JP4944384B2 - Meshimakobu liquid culture medium - Google Patents

Description

The present invention relates to a liquid culture medium composition of Meshima Cobb.

Technical background

  Meshimakobu is a woody mushroom like a moss mushroom that infests specifically with mulberry and has a semicircular shape with a bright yellow or tan color on the back of the fruit body. Meshimakobu is called mulberry yellow in Kampo, and in Japan it is widely distributed and grown naturally in the gender archipelago of Nagasaki Prefecture, and in temperate regions of Southeast Asia such as Korea, North America, Australia, and the Philippines. However, in recent years, antitumor polysaccharides have been extracted from Meshimakobu as a physiologically active ingredient and proved to have an excellent immunostimulatory effect, increasing demand, leading to overexploitation in the country of origin and making it difficult to obtain. Yes.

Therefore, in recent years, a method for artificially cultivating the mycelium of Meshimakobu has been proposed (for example, JP-A-2001-178448). This culture method is inoculated with Mesima Kob mycelium in a liquid medium and cultured at 22 to 35 ° C., and the carbon source of the liquid medium is selected from glucose, mannose, galactose, sucrose, trehalose, cellobiose, maltose, lactose, and raffinose The culture conditions include use of one or more saccharides, culturing under aerobic conditions, and adjusting the pH of the medium at the start of culture to 4.5 to 6.5.

Japanese Patent Application Laid-Open No. 2003-259857 describes a method of liquid-culturing Meshima Kob mycelium by adding an extract obtained by hot water extraction from trunks, branches, leaves, and roots of Yamaguchi and mulberry trees.

Furthermore, Japanese Patent Application Laid-Open No. 2002-70 describes a method of adding okara to a culture medium for artificial cultivation of Hatake Shimeji. However, the culture medium described in this publication is a solid culture medium for generating fruit bodies of basidiomycetes, and is not the liquid culture medium of the present invention. In general, the nutrients necessary for the growth of basidiomycetes and the nutritional conditions necessary for the formation of fruiting bodies are greatly different. In addition, the components in the fruit body, the components in the mycelium obtained by liquid culture, and the components in the mycelium obtained by solid culture or fungal bed culture are often very different from each other. A nutrient component effective for a solid culture medium is not necessarily effective for a liquid culture medium as it is.
JP 2001-178448 A JP 2003-259857 A Japanese Patent Laid-Open No. 2002-70

The problem to be solved is that Mesima Kobu has a slow growth of mycelia, and the yield of mycelium obtained by the conventional liquid culture method is small, and a sufficient mycelium cannot be obtained.

The present inventors have found that the yield of Meshimakob mycelium is dramatically increased by adding okara powder or soy milk to a conventional liquid medium, and the present invention has been completed. That is, according to the present invention, the growth of Mesima Kob mycelium is drastically increased by inoculating Mesima Kob mycelium in a liquid medium to which Okara powder or soymilk is added and culturing the same.

According to the present invention, according to the method for liquid culture of mesimacob mycelium according to the present invention, the yield of mesimacob mycelium is dramatically increased in a short period of time by culturing in a liquid medium containing okara powder and soymilk. Is possible.

The okara powder in the present invention is a known drying means such as an air-drying method or a drum drying method for producing okara as a by-product produced by grinding and heating and squeezing a soybean soaked in water. Dry by using. In addition, soy milk is a juice obtained by grinding and squeezing soybeans soaked in water. Okara powder and soy milk contain components such as proteins, lipids, and minerals, so they are good sources of nitrogen for basidiomycetes.

The okara powder is preferably added in the range of 0.1 to 10% by weight with respect to the basic medium. Okara powder is added as a solid in the liquid medium and dispersed. Thereafter, sterilization is performed to obtain a culture medium. Ocara powder is decomposed by protease produced by hyphae as Meshimakobu is cultured, and is supplied to the mycelia as a nitrogen source, thereby promoting growth of Meshimakob mycelium. In particular, the effect is remarkably exhibited when 0.5 to 5.0% by weight is added to the basic medium.

In addition, it is desirable to add soy milk in the range of 1.0 to 30% by weight with respect to the basic medium, and the effect is particularly remarkable when 5.0 to 20.0% by weight is added to the basic medium. Demonstrated.

The basic medium used in the liquid culture according to the present invention contains nutrients necessary for growth when cultivating the mycelium of Meshimakobu, such as a carbon source and a nitrogen source, and may further contain vitamins and minerals.

As the carbon source, glucose, sucrose, mannitol, galactose, trehalose, maltose, lactose, dextrin and the like are added.

As the nitrogen source, yeast extract, malt extract, peptone, polypeptone, casamino acid, calcium nitrate, ammonium sulfate and the like are added.

As vitamins and minerals, thiamine, biotin, folic acid, calcium chloride, magnesium sulfate and the like are added.

The basic medium in the present invention is composed of, for example, a medium composition to which 2.0% glucose and 0.5% yeast extract are added. Moreover, even if it is a culture medium composition other than the above, as long as it is a culture medium suitable for Meshimakob mycelium culture, of course, it is applied to this invention.

In the liquid culture method of mesimacob in the present invention, it is desirable to inoculate the liquid medium supplemented with the above-mentioned okara powder with the inoculum of mesimacob and cultivate under aerobic conditions at room temperature of about 20 to 30 ° C. for about 21 to 28 days. The culture may be shake culture or aeration culture.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto. In addition, culture and operation methods shall be performed aseptically.

[Example 1] Effect test of okara powder added to medium Inoculate a portion of the cultivated strain of Meshima Kobu (preserved strain of Naris Cosmetic Co., Ltd.) on an agar medium of potato dextrose (DIFCO), After 4 weeks of planar culture, this is used as an inoculum.

A basic liquid medium was made by adding 2.0% glucose and 0.5% yeast extract to 100 ml water. Also, a liquid medium is prepared by adding the amounts of dry powder of okara, mulberry extract (4.0% concentration aqueous solution), bran and rice bran as additives in Table-1. 100 ml of these liquid media are dispensed into 200 ml Erlenmeyer flasks and then autoclaved using an autoclave. The mulberry extract is cut into 5 mm with pruning shears and the mulberry tree trunks and branches, extracted with hot water for about 60 minutes in addition to water, solids removed after cooling, and the above evaporation residue. What was concentrated so that it might become a minute was used.

After allowing the above-mentioned liquid medium to cool to room temperature, take the Meshimakobu culture that has been cultured on the agar medium, and inoculate each flask with 1 ml of a well-homogenized basic medium consisting of glucose and yeast extract. After inoculation, shaking culture was performed under dark conditions at 24 ° C. and 150 rpm.

About 21 days after the start of culture, mycelium grows in the liquid medium. Therefore, the grown mycelium was collected, washed thoroughly with water, and then air-dried at 60 ° C. for 5 hours.

The growth ratio of the collected mycelium is shown in Table-1. The growth ratio was calculated by calculating the ratio with respect to 100 when the yield when cultured in the basic medium of Example-1 was 100.

According to Table-1, compared with what was cultured with the basic culture medium of the comparative example-1, Example-1 which added 1% of okara powder showed the growth rate of 328%. On the other hand, the growth rate of Comparative Example-2 to which mulberry extract was added was 109%, and it was revealed that the effect of adding okara powder was remarkable. In addition, the growth rates of Comparative Example-3 added with bran commonly used in solid media and Comparative Example-4 added with rice bran were 165% and 149%, respectively. Did not reach. From the above results, it is clear that the required substances are not the same in the liquid culture and the solid culture. In addition, the growth rates of Example-2, Comparative Example-5, and Comparative Example-6, in which Mulberry extract was added to Example-1, Comparative Example-3, and Comparative Example-4, respectively, were compared with those in which Mulberry extract was not added. There was no big difference, and it became clear that Okara powder had a great influence on the growth of Meshimakobu hyphae.

[Example 2] Addition amount of okara powder to be added to the medium In order to examine the addition amount of okara powder to be added to the medium, a medium as shown in Table-2 was prepared, and the same method as in Example 1 was used. The amount of cells was measured.

Table 2 shows the results. When the amount of okara powder added was 0.05%, the growth rate was 156%. The proliferation rate increased, but no significant proliferation effect was shown. On the other hand, when 0.1% to 5.0% was added, a remarkable proliferation effect of Meshimakob mycelium was recognized.

[Example 3] Amount of soy milk added to the medium In order to examine the amount of soy milk added to the medium, a medium as shown in Table 3 was prepared, and the same method as in Example 1 was used. The amount was measured.

The results are shown in Table-3. When the amount of soy milk added was 1.0%, the growth rate was 181%. On the other hand, when 5.0% to 20.0% was added, the proliferation rate was 281% to 437%, and a remarkable proliferation effect of Meshimakob mycelium was recognized. Further, even when the addition amount was increased to 30%, no growth proportional to the addition amount was observed.

According to the liquid culture method of Meshimakob mycelium according to the present invention, it is possible to dramatically increase the yield of Meshimakob mycelium in a short period of time by culturing it in a liquid medium containing okara powder and soymilk. Wide application can be expected in the fields of cosmetics, pharmaceuticals, and foods.

Claims (1)

A basal medium containing glucose at least as a carbon source, a liquid culture medium of Phellinus, characterized in that blended with soy milk 20.0 to 30.0 by weight%.