KR20050012053A - Mushroom culture method - Google Patents

Abstract

PURPOSE: A method of culturing mushrooms by culturing mushroom spawn in a culture medium containing Artemisia capillaris Thunb., Ginseng Radix, Angelica keiskei, kale, mulberry leaves, Eucommiae Cortex, pine needles, green tea leaves, Gastrodiae Rhizoma, Pueraria lobata and unpolished rice is provided. It shortens the culturing time and remarkably improving the culturing yield. CONSTITUTION: The mushroom culturing method comprises the steps of: preparing solid culture medium by adding water to dried herbs and sterilizing to a moisture content of 57 to 67%; controlling the pH of the solid culture medium and inoculating with 5 to 15% by weight of mushroom spawn; and culturing the inoculated culture medium at 23 to 33deg.C for 30 to 35 days. The solid culture medium contains one or more selected from the group consisting of: Artemisia capillaris Thunb., Ginseng Radix, Angelica keiskei (Miq.) Koidz., kale, mulberry leaves, Eucommiae Cortex, pine needles, green tea leaves, Gastrodiae Rhizoma, Pueraria lobata and unpolished rice.

Description

Mushroom cultivation method {MUSHROOM CULTURE METHOD}

[TECHNICAL FIELD OF THE INVENTION]

The present invention relates to a mushroom culturing method, and more particularly, to a cultivation method and conditions that can shorten the mushroom cultivation period and improve the mushroom cultivation yield as compared to the conventional method.

[Private Technology]

Mushrooms (mushrooms) belonging to basidiomycetes are important food resources and medicinal plants that decompose and grow various organic matters. Mushrooms have unique tastes and aromas, and contain a lot of proteins, vitamins, and other inorganic nutrients. In particular, recent studies have shown that anticancer and antiviral effects caused by the glucan derivatives, the polysaccharides in mushrooms, It has been reported and attracted worldwide attention as a health food.

Situation mushrooms belong to Aphylloporales, Hymnenochaetaceae and Phellinus. The scientific name is Phellinus linteus . Situation mushrooms are mainly distributed in Northeast Asia, and wild in Korea is not so common, and it is wild in some parts of Gangwon-do, and grows on stems of mulberry and hardwoods. The surface of the situation mushroom is dark brown, covered with the initial thin hairs, but it is soon browned to blackish brown, and the outer surface is solid wood tissue, forming a clear concentric round hole and a vertically cracked rectangular crack. Situation mushroom is known to have an excellent effect on gastrointestinal cancer, esophageal cancer, duodenal cancer, colon cancer and rectal cancer, which are diseases of the digestive system, and promote immune function. Situation mushroom cultivation method is a common method of cultivating wood, but methods and media for liquid culture of mycelium have been developed. Among them, Korean Patent Laid-Open Publication No. 2001-113057 discloses a method for feeding glucose or glucose and yeast extracts intermittently or continuously during cultivation, which is a fed-batch culture method of the situation mushroom mycelium.

In addition, the mushroom mycelium cultivation method used in the prior art is different depending on the type of mushroom used, but is cultured in a representative mushroom medium shown in Table 1 below. However, the mushroom medium has an expensive problem, there is a disadvantage that the culture period of the mycelium takes a long time at least 40 days or more.

Nutrition Badge type (g / L) MCM MYPA PDA ME YM YMPG YMG PYG GP GPY potato 250.0 K ₂ HPO ₄ 1.0 10 KH ₂ PO ₄ 0.46 2.0 MgSO _{4 7} H ₂ O 0.5 1.0 0.7 0.5 FeSO ₄ 7H ₂ O Glucose 20.0 20.0 10.0 10.0 4.0 14 30 20 Thiamine-HCl 1.0 *) DL-asparagine 1.0 peptone 2.0 1.0 5.0 5.0 2.0 1.25 3 2 Malt extract 30.0 20.0 3.0 10.0 10.0 Yeast extract 2.0 2.0 3.0 2.0 4.0 1.25 2 Agar 20.0 20.0 20.0 20.0 20.0 20.0 20.0 20.0 20.0 20.0

It is an object of the present invention to provide a method for culturing mushroom mycelium which can shorten the incubation time.

Another object of the present invention is to provide a method for mass culturing mycelium of situation mushrooms.

It is another object of the present invention to provide a mushroom mycelium or fruiting body cultured by the above method.

Figure 1 shows the yield according to the effect of the temperature at the time of spawning mushroom spawn culture.

Figure 2 shows the yield according to the pH of the spawn mushroom culture.

Figure 3 shows the yield according to the stirring speed during the spawn culture of the situation mushroom.

Figure 4 shows the yield according to the culture time (days) when the spawn culture of the mushrooms.

Figure 5 shows the degree of mycelial growth according to the incubation time (day) when cultivating the situation mushroom in cheonma solid medium.

6 is a photograph of the upper (a) and lower (b) of the mycelium cultured by inoculating situation mushrooms on cheonma solid medium.

In order to achieve the above object, the present invention provides a moisture content of 57 to 67% in the dried material of a plant selected from the group consisting of (a) ginseng, ginseng, sinchocho, kale, upper leaf, tofu, pine needle, green tea leaf, cheonma, 칡 and brown rice Adding water and sterilizing the same to prepare a solid medium, (b) titrating the solid medium to pH 4.5 to 6.5, and then inoculating 5 to 15% by weight of the mushroom spawn based on the weight of the solid medium, and (c) the inoculation. It provides a mushroom culture method comprising the step of culturing mushroom mycelia by placing one solid medium at 23 to 33 ℃ 30 to 35 days.

In another aspect, the present invention provides a mushroom mycelium cultured by the mushroom culture method.

In another aspect, the present invention provides a mushroom fruiting body cultured by the method further comprising the step of forming the fruiting body by secondary culture of the mycelium for 10 to 15 days at 15 to 20 ℃ after the mushroom culture method.

In another aspect, the present invention provides a mushroom extract obtained by adding one or more solvents selected from the group consisting of distilled water, alcohol, hexane, chloroform, ethyl acetate and chloroform to the mushroom mycelium or fruiting body prepared by the above method.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention are studying a method of improving the cultivation efficiency of mushrooms by using medicinal plants as a solid medium, and a method for maximizing mushrooms by optimizing the water content in the medium, the pH of the medium, the culture temperature and time. Established.

Mushroom culture method of the present invention is to inoculate the mushroom spawn in a solid medium, and to culture it. The mushroom may be a conventional mushroom, preferably a situation mushroom.

The solid medium of the present invention includes a medicinal plant as an active ingredient, examples of medicinal plants include ginseng mugwort, ginseng, sinchocho, kale, upper leaf, tofu, pine needles, green tea leaves, cheonma, 칡 and brown rice. The medicinal plants may be used as a single species, or may be used by mixing two or more kinds.

Water content in the solid medium is 57 to 67 (v / w)%, the mushroom mycelium yield may be reduced if the water content is out of the above range. Solid medium may be added to the medicinal plant dried water to have a water content of 57 to 67 (v / w)%, it can be prepared by sterilization at high temperature. Preferred moisture content is 65 (v / w)%.

In addition, the pH of the solid medium is 4.5 to 6.5. If the pH of the solid medium is out of the above range, mycelial yield can be significantly reduced compared to the incubation time. Preferred pH is pH5.

The spawn seed inoculated into the solid medium is prepared according to the type of mushroom in a preferred manner. That is, a commonly used liquid medium, YMPG, MCM, PDA or soybean meal can be used. In one embodiment of the present invention, the spawn culture method of the situation mushroom is described.

Situation mushroom spawn culture method is to inoculate the situation mushroom spawn in the liquid culture medium for spawn, it is cultured shaking for 5 to 10 days at 23 to 33 ℃, pH 4 to 10, agitation speed 100 to 250 rpm. The above range is also an optimally established range for the situation mushroom, but preferably incubation temperature of 25 to 30 ℃, medium pH 6.0 to 8.0, stirring speed 120 to 200 rpm and incubation period 6 to 10 days.

The spawn prepared by the mushroom spawn culture method is inoculated at 5 to 15% by weight based on the weight of the solid medium, and placed at 23 to 33 ° C. for 30 to 35 days to grow the mushroom mycelium. Preferably, the situation mushroom is incubated for 30 days at 30 ℃.

Referring to the mushroom mycelium culture method of the present invention mentioned above is as follows.

(a) Add water to the dry matter of at least one plant selected from the group consisting of ginseng, ginseng, fresh vinegar, kale, upper leaf, tofu, pine needles, green tea leaf, cheonma, 칡 and brown rice with water content of 57 to 67%, and sterilize it. To prepare a solid medium.

(b) The solid medium is titrated to pH 4.5 to 6.5 and then the mushroom seed is inoculated 5 to 15% by weight based on the weight of the solid medium.

(c) The inoculated solid medium is placed at 23 to 33 ° C. for 30 to 35 days to culture mushroom mycelium.

In addition, the mushroom mycelium may form a fruiting body by secondary culture for 10 to 15 days at 15 to 20 ℃.

The mushroom mycelium culturing method of the present invention is an effective method that can harvest a large amount of mushroom mycelium and fruiting body while significantly shortening the culture period compared to the conventional method.

Mushroom mycelium or fruiting body grown by the method of the present invention can be used as a food as well as to prepare a mushroom extract by a conventional method and can be used as food or food additives.

The preparation method of the mushroom extract may be a method of extracting heat or room temperature by adding one or more solvents selected from the group consisting of water, alcohol, hexane, chloroform, ethyl acetate and chloroform to the mushroom culture, mushroom mycelium or fruiting body. However, the mushroom extract manufacturing method is preferably used according to the effective method to be extracted. The alcohol is preferably ethanol, methanol, butanol or isopropanol.

Preferred mushroom extract preparation method is hot water extract by adding 200 to 600 parts by weight of water based on 100 parts by weight of the mushroom culture, hot water extract after filtration, F / D vacuum freeze drying method, V / D vacuum pressure drying method, hot air drying method or spraying It is powdered by carrying out by the drying method. The F / D vacuum freeze drying method is a method of quenching the extract and then drying it in a vacuum state of -20 to -80 ° C. The V / D vacuum reduction drying method concentrates the extract solution and then vacuums the extract to 60 to 65 ° C. The drying method is a method of drying the extract by concentrating the extract, and then blowing by heating a heated wind of 60 ° C. or more. The spray drying method is a method of concentrating and spraying the extract solution and drying it by hot air of 100 ° C. or more.

Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention and the present invention is not limited to the following examples.

Example 1 Situation Mushroom Hyphae Culture Using Chunma Solid Medium

1-1. Spawn culture

end. Influence of temperature

S. mushrooms were inoculated in YMPG, MCM, PDA and soybean meal medium at pH 6, respectively, and cultured at 15, 20, 25, 30, 35, and 40 ° C for 8 days, respectively, and the mycelia yield (g / L) was measured. Mycelial yields are shown in Table 2 below and FIG. 1.

Temperature 20 ℃ 25 ℃ 30 ℃ 35 ℃ 40 ℃ Mycelial yield (g / L) 7 9.3 10 2 0.4

In Table 2, the situation mushroom showed a big difference in growth according to the culture temperature, the culture temperature of 25 to 30 ℃ was appropriate, it was most effective to incubate at 30 ℃.

I. Effect of pH in Medium

Situation mushrooms were inoculated into appropriate media at pH 4, 6, 8, 10, and 12, respectively, and cultured with rotation shaking at 30 ° C for 8 days. Since mycelial yield (g / L) was measured, the results are shown in Table 3 and FIG.

pH 4 6 8 10 12 Mycelial yield (g / L) 7.5 10 9 8.2 0.2

In Table 3 and Figure 2, the situation mushroom showed a relatively good growth rate at pH 4 to 10, pH 10 was the most suitable pH for the situation mushroom culture.

All. Influence of the stirring speed

Situation mushrooms were inoculated into a suitable medium at pH 6, and cultured with agitation at 30 ° C. for 8 days. At this time, the stirring speed was incubated at 0, 100, 120, 150, 200 and 250 rpm, respectively, and the mycelia yield (g / L) was measured according to the stirring speed. The results are shown in Table 4 and FIG. 3.

Stirring Speed (rpm) 0 100 120 150 200 250 Mycelial yield (g / L) 2.1 6.5 8.8 10.3 9.2 8.5

In Table 4 and Figure 3, the situation mushroom showed an excellent growth rate at agitation speed 120 to 250 rpm.

la. Influence of days of culture

Situation mushrooms were inoculated into a suitable medium at pH 6, and cultured by rotating shaking at 150 rpm for 30 days at 30 ° C. Since mycelial yield (g / L) was measured according to the culture days, the results are shown in Table 5 and FIG.

Days of Culture 0 2 4 6 8 10 Mycelial yield (g / L) 0 One 6 9 10 9.5

In Table 5 and Figure 4, the situation mushroom showed a tendency to increase gradually with the incubation time and then gradually decrease after reaching the peak at 8 days of culture. Therefore, it is appropriate to culture the situation fungi 6 to 10 days.

Based on the above results,

Situation mushrooms were inoculated in YMPG, MCM, PDA and soybean meal at pH 6, and cultured for 8 days with stirring at 150 rpm at 30 ° C. to obtain a situation mushroom spawn.

1-2. Situation Mushroom Mycelial Growth

Situation of mycelial growth according to medium moisture content, culture temperature, medium acidity, and culture days in cultivation of mycelium mushroom mycelium using cheonma solid medium.

end. Water content of the medium

Water was added to each of the dried Cheonma dry matter to 40, 45, 50, 55, 60, 65, 70 and 75%, and sterilized at 121 ° C. for 3 hours to prepare a solid medium.

The solid medium was titrated to pH 5.5, inoculated with 10% by weight of the situation mushroom seedlings and observed mycelial growth while incubated at 30 ° C. for 70 days. Table 6 shows the mycelial growth according to the media moisture content. Incubation days is the number of days it takes to grow completely from the inoculation day to the bottom.

Medium moisture content (%) Mycelial density Incubation days 40 * 50 45 * 47 50 * 45 55 * 40 60 *** 33 65 **** 31 70 * 35 75 * 42 *: Lean **: somewhat lean ***: somewhat dense ****: dense

In Table 6, as a result of observing the mycelial density and the number of culture days according to the media moisture content, when the media moisture content was 65%, it showed a significant mycelial growth effect even in a short time of culture.

I. Effect of incubation temperature

Water was added to the dried Cheonma water content of 65%, sterilized for 3 hours at 121 ℃ to prepare a solid medium.

The solid medium was titrated to pH 5.5, inoculated with 10% by weight of the situation mushroom seedlings and then observed mycelial growth while incubated at 15 to 40 ℃ for 70 days. Table 7 shows mycelial growth according to the culture temperature.

Incubation temperature (℃) Mycelial density Incubation days 15 * 66 20 ** 50 25 *** 35 30 **** 28 35 * 43 40 * 59 *: Lean **: somewhat lean ***: somewhat dense ****: dense

In Table 7, the results of observing the mycelial density and the number of days of culture according to the culture temperature, when the culture temperature is 30 ℃ showed a significant mycelial growth effect even for short time culture.

All. Effect of pH

Water was added to the dried Cheonma water content of 65%, sterilized for 3 hours at 121 ℃ to prepare a solid medium.

The solid medium was titrated to pH 4 to 9, respectively, and inoculated with 10% by weight of situation mushroom seedlings, followed by incubation at 30 ° C. for 70 days, and observed mycelial growth. Table 8 shows mycelial growth according to the culture temperature.

Medium pH Mycelial density Incubation days 4 * 40 5 **** 33 6 *** 34 7 * 49 8 * 51 9 * 60 *: Lean **: somewhat lean ***: somewhat dense ****: dense

In Table 8, as a result of observing the mycelial density and the number of culture days according to the acidity of the medium, when the pH of the medium is 5, the mycelial growth was remarkable even in a short time of culture.

la. Effect of culture days

Water was added to the dried Cheonma water content of 65%, sterilized for 3 hours at 121 ℃ to prepare a solid medium.

The solid medium was titrated to pH 5, inoculated with 10% by weight of situation mushroom seedlings, and then cultured at 30 ° C. for 70 days to observe mycelial growth according to the number of days of embryos. Table 9 and Figure 5 shows the mycelial growth according to the culture days.

Days of Culture 10 15 20 25 30 35 40 Mycelial growth (%) 15 42 68 85 100 100 100

In Table 9 and Figure 5, the mycelial growth gradually increased with the number of days of culture, indicating the growth of 100% at 30 days of culture.

As a result of the above, the most optimal culture conditions for mycelial culture of situation mushrooms are as follows.

Moisture content of solid medium: 65%

PH of solid medium: pH 5

Incubation temperature: 30 ℃

Cultivation date: 30 days

Example 2: Situation mushroom hyphae, fruiting body culture and mushroom extract preparation using medicinal plant solid medium

2-1. Culture of spawn mushrooms

Situation mushrooms were inoculated in YMPG, MCM, PDA and soybean meal at pH 6, and cultured for 8 days with stirring at 150 rpm at 30 ° C. to obtain Ganoderma lucidum spawn.

2-2. Culture of Situation Mushroom Mycelia

Purchase dried each of Injin mugwort, Ginseng, Sinchocho, kale, upper leaf, Tofu, pine needle, green tea leaf, green tea and brown rice, add water with 65% water content and sterilize at 121 ℃ for 3 hours Prepared.

Each of the solid medium was titrated to pH 5, respectively, inoculated with 10% by weight of mushroom mushroom spawn and incubated at 30 ° C. for 30 days to obtain mushroom mushroom mycelia.

6 is a photograph of the upper (a) and lower (b) of the mycelium cultured by inoculating situation mushrooms on cheonma solid medium.

2-3. Fruiting body culture

The situation mushroom mycelium cultured in the above was second cultured at a culture temperature of 17 ° C. for 10 to 15 days to obtain fruiting bodies.

2-4. Mushroom extract manufacturer

4 times the weight of water was added to the mycelium mushroom mycelium or fruiting body and extracted with the extractor at 100-120 ° C. for 10 hours or more. The extract was filtered, the filtrate was liquefied and then powdered by spray drying through a concentration and filtration process.

Or it was powdered by vacuum freeze drying at -20 to -80 ℃ instead of spray drying.

Comparative Example: Culture of Roebuck Mushroom

end. Spawn culture

The seedling fungus was inoculated in YMPG or MCM medium adjusted to pH 5.5, and shaken for 7 days at a culture temperature of 25 ° C. and a stirring speed of 120 rpm to incubate the seed.

I. Mycelium and Fruiting Body Culture

Water corresponding to twice the total weight of fresh vinegar was added to fresh vinegar, which was sterilized at 120 ° C. for 2-4 hours to prepare a solid medium.

The seed was added at 10% by weight based on the weight of the solid medium, the mycelia were first grown at 25 ° C. for 40 to 50 days, and then secondary growth was performed at a culture temperature of 15 to 20 ° C. for 10 to 15 days. Fruiting bodies were grown.

Compared to the method for culturing mycelium and fruiting bodies of Examples 2 and 4, which are examples of the present invention, the method of the present invention shortens the mushroom growing period from 10 to 15 days as compared to the comparative example.

Example 3-17 Mushroom Preparation

The mushroom mycelium cultures were obtained in the same manner as in Example 2, except that the upper leaves, green tea leaves, larvae leaves, pine needles, and cheonma, locust mushrooms, and mushrooms were used.

1500 g of water was added to 500 g of the mushroom mycelium culture, heated at 121 ° C. for 3 hours and 30 minutes, and filtered through 325 mesh to obtain a filtrate. Each filtrate was brixified with ATAGO refactometer PR-32 (0-32%) and lyophilized to determine the yield (Table 10).

Example product name Amount collected (g, based on culture 500g, out of total 2,000g with water) Brix (%) Collected solids (g) Final yield (%) 3 Roe Deer-Upper Leaf 1,260 3.2 40.3 8.1 4 Manor 1,427 2.5 35.7 7.1 5 Situation-upper lobe 1,284 2.3 29.5 5.9 6 Roe Deer Bee-Green Tea Leaf 1,004 2.8 28.1 5.6 7 Ganoderma-green tea leaves 1,112 1.0 11.1 2.2 8 Situation-green tea leaves 1,148 1.7 19.5 3.9 9 Roe Deer Beetle 1,127 2.7 30.4 6.1 10 Ganoderma 1,118 1.7 19.0 3.8 11 Situation-Tomb Leaf 1,236 1.4 17.3 3.5 12 Roe Deer Bee-Pine Needle 1,379 1.4 19.3 3.9 13 Ganoderma-pine needles 1,307 0.5 6.5 1.3 14 Situation-Pine Needles 1,384 0.7 9.7 1.9 15 Roe Deer Bee 1,359 4.1 55.7 11.1 16 Manor 1,356 4.9 66.4 13.28 17 Situation 1,198 4.8 57.5 11.5

As described above, the present invention can provide a mushroom cultivation method using a medicinal plant as a solid medium to shorten the existing mushroom cultivation period, it is possible to improve the mushroom cultivation yield. In addition, the cultured mushroom mycelium and fruiting body of the present invention can be provided as a food, it can be used to extract the bioactive material.

Claims (8)

(a) water of 57-67% water content is added to the dried material of the plant selected from the group consisting of ginseng, ginseng, sinchocho, kale, upper leaf, tofu, pine needle, green tea leaf, cheonma, 칡 and brown rice, and sterilized it to solid medium. Preparing a; (b) titrating the solid medium to pH 4.5 to 6.5 and then inoculating the mushroom spawn 5 to 15% by weight based on the weight of the solid medium; And (c) culturing the mushroom mycelium by placing the inoculated solid medium at 23 to 33 ° C. for 30 to 35 days Mushroom culture method comprising a. The method of claim 1, Step (b) is the step of titrating the solid medium to pH 5 and then inoculating 10% by weight of the situation mushroom spawn based on the solid medium weight; The step (c) is the step of culturing the situation mushroom mycelia by placing the inoculated solid medium at 30 ℃ 30 days; Phosphorus culture method. The method of claim 1, wherein the mushroom spawn is obtained by inoculating the situation mushroom bacteria in a liquid culture medium for spawn, and shaking it for 5 to 10 days at 23 to 33 ℃, pH 4 to 10, agitation speed 100 to 250 rpm Culture method. The method of claim 1, wherein the culturing method is The culture method further comprises the step of secondary culture of the mycelium at 15 to 20 ℃ for 10 to 15 days to form a fruiting body. Mushroom mycelium cultured by the method according to any one of claims 1 to 3. Mushroom fruiting body cultured by the method according to claim 4. A mushroom extract obtained by adding a distilled water, alcohol, hexane, chloroform, ethyl acetate and chloroform to the mushroom mycelium of claim 5 by adding one or more solvents selected from the group. The mushroom extract extracted by heat or room temperature by adding at least one solvent selected from the group consisting of water, alcohol, hexane, chloroform, ethyl acetate and chloroform to the mushroom fruiting body of claim 6.