixture thereof, corn flour, corn, peptone and mixtures thereof is added to 100 parts by weight of water. To 0.5 parts by weight of an inorganic filler composed of K ₂ HPO ₄ , K ₃ PO ₄ , NaH ₂ PO ₄ , Na ₂ HPO ₄ , Na ₃ PO ₄ , KHCO ₃ , K ₂ CO ₃ , NaHCO ₃ , Na ₂ CO ₃ , 0.03 to 0.1 part by weight of MgSO ₄ .7H ₂ O and 0.02 to 0.2 part by weight of NaNO ₃ , Ca (NO ₃ ) ₂ and mixtures thereof, , Sterilized, cooled in an aseptic state, placed in a flask or a Ringer's bottle, inoculated with 2 to 5 pieces of an average sized 5 to 10 mm flanking section obtained with the above-mentioned flat plate medium, Lt; / RTI > for 20 to 25 days. Scale it up again and mass-cultivate liquid seeds in the tank. It is preferable to pressurize the tank by 5 to 30% in order to sufficiently inject air into the tank.

Further, it is preferable that the liquid medium is further added with 0.002 to 0.05 part by weight of a defoamer or cooking oil per 100 parts by weight of water to remove the bubbles generated during the culture.

It is preferable to inspect the culture solution under a microscope at 18 to 23 days after the tank culture to check whether the bacteria are present and the clamp condition of the seed bacterium is appropriate.

The thus-prepared liquid seed culture, i.e., mushroom culture liquid, is inoculated in a sterilization chamber in a solid medium in the above-mentioned area by 5 to 15 parts by weight, based on 100 parts by weight of the sawdust, and cultured at 15 to 25 DEG C for 40 to 55 days. The concentration of carbon dioxide in the incubation chamber increases with the progress of cultivation in such a culture, and it is preferable that the concentration is controlled not to exceed 1000 to 3000 ppm.

These cultured cultures, that is, the mixture of the solid medium and the shiitake mushroom mycelia, are taken out from the above-mentioned vials, mixed and then put in a porous bag and incubated at 15 to 25 ° C for 45 to 60 days. By mixing the solid medium once, oxygen is sufficiently supplied to the medium to allow the cells to vigorously grow without restriction due to oxygen in the subsequent culture.

Here, it is preferable that the humidity when the cultured culture is put back into the porous bag is 1 to 60%. If the humidity is more than 60%, the possibility of contamination by germs increases. Also, the temperature of 10 to 20 DEG C is preferable for growing shiitake without lag time. In addition, the illuminance of the light culture is preferably 150 to 400 Lux, and the humidity during light culture is preferably 50 to 70%. In addition, as in the case of the dark culture, the concentration of carbon dioxide in the culture chamber increases with the progress of culture, and it is preferable to control the concentration of the culture medium so as not to exceed 1000 to 3000 ppm.

Hereinafter, embodiments of the present invention will be described.

Example

Example  - Tissue separation

A plate medium containing a PDA (Potato dextrose agar) was placed in a petri dish, and shiitake mushrooms having a diameter of 5 cm were cut to have an average edge length of 5 mm, and the pieces were transplanted into the plate medium aseptically. The tissue-implanted Petri dishes were incubated at 25 ° C. for 12 days, and the grown mycelia of mushrooms were cut with a plate medium to an average length of 7 mm, transferred to a new PDA plate medium and cultured for one time Further, a shiitake mushroom was obtained.

Example  - Liquid culture (1)

1, distilled water 500 ml in l vessel, glucose 10 g, okbun (Lotte, _{Korea) 2 g, Na 2 CO 3} 0.4 g, MgSO 4 · 7H 2 O 0.4 g, Ca (NO 3) the addition of ₂ 0.6 g, was mixed The medium was sterilized in an autoclave (120 ° C) for 1 hour, cooled in an aseptic state, and then inoculated with 4 pieces of the average size 7 mm pieces of the virus obtained together with the plate medium, And cultured in a shaker for 25 days.

Example  - Liquid culture (2)

500 l of distilled water, 10 kg of glucose, 2 kg of lactose (Korea), 400 g of Na ₂ HPO _4, 400 g of MgSO ₄ .7H ₂ O, 600 g of NaNO ₃ , 0.3 ml of cooking oil (CJ, Korea) And the mixture was sterilized at 120 DEG C for 2 hours. After cooling in an aseptic state, 500 mL of the obtained mushroom culture was inoculated and cultured at 25 DEG C for 25 days.

Example  - Cancer culture

10 kg of oak sawdust, 1.5 kg of rice bran, 1.5 kg of bran, 20 kg of water was added to the above mixture, 700 g of the mixture was added to the vial bottle, sterilized at 120 ° C. for 100 minutes, Medium was prepared. In the sterilization chamber, 20 cc of the mushroom culture medium was inoculated into one solid medium of the above-mentioned pore bottle and incubated for 45 days at 20 ° C.

Example  - Cultivation

The above mushroom cultures mixed with the mushroom mycelium and the solid medium were taken out from the mosaic bottle and mixed. The mushroom cultures were mixed again in 70 pores with a through hole and cultured at 20 ° C for 50 days.

It was confirmed that the cultured mushroom culture of the present invention requires only one to two weeks for fully spreading the hyphae throughout the solid medium, and that the commercial cultivation can be performed at least 11 times to 17 times in one solid medium.

Comparative Example  One: Solid seed  culture

After preparing a plate culture medium containing PDA in a Petri dish, shiitake mushrooms having a diameter of 5 cm were cut to have an average edge length of 5 mm, and the pieces were transplanted into the plate medium aseptically. The tissue-implanted Petri dishes were incubated at 25 ° C. for 12 days, and the grown mycelia of mushrooms were cut with a plate medium to an average length of 7 mm, transferred to a new PDA plate medium and cultured for one time Further, a shiitake mushroom was obtained.

Then, 20 kg of water was added to 10 kg of oak sawdust, 1.5 kg of rice bran, and 1.5 kg of bran, and sterilized at 120 ° C. for 100 minutes and cooled to prepare a solid medium. The solid medium was placed in a bag having 70 through-holes in a sterilization chamber, and 4 pieces of the sliced mushroom slices having an average length of 7 mm obtained together with the plate medium were inoculated and incubated at 20 ° C for 70 days.

The cultivated shiitake mushroom cultures were found to be commercially available for up to 6 times on a single solid medium, requiring 3-4 months to completely spread mycelium throughout the solid medium.

Comparative Example  2: Liquid seed  culture

A plate medium containing a PDA (Potato dextrose agar) was placed in a petri dish, and shiitake mushrooms having a diameter of 5 cm were cut to have an average edge length of 5 mm, and the pieces were transplanted into the plate medium aseptically. The tissue-implanted Petri dishes were incubated at 25 ° C. for 12 days, and the grown mycelia of mushrooms were cut with a plate medium to an average length of 7 mm, transferred to a new PDA plate medium and cultured for one time Further, a shiitake mushroom was obtained.

1 l To a container, 500 ml of distilled water, 17 g of sugar, 2 g of soybean meal, 0.4 g of KH ₂ PO _4, 0.4 g of MgSO ₄ .7H ₂ O and 0.6 g of KNO ₃ were added and mixed to prepare a liquid medium, (120 DEG C) for 1 hour, cooled in an aseptic state, and then inoculated with 4 pieces of the average size 7 mm pieces of the virus obtained together with the plate medium, and cultured in a shaker at 25 DEG C for 25 days.

10 kg of oak sawdust, 1.5 kg of rice bran, 1.5 kg of bran, 20 kg of water was added to the above mixture, sterilized at 120 캜 for 100 minutes, and cooled to prepare a solid medium. The solid medium was placed in a bag having 70 through-holes formed in a sterilization chamber, 100 cc of the shiitake medium was inoculated, and incubated at 20 DEG C for 70 days.

The cultivated shiitake cultivate is more than half contaminated by other bacteria before the mycelium completely spreads throughout the solid medium, and the amount of shiitake produced in one solid medium is less than 10% of the embodiment of the present invention I can not.

While the present invention has been described in connection with what is presently considered to be practical exemplary embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, Of course it is possible. Accordingly, the scope of the present invention should not be construed as being limited to the above-described embodiments, but should be determined by equivalents to the appended claims, as well as the following claims.

Claims (5)

(A) 100 parts by weight of sawdust and 10 to 40 parts by weight of rice bran, and adding 150 to 250 parts by weight of water to prepare a solid medium,
(B) sterilizing the solid medium at a temperature of 100 to 135 캜 for 2 to 3 hours and cooling in a sterilization chamber,
(C) inoculating the solid medium with 5 to 15 parts by weight of the mushroom culture on the basis of 100 parts by weight of the sawdust in a sterilization chamber, incubating the mixture at 15 to 25 DEG C for 40 to 55 days, and
(D) removing the cultured cultures from the vials, re-introducing them into porous bags, culturing them at 15 to 25 DEG C for 45 to 60 days
Lt; / RTI >
The mushroom culture of step (C)
100 parts by weight of water,
1 to 5 parts by weight of glucose,
0.2 to 0.5 parts by weight of a powder selected from the group consisting of corn flour, corn, peptone and mixtures thereof,
Selected from the group consisting of K ₂ HPO ₄ , K ₃ PO ₄ , NaH ₂ PO ₄ , Na ₂ HPO ₄ , Na ₃ PO ₄ , KHCO ₃ , K ₂ CO ₃ , NaHCO ₃ , Na ₂ CO ₃ and mixtures thereof 0.01 To 0.1 part by weight,
0.03 to 0.1 part by weight of MgSO ₄ .7H ₂ O, and
NaNO ₃ , Ca (NO ₃ ) ₂ and mixtures thereof 0.02 to 0.2 parts by weight
Wherein the mushroom seed culture is cultivated in a liquid medium containing the mushroom culture medium. The method according to claim 1,
Wherein the solid medium of step (A) further comprises 10 to 20 parts by weight of wheat bran per 100 parts by weight of sawdust. The method according to claim 1,
Wherein the solid medium of step (A) further comprises 5 to 10 parts by weight of cotton seed hull per 100 parts by weight of sawdust. The method according to claim 1,
Wherein the liquid medium of step (C) further comprises 0.002 to 0.05 parts by weight of a defoamer or cooking oil per 100 parts by weight of the water. The method according to claim 1,
Wherein the humidity at the time when the culture incubated in step (D) is put back into the porous bag is 1 to 60%.

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