JP3509736B2 - A method for preparing a fungus bed of Physcomitrella patens having physiological functions - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for preparing a fungal bed of Phellinus linteus (Sparassis Crispa Rr.) Having physiological function activity.

[0002]

2. Description of the Related Art Hanabitake mushrooms are extremely rare mushrooms that grow on Japanese larch and are also called phantom mushrooms. In addition to having a good chewy texture, the Hanabitake mushroom is also prized in the Japanese food industry as a valuable food because of its pale yellow color close to pure white and its shape like leaf peony.

[0003] Furthermore, Hanabitake mushroom contains a large amount of useful physiologically active substances, and attention is paid also in this respect.
Particularly, the content of total β-glucan, which is known as a physiologically active substance having an excellent immunostimulatory action, is 43.100 g per 100 g.
It is 6 g . Even Agaricus or Maitake mushrooms, which are known as mushrooms with high total beta-glucan content, their total beta-glucan content is 11.6 g and 18.1 g per 100 g, respectively.
Compared with the above, it can be seen that the content contained in Hanabi-ratake is surprisingly large. In addition, among the total beta glucans, the antitumor effect has been proven especially for β-1,3-D-glucans. Therefore, Hanabitake mushroom is not only an excellent food material as a health food, but also a valuable raw material for producing an antitumor agent.

Despite having the excellent properties as described above, Hanabitake mushrooms are extremely rare, as is apparent from the fact that they are called phantom mushrooms, and are too expensive to be used as general foods. There is a problem. Although various attempts have been made on artificial cultivation, artificial cultivation itself is difficult because it has the property that other mushrooms are weak against molds, and even if the artificial cultivation is fortunately successful, the growth is slow. Therefore, the current situation is that we have not yet achieved sufficient results.

Furthermore, in the conventional artificial cultivation method of Hanabi-take mushrooms, larch large sawdust used for the culture medium is prepared in advance with hot water boiling temperature of 120 to 121 ° C. and pressure of 2 when the fungus bed is prepared.
The medium was treated by exposing it to high-temperature steam at atmospheric pressure for 60 minutes and then washing it with water. This is because the inhibitor contained in larch large sawdust inhibits the growth of Hanabi-take mushrooms, so that unless the inhibitor is removed, the Hanabi-take mushrooms do not grow smoothly. However, this method for removing the inhibitory substance is complicated in operation and requires a large amount of cost, which has been an obstacle to the preparation of the bacterial bed.

[0006]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and it is an object of the present invention to make it possible to reliably artificially grow Hanabi-take mushrooms, and to omit the conventional complicated and expensive medium treatment. It is to provide a method for producing a fungal bed of Bamboo shoots capable of performing.

[0007]

[Means for Solving the Problems] The above-mentioned problems have been solved by using a culture medium containing sawdust as a main component in the Hanabitake mushroom MH-3 (accession number F).
ERM P-17221) is inoculated, which is achieved by a method for producing a fungal bed for artificial cultivation of Ganoderma lucidum.

Hanabitake MH-3 used in the present invention
Was identified by the inventor as a strain suitable for artificial cultivation.
It has been deposited with the Institute of Biotechnology, Institute of Industrial Science and Technology under the MP-17221. By using a fungal bed prepared by inoculating this strain into a medium containing sawdust as a main component, it is possible to artificially grow Hanabi-take mushrooms, which has been difficult in the past. Inoculation Hanabitake MH-3
It is preferable to use the spores or mycelia.

The bacterial bed obtained by the present invention has an anti-cancer effect,
Β-1,3-D having hypoglycemic action and hypotensive action
It is suitable for use in the industrial artificial cultivation of Hanabitake mushrooms (mushrooms) that contain large amounts of glucan.

In the present invention, it is preferable to add 5 to 10% of wheat flour to the medium containing sawdust as the main component.

In the present invention, it is possible to use without removing the Hanabi-take mushroom growth-inhibiting substance of the above larch large sawdust.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION It is technically extremely difficult to artificially cultivate Hanabitake mushrooms as compared with mushrooms that are conventionally artificially cultivated, that is, Mediterranean shimeji mushrooms, shiitake mushrooms, and oyster mushrooms. However, as a result of diligent research, the inventor of the present application cultivates mushrooms containing a large amount of β-1,3-D glucan of interest by the selective use of bacteria (MH-3), and requires conventional medium treatment. We were able to reach it in a short period of time and economically. Also, by improving the cultivation room and improving the medium composition,
Further improved results were obtained.

The medium used in the present invention is mainly composed of large sawdust of Tsuga, fir, pine, beech, and shii. It is preferable to use these single sawdust or a mixture, or the chip thereof, mixed with the nutrients required for cultivation of Hanabitake mushrooms (mushrooms).

Hanabitake MH-3 inoculated into the above medium
The spores or hyphae can be used as. In the present invention, for example, tsuga, fir, pine, beech, Japanese sawdust, or chips are mixed with nutrients necessary for the growth of Hanabitake mushroom, which is filled in a polyethylene bottle of 500 mL and sterilized, and the medium is kept at room temperature. After cooling, M
H-3 spores or hyphae are inoculated aseptically. By cultivating fruiting bodies from the bacterial bed thus prepared, β-1, which is effective for cancer prevention, treatment, diabetes, and treatment of hypertension,
It is possible to cultivate a bamboo shoot containing a large amount of 3-D glucan.

The first aspect of the present invention is to provide the Hanabitake MH.
The use of -3 made it possible to prepare an artificially cultivated fungus bed of Bamboo shoots without the need for conventional complicated and expensive medium treatment such as high temperature steam treatment and washing with water. That is, the inventor has succeeded in successfully growing Hanabi-take mushrooms by using the MH-3 bacterium without the need for the above-mentioned inhibitor removal operation. As a result, it has become possible to produce a Hanabitake mushroom bed extremely economically and easily. In addition, until now, it was considered impossible to cultivate Hanabitake mushrooms without mixing glass bottles as a container for the fungus bed, which made it impossible to cultivate Hanabitake mushrooms.
A polyethylene container can be used by using H-3 bacteria,
It has become possible to carry out industrialized mass cultivation that allows the use of large automatic machines.

The second aspect of the present invention is directed to improvement of the bed culture medium. As described above, in the present invention, sawdust is used as a main component as a medium for preparing a fungal bed of Hanabi-ratake MH-3, but by adding wheat as another nutrient to be added thereto, shortening of cultivation period and harvesting It has become possible to achieve an increase in quantity.

It is confirmed that MH-3 agaricus, which is artificially cultivated using the above-prepared bacterial bed, has a strong anti-cancer action, blood glucose level lowering action, immunostimulating action, and anti-hypertensive action. The inventor discovered. This experiment will be described below.

The test period was 35 days, and the weight of the mouse used was about 30 g. Sarcoma 180 type solid liver cancer was transplanted to this mouse. To this liver cancer-transplanted mouse, 10 times of β-1,3-D glucan extracted from the hot water extract of Hanabi-take (MH-3) was used only 3 times on the 7th, 9th, and 11th days from the start of the test.
0 ppm and 500 ppm were orally administered. There was no cancer occurrence at 100% at any dose, and healing was achieved.

NOD, an autoimmune diabetes model
Β-1,3-D glucan (100 ppm, 500 pp) extracted from Bamboo shoots mushroom (30 g)
When each of m) was orally administered, the onset was delayed or cured at any dose.

M in systemic autoimmune disease diabetes model
Symptoms were ameliorated by orally administering β-1,3-D glucan (100 ppm, 500 ppm) of Hanabitake mushroom extract (MH-3) to RL-1PL-1Pr mice (about 30 g).

Blood pressure returned to normal when hypertensive mice (about 30 g) were administered with hot water-extracted β-1,3-D glucan of Hanabitake (MH-3) at 50 ppm and 100 ppm,
Prevented mouse death.

[0022]

Example 1 6 g of powdered banana, 45 g of Ebios, 1 g of peptone, 0.6 g of calcium, 0.5 g of magnesium chloride, 2 g of powdered honey and 100 g of wheat flour were mixed. Then this is 1k
g of larch large sawdust (water content about 20%), and 1.5 L of normal water was added and mixed.

Next, the above-obtained mixture was filled in a polyethylene 500 mL bottle, a hole for air circulation having a diameter of 1.5 cm was opened at the center of the bottle and the lid was closed, and then placed in a steam sterilizer at 100 ° C. It was put in and sterilized for 5 hours. Since this method can prevent the damage of larch large sawdust caused by the Hanabitake mushroom growth-inhibiting substance, the heating of the large sawdust conventionally performed for treating the Hanabitake mushroom growth-inhibiting substance is not required.

When the sterilized fungal bed was cooled to room temperature, it was aseptically inoculated with the mycelium of MH-3 fungus (Agaricus edulis). After inoculation, primordia developed after 1 month of incubation at 20 ° C. Remove the lid of the container
When left in a dark room at 80 ° C and 80% humidity for 1 month, it was possible to harvest Hanabi-take mushrooms containing a large amount of the target β-1,3-D glucan.

Example 2: Effect of adding wheat to the medium A bacterial bed (medium A) prepared without adding wheat to the medium,
A bacterial bed (medium B) prepared by adding wheat was prepared. Perform artificial cultivation of Hanabi-take mushrooms using both of these fungus beds,
The results were compared. The medium B is the same as the medium used in Example 1, and the method for producing the bacterial bed is the same as that described in Example 1.

<Composition of medium (fungal bed)> Medium A (without addition of wheat); powdered banana 6 g, brewer's yeast 45 g, peptone 1 g, calcium chloride 0.6 g, powdered honey 2 g, larch large sawdust 1 kg Medium B (with wheat added) ); Powdered banana 6 g, brewer's yeast 45 g, peptone 1 g, calcium chloride 0.6 g, powdered honey 2 g, magnesium chloride 0.5 g, wheat flour 100
g, larch large sawdust 1 kg <Results> Cultivation progress Table 1 below shows the progress in the cultivation with the medium A and the cultivation with the medium B. As can be seen from this process, the generation period of the primordia from the bacterial inoculation is 1 month in the medium B compared with 2 months in the case of the medium A, and the generation period can be shortened to 1/2 by using the medium B I was able to.

On the other hand, the harvest period from the development of fruiting bodies was one month in both medium A and medium B, but the period from inoculation of bacteria to harvest was 3 months in medium A. In contrast, medium B was 2 months old. As a result, the cultivation period could be shortened by using the medium B.
This improves economic efficiency by shortening the cultivation cycle,
It means that profit efficiency can be improved.

[0028]

[Table 1]

Yield amount Between the cultivation using the medium A and the cultivation using the medium B, the amount of the raw bamboo shoots produced at the time of harvest and β-1,3-D.
Glucan weights were compared. The results are shown in Table 2.

On average per 100 strains at the time of harvest, the medium A was 50.5 g of fresh Hanabitake mushroom, whereas the medium B was 10
The amount of harvest of medium B was about twice as much as that of medium A at 0.5 g / 1 strain.

[0031]

[Table 2]

Cultivation conditions in Tables 1 and 2: ・ Larch large sawdust ・ Strain (Mushroom mushroom) MH-3 ・ Temperature after inoculation of the strain until generation of primordia ・ ・ ・ 18-25 ° C, Humidity ・ ・ ・ 55-75% ・ Fruit body Temperature from generation to harvest: 18 to 25 ° C., humidity: 75 to 85% When the production amounts of β-1,3-D glucan are compared between the media A and B, A is 35.8 g / 1 strain, B is B 45.2g / 1
As a strain, medium B was able to produce about 1.3 times as much as medium A, and the economic effect was significantly improved by medium B.

[0033]

The following effects were obtained by using MH-3 bacteria and using wheat flour as an added nutrient in the medium.

(1) The larch large sawdust used to prepare the bacterial bed has been conventionally boiled with hot water, treated with high temperature steam, and washed with water for the purpose of preventing the growth inhibition of Bamboo shoots.
According to the method of the present invention, these conventional processes are not required,
The economic effect has been greatly improved by omitting one step for producing the bacterial bed.

(2) By adding 5 to 10% wheat flour as a nutrient to the B formulation of the present invention, that is, the conventional formulation, the growth of Hanabi-take mushrooms is promoted, and the desired β-1,
The amount of 3-D glucan was increased by about 1.3 times compared with the conventional method (formulation A), and the economic effect was remarkably improved.

(3) Conventionally, it has been said that Hanabitake mushrooms cannot be cultivated by mixing various bacteria unless a glass bottle is used as a container for the fungus bed. However, by using MH-3 bacteria, a polyethylene container can be used, which is large. It has become possible to carry out industrial mass cultivation that allows the use of automatic machines.

Claims (4)

(57) [Claims] 1. A medium containing sawdust as a main component, the medium pair
Without performing high-temperature steam treatment and washing with water, Hanabitake MH-3 (accession number FERM P-1722
1) Inoculation with a method for producing a fungal bed for artificial cultivation of Ganoderma lucidum. 2. The method for producing a fungal bed for artificial cultivation of Hanabi-take mushroom according to claim 1, wherein a plastic container is used as the fungal bed container. 3. The method for producing a fungal bed for artificial cultivation of Hanabitake mushrooms according to claim 1 or 2, wherein the medium contains 5 to 10% of wheat flour. 4. The method for producing a fungal bed for artificial cultivation of Hanabi-taketake according to claim 1, wherein the medium contains powdered honey.