JP2002125460A - Method for making mushroom bed of sparassis crispa rr. having physiologically functional activity - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for making a mushroom bed for Sparassis Crispa Rr., capable of artificially cultivating Sparassis Crispa Rr. without fail and omitting a conventional complicated and costly culture medium treatment. SOLUTION: This method for making a mushroom bed for artificially cultivating Sparassis Crispa Rr. is characterized by vaccinating Sparassis Crispa Rr. MH-3 (FERM P-17221) to a culture medium having sawdust as the main ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for preparing a bacterial bed of Physcomitrella patens (Sparassis Crispa Rr.) Having a 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 its good chewyness, Hanabitake mushrooms are also prized in the Japanese cuisine world as valuable foodstuffs because of their pale yellow color close to pure white and forms like leaf peony.

Further, Hanabitake mushrooms contain a large amount of useful physiologically active substances, and attention has been paid to this point as well.
In particular, the content of total β-glucan known as a physiologically active substance having an excellent immunostimulating action is 43.
6 mg. Agaricus and maitake, also known as mushrooms with a high total beta-glucan content, have a total beta-glucan content of 11.6 g per 100 g and 18.1 respectively.
g, it can be seen that the content of Hanabitaketake is surprisingly high. In addition, among the total beta-glucan, in particular, β-1,3-D-glucan has been proven to have an antitumor effect. Therefore, Hanabiratake is not only an excellent food material as a health food, but also a valuable raw material for producing an antitumor agent.

[0004] Despite having the above excellent properties, Hanabitake mushrooms are very rare, as evident from being 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 a property that is not susceptible to fungi, which is not found in other mushrooms, and it is slow to grow even if it is lucky to succeed in artificial cultivation Therefore, at present, sufficient results have not yet been obtained.

Further, in the conventional artificial cultivation method of Hanabiratake, larch sawdust used as a culture medium is previously heated to a boiling water temperature of 120 to 121 ° C. and a pressure of 2 at the time of preparing the bacterial bed.
After being exposed to high-pressure steam at atmospheric pressure for 60 minutes, the medium was treated by washing with water. This is based on the reason that the inhibitor contained in the larch sawdust inhibits the growth of Hanabitaketake, and unless this inhibitor is removed, Hanabitaketake does not grow smoothly. However, this method for removing the inhibitor is complicated in operation and requires a large amount of cost, which has been an obstacle in the preparation of a bacterial bed.

[0006]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above circumstances, and an object of the present invention is to make it possible to reliably cultivate Hanabitake mushrooms and to omit the conventional complicated and expensive medium treatment. It is an object of the present invention to provide a method for preparing a fungus bed of C. versicolor.

[0007]

Means for Solving the Problems The object of the present invention is to provide a medium containing sawdust as a main component, Hanabitaketake MH-3 (accession number F).
ERM P-17221), which is achieved by a method for producing a bacterial bed for artificial cultivation of Hanabitaketake.

[0008] Hanabitaketake MH-3 used in the present invention
Was identified by the inventor as a strain suitable for artificial cultivation, and on February 17, 1999, the accession number FER
Deposited with the Institute of Biotechnology and Industrial Technology under the MP-17221. By using a bacterial bed prepared by inoculating this strain into a medium mainly composed of sawdust, artificial cultivation of Hanabitake mushrooms, which has been difficult in the past, can be made possible. Inoculated Hanabitaketake MH-3
It is preferable to use the spores or hypha.

The bacterial bed obtained by the present invention has an anticancer effect,
Β-1,3-D having blood glucose lowering action and blood pressure lowering action
It is suitable for industrial artificial cultivation of Hanabitake mushroom (mushroom) containing a large amount of glucan.

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

In the present invention, the larch sawdust can be used without removing the growth inhibitory substance of the Japanese larch sawdust.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The artificial cultivation of Hanabitake mushrooms has been extremely difficult technically in comparison with mushrooms conventionally subjected to artificial cultivation, namely Mediterranean shimeji, shiitake and oyster mushrooms. However, as a result of intensive research, the inventor of the present application has found that cultivation of mushrooms containing a large amount of the desired β-1,3-D glucan requires conventional medium treatment by selective use of bacteria (MH-3). Without any short and economical consequences. In addition, 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 sawdust of hemlock, fir, pine, beech, and shii. It is preferable to use nutrients required for cultivating Hanabitake mushrooms in these single sawdust or mixture or chips thereof.

[0014] Hanabitaketake MH-3 inoculated into the above medium
The spores or hyphae can be used. In the present invention, for example, sawdust, fir, pine, beech, shii sawdust, or chips are mixed with nutrients necessary for the growth of Hanabitake mushrooms, filled into a 500 mL polyethylene bottle, sterilized, and the medium is cooled to room temperature. After cooling, M
H-3 spores or hyphae are inoculated aseptically. By cultivating fruiting bodies from the fungal bed thus prepared, β-1, effective for cancer prevention, treatment, diabetes and hypertension treatment.
Hanabitake mushrooms containing a large amount of 3-D glucan can be grown.

[0015] The first aspect of the present invention relates to Hanabitaketake MH.
By using -3, it is possible to produce an artificially cultivated bacterial bed of Hanabitake mushroom without the need for conventional complicated and expensive medium treatment such as high-temperature steam treatment and water washing. That is, the inventor succeeded in successfully growing Hanabitaketake without using the above-mentioned inhibitory removal operation by using the MH-3 bacteria. As a result, it became possible to prepare the Hanabitake mushroom bed extremely economically and easily. In addition, conventionally, it was said that cultivation of Hanabiratake was impossible if various bacteria were mixed unless a glass bottle was used as a container for the fungus bed.
A polyethylene container can be used by using H-3 bacteria,
Industrialized mass cultivation that allows the use of large automatic machines has become possible.

[0016] A second aspect of the present invention is directed to improvement of a bacterial bed medium. As described above, in the present invention, sawdust is used as a main component as a medium for preparing a fungus bed of Hanabitaketake MH-3, but by adding wheat as another nutrient to be added thereto, the cultivation period can be shortened and the harvest can be shortened. It has become possible to achieve increased amounts.

The MH-3 mushrooms selected by artificial cultivation using the bacterial bed prepared as described above show that the anti-cancer, hypoglycemic, immunostimulatory and anti-hypertensive effects are strong. The inventor has discovered. This experiment is described below.

The test period was 35 days and the weight of the mice used was about 30 g. Sarcoma 180 type solid liver cancer was transplanted to this mouse. To this liver cancer transplanted mouse, β-1,3-D glucan extracted from Hanabitake mushroom (MH-3) was extracted 10 times only on days 7, 9, and 11 from the test start date.
0 ppm and 500 ppm were orally administered. At all doses, 100% did not develop cancer and healed.

NOD, an autoimmune diabetes model
For a mouse (about 30 g), β-1,3-D glucan extracted from Hanabitaketake (100 ppm, 500 pp)
When m) was orally administered, onset was delayed or cured at any dose.

M of the diabetes model for systemic autoimmune diseases
The symptom was improved by orally administering β-1,3-D glucan (100 ppm, 500 ppm) of Hanabitaketake (MH-3) to RL-1PL-1Pr mice (about 30 g).

When hypertensive mice (approximately 30 g) were administered 50 ppm and 100 ppm of hot-water-extracted β-1,3-D glucan of Hanabitaketake (MH-3), the blood pressure returned to normal.
Mouse death was prevented.

[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 flour were mixed. Then, this is 1k
g of larch sawdust (water content: about 20%), and 1.5 L of ordinary water was poured and mixed.

Next, the mixture obtained above was filled in a 500 mL bottle made of polyethylene, a hole for air circulation having a diameter of 1.5 cm was formed at the center of the bottle, and the bottle was covered with a steam sterilizer at 100 ° C. And sterilized for 5 hours. By this method, the larch sawdust can be prevented from being hindered by the growth inhibitor of Hanabitaketake, so that the heating of sawdust conventionally performed for treating the growth inhibitor of Hanabitaketake is not required.

When the sterilized bed was cooled to room temperature, the mycelium of MH-3 (mushroom Hanabitaketake) was aseptically inoculated. After inoculation, primordia developed after incubation at 20 ° C. for one month. Remove the container lid and allow
When left in a dark room at 80 ° C. and a humidity of 80% for one month, it was possible to harvest Hanabitake mushrooms containing a large amount of the desired β-1,3-D glucan.

Example 2: Effect of the addition of wheat to the medium A bacterial bed (medium A) prepared without adding wheat to the medium,
A bacterial bed (medium B) was prepared by adding wheat. Artificial cultivation of Hanabiratake is performed using both of these bacterial beds,
The results were compared. The medium B is the same as the medium used in Example 1, and the method for preparing the bacterial bed is the same as that described in Example 1.

<Composition of Medium (Bacterial Bed)> Medium A (without wheat); powdered banana 6 g, brewer's yeast 45 g, peptone 1 g, calcium chloride 0.6 g, powdered honey 2 g, larch sawdust 1 kg Medium B (with wheat added) ); Powdered banana 6g, brewer's yeast 45g, peptone 1g, calcium chloride 0.6g, powdered honey 2g, magnesium chloride 0.5g, flour 100
g, larch sawdust 1 kg <Results> Cultivation progress Table 1 below shows the progress in cultivation using medium A and cultivation using medium B. As can be seen from this process, the development period of the primordium from the bacterial inoculation is one month in the medium B for two months in the case of the medium A, and the development period is reduced to half by using the medium B. Was completed.

On the other hand, the harvesting period from the emergence of fruiting bodies was 1 month in both the medium A and the medium B. However, the period from the inoculation to the harvest was 3 months in the medium A. In contrast, medium B was 2 months. As a result, the cultivation period could be shortened by using the medium B.
This increases economy by shortening the cultivation cycle,
This means that profit efficiency can be improved.

[0028]

[Table 1]

Amount of Harvest Between the cultivation using the medium A and the cultivation using the medium B, the production amount of the raw mushroom at the time of harvest and the β-1,3-D
The glucan weights were compared. Table 2 shows the results.

On average, the medium A was harvested at a rate of 50.5 g / raw strain of A. mushroom per 100 strains at the time of harvesting, and the medium B was 10
The yield of the medium B was about twice as large as that of the medium A at 0.5 g / 1 strain.

[0031]

[Table 2]

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

[0033]

The following effects were obtained by using MH-3 bacteria and using flour as a nutrient added to the medium.

(1) Larch sawdust used for the preparation of the fungus bed has been conventionally subjected to boiling with hot water, high-temperature steam treatment and washing with water for the purpose of preventing growth inhibition of Hanabitake mushroom.
According to the method of the present invention, these conventional processes are not required,
The economic effect was greatly improved by eliminating one step for the preparation of the bacterial bed.

(2) The addition of 5 to 10% flour as a nutrient to the B-formulation of the present invention, ie, the conventional formula, promotes the growth of C. versicolor and further improves the desired β-1,
The amount of 3-D glucan was increased about 1.3 times as compared with the conventional method (formulation A), and the economic effect was remarkably improved.

(3) Conventionally, it has been considered that cultivation of Hanabitake mushrooms is impossible due to contamination of various bacteria unless glass bottles are used as a container for the fungus bed, but polyethylene containers can be used by using MH-3 bacteria. Industrial mass cultivation that can use automatic machines has become possible.

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[Procedure amendment]

[Submission date] July 11, 2001 (2001.7.1)
1)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0003

[Correction method] Change

[Correction contents]

Further, Hanabitake mushrooms contain a large amount of useful physiologically active substances, and attention has been paid to this point as well.
In particular, the content of total β-glucan known as a physiologically active substance having an excellent immunostimulating action is 43.
6 g . Agaricus and maitake, which are known as mushrooms having a high total beta-glucan content, have a total beta-glucan content of 11.6 g and 18.1 g per 100 g, respectively.
It can be seen from the comparison with that that the content of Hanabitaketake is surprisingly high. In addition, among the total beta-glucan, in particular, β-1,3-D-glucan has been proven to have an antitumor effect. Therefore, Hanabitake mushrooms are not only excellent foods as health foods, but also valuable raw materials for producing antitumor agents.

Claims (4)

[Claims] 1. A method for producing a fungus bed for artificial cultivation of Hanabitake mushrooms, which comprises inoculating Hanabitaketake MH-3 (Accession No. FERM P-17221) into a medium mainly containing sawdust. 2. The method for producing a fungus bed for artificial cultivation of Hanabitake mushrooms according to claim 1 or 2, wherein a high-temperature steam treatment and a washing step for removing the growth inhibitory substances of Hanabitaketake in the culture medium are not performed. Method. 3. The method according to claim 1, wherein a plastic container is used as the bacterial bed container. 4. The method according to claim 1, wherein the culture medium contains 5 to 10% of flour.