A composite culture medium and its preparation method and application
By using a compound culture medium in Ganoderma lucidum cultivation, combining chitosan and water-soluble fish protein, the problems of long growth cycle and pollution have been solved, resulting in improved yield and quality of Ganoderma lucidum and meeting the needs of the high-end market.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- ZHEJIANG CASNOVO MATERIALS
- Filing Date
- 2026-02-04
- Publication Date
- 2026-05-15
AI Technical Summary
Existing Ganoderma lucidum cultivation techniques suffer from long growth cycles, susceptibility to contamination, and unstable quality, making it difficult to meet the demands of the high-end market.
A composite culture medium was used, including a basal medium, chitosan, and water-soluble fish protein. Chitosan served as a functional additive to stimulate mycelial growth potential and inhibit contaminating bacteria, while fish protein served as an efficient nitrogen source to promote rapid mycelial proliferation and fruiting body development.
It shortens the time for mycelium to fully cover the fruiting cycle, increases the yield and content of functional components in Ganoderma lucidum, reduces the risk of contamination, and enhances the economic value and quality stability of the product.
Smart Images
Figure CN121621178B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of composite culture medium technology, specifically to a composite culture medium, its preparation method, and its application. Background Technology
[0002] Reishi mushroom, a precious medicinal and edible fungus, is highly regarded for its rich content of polysaccharides, triterpenoids, and other bioactive components. With the increasing demand for health-related products, the market is experiencing a surge in demand for high-quality, safe, and consistently effective reishi mushroom products.
[0003] Currently, artificial cultivation is the main way to supply Ganoderma lucidum. Chinese invention patent application CN107056404A provides a mother culture carrier culture medium and cultivation method for Ganoderma lucidum cultivation, including 2500 g of corn grits soaked for 12 hours and filtered through gauze, 2 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate, 10 mg of vitamin B1, 7.5 g of alumina, 1.5 g of allantoin, and 500 ml of water at 60-70℃. However, the current cultivation model, whether it is scattered planting by farmers or conventional industrial production, still faces many challenges: First, the cultivation cycle is long, and the mycelial growth rate and fruiting body yield are limited by the nutritional composition of the culture medium; second, it is easy to be contaminated by miscellaneous bacteria during the cultivation process, resulting in a decrease in the yield of finished products; third, the unbalanced nutrition of the culture medium may affect the effective accumulation of Ganoderma lucidum active ingredients, resulting in inconsistent product quality and difficulty in meeting the requirements of the high-end market.
[0004] Therefore, developing a new culture medium that can shorten the growth cycle, enhance pollution resistance, and significantly increase the yield and functional component content of Ganoderma lucidum is of great significance for promoting the modernization and standardization of the Ganoderma lucidum industry. Summary of the Invention
[0005] The present invention aims to overcome the shortcomings of the prior art and provide a composite culture medium, its preparation method and application, so as to solve the problems of long growth cycle, easy contamination and unstable quality in the existing Ganoderma lucidum cultivation technology.
[0006] To achieve the above objectives, the first aspect of the present invention provides a composite culture medium comprising a basal culture medium, chitosan, and fish protein;
[0007] The amount of chitosan added is 0.1-2% of the total weight of the basal culture medium;
[0008] The amount of fish protein added is 1-5% of the total weight of the basal culture medium;
[0009] The raw materials of the basal culture medium, by weight, include: 400-600 parts hardwood chips, 100-200 parts wheat bran, 20-50 parts corn flour, 5-15 parts gypsum powder, and 5-15 parts sugar.
[0010] In this composite culture medium, chitosan, as a functional additive, not only supplements nutrition but also utilizes its biological activity to stimulate mycelial growth potential and inhibit contaminating bacteria; water-soluble fish protein, as a highly efficient composite nitrogen source, supplements the key amino acids and trace elements required for rapid mycelial proliferation and fruiting body development.
[0011] In one embodiment, the fish protein is a water-soluble fish protein.
[0012] Water-soluble fish protein is rich in specific small molecule peptides and amino acids, which can provide a high-quality nitrogen source that is easily absorbed and utilized for the growth of Ganoderma lucidum mycelium, thereby ensuring the healthy growth of mycelium.
[0013] In one embodiment, the raw materials of the basal culture medium, by weight, include: 450-550 parts hardwood chips, 130-180 parts wheat bran, 30-50 parts corn flour, 10-15 parts gypsum powder, and 10-15 parts sugar. Preferably, the raw materials of the basal culture medium, by weight, include: 500 parts sawdust, 150 parts wheat bran, 30 parts corn flour, 10 parts gypsum powder, and 10 parts sucrose.
[0014] In one embodiment, the hardwood chips are dried hardwood chips sourced from Pinshuo Trading Co., Ltd.
[0015] A second aspect of the present invention provides a method for preparing a composite culture medium, comprising at least the following steps:
[0016] (1) Mix the raw materials of the basal culture medium evenly;
[0017] (2) Dissolve chitosan and fish protein in water to prepare a mixed nutrient solution;
[0018] (3) Add the mixed nutrient solution to the raw materials that were mixed evenly in step (1), adjust the water content to 55-65%, and stir evenly.
[0019] (4) Dispense and sterilize the well-mixed culture medium to obtain the finished product.
[0020] A third aspect of the present invention provides an application of a composite culture medium, which is used in the cultivation of Ganoderma lucidum.
[0021] In one embodiment, the method of applying the composite culture medium to Ganoderma lucidum cultivation includes the following steps:
[0022] S1. Provide the composite culture medium;
[0023] S2, Ganoderma lucidum ( Ganoderma lucidum The strain is inoculated onto the composite culture medium for mycelial culture and fruiting body induction and growth until the Ganoderma lucidum matures and is harvested.
[0024] In one embodiment, the conditions for mycelial culture in step S2 are: temperature 22-28℃, dark environment, and relative humidity of 60-70%.
[0025] In one embodiment, the conditions for inducing and growing fruiting bodies in step S2 are: temperature 24-30℃, diffused light 500-1000 lux, relative humidity 85-95%, and carbon dioxide concentration maintained at 2000-3000 ppm during the bud induction period and 800-1000 ppm during the cap differentiation period.
[0026] Beneficial effects
[0027] 1. This invention combines chitosan with fish protein from a specific source. Chitosan can stimulate the defense response and growth potential of Ganoderma lucidum mycelium, while fish protein provides easily absorbed short-chain polypeptides and essential amino acids, serving as a highly efficient nitrogen source to strongly support rapid colonization and robust growth of mycelium. The synergistic effect of the two can effectively shorten the time for mycelial full coverage and the fruiting cycle.
[0028] 2. The composite culture medium provided by this invention contains chitosan, which has natural broad-spectrum antibacterial activity. It can form a biological defense line in the culture medium, effectively inhibit the growth of a variety of common bacteria, significantly reduce the risk of contamination in high-density industrial cultivation, and improve the cultivation success rate and economic benefits.
[0029] 3. This invention controls the amount of basal culture medium, chitosan and fish protein in the composite culture medium. When it is used for Ganoderma lucidum cultivation, the mycelium grows more robustly and absorbs nutrients more fully. The cultivated Ganoderma lucidum fruiting bodies are thicker and have a denser texture. Its biological efficiency (i.e. yield) is significantly improved compared with conventional culture medium, thereby enhancing the overall economic value of the product.
[0030] 4. This invention utilizes chitosan and fish protein, byproducts of marine biological processing, to provide a new and sustainable source of nutrition for Ganoderma lucidum cultivation. This helps reduce over-reliance on traditional agricultural and forestry resources and aligns with the development direction of green circular agriculture. Attached Figure Description
[0031] Figure 1 This is a schematic diagram showing the growth status of Ganoderma lucidum fruiting bodies after cultivation using the composite culture medium of the present invention. Detailed Implementation
[0032] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. Experimental methods not specifying specific conditions in the embodiments were performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.
[0033] Example 1
[0034] The first aspect of this example provides a composite culture medium comprising 700 kg of basal culture medium, 7 kg of chitosan, and 21 kg of water-soluble fish protein.
[0035] The raw materials for the basic culture medium include: 500 kg of dried hardwood chips, 150 kg of wheat bran, 30 kg of corn flour, 10 kg of gypsum powder, and 10 kg of sucrose.
[0036] The dried hardwood chips were sourced from Pinshuo Trading Co., Ltd.
[0037] The chitosan, with a degree of deacetylation ≥90%, is sourced from Zhengzhou Mingze Biotechnology Co., Ltd.
[0038] The water-soluble fish protein comes from the root-promoting fish (fish protein rooting type) amino acid-containing water-soluble fertilizer of Zhejiang Ougenake Marine Biotechnology Co., Ltd.
[0039] The second aspect of this example provides a method for preparing a composite culture medium, comprising the following steps:
[0040] (1) Place the raw materials of the basal culture medium in a large mixer and premix them evenly;
[0041] (2) Dissolve chitosan and fish protein in 1000L of water to prepare a mixed nutrient solution;
[0042] (3) Add the mixed nutrient solution to the premixed raw materials in step (1), start the mixer to stir thoroughly, so that the liquid is mixed evenly, and finally control the water content at 60%;
[0043] (4) Pack the mixed culture medium into special polypropylene cultivation bags, each bag containing 1.5 kg of material. After compacting and sealing, put the bags into an autoclave and sterilize them at 121°C for 110 min. Then let them cool naturally to room temperature to obtain the finished product.
[0044] The third aspect of this example provides an application of a composite culture medium used in Ganoderma lucidum cultivation.
[0045] The method for applying the composite culture medium to Ganoderma lucidum cultivation includes the following steps:
[0046] S1. Provide the composite culture medium;
[0047] S2. In a sterile operating table, place the Ganoderma lucidum ( Ganoderma lucidum After the bacterial strain is inoculated into the composite culture medium, it is transferred to a dark, temperature- and humidity-controlled incubation room, with the temperature set at 25°C and the relative humidity at 65%. Incubate for 25-30 days, until the mycelium has completely covered the composite culture medium.
[0048] The mycelium-covered spawn bags (compound culture medium) were transferred to the fruiting room, the opening was made, and stimulating conditions were created. The ambient temperature was set to 26℃, the relative humidity was increased to 95%, the carbon dioxide concentration was maintained at 2500ppm, and 500 lux of diffused light was provided to stimulate primordium formation.
[0049] After the primordia form and differentiate into stipes and caps, adjust the environmental parameters, reduce the relative humidity to 90%, increase ventilation, reduce the carbon dioxide concentration to 900 ppm, and maintain suitable (500 lux) diffused light.
[0050] When the Ganoderma lucidum cap is fully expanded, the white growth ring at the edge disappears, and the color turns into a reddish-brown consistent with the center of the cap, and before a large amount of spores are released, cut the fruiting body from the base of the stipe with sterile scissors.
[0051] After harvesting, the Ganoderma lucidum is dried using a low-temperature slow drying process. The specific process parameters are as follows: the process is carried out in three stages. In the first stage, the temperature is raised to 50℃ for 4 hours. In the second stage, the temperature is slowly raised to 60℃ and dried for 2 hours. In the third stage, the temperature is lowered to 55℃ and dried until the dry weight of the fruiting body is constant. The final moisture content is controlled below 12%. Then, the fruiting bodies are graded, vacuum-packed, and stored in a warehouse.
[0052] The Ganoderma ( Ganoderma lucidum The strains were sourced from Hansong Edible Fungus Business Department in Hongshan District, Wuhan City.
[0053] Figure 1 This is a schematic diagram showing the growth status of Ganoderma lucidum fruiting bodies after cultivation using the composite culture medium of the present invention.
[0054] Example 2
[0055] The specific implementation method in this example is the same as in Example 1, except that the amount of chitosan used in this example is 0.7 kg.
[0056] Example 3
[0057] The specific implementation method in this example is the same as in Example 1, except that the amount of chitosan used in this example is 14 kg.
[0058] Example 4
[0059] The specific implementation method of this example is the same as that of Example 1, except that the amount of water-soluble fish protein used in this example is 7 kg.
[0060] Example 5
[0061] The specific implementation method in this example is the same as in Example 1, except that the amount of water-soluble fish protein used in this example is 35 kg.
[0062] Example 6
[0063] The specific implementation method of this example is the same as that of Example 1, except that the amount of chitosan used in this example is 3.5 kg and the amount of water-soluble fish protein used is 14 kg.
[0064] Comparative Example 1
[0065] The specific implementation method of this example is the same as that of Example 1, except that the water-soluble fish protein is replaced with 21 kg of untreated raw soybean flour, which comes from Gao'an Phoenix Peanut Grain and Oil Store.
[0066] Comparative Example 2
[0067] The specific implementation method of this example is the same as that of Example 1, except that the amount of chitosan used in this example is increased to 70 kg.
[0068] Comparative Example 3
[0069] The specific implementation method in this example is the same as in Example 1, except that chitosan and water-soluble fish protein are not added in this example, and only the basic culture medium is used.
[0070] Comparative Example 4
[0071] The specific implementation method of this example is the same as that of Example 1, except that the dried hardwood chips in the basal culture medium are replaced with an equal amount of fresh pine chips (from Chuna Trading Company).
[0072] Comparative Example 5
[0073] The specific implementation method in this example is the same as in Example 1, except that the amount of chitosan used in this example is increased to 35 kg.
[0074] Comparative Example 6
[0075] The specific implementation method of this example is the same as that of Example 1, except that the amount of water-soluble fish protein used in this example is increased to 70 kg.
[0076] Comparative Example 7
[0077] The specific implementation method of this example is the same as that of Example 1, except that the raw materials of the basic culture medium include: 640 kg of sawdust, 30 kg of wheat bran, 10 kg of corn flour, 10 kg of gypsum powder, and 10 kg of sucrose.
[0078] Comparative Example 8
[0079] The specific implementation method of this example is the same as that of Example 1, except that the composite culture medium includes 700 kg of basal culture medium and 7 kg of chitosan.
[0080] Comparative Example 9
[0081] The specific implementation method in this example is the same as in Example 1, except that the composite culture medium includes 700 kg of basal culture medium and 21 kg of water-soluble fish protein.
[0082] Performance testing
[0083] The mycelium bags (compound culture medium) of each embodiment and comparative example were cultivated under the same environment, with 1000 mycelium bags set as replicates for each group. The following key performance indicators were recorded, and the results are detailed in Table 1.
[0084] 1. Time for mycelium to fully fill the package:
[0085] The specific calculation method is as follows: Time to full mycelial coverage (days) = Date of full mycelial coverage - Inoculation date.
[0086] The mycelial coverage time refers to the number of days from the day the inoculum is introduced into the culture medium until the mycelium (white filamentous material) visually completely covers the entire surface of the culture medium.
[0087] Evaluation criteria: The shorter the time, the better. A shorter time indicates strong bacterial activity, suitable culture medium nutrition, and proper environmental control.
[0088] Excellent: A full enclosure typically completed within 25-30 days is considered a sign of fast growth.
[0089] Generally, 30-40 days is within the normal range.
[0090] Poor: Growth exceeding 40 days indicates slow growth, potentially due to strain degeneration, malnutrition, or unsuitable environment. In the comparative examples of this application, the failed groups even exceeded 90 days or failed to reach full growth.
[0091] 2. Mushroom emergence time:
[0092] The specific calculation method is as follows: Mushroom emergence time (days) = Date of observation of the first batch of primordia - Date of opening of the spawn bag.
[0093] The fruiting time refers to the number of days required for the first batch of effective and healthy fruiting body primordia (commonly known as "mushroom buds") to form after the mushroom bag meets the fruiting conditions (such as opening, changing temperature, humidity and light).
[0094] Evaluation criteria: The shorter the time, the better. A shorter time indicates that the mycelium has accumulated sufficient nutrients, is physiologically mature, and that the conditions for fruiting are appropriate.
[0095] Excellent: Mushrooms appear within 7-10 days, indicating that the mycelium is very robust and the conversion rate is fast.
[0096] Generally, 10-15 days is within the normal range.
[0097] Poor: If no mushrooms appear after more than 15 days, or if the mushrooms are sparse or deformed, it indicates that the mycelium may be weak or under stress.
[0098] 3. Biological efficiency:
[0099] The specific calculation method is as follows: Biological efficiency (%) = (Total dry weight of all harvested Ganoderma lucidum fruiting bodies / Total dry weight of culture medium) × 100%.
[0100] Biological efficiency is a core indicator for measuring yield and culture medium conversion rate. When calculating, the following should be noted:
[0101] Dry weight of culture medium: Wet weight of culture medium when packaged × (1 - moisture content of culture medium). For example, a 1.5kg bag with a moisture content of 60% has a dry weight of 1.5kg × (1 - 0.6) = 0.6kg.
[0102] Dry weight of Ganoderma lucidum: The weight of all freshly harvested Ganoderma lucidum after drying in an oven to constant weight.
[0103] Evaluation criteria: The higher the value, the better. It directly reflects the output capacity per unit of material.
[0104] Excellent: For Ganoderma lucidum cultivation, a biological efficiency of over 20% is an ideal level for commercial production.
[0105] Generally, 15-20% is considered good or normal.
[0106] Poor: Below 15% indicates low output and poor input-output efficiency.
[0107] 3. Pollution rate:
[0108] The specific calculation method is as follows: Contamination rate (%) = (Number of spawn bags contaminated by miscellaneous bacteria / Total number of spawn bags cultivated in the same batch) × 100%.
[0109] This indicator is used to assess the cleanliness and stability of the production process. It measures the percentage of spawn bags that are discarded due to contamination by fungi such as Penicillium and Mucor throughout the entire cultivation cycle (from inoculation to harvest).
[0110] Evaluation criteria: The lower the value, the better. This is key to measuring production management level and process stability.
[0111] Excellent: The contamination rate is controlled within 1-3%, which is an ideal target for modern factory production. In the embodiments of this application, the contamination rate is even lower than 1.5%;
[0112] Acceptable: Below 5% is generally considered to be within an acceptable range;
[0113] Serious problem: If the contamination rate is consistently higher than 8-10%, it indicates serious loopholes in the processes from raw material handling, sterilization, inoculation to environmental management, which must be investigated and corrected.
[0114] Table 1
[0115]
[0116] As can be seen from the table above, the results of each comparative example define the necessary conditions for the successful implementation of this technical solution from different aspects, proving that the selection and dosage of nitrogen source, chitosan, and basic raw materials are key prerequisites for successful Ganoderma lucidum cultivation. For example, Comparative Example 1, due to the use of raw soybean powder as a nitrogen source, resulted in more than 70% of the spawn bags being severely contaminated by miscellaneous bacteria, making it impossible to complete the cultivation cycle; Comparative Example 2, due to the use of extremely high concentrations of chitosan, suffered severe inhibition of mycelial growth, failing to fully colonize the spawn bags after 90 days and failing to produce fruiting fruits. Comparative Example 4, due to the use of pine sawdust, failed to allow mycelial growth to proceed normally. Compared with the comparative examples, Examples 1-6 of this application show significant advantages in terms of spawn bag full coverage time, fruiting time, biological efficiency (yield), and contamination rate control, proving that the composite culture medium provided by this application can shorten the growth cycle, enhance contamination resistance, and significantly increase Ganoderma lucidum yield.
Claims
1. A compound culture medium for Ganoderma lucidum cultivation, characterized in that, The composite culture medium includes a basal culture medium, chitosan, and fish protein; The amount of chitosan added is 0.1-2% of the total weight of the basal culture medium; The amount of fish protein added is 1-5% of the total weight of the basal culture medium; The raw materials of the basal culture medium, by weight, include: 450-550 parts hardwood chips, 130-180 parts wheat bran, 30-50 parts corn flour, 10-15 parts gypsum powder, and 10-15 parts sugar; the fish protein is water-soluble fish protein.
2. A method for preparing a composite culture medium according to claim 1, characterized in that, At least the following steps are included: (1) Mix the raw materials of the basal culture medium evenly; (2) Dissolve chitosan and fish protein in water to prepare a mixed nutrient solution; (3) Add the mixed nutrient solution to the raw materials that were mixed evenly in step (1), adjust the water content to 55-65%, and stir evenly. (4) Dispense and sterilize the well-mixed culture medium to obtain the finished product.
3. A method for applying the composite culture medium according to claim 1 to Ganoderma lucidum cultivation, characterized in that, Includes the following steps: S1. Provide the composite culture medium; S2. Inoculate the Ganoderma lucidum strain onto the compound culture medium for mycelial culture and fruiting body induction and growth until the Ganoderma lucidum matures and is harvested.
4. The method according to claim 3, characterized in that, The conditions for mycelial culture in step S2 are: temperature 22-28℃, dark environment, and relative humidity 60-70%.
5. The method according to claim 3, characterized in that, The conditions for fruiting body induction and growth in step S2 are: temperature 24-30℃, diffused light 500-1000 lux, relative humidity 85-95%, and carbon dioxide concentration maintained at 2000-3000 ppm during the bud induction period and 800-1000 ppm during the cap differentiation period.