TWI628278B - Culture method for improving amount of polysaccharides in inonotus obliquus - Google Patents

Abstract

The present invention provides a solid-state culture method for increasing the content of polysaccharides in Chaga, comprising an inoculation step, a first culture environment adjustment step, a nutrient addition step, a second culture environment adjustment step, and a harvesting step. First, the Chaga strain was inoculated to a solid medium to culture the Chaga mycelium. When the pyriflora mycelium reaches the first predetermined area, the brightness and temperature of the culture environment are lowered, and a culture time is cultured in a dark environment. Then, after the nutrient solution is added, the brightness and temperature of the culture environment are restored, and the birch mycelium is harvested when the second predetermined area is reached. Thereby, the invention can effectively increase the content of triterpenoids in the mycelium product of the beetle.

Description

Solid-state culture method for increasing polysaccharide content in Chaga

The present invention relates to a method for cultivating a Chaga, and more particularly to a method for cultivating a polysaccharide content in Chaga.

Inonotus obliquus is a folk medicinal fungus mainly distributed in Russia and Eastern Europe. Chinese aliases include white birch, birch mushroom, betulin, Siberian Ganoderma lucidum, birch hole and tree mushroom. The classification belongs to the fungi community, Basidiomycota, Agaricomycetes, Hymenochaetales, Hymenochaetaceae, and Inonotus.

In recent years, studies have shown that polysaccharides and triterpenoids contained in Chaga can effectively activate the immune system and inhibit the proliferation of viruses, and prevent breast cancer, liver cancer, uterine cancer, stomach cancer, diabetes and hypertension. Effect, its nutritional and medicinal value gradually Arouse the attention of the people.

The early use of Chaga is mainly based on the naturally occurring wild Chaga. The areas that can be collected are Russia's Siberia, Japan's Hokkaido, Finland, China's Jilin Changbai area, and Heilongjiang's large and small Xing'an Mountains. However, due to the high yield and high collection cost of naturally occurring Chaga, the wild Chaga is expensive, and the wild resources are limited and increasingly exhausted, which is difficult to meet the market demand.

Accordingly, at present, the production methods of Chaga are mostly produced by artificial fermentation such as liquid fermentation culture or solid state culture. Among them, the liquid fermentation culture technology, although the speed of cultivation of Chaga is fast, suitable for industrial mass production, but must use a large amount of culture solution, or use a larger capacity of the fermentation tank, and the growth environment and natural Chaga differ greatly, the product The nutrients are also different from natural products. The solid culture will be close to the growth environment and conditions of natural Chaga, so the products and nutrients are closer to the natural Chaga. However, at this stage, the product of Chaga solid-state culture technology has a lower yield, a slower growth rate, and a lower ratio of the active ingredient polysaccharides than the naturally occurring wild Chaga.

Therefore, how to improve the proportion of polysaccharides in artificially cultivated Chaga is a hot topic in the research of Chaga cultivation.

In view of the above, the present invention provides a novel method for cultivating a beetle, thereby effectively cultivating the mycelium product of the Chaga, and simulating the natural growth environment of the Chaga by solid state culture, thereby increasing the content of the polysaccharide in the product.

In view of the above, an embodiment of the present invention provides a solid-state culture method for improving the content of polysaccharides in Chaga, comprising: performing an inoculation step, performing a first culture environment adjustment step, and performing a nutrient addition step, A second culture environment adjustment step is performed and a recovery step is performed. In the inoculation step, at least one Chaga strain is inoculated to a solid medium and the Chaga strain is cultured in a culture environment to produce at least one Chaga mycelium. In the first culture environment adjustment step, when the Chaga mycelium is cultured to reach a first predetermined area, one brightness of the culture environment and a temperature are lowered to culture the Chaga mycelium for a culture time. In the step of adding the nutrient solution, a nutrient solution is added to the solid medium. In the second culture environment adjustment step, the brightness and temperature of the culture environment are restored. In the harvesting step, when the Chaga mycelium is cultured to reach a second predetermined area, the Chaga mycelium is harvested.

A solid-state culture method for improving the content of polysaccharides in Chaga according to the above embodiment, wherein a preparation method of the solid medium in the inoculation step may comprise mixing a base, a sugar, birch shavings, an agar and water And the aforementioned base material may comprise a cereal or a bean class. Next, a high temperature sterilization step is performed to prepare a solid medium.

The solid culture method for improving the polysaccharide content in the chafer according to the foregoing embodiment, wherein the solid medium can be placed in a solid culture flask during the inoculation step, and the bottom area of one of the solid culture flasks can be greater than or equal to 200 Square centimeters.

Solid-state culture method for improving polysaccharide content in Chaga by increasing polysaccharide content in Chaga according to the above embodiment, solid-state culture method for improving polysaccharide content in Chaga by increasing polysaccharide content in Chaga, wherein the nutrient solution It can be a Chinese herbal extract and can be extracted from a group of free birch leaves, safflower, peach kernel and radix isatidis.

The solid-state culture method for improving the polysaccharide content in the Chaga according to the foregoing embodiment for improving the polysaccharide content in the Chaga cultivar to increase the polysaccharide content in the Chaga, wherein the first preset in the first culture environment adjustment step is performed. The area may be greater than or equal to 50 square centimeters and less than or equal to 100 square centimeters.

The solid-state culture method for improving the polysaccharide content in the white birch to increase the polysaccharide content in the white birch according to the foregoing embodiment, wherein the second predetermined area in the foregoing harvesting step may be greater than or equal to 150 square centimeters and less than or equal to 180 square centimeters.

a solid-state culture method for improving the content of polysaccharides in Chaga by increasing the polysaccharide content in Chaga according to the foregoing embodiment, The temperature of the aforementioned culture environment can be lowered to between 11 ° C and 15 ° C in the aforementioned first culture environment adjustment step.

The solid-state culture method for improving the polysaccharide content in the white birch to increase the polysaccharide content in the chafer according to the foregoing embodiment, wherein the step of performing the second culture environment adjustment step and the foregoing harvesting step further comprises repeating the foregoing addition The nutrient solution step, and the frequency of adding the aforementioned nutrient solution is 20 ml per day to 1000 ml per day.

The Summary of the Invention is intended to provide a simplified summary of the present disclosure in order to provide a basic understanding of the disclosure. This Summary is not an extensive overview of the disclosure, and is not intended to be an

S100, S102, S104, S106, S108, S110‧‧ steps

The above and other objects, features, advantages and experimental examples of the present invention will be more apparent and understood. The description of the drawings is as follows: Figure 1 shows an embodiment of the present invention for improving the content of polysaccharides in Chaga A flow chart of a solid state culture method for increasing the polysaccharide content in Chaga.

Please refer to FIG. 1 , which illustrates an embodiment of the present invention for improving polysaccharide content in Chaga cultivars and improving polysaccharides in Chaga A flowchart of the solid state culture method of the content, and the aforementioned culture method includes the step S100, the step S102, the step S104, the step S106, and the step S108.

Step S100 is to perform an inoculation step. In step S100, at least one Chaga strain is inoculated onto a solid medium, wherein the solid medium can be placed in a solid culture bottle, and the bottom area of one of the solid culture bottles can be greater than or equal to 200 square centimeters, but The invention is not intended to be limited to this. Further, although not shown, in the present invention, the solid medium may be prepared by previously mixing a base material, a saccharide, birch wood chips, an agar and water, and then preparing it through a high temperature sterilization step. Preferably, the aforementioned binder may comprise a cereal or a legume. Next, the Chaga strain is cultured in a culture environment to produce at least one Chaga mycelium, and the aforementioned culture environment is specifically a normal temperature light environment.

Step S102 is to perform a first culture environment adjustment step. In step S102, when the Chaga mycelium is cultured in the culture environment to reach a first predetermined area, the step S102 is used to reduce the brightness and temperature of the culture environment to make the Chaga mycelium follow-up. Culture one culture time in a dark environment. Specifically, the aforementioned culture time may be between 6 hours and 24 hours, and the temperature is adjusted to be between 11 ° C and 15 ° C. More specifically, the aforementioned first predetermined area is greater than or equal to 50 square centimeters and less than or equal to 100 square centimeters.

Step S104 is a step of adding a nutrient solution. In detail, in step S102, the white birch mycelium is cultured in a light-proof environment at a low temperature. After hours to 24 hours, nutrient solution can be further added to the solid medium to serve as a nutrient source for the subsequent growth of the Chaga mycelium. Specifically, the nutrient solution is also a Chinese herbal medicine extract, and the extract is a group consisting of birch leaves, safflower, peach kernel and Banlangen.

Step S106 is to perform a second culture environment adjustment step. After step S104, the Chaga mycelium is continuously cultured for another incubation time, and the culture time is from 1 hour to 10 hours. Next, in step S108, the brightness and temperature of the culture environment are restored, and even if the culture environment is restored to the normal temperature illumination environment.

Step S108 is a harvesting step. In detail, after the step S106, the Chaga mycelium is continuously cultured in a normal temperature environment, and when the Chaga mycelium is cultured to reach a second predetermined area, the harvesting can be performed. Specifically, the second predetermined area is greater than or equal to 150 square centimeters and less than or equal to 180 square centimeters.

Further, as described above, after the step S108, the Chaga mycelium system is continuously cultured in a normal temperature light environment, so that the step S106 may be repeated during the cultivation to supplement the nutrient solution to the solid medium. Preferably, the frequency of the nutrient solution is from 20 ml per day to 1000 ml per day.

The details of the culture method of the present invention for improving the polysaccharide content in Chaga, and the effects thereof, will be described in detail below by way of an embodiment and a comparative example of an embodiment of the present invention. However, this The embodiments may be applied to various inventive concepts and may be embodied in various specific ranges. The specific embodiments are for illustrative purposes only and are not limited by the scope of the disclosure.

[Examples]

As shown in step S100 of Fig. 1, in the present embodiment, the Chaga strain is first inoculated onto a solid medium, wherein the solid medium is placed in a solid culture flask, and the bottom area of the solid culture flask is 220 square. Centimeters. Further, the solid medium is preferably mixed with mung bean powder, glucose, birch shavings, agar and water in the following ratios: 11.8% of mung bean powder, 9.7% of birch shavings, 1.3% of glucose, 9.5% of agar And 67.7% of water, and prepared by high temperature sterilization. Next, the Chaga strain is cultured in a normal temperature light environment to produce a Chaga mycelium, wherein the normal temperature means that the temperature of the culture environment is controlled between 15 ° C and 25 ° C, and preferably 25 ° C.

Subsequently, as shown in step S102 of Fig. 1, when the beetle mycelium is cultured to an area of from 50 square centimeters to 100 square centimeters, and preferably 80 square centimeters, the aforementioned white birch hyphae The brightness and temperature of the culture environment of the body are lowered to provide a light-proof low temperature environment, and the Chaga mycelium is continuously cultured for one culture time in the adjusted culture environment. In the present embodiment, the temperature of the aforementioned culture environment is lowered to between 10 ° C and 15 ° C, and preferably 11 ° C. In addition, the Chaga mycelium system is continuously cultured in the above-mentioned light-proof and low temperature environment. The raising time can be between 6 hours and 24 hours, preferably 12 hours.

Next, as shown in step S104 of Fig. 1, the nutrient solution is added to the solid medium as a nutrient source for the subsequent growth of the Chaga mycelium. In this embodiment, the nutrient solution is a herbal extract, and is obtained by the following steps. First, the birch leaves are mixed with a 95% ethanol solution in a ratio of 1:25, and reacted at 60 ° C for at least one hour. After collecting the extract, a safflower with a volume of 1:50 and a peach of 1:40 volume are added. The solution was prepared in a volume of 1:50 from Radix Isatidis to the extract, heated and continuously stirred for 2 hours, and filtered to remove the residue to obtain the nutrient solution. After the addition of the nutrient solution, the Chaga mycelium is continuously cultured for another incubation time, and the culture time is 1 to 10 hours, preferably 5 hours.

Then, as shown in step S106 of FIG. 1 , the brightness and temperature of the culture environment are restored, and even if the culture environment is restored from a low-temperature environment to a normal temperature environment, and the beetle mycelium is exposed to normal temperature. Continue to cultivate. In addition, during the cultivation of the Chaga mycelium in a normal temperature environment, 20 ml to 1000 ml of the nutrient solution may be added to the solid medium every day, preferably 100 ml per day.

Further, as shown in step S108 of Fig. 1, when the birch mycelium is cultured to an area of from 150 square centimeters to 180 square centimeters, the harvesting of the chamomile product can be carried out.

Finally, the polysaccharide is heated and extracted according to the following steps. Calculate the polysaccharide content of the product: 11.5kg dry and impurity-free, crushed white birch product placed in the container; 2 take anhydrous propionic acid 0.9L, add 0.1L water to form propionic acid solution 1L; 3 take the above solution 1L, add Into the container, fully stirred to evenly wet; 4 continue to stir the above materials for 1h, vacuum distillation under reduced pressure, remove the organic acid solution, to the container without liquid, to complete the organic solvent treatment of the Chaga raw materials; 5 take the above steps 4 g of the organic solvent-treated Chaga product was placed in a 1 L beaker, 500 ml of distilled water was added, and extracted in a hot water bath at 75 ° C for 1.5 h, filtered, and the above process was repeated once, and the filtrate was combined twice to obtain An aqueous solution containing a polysaccharide of Chaga, 5 L of 95% (V/V) ethanol was added to the aqueous solution, placed for 3 hours, centrifuged, and the precipitate was freeze-dried to obtain an active polysaccharide; 6 The content of the polysaccharide was listed in Table 1. .

[Comparative Example 1]

Compared with the examples, Comparative Example 1 was carried out by culture of Chaga mycelium in a solid culture method without adding a nutrient solution, and after the harvest, the polysaccharide of the product was extracted and the content in the product was estimated. As shown in Table 1.

[Comparative Example 2]

Compared with the examples, Comparative Example 2 was carried out by culture of Chaga mycelium in a solid culture method without an environmental adjustment step, and after the harvesting, the polysaccharide of the product was extracted and the content in the product was estimated. Organize as shown in Table 1.

[Comparative Example 3]

Compared with the examples, Comparative Example 3 was carried out by culturing the Chaga mycelium with a solid culture method without adding a nutrient solution and without an environmental adjustment step, and extracting the polysaccharide of the product after harvesting and estimating the The content in the product was as follows.

[Comparative Example 4]

Unlike the examples and other comparative examples, the one used in Comparative Example 2 was wild Chaga. After extracting the polysaccharide and collecting the content in wild Chaga after harvesting, they were combined as shown in Table 1.

Table 1 shows the dry weight and polysaccharide content of the Chaga products of the respective Examples and Comparative Examples.

It can be seen from Table 1 that the chamomile product obtained by the solid state culture method provided by the present invention is obtained by extracting the chamomile product or the wild Chaga extract which has not been added with the nutrient solution step, without the loop adjustment step. Comparing the polysaccharide content, it is obvious that the Chaga mycelium cultured by the culture method provided by the present invention has a polysaccharide content higher than that of the solid culture method without the nutrient solution step and without the loop adjustment step. Product, and its content is closer to wild natural Chaga, obvious Compared with the conventional solid culture method, the present invention can effectively increase the content of polysaccharides in Chaga.

In summary, the improved culture method and the addition of the Chinese herbal medicine extract as the nutrient source of the culture body of the Chaga mycelium can effectively improve the polysaccharide content of the active ingredients in the Chaga. In addition, the operation method of the culture method provided by the invention is simple, and the effect of reducing the labor and time cost of the culture can be achieved.

Although the present invention has been disclosed in the above embodiments, it is not intended to limit the present invention, and the present invention can be modified and modified without departing from the spirit and scope of the present invention. The scope is subject to the definition of the scope of the patent application attached.

Claims (5)

A solid-state culture method for increasing the content of polysaccharides in Chaga, comprising: performing an inoculation step of inoculating at least one Chaga strain into a solid medium and cultivating the Chaga strain in a culture environment to produce at least a white birch mycelium; performing a first culture environment adjustment step, wherein when the white birch mycelium is cultured to reach a first predetermined area, the brightness of one of the culture environments is lowered, and the temperature is lowered to 11 ° C Between 15 ° C, so that the white birch mycelium is cultured for a culture time, wherein the first predetermined area is greater than or equal to 50 square centimeters and less than or equal to 100 square centimeters; a step of adding a nutrient solution is added a nutrient solution made of birch leaves to the solid medium, wherein the nutrient solution is a Chinese herbal extract, and the nutrient solution is a botanical leaf with a volume of 1:25, a volume of 1:50 safflower, a volume of 1:40 peach kernel and a volume of 1:50 radix is added to a group of 95% ethanol solution; a second culture environment adjustment step is performed to restore the brightness of the culture environment and recover the temperature Between 15 ° C and 25 ° C; and performing a harvesting step, wherein when the Chaga mycelium is cultured to reach a second predetermined area, the Chaga mycelium is harvested, wherein the second predetermined area is Greater than or equal to 150 square centimeters and less than or equal to 180 square centimeters. The solid-state culture method for improving the content of the polysaccharide in the chafer according to the first aspect of the patent application, wherein the preparation method of the solid medium in the inoculating step comprises: mixing a base material and a sugar , birch shavings, agar and water, wherein the base comprises a cereal or a bean; and a high temperature sterilization step is performed to prepare the solid medium. Solid-state culture for improving the content of polysaccharides in Chaga, as described in Item 1 of the patent application The method wherein the solid medium is placed in a solid culture flask during the inoculation step, and one of the solid culture flasks has a bottom area of greater than or equal to 200 square centimeters. The solid culture method for improving the content of polysaccharides in Chaga, as described in claim 1, wherein the nutrient solution is a Chinese herbal extract, and the nutrient solution is extracted from birch leaves, safflower, peach kernel and Banlangen. Form a group. The solid culture method for improving the content of the polysaccharide in the chafer according to the first aspect of the patent application, wherein the step of adjusting the second culture environment and the harvesting step further comprises: repeating the adding nutrition The liquid step, wherein the nutrient solution is added at a frequency of 20 ml per day to 1000 ml per day.