A culture medium for Antrodia camphorata mycelium, its preparation method and application
By using a plant hormone-prepared Antrodia camphorata mycelium culture medium, the problems of low biomass and unsatisfactory yield of active ingredients in Antrodia camphorata cultivation technology have been solved, achieving efficient mycelial growth and synthesis of various active products, thus promoting the large-scale production of Antrodia camphorata and the stability of product quality.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- GUANGZHOU BUQIAN BIOTECHNOLOGY CO LTD
- Filing Date
- 2026-05-14
- Publication Date
- 2026-06-30
AI Technical Summary
Existing Antrodia camphorata mycelium culture technologies suffer from low biomass and unsatisfactory yields of active ingredients, and it is difficult to simulate the natural growth environment under artificial conditions, resulting in low production efficiency and difficulty in standardizing product quality.
An agaricus camphoratus mycelium culture medium containing plant hormones such as indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide was used. By combining solid and liquid culture media, the growth of agaricus camphoratus mycelium and the efficient synthesis of various active products were promoted.
It significantly improved the yield of crude polysaccharides and crude triterpenes in Antrodia camphorata mycelium, enabling stable and reliable large-scale production and improving the content of effective ingredients and quality consistency of the product.
Smart Images
Figure CN122303047A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of artificial culture technology for edible and medicinal fungi, particularly to the deep fermentation technology for the precious medicinal fungus *Antrodia camphorata*. It aims to overcome the technical bottlenecks in existing *Antrodia camphorata* mycelial culture techniques, such as low biomass and unsatisfactory yields of active ingredients (polysaccharides, triterpenoids), and to provide a novel *Antrodia camphorata* mycelial culture medium, its preparation method, and its applications. Background Technology
[0002] Antrodia cinnamomea, also known as camphor mushroom, is a rare medicinal fungus endemic to Taiwan, my country, with a complex composition. It mainly contains macromolecular compounds such as proteins, nucleic acids, and polysaccharides; small molecular compounds such as lignin and aromatic compounds; terpenoids; nucleotides; fatty acids; and fatty acid esters. The active ingredients in Antrodia cinnamomea include triterpenoids, polysaccharides, superoxide dismutase (SOD), adenosine, proteins, amino acids, vitamins, trace elements, nucleic acids, and lectins, with triterpenoids, polysaccharides, adenosine, and SOD being the most prevalent. Traditional uses of Antrodia cinnamomea include relieving headaches, diarrhea, abdominal pain, alcohol poisoning, and hypertension. Current research has revealed that it possesses various pharmacological effects, including anti-tumor, liver-protective, anti-inflammatory, and neuroprotective properties.
[0003] Due to its unique bioactivity, *Antrodia camphorata* has gained widespread market attention. Although its significant pharmacological functions, such as anti-cancer and antioxidant effects, have become a hot topic in academic research, the efficient cultivation technology for *Antrodia camphorata* has lagged behind in the process of industrialization. Current bottlenecks are mainly concentrated in the following aspects: First, host resources are unsustainable; wild or traditional cultivation methods of *Antrodia camphorata* are highly dependent on scarce *Cinnamomum camphora* wood resources. Second, growth conditions are demanding; the natural growth environment is difficult to completely simulate under artificial conditions. Third, growth and reproduction efficiency is low; mycelial growth is slow, and fruiting bodies are difficult to artificially induce reproduction. Fourth, while previous methods have explored adding specific microbial fermentation products and food and medicinal extracts (such as bran extract and tangerine peel extract) to the culture medium to promote growth can improve the cultivation effect to some extent, the introduction of exogenous complex components may lead to uncontrollable product composition, increased difficulty in downstream classification and purification, and increased production costs. More fundamentally, existing research on cultivation technologies focuses on increasing mycelial biomass or the yield of a single target metabolite, lacking a comprehensive solution that can coordinate and efficiently enhance multiple core active ingredients. The limitations of this technology have led to widespread problems in the current market for Antrodia camphorata-related products (most of which are crude extracts of fermentation broth or fruiting body powder), such as unclear effective components, large fluctuations in content, and difficulty in standardizing quality. These issues have severely hampered the development of high-value-added Antrodia camphorata and the progress of clinical application research.
[0004] Therefore, how to overcome the limitations of wild hosts and establish a culture system that can simultaneously promote the growth of Antrodia camphorata mycelium and the efficient synthesis of multiple active products without introducing exogenous complex bioactive substances, and achieve stable, reliable, and large-scale production management, has become a key issue that urgently needs to be addressed in the current industrial and technological development. Summary of the Invention
[0005] In order to achieve large-scale production of Antrodia camphorata, in-depth research has been conducted on the separation and purification of Antrodia camphorata monomers, laying the foundation for the development of Antrodia camphorata drugs. This invention provides an Antrodia camphorata mycelial culture medium, characterized in that the Antrodia camphorata mycelial culture medium includes a solid culture medium and a liquid culture medium, wherein the raw materials of the liquid culture medium include: glucose, peptone, anhydrous magnesium sulfate and plant hormones. The plant hormones include one or more of indolebutyric acid (IBA), lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide.
[0006] In one embodiment, the liquid culture medium contains glucose at a concentration of 15-25 g / L, peptone at a concentration of 5-15 g / L, anhydrous magnesium sulfate at a concentration of 2-3 g / L, indolebutyric acid at a concentration of 0-8 mg / L, lignin at a concentration of 0-8 g / L, gibberellin at a concentration of 0-8 mg / L, 6-benzylaminopurine at a concentration of 0-8 mg / L, and 24-epibrassinolide at a concentration of 0-8 mg / L. The concentrations of indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide are not all zero at the same time.
[0007] In one embodiment, the liquid culture medium contains indolebutyric acid at a concentration of 0.5-8 mg / L, lignin at a concentration of 0.5-8 g / L, gibberellin at a concentration of 0.5-8 mg / L, 6-benzylaminopurine at a concentration of 0.5-8 mg / L, and 24-epibrassinolide at a concentration of 0.5-8 mg / L.
[0008] In one embodiment, the raw materials of the solid culture medium include: carrot powder, potato powder, glucose and agar powder.
[0009] In one embodiment, the concentration of carrot powder in the solid culture medium is 35-45 g / L, the concentration of potato powder is 35-45 g / L, the concentration of glucose is 15-25 g / L, and the concentration of agar powder is 15-25 g / L.
[0010] In one embodiment, the raw materials of the solid culture medium further include plant hormones, which include one or more of indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide.
[0011] In one embodiment, the concentration of indolebutyric acid in the solid culture medium is 0-8 mg / L, the concentration of lignin is 0-8 g / L, the concentration of gibberellin is 0-8 mg / L, the concentration of 6-benzylaminopurine is 0-8 mg / L, and the concentration of 24-epibrassinolide is 0-8 mg / L. The concentrations of indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide are not all zero at the same time.
[0012] In one embodiment, the solid culture medium contains indolebutyric acid at a concentration of 0.5-8 mg / L, lignin at a concentration of 0.5-8 g / L, gibberellin at a concentration of 0.5-8 mg / L, 6-benzylaminopurine at a concentration of 0.5-8 mg / L, and 24-epibrassinolide at a concentration of 0.5-8 mg / L.
[0013] In a second aspect, the present invention also provides a method for preparing the above-mentioned Antrodia camphorata mycelium culture medium, including the preparation of a solid culture medium and the preparation of a liquid culture medium; the method for preparing the liquid culture medium includes the following steps: taking glucose, peptone and anhydrous magnesium sulfate, dissolving them in water, adding plant hormones, sterilizing, and cooling to obtain the culture medium.
[0014] In one embodiment, the preparation method of the solid culture medium includes the following steps: taking carrot powder, potato powder, glucose and agar powder, dissolving them in water, adding plant hormones, sterilizing, and cooling to solidify.
[0015] A third aspect of the present invention also provides the application of the above-described Antrodia camphorata mycelium culture medium or the Antrodia camphorata mycelium culture medium prepared by the above-described method in the culture of Antrodia camphorata mycelium.
[0016] In addition, the present invention also provides a method for culturing Antrodia camphorata mycelium, comprising the following steps: Antrodia camphorata mycelium was inoculated into the solid culture medium of the above-mentioned Antrodia camphorata mycelium culture medium and cultured to obtain a well-cultivated strain. The strain was then inoculated into the liquid culture medium of the above-mentioned Antrodia camphorata mycelium culture medium for expansion culture.
[0017] Compared with the prior art, the present invention has the following beneficial effects: This invention, by using plant hormones to prepare the culture medium, significantly increases the yield and the production rate of the active ingredients, crude polysaccharides and crude triterpenes, compared to current Antrodia camphorata mycelium culture methods. Attached Figure Description
[0018] Figure 1 This is a picture of a solid culture specimen. Figure 2 This is a picture of a liquid culture specimen. Detailed Implementation
[0019] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.
[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.
[0021] Unless otherwise specified, all reagents, materials, and equipment used in this embodiment are commercially available; unless otherwise specified, all test methods are conventional test methods in this field.
[0022] Example 1 The formula for solid culture medium for Antrodia camphorata mycelium is as follows: 40 g / L carrot powder, 40 g / L potato powder, 20 g / L glucose, and 20 g / L agar powder.
[0023] In this embodiment, the method for preparing the solid culture medium formula for Antrodia camphorata mycelium includes the following steps: Step 1: Weigh out 10g of carrot powder, 10g of potato powder, 5g of glucose and 5g of agar powder, pour them into a 500mL Erlenmeyer flask, add 250mL of ultrapure water, heat in a water bath to 80℃ and stir to dissolve. Step 2: Seal the mouth of the triangular flask with a breathable membrane and tie it tightly with a thin rope to prevent it from bursting open, then put it into a high-temperature sterilizer for sterilization. Step 3: After sterilization, cool to about 55°C and dispense into petri dishes in a clean bench to cool and solidify for later use.
[0024] In this embodiment, a basic solid culture medium was prepared using carrot powder, potato powder, glucose, and agar powder for the basic cultivation of Antrodia camphorata mycelium.
[0025] Example 2 The formula for liquid culture medium of Antrodia camphorata mycelium is as follows: glucose 20g / L, peptone 10g / L, and anhydrous magnesium sulfate 2.4g / L.
[0026] In this embodiment, the method for preparing the liquid culture medium formula for Antrodia camphorata mycelium includes the following steps: Step 1: Weigh out 5g of glucose, 2.5g of peptone, and 0.6g of anhydrous magnesium sulfate, pour them into a 500mL Erlenmeyer flask, and then add 250mL of ultrapure water to dissolve them. Step 2: Seal the mouth of the triangular flask with a breathable membrane and tie it tightly with a thin rope to prevent it from bursting open, then put it into a high-temperature sterilizer for sterilization. Step 3: After sterilization, cool to room temperature for later use.
[0027] In this embodiment, a basic liquid culture medium for Antrodia camphorata mycelium was prepared using glucose, peptone, and anhydrous magnesium sulfate as a control group.
[0028] Example 3 The formula for liquid culture medium of Antrodia camphorata mycelium is as follows: glucose 20g / L, peptone 10g / L, anhydrous magnesium sulfate 2.4g / L, and IBA indolebutyric acid 0.5mg / L or 4mg / L.
[0029] In this embodiment, the method for preparing the liquid culture medium formula for Antrodia camphorata mycelium includes the following steps: Step 1: Weigh out 5g of glucose, 2.5g of peptone, and 0.6g of anhydrous magnesium sulfate, pour them into a 500mL Erlenmeyer flask, then add 250mL of ultrapure water to dissolve them. Repeat twice. Step 2: Add 0.125 mg and 1 mg of IBA indolebutyric acid to two conical flasks respectively. The IBA indolebutyric acid is weighed quantitatively, dissolved in anhydrous ethanol, and then added quantitatively. Step 3: Seal the mouth of the triangular flask with a breathable membrane and tie it tightly with a thin rope to prevent it from bursting open, then put it into a high-temperature sterilizer for sterilization. Step 4: After sterilization, cool to room temperature for later use.
[0030] In this embodiment, two concentrations of IBA (indolebutyric acid) were added to the basic liquid culture medium of Antrodia camphorata mycelium as experimental groups, named medium concentration IBA and high concentration IBA.
[0031] Example 4 The preparation method of the liquid culture medium for Antrodia camphorata mycelium in this embodiment is the same as that in Example 3. The only difference is that the liquid culture medium for Antrodia camphorata mycelium in this embodiment is as follows: glucose 20g / L, peptone 10g / L, anhydrous magnesium sulfate 2.4g / L, and lignin 0.5g / L.
[0032] The experimental group was named lignin.
[0033] Example 5 The preparation method of the liquid culture medium for Antrodia camphorata mycelium in this embodiment is the same as that in Example 3. The only difference is that the liquid culture medium for Antrodia camphorata mycelium in this embodiment is as follows: glucose 20g / L, peptone 10g / L, anhydrous magnesium sulfate 2.4g / L, and gibberellin 1mg / L or 8mg / L.
[0034] The experimental groups were named low-concentration gibberellin and high-concentration gibberellin.
[0035] Example 6 The preparation method of the liquid culture medium for Antrodia camphorata mycelium in this embodiment is the same as that in Example 3. The only difference is that the liquid culture medium for Antrodia camphorata mycelium in this embodiment is as follows: glucose 20g / L, peptone 10g / L, anhydrous magnesium sulfate 2.4g / L, and 6-benzylaminopurine 0.5mg / L.
[0036] The experimental group was named 6-benzylaminopurine.
[0037] Example 7 The preparation method of the liquid culture medium for Antrodia camphorata mycelium in this embodiment is the same as that in Example 3. The only difference is that the liquid culture medium for Antrodia camphorata mycelium in this embodiment is as follows: glucose 20g / L, peptone 10g / L, anhydrous magnesium sulfate 2.4g / L, and 24-epibrassinolide 0.5mg / L.
[0038] The experimental group was named 24-epibrassinolide.
[0039] Example 8 The preparation method of the liquid culture medium formula for Antrodia camphorata mycelium in this embodiment is the same as that in Example 3. The only difference is that the plant hormones added to the liquid culture medium formula for Antrodia camphorata mycelium in this embodiment are composed of three combinations of IBA indolebutyric acid 0.5 mg / L, gibberellin 0.5 mg / L, and lignin 0.5 g / L.
[0040] The experimental groups were named IBA+gibberellin, gibberellin+lignin, and IBA+lignin.
[0041] Implementation effect evaluation Table 1
[0042] Examples 2-8 above are summarized in Table 1, totaling eleven treatment combinations, with Example 2 serving as the control group. Each treatment was inoculated at a depth of 2 cm. 2 The mycelium cultured from the solid culture medium of Antrodia camphorata mycelium in Example 1 was placed in a constant temperature shaking incubator at 26°C for 14 days, and the effective components of the resulting mycelium were then measured.
[0043] Table 2. Crude polysaccharide content of mycelia obtained from different culture media
[0044] As shown in Table 2, medium concentrations of IBA, lignin, high concentrations of gibberellin, 6-benzylaminopurine, and 24-epibrassinolide can all increase the crude polysaccharide content of Antrodia camphorata mycelium when used alone, with medium concentrations of IBA, high concentrations of gibberellin, and 24-epibrassinolide being the most significant. The combined use of lignin, IBA, and gibberellin in pairs was not very effective.
[0045] Table 3. Crude triterpenoid content of mycelia obtained from different culture media
[0046] Table 3 shows that medium concentrations of IBA, lignin, high concentrations of gibberellin, 6-benzylaminopurine, and 24-epibrassinolide can all increase the crude triterpenoid content of Antrodia camphorata mycelium when used alone, with 24-epibrassinolide, high concentrations of gibberellin, and 6-benzylaminopurine being more significant. The combined use of IBA with gibberellin and lignin is not very effective, while the combined use of gibberellin and lignin is relatively effective.
[0047] Analysis of Tables 2 and 3 shows that the optimal liquid culture medium for Antrodia camphorata mycelium is 20 g / L glucose, 10 g / L peptone, 2.4 g / L anhydrous magnesium sulfate, and 8 mg / L gibberellin.
[0048] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0049] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.
Claims
1. A culture medium for Antrodia camphorata mycelium, characterized in that, The Antrodia camphorata mycelium culture medium includes a solid culture medium and a liquid culture medium. The raw materials of the liquid culture medium include glucose, peptone, anhydrous magnesium sulfate and plant hormones. The plant hormones include one or more of indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassin lactone.
2. The Antrodia camphorata mycelium culture medium according to claim 1, characterized in that, The liquid culture medium contains glucose at a concentration of 15-25 g / L, peptone at a concentration of 5-15 g / L, anhydrous magnesium sulfate at a concentration of 2-3 g / L, indolebutyric acid at a concentration of 0-8 mg / L, lignin at a concentration of 0-8 g / L, gibberellin at a concentration of 0-8 mg / L, 6-benzylaminopurine at a concentration of 0-8 mg / L, and 24-epibrassinolone at a concentration of 0-8 mg / L. The concentrations of indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide are not all zero at the same time.
3. The Antrodia camphorata mycelium culture medium according to claim 1, characterized in that, The raw materials for the solid culture medium include: carrot powder, potato powder, glucose and agar powder.
4. The Antrodia camphorata mycelium culture medium according to claim 3, characterized in that, In the solid culture medium, the concentration of carrot powder is 35-45 g / L, the concentration of potato powder is 35-45 g / L, the concentration of glucose is 15-25 g / L, and the concentration of agar powder is 15-25 g / L.
5. The Antrodia camphorata mycelium culture medium according to claim 3, characterized in that, The raw materials for the solid culture medium also include plant hormones, which include one or more of indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide.
6. The Antrodia camphorata mycelium culture medium according to claim 5, characterized in that, In the solid culture medium, the concentration of indolebutyric acid is 0~8 mg / L, the concentration of lignin is 0~8 g / L, the concentration of gibberellin is 0~8 mg / L, the concentration of 6-benzylaminopurine is 0~8 mg / L, and the concentration of 24-epibrassinolone is 0~8 mg / L. The concentrations of indolebutyric acid, lignin, gibberellin, 6-benzylaminopurine, and 24-epibrassinolide are not all zero at the same time.
7. The method for preparing the Antrodia camphorata mycelium culture medium according to any one of claims 1-6, characterized in that, The preparation includes the preparation of solid culture medium and liquid culture medium; the preparation method of the liquid culture medium includes the following steps: taking glucose, peptone and anhydrous magnesium sulfate, dissolving them in water, adding plant hormones, sterilizing under high temperature and high pressure, and cooling to obtain the liquid culture medium.
8. The preparation method according to claim 7, characterized in that, The preparation method of the solid culture medium includes the following steps: take carrot powder, potato powder, glucose and agar powder, dissolve them in water, add plant hormones, sterilize under high temperature and high pressure, and cool down to solidify.
9. The application of the Antrodia camphorata mycelium culture medium according to any one of claims 1-6 or the Antrodia camphorata mycelium culture medium obtained by the preparation method according to any one of claims 7-8 in the culture of Antrodia camphorata mycelium.
10. A method for culturing Antrodia camphorata mycelium, characterized in that, Includes the following steps: Antrodia camphorata mycelium is inoculated into the solid culture medium of the Antrodia camphorata mycelium culture medium according to any one of claims 1-6 for cultivation to obtain a cultured strain. The cultured strain is then inoculated into the liquid culture medium of the Antrodia camphorata mycelium culture medium according to any one of claims 1-6 for expansion culture.