TW201932592A - An Taiwanofungus camphoratus culture medium for increasing the fruiting bodies and triterpenoids - Google Patents

Abstract

An Taiwanofungus camphoratus culture medium for increasing the fruiting bodies and triterpenoids is disclosed. The culture medium includes agar, D-glucose, malt extract, and 0.5 to 1% of peptone which effectively increases the quality of Antrodia camphorata.

Description

Antrodia cinnamomea medium capable of increasing Antrodia cinnamomea fruit body and enhancing triterpenes

The present invention relates to the cultivation of medicinal fungi; in particular, it refers to an Antrodia cinnamomeus medium component that can increase the fruit body of Antrodia camphorata and enhance triterpenes.

Antrodia cinnamomea, also known as Antrodia camphorata, is Taiwan’s six largest medicinal fungi in the world. It grows only on the decayed inner wall of the trunk of Antrodia camphorata, which is endemic to Taiwan for more than 100 years, or on the moist surface of dead and fallen Antrodia camphora wood. The wild Antrodia camphorata has been listed as one of the conservation creatures by the Taiwan government in recent years and is not allowed to be exported. Currently only in Taiwan, it is extremely precious and is regarded as the treasure of Taiwan, the ruby in the forest, and the king of Ganoderma.

The medicinal properties of Antrodia camphorata include:

1. Detoxification and detoxification;

2. Resist virus infestation, prevent influenza, and improve immunity. The polysaccharides contained in Antrodia camphorata can improve immune regulation in the human body.

3. Prevent cancer and inhibit the growth of cancer cells;

Fourth, the treatment of hepatitis, cirrhosis, liver cancer, because Antrodia camphorata is rich in triterpenes and polysaccharides, has the effect of protecting the liver, promoting liver cell regeneration, and fighting liver cancer.

5. Treatment of various cardiovascular and cerebrovascular diseases. Among them, Antrodia cinnamomea can reduce the cholesterol and fat content in the blood, and its adenosine can reduce the clotting function of platelets, so it can treat various cardiovascular and cerebrovascular diseases, such as hypertension, hypotension, Arteriosclerosis and so on.

As people's awareness of Antrodia camphorata and its medicinal effects improves, its health care needs are increasing. However, the current cultivation methods: segment wood cultivation method, individual cultivation method and liquid fermentation method can not produce high quality, mass production and Antrodia cinnamomea with short cycle. That is, there is still a lack of cultivation methods that can produce high quality and short cycle Antrodia camphorata.

In view of this, the purpose of the present invention is to provide an Antrodia cinnamomea medium that can increase Antrodia cinnamomea fruit bodies and enhance triterpenes, and establish an optimal medium for Antrodia cinnamomea and various component indexes.

In order to achieve the above object, the present invention provides an Antrodia cinnamomea media that can increase the fruit body of Antrodia camphorata and enhance triterpenes, including agar, D-glucose, malt extract, and 0.5 to 1% protein hydrazone.

The effect of the present invention is to add plant protein pouches to Antrodia camphorata culture medium, and effectively improve the quality of Antrodia camphorata.

In order to explain the invention more clearly, the following preferred embodiments are described in detail as follows.

Example 1 Exploration of the best cultivation conditions of Antrodia camphorata and its component analysis

Source of strain

The fungus and plant biotechnology team from the University of Science and Technology of China collected the fresh Antrodia camphorata on the mountain and separated them.

Explore the best cultivation conditions

The MEA medium is mainly used for the separation, counting and cultivation of fungi (mould, yeast, etc.), and the medium of the present invention contains the basic ingredients: grains (corn, rice or wheat, etc.), which contains 10-15% of purple rice Therefore, the color is reddish, and 0.1-1% protein peptone is added. Among them, the protein peptone slows the growth rate, so it can grow a similar fruiting body structure.

The cultivation device of Antrodia cinnamomea fruit body mainly includes: a bag body, which is composed of a ring wall and a top wall, so that an opening is formed on the top side, and an accommodating space is formed inside. The aforementioned culture medium is accommodated in the accommodating space of the bag body to cultivate Antrodia camphorata. Using this cultivation method can greatly increase production and reduce costs.

The experiments of the present invention are mainly based on solid culture medium, and are cultured under the conditions of five kinds of culture mediums, which are respectively represented by codes MEA, P0.1, P0.5, A0.1, A0.5, and the control group is MEA For the culture medium, the experimental group is supplemented with 0.1% and 0.5% animal and vegetable protein peptone, as shown in Table 1. Table 1. Composition of each group of culture medium

The configuration method in the above table is as follows: agar, D-glucose, malt extract and protein hydrazone are added to distilled water to prepare a 1 liter solution, which is sterilized at 121℃ for 15 minutes, and then the pH value is adjusted to 5.6 ± 0.2. Among them, the preferred weight percent concentrations of agar, D-glucose and malt extract are 2%, 4% and 2%, respectively.

Comparison of diameter growth and dry weight

During the inoculation of Antrodia camphorata, a hole is drilled to control the diameter of each inoculated strain to be the same. Then take a pick rod to take a piece and put it in the center of the medium, and then put it into a 26 ℃ incubator for 1-2 Measure and record the diameter of mycelium growth in a month, and then dry it in an oven for 46 hours. After drying, cut the portion of the culture medium that does not contain mycelium of Antrodia camphorata with scissors, and weigh the record. The results are shown in Table 2 below. Table 2. Diameter growth and dry weight comparison of Antrodia camphorata under five culture media

The P0.5 and A0.5 groups with the most significant increase in diameter and dry weight (after 2 months of cultivation) were analyzed by high-performance liquid chromatography (HPLC). The results are shown in Figure 1, where A, Peak B exists alone in P0.5, and peak C is higher. Therefore, determine the best solid culture conditions of Antrodia camphorata as the components contained in P0.5 group.

Index component analysis

The culture medium (P0.5) of the present invention is characterized by the addition of 0.5-1% of vegetable protein hydrazone. The content of its index components is shown in Table 3 below. Table 3. The content of index components of the medium of the present invention

Example 2 　　Antrodia cinnamomea to repair liver damage

Alcohol-induced acute alcohol liver injury in mice

After a week of adaptation, C57BL/6 mice were randomly divided into three groups, with six mice in each group. The grouping method is as shown in Table 4 below: Table 4: Group of Antrodia camphorata for liver injury repair test

Among them, physiological saline and Antrodia cinnamomea powder were given every day at 9 am. After continuous oral gavage for 30 days, the mice were fasted for 6 hours and then gavage of alcohol (5.0 g/kg, 60%). After 16 hours, the mice were sacrificed and blood was taken. The serum was used to measure various biochemical parameters. Another part of the liver of the mice was added with quantitative physiological saline to prepare liver tissue homogenate, and the degree of liver injury was evaluated.

Effect of Antrodia Cinnamomea on Acute Alcoholic Liver Injury

Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are the hallmark enzymes in the liver. When the liver is damaged, both of them will abnormally increase. This experiment investigated the effects of Antrodia camphorata powder on the content of alanine aminotransferase and aspartate aminotransferase in mice with acute alcohol liver injury. The results are shown in Figure 2.

Compared with the control group, the alanine aminotransferase and aspartate aminotransferase contents of the model group were significantly increased. Antrodia cinnamomea powder can significantly inhibit the increase of alanine aminotransferase and aspartate aminotransferase content in mice with acute alcohol liver injury. Among them, compared with the blank group, ***p <0.001; compared with the model group, # # #p ＜ 0.001, # #p ＜ 0.01.

Effect of Antrodia camphorata on antioxidant activity of acute alcohol liver injury

MDA is a secondary product produced by lipid peroxidation. SOD and GSH-Px are the key enzymes for the liver to eliminate peroxides and prevent oxidative damage. The liver MDA content and the activities of SOD and GSH-Px enzymes can reflect lipid peroxidation in the liver And the degree of oxidative damage. This experiment investigated the effects of Antrodia cinnamomea powder on the liver MDA content and SOD and GSH-Px enzyme activities in mice with acute alcohol liver injury. The results are shown in Figure 3.

Compared with the control group, the MDA content of the model group was significantly increased, while the activities of SOD and GSH-Px were significantly reduced. Antrodia camphorata powder can significantly inhibit the increase of MDA content and the decrease of SOD and GSH-Px activity in mice with acute alcohol liver injury. Among them, compared with the blank group, ***p <0.001; compared with the model group, # # p <0.01.

The above experiments show that Antrodia camphorata powder has a certain protective effect on acute alcoholic liver in mice.

In summary, the composition of the Antrodia camphorata culture medium of the present invention can indeed produce high-quality and short-cycle Antrodia camphorata.

The above is only the preferred and feasible embodiments of the present invention, and any equivalent changes in applying the description of the present invention and the scope of patent application should be included in the patent scope of the present invention.

Figure 1 shows high-performance liquid chromatography (HPLC) analysis of medium A0.5 and medium P0.5. Figure 2 shows the effect of Antrodia camphorata on ALT and AST in mice with acute alcohol liver injury. Figure 3 shows the effect of Antrodia camphorata on MDA, GSH-Px and SOD levels in mice with acute alcohol liver injury.

Claims (8)

An Antrodia cinnamomea medium capable of increasing Antrodia cinnamomea fruit body and enhancing triterpenes, which contains agar, D-glucose, malt extract and 0.5 to 1% protein hydrazone. As mentioned in claim 1, the Antrodia camphorata culture medium which can increase the fruit body of Antrodia camphorata and enhance the triterpenes has a PH value range of 5.6 ± 0.2. The Antrodia camphorata culture medium which can increase the fruit body of Antrodia camphorata and enhance the triterpenoids as described in claim 1, wherein the protein paeonia is a vegetable protein paeonia. The Antrodia camphorata culture medium which can increase the fruit body of Antrodia camphorata and enhance the triterpenes as described in claim 1, wherein the vegetable protein pouch is soybean protein pouch. The Antrodia cinnamomea medium which can increase Antrodia cinnamomea fruit body and enhance triterpenes as described in claim 1, contains grains of grains. The Antrodia cinnamomea medium which can increase Antrodia camphorata fruit body and enhance triterpenes as described in claim 5, wherein the grains contains 10-15% of purple rice. Antrodia camphorata culture medium capable of increasing Antrodia cinnamomea fruit body and enhancing triterpenes as described in claim 1, wherein the weight percentage concentrations of the agar, D-glucose and malt extract are 1.5-2%, 3-5% and 2-3%. The Antrodia cinnamomea medium capable of increasing Antrodia cinnamomea fruit body and enhancing triterpenes as described in claim 7, wherein the weight percentage concentrations of the agar, D-glucose and malt extract are 2%, 4% and 2%, respectively.