KR20010063868A - Grain Containing Mycellium of Mushroom and Preparation Method thereof - Google Patents

Abstract

PURPOSE: A method for preparing various kinds of cereals such as brown rice or wheat containing mushroom mycelium by using a bioreactor is provided to mass produce the nutritious cereals in a short time. CONSTITUTION: The method is characterized by the following steps of: (i) inoculating mushroom mycelium to a high pressure sterilized culture medium board of which pH is adjusted to 5-8 aseptically and pre-cultivating for 2-10days; (ii) aseptic inoculating the precultured mycelium to a liquid medium of which ingredients are same with the pre-culture medium and culturing the mycelium at 20-35deg.C for 10-15days with stirring and ventilating; (iii) heating cereals at 90-100deg.C for 0.5-1hr by mixing the cereals with water 50-60wt% in a rotary drum type bioreactor(100) and supplying steam through a steam supply line(125); (iv) adding calcium sulfate of 0.6-2.0wt% based on the weight of the cereals, adjusting the pH of the cereals to 5-8 and sterilizing the cereals at 90-130deg.C with high temperature steam from the steam supply line; and (v) supplying the cultivated mushroom mycelium to the cereals in the bioreactor to be grown at the cereals aseptically at constant temperature for 10-15days before dehydrating the resulting cereals.

Description

Grain containing mushroom mycelium and its preparation method {Grain Containing Mycellium of Mushroom and Preparation Method}

The present invention relates to grains containing mushroom mycelium and a method for producing the same, and more particularly, to a method for rapidly mass production of grains containing mushroom mycelium.

Mushrooms are microorganisms belonging to basidiomycetes in botany. Mushrooms have been widely used in Korea as a food of Buljangjang, which has a unique aroma and taste.

Mushrooms are symbiotic and by-products that maintain ecosystems in forests, and the distribution of these species is unusual and diverse, so that each species grows slightly differently.

About 230 kinds of mushrooms growing in Korea are reported as edible and medicinal mushrooms. Among the mushrooms that are widely used for edible and medicinal mushrooms, silver mushrooms, which are artificially cultivated, include shiitake, oysters, tops, ganoderma lucidum, fingerlings and cordyceps. It's just a species.

In order to artificially cultivate mushrooms, spawns, which are secondary mycelia of seeds, are used as seeds. The spawn used in mushroom cultivation should be easily decomposed by mushroom mycelium, so that the mycelial growth rate is fast and its adaptability to cultivation medium should be excellent. Most of the spawns currently in use were developed at the early stage of the introduction of artificial cultivation technology, using solid media such as spawns and sawdust spawns. Seedlings are removed from wood where mushroom mycelia are infested and inserted into logs. Mushroom mycelia erode the logs, and when mushrooms are produced, they are collected and edible. To date, various additives have been used for mushroom cultivation. .

Sawdust spawn is the most commonly used spawn for mushroom cultivation, and is used as mushroom spawn by adding various nutrients to sawdust.

In addition, pulverulent spawn collected from wild and grown in compost medium box, manure of livestock mixed with block spawn inoculated with mycelium fragment, inoculated with mycelium fragment, and mycelia separated from fruiting body or mycelium of mushroom Pure cultured seedlings inoculated into bottles are known.

To date, the spawns used for the cultivation of mushrooms are generally a mixture of sawdust and bran, sterilized in a polypropylene pot as a spawn medium, inoculated with mycelium grown on an agar plate and over several days at the optimum temperature for growth. The mycelium was pulled and used for the cultivation of mushrooms.

However, the production of spawns in this way requires not only a huge spawn production facility, but also entails heat loss and other labor costs for the optimal temperature maintenance for long-term culture, and overhead costs for transportation. There was a risk of contamination.

In addition, the method of cultivating a solid spawn is to first isolate the tissue of the mushroom or mycelium first and incubate it in a test tube, then incubate in a medium to be used as an inoculum, and then inoculate into a spawn medium and use it as a spawn for mushroom cultivation. is average. In other words, in order to obtain a solid spawn, at least four pass passages must be passed, and in the process, the force of the bacterium is weakened, and there is a problem that the productivity of the mushroom, which is the final product, is not very high.

However, in recent years, mushroom mycelium has nutritional and pharmacological value as much as mushroom fruiting bodies, and drinks using mushroom fruiting bodies have been produced and marketed.

In addition, recently, it has been reported that a technique for growing spawn in rice using the situation mushroom spawn is developed. According to this method, it is adopted to grow the situation spawn in the rice by flat-cultivating the situation spawn, placing rice in a large container, and transplanting the situation spawn on it. However, according to this method, it takes a long time to get the situation seed and political culture, so it takes a long time for the situation seed to penetrate into the rice in the lower part of the container. Mixing of the grown grains to mix evenly throughout the medium had to be carried out several times during the incubation period. In addition, there is a possibility that the product is contaminated by other microorganisms due to the penetration of foreign substances in the process of injecting the seed according to the progress of the growth of the seed.

Accordingly, the present inventors have developed a method of mass-producing mushroom mycelium-containing grains in a short time by liquid culture of the mushroom mycelium and allowing the obtained mushroom mycelium to grow in cereals in a bioreactor.

Accordingly, the present invention provides a method for rapidly and rapidly producing grains containing mushroom mycelium using a bioreactor.

1 is a block diagram showing a rotary drum-type bioreactor for culturing cereals containing mushroom mycelium according to the present invention.

♣ Explanation of symbols for the main parts of the drawing

80: air floating bioreactor 90: level window

100: rotary drum type bioreactor 101: spur gear

105: roller guide rail 106: drum body

107, 126, 127: rotary joint 108: air return line

109: Air medium supply line 110: Air return solenoid

113: feed discharge cover 116, 131: rotary roller

117: pivoting pin 118: anchor

119: motor 120: drive gear

121: hydraulic cylinder 123: steam discharge line

124: cooling water supply line 125: steam supply line

130: bass

In order to achieve the above object, the method for preparing grains containing the mushroom mycelium of the present invention is inoculated with mushroom mycelium aseptically on autoclaved plate medium adjusted to pH 5 to 8 to incubate for 2 to 10 days before incubation. step; Sterilely inoculating the precultured mycelium on a liquid medium of the same component as the preculture medium, and culturing the cancer while stirring and aeration at 20 to 35 ° C. for 10 to 15 days; Putting the grains into the rotary drum bioreactor 100, mixing 50 to 60% by weight of water, and then adding steam through a steam supply line 125 for 30 minutes to 1 hour at 90 to 100 ° C .; Adding 0.6 to 2.0% by weight of the calcium sulfate and then adjusting the pH to 5 to 8 to sterilize at a temperature of 190 to 130 ° C. by adding high temperature steam through a steam supply line 125; Supplying the mushroom mycelium obtained through the liquid cancer culture through a line (60) to sterile cancer culture under constant temperature conditions; And drying the grains in which the mushroom mycelium is grown.

In the production method of cereal grains containing mushroom mycelium according to the present invention, as grains, any food similar to grains such as potatoes, sweet potatoes, etc. may be used in addition to rice, barley, beans and wheat.

Medium used in the method for producing a grain containing mushroom mycelium according to the present invention can be used as long as it is generally used as a liquid medium, for example, GPM, GA, Oyster-mushroom Culture, oak- Oak-mushroom culture, PG, MYG and the like can be used, oyster-mushroom medium, oak-mushroom medium, lyofilum-mushroom medium, PG medium are preferred, and oak-mushroom medium is particularly preferred.

In the production method of cereal grains containing mushroom mycelium according to the present invention, the structure of the rotary drum type bioreactor 100 is a rotary drum type driven by a motor together with a conventional air floating bioreactor as shown in FIG. The bioreactor is installed in the base 130 and the pivot 118 around the pivot pin 117 by the operation of the hydraulic cylinder 121 and the base 130 is disposed in the vertically extending in the vertical direction in the center. It is rotatably mounted on the frame 111 by the frame 111 and the plurality of rotating rollers 116, the steam supply line 125, the cooling water supply line 124 is connected The air supply which supplies the air which flowed in from the air tank 58 to the said body 106 while supplying the body 106 which has the body 106 and the medium which flowed in from the air floating bioreactor 80 to the said body 106, Badge And a discharging means and a rotating means for rotating the body 106, the rotating means being fixed to the frame 111 and the spur gear 101 attached on the outer circumference of the body 106. Motor 119 and a drive gear 120 engaged with the spur gear 101 in a state of being fixed to the end of the motor shaft of the motor 119.

Referring to the culture process using the rotary drum-type bioreactor 100 of the present invention in more detail, first, the mushroom mycelium culture solution in the air floating bioreactor 80 is put into the air floating bioreactor 80 by the operation of the solenoid The supply of oxygen begins. Next, the steam solenoid is opened to supply steam into the body 80a to raise the temperature. When the proper temperature is reached, the steam solenoid is turned off to stop the supply of steam, and the mushroom mycelium broth is supplied to the rotary drum type bioreactor 100 through the line 60, and the surplus is returned to the body 80a through the return line 62. ) Is recovered.

In the method for producing a grain containing mushroom mycelium according to the present invention, the addition of calcium sulfate to the grain heated by the steam in the bioreactor is the residual moisture remaining after the water added in the bioreactor penetrates into the grain At the same time to eliminate the persistence of the grain itself to remove the grains to prevent the phenomenon of coagulation of grains to allow the mushroom mycelium to evenly penetrate the grain surface.

Rotary drum type bioreactor 100 used to produce grains containing mushroom mycelium according to the present invention is maintained by a constant temperature and aeration conditions while the mushroom mycelium supplied by the motor 119 evenly contacts the grain To make it possible. In addition, through the monitoring window 103 to determine the growth of the mycelium so that the mushroom mycelium can be additionally supplied.

In the production method of grains containing mushroom mycelium according to the present invention, the temperature of the mushroom culture by feeding the mushroom mycelium to the rotating drum-type bioreactor 100, the optimum growth temperature varies depending on the type of mushroom mycelium used Can be.

In the method for producing grains containing mushroom mycelium according to the present invention, if the mushroom mycelium is supplied by stirring the mushroom mycelium to the rotary drum-type bioreactor 100, the white mushroom mycelium grows in the grain after about 10 to 15 days after cultivation. Done.

In the production method of grains containing mushroom mycelium according to the present invention, the mycelia obtained when the liquid culture in the rotary drum bioreactor 100 in the process of obtaining the mushroom mycelium by cancer culture is not only very excellent activity The growth rate of the mycelium was also confirmed to be very fast.

In the method for producing a grain containing mushroom mycelium according to the present invention, the grain obtained by growing the mushroom mycelium in the cereal grains in the bioreactor 100 using a freeze-drying method or a heat-drying method commonly used for drying foods To dry.

In the production method of grains containing mushroom mycelium according to the present invention, in order to have the inherent aroma of mushroom mycelium, it is preferable to use pine needles, pine, hardwood extract instead of water when controlling moisture.

At this time, the extract is preferably used to extract the stem or leaves extracted by heating the steam or more than 100 ℃, the ratio of pine needles or pine extract and hardwood is preferably mixed in a ratio of 3: 7.

Hereinafter, the method for preparing grains containing mushroom mycelium according to the present invention will be described in more detail. However, the following examples are presented as an illustration of the present invention, and the present invention is not limited to the following examples.

Examples 1-9

(1) Preparation of mushroom tissues: Tissue isolates from fresh fresh tissues of Matsutake mushrooms and Twill mushrooms were used.

Badge Name Liquid medium composition With agar additives on flat plate Example 1 Modified Hamada Glucose (10 g), yeast (5 g), KH ₂ PO ₄ (1 g), MgSO ₄ · 7H ₂ O (0.5 g), distilled water (1 l ), pH5.1 20 g Example 2 PD / PDA 200 g potato boiled water (1 l ), glucose (dextrose) (20 g), pH6.1 PDA 39g Example 3 GPM Glucose (20 g), peptone (1 g), malt extract (20 g), distilled water (1 l ), pH5.9 20 g Example 4 GA Glucose (10 g), ammonium tartarate (1 g), KH ₂ PO ₄ (1 g), iron (II) citrate (5 mg), ZnSO ₄ 7H ₂ O (4.4 mg), MnSO ₄ 4H ₂ O (5 mg ), CaCl ₂ · 2H ₂ O (55.5 mg), vitamin B ₁ (10 mg), nicotinic acid (10 mg), distilled water (1 l ), pH5.3 20 g Example 5 Oyster-mushroomCulture Sulfur Sugar (30g), Yeast Extract (3.0g), KH ₂ PO ₄ (1g), MgSO ₄ · 7H ₂ O (1g), Distilled Water (1 l ), pH5.2 20 g Example 6 Czapex-dox Sucrose (30g), NaNO ₃ (2.0g), K ₂ HPO ₄ (1g), MgSO ₄ · 7H ₂ O (0.5g), FeSO ₄ · 7H ₂ O (0.01g), distilled water (1 l ), pH5 .2 20 g Example 7 MYG Malt extract (5g), yeast extract (5g), glucose (20g), distilled water (1 l ), pH4.8 20 g Example 8 Oak-mushroom Culture Glucose (50 g), Peptone (2.5 g), Yeast extract (2.5 g), KH ₂ PO ₄ (1 g), MgSO ₄ · 7H ₂ O (0.5 g), CaCl ₂ · 2H ₂ O (0.5 g), FeSO ₄ 6H ₂ O (10 mg), MnCl ₂ 4H ₂ O (7.2 mg), ZnCl ₂ (4 mg), CuSO ₄ 5H ₂ O (1 mg), distilled water (1 l ), pH5.3 20 g Example 9 Lyophyllum-mushroom Culture Glucose (20 g), L-valine (1.1 g), KH ₂ PO ₄ (0.1 g), MgSO ₄ · 7H ₂ O (0.1 g), ZnSO ₄ · 7H ₂ O (1.0 mg), FeSO ₄ · 7H ₂ O (1.0 mg), CuSO ₄ · 5H ₂ O (0.2 mg), MnCl ₂ · 5H ₂ O (0.2 mg), nicotinic acid (0.1 mg), thiamine (0.2 mg), distilled water (1 l ), pH5.6 20 g Example 10 PG Glucose (30 g), Peptone (6 g), KH ₂ PO ₄ (0.5 g), MgSO ₄ · 7H ₂ O (0.5 g), CaCl ₂ · 2H ₂ O (0.1 g), FeSO ₄ · 6H ₂ O (10 mg) , MnCl ₂ · 4H ₂ O (3 mg), ZnCl ₂ (3 mg), CuSO ₄ · 5H ₂ O (1 mg), thiamin (10 mg), MoO ₃ · H ₂ O (3 mg) distilled water (1 l ), pH4.6 20 g

(2) Media Preparation: The liquid medium of the composition shown in Table 1 was sterilized by adjusting the pH to about 5. Plate medium to be used in the pre-cultivation step was prepared by sterilizing the pH of the composition and the agar of the composition of Table 1 adjusted to about 5.

(3) pre-stage culture: Aseptically inoculated each of the clustered and fungus mycelium on the prepared plate medium and incubated at about 22.5 ° C for about 7 days to visually check the activity of the fungus and fungus mycelium, and then if the activity is confirmed Incubate for 2 more days.

(4) Cancer culture step: Activated clusters and sterile mycelium were inoculated into the liquid medium according to the present invention, respectively, and cultured for about 13 days while stirring in the bioreactor 100 at about 22.5 ° C. and aeration at 1.5vvm. . 40 ml of the clustered and sterile mycelium culture broth obtained at this time were taken, and the dry weight thereof was measured. In addition, as a control, the same weight of the clustered and sterile mycelium cultured in the same medium was taken to obtain a dry weight and the growth ratio was calculated and described together.

(5) Sterilization of grains: Brown rice or wheat in a rotating drum bioreactor 100, put about 50 to 60% by weight of water and mix, then put steam through the steam supply line 125 at about 100 ℃ 40 Heated for minutes. Calcium sulfate was added to 0.8% by weight of brown rice, and then the pH was adjusted to 5 to 8 to sterilize at a temperature of about 121 ° C. by putting steam of high temperature through the steam supply line 125.

(6) Cultivation and drying of grains: The bioreactor in which the mycelium used in step (4) was cultured was separated from the frame 11, stood vertically, and supported by the legs 92, and then line 60 and return to each inlet. While connecting the line 62, the rotating drum-type bioreactor 100 is put brown rice and the mushroom cultured mycelium is supplied through the line 60 to rotate the stirring at a speed of about 6rmp at a constant temperature of 22.5 ℃ Cancer culture was aseptically for 15 days.

(6) Drying step: The grains in which the mushroom mycelium obtained in (5) was grown were dried using a conventional freeze drying method.

badge cluster Abilities Dry weight (mg / 40ml) Dry weight of control group (mg / 40ml) Dry weight (mg / 40ml) Dry weight of control group (mg / 40ml) Example 1 Modified Hamada 27.7 26.0 24.0 23.0 Example 2 PD / PDA 15.6 14.5 20.4 19.9 Example 3 GPM 18.3 17.1 28.3 26.9 Example 4 GA 20.2 18.9 23.8 21.6 Example 5 Oyster-mushroomCulture 28.8 26.8 37.0 34.0 Example 6 Czapex-dox 19.4 18.0 16.0 15.2 Example 7 MYG 29.0 27.3 26.6 25.3 Example 8 Oak-mushroom Culture 42.4 39.9 49.8 47.3 Example 9 Lyophyllum-mushroom Culture 24.0 22.3 33.8 31.8 Example 10 PG 25.0 23.3 35.8 34.0 Average 23.5 23.4 28.0 27.9

As can be seen from the above results, the mushroom mycelium which has undergone the preculture step and the stirred cancer culture step according to the present invention can be confirmed that the growth rate is higher than that of the control group which has just been liquid culture. It can be seen that the mycelial growth is vigorous in the medium, lyophilum-mushroom medium, and PG medium.

In addition, as a result of developing mushroom mycelium on rice using a rotary drum bioreactor, it was confirmed that mushroom mycelium was sufficiently grown in brown rice in about 15 days, which allowed mushroom mycelium to remain in brown rice without using a rotary drum bioreactor. In the case of development, it can be seen that the mycelium grows more than twice as fast as it takes about 30 days or more.

It can be seen that the mushroom mycelium obtained according to the grains containing the mushroom mycelium of the present invention and the preparation method thereof is very active, and the growth conditions in the bioreactor are optimal conditions.

As described above, the method for preparing grains containing mushroom mycelium according to the present invention is not only able to produce a large amount of mushroom mycelium, but also nutritionally by allowing a rapid and mass production of grains containing mushroom mycelium. Very good mushrooms can be easily eaten into the diet.

Claims (8)

aseptically inoculating mushroom mycelium on autoclaved plate medium adjusted to pH 5 to 8 for 2 to 10 days incubation; Sterilely inoculating the precultured mycelium on a liquid medium of the same component as the preculture medium, and culturing the cancer while stirring and aeration at 20 to 35 ° C. for 10 to 15 days; Putting the grains into the rotary drum bioreactor 100, mixing 50 to 60% by weight of water, and then adding steam through a steam supply line 125 for 30 minutes to 1 hour at 90 to 100 ° C .; Adding 0.6 to 2.0% by weight of the calcium sulfate and then adjusting the pH to 5 to 8 to sterilize at a temperature of 190 to 130 ° C. by adding high temperature steam through a steam supply line 125; Supplying the mushroom mycelium obtained through the liquid cancer culture through a line (60) to sterile cancer culture under constant temperature conditions; And a step of drying the cereal grains in which the mushroom mycelium is grown. The method of claim 1, Method for the culture of the pre-cultured mycelia in a liquid medium, characterized in that the progress in the rotary drum bioreactor (100). The method according to claim 1 or 2, Said grains being brown rice or wheat. The method according to claim 1 or 2, The medium is oyster-mushroom medium, oak-mushroom medium, lyophyllum mushroom medium, characterized in that the PG medium. The method of claim 4, wherein Wherein said medium is an oak-mushroom medium. The method according to claim 1 or 2, The period of incubation of grains containing mushroom mycelium in the rotary drum bioreactor (100) is about 10 to 15 days. The method according to claim 1 or 2, Method for supplying pine needles, pine, hardwood extract instead of water in the step of supplying water to the cereal drum bioreactor (100). The method of claim 7, wherein The mixing ratio of the pine needles or pine extract and hardwood is characterized in that the mixture in a ratio of 3: 7.