Method for cultivating cordyceps in tremella culture medium

Abstract

This invention discloses a method for cultivating Cordyceps using Tremella fuciformis culture medium. The method uses Tremella fuciformis as the main ingredient to prepare a culture medium for cultivating Cordyceps. The culture medium comprises the following raw materials in parts by weight: 10-20 parts Tremella fuciformis; 8-12 parts Job's tears; 5-6 parts Chinese yam powder; 0.02-0.03 parts glucose; 0.003-0.005 parts lactose; 0.02-0.03 parts peptone; 0.01-0.03 parts potassium dihydrogen phosphate; 0.003-0.005 parts glycine; 0.01-0.02 parts magnesium sulfate; and 32-50 parts water. The method includes steps such as preparing the culture medium, sterilization, inoculation with Cordyceps spores, dark culture, light-induced color change culture, and fruiting body growth culture. The method of this invention is simple and low in cost. Using this method to cultivate cordyceps can greatly increase the content of cordycepin, cordycepic acid, cordyceps polysaccharide and adenosine in cordyceps. The cultivated cordyceps has high yield and good quality, contains rich polysaccharides, cordycepin and other substances, has high nutritional content, and has good medicinal and health care effects.

Description

Technical Field

[0001] This invention relates to the field of cordyceps technology, and specifically to a method for cultivating cordyceps using tremella culture medium. Background Technology

[0002] Cordyceps sinensis, also known as caterpillar fungus, is a traditional and precious Chinese medicinal herb. It is formed when the fungus *Cordyceps sinensis*, belonging to the family Clavicipitaceae, parasitizes the larvae of the ghost moth in alpine meadows, causing the larvae to stiffen. Under suitable conditions, a long, club-shaped fruiting body emerges from the head of the stiffened larva in summer, forming a complex (i.e., a composite of the fruiting body of the *Cordyceps sinensis* fungus and the sclerotium of the stiffened larva). It is mainly produced in the high-altitude and snow-capped grasslands of Qinghai, Tibet, Sichuan, Yunnan, Gansu, and Guizhou provinces and autonomous regions of China. Cordyceps sinensis has high medicinal value. Its main active ingredients are cordycepin, cordycepic acid, and cordyceps polysaccharides, which are beneficial for human absorption. It possesses various effects, including tonifying the kidneys and replenishing essence, stopping bleeding and resolving phlegm, tonifying deficiency, enhancing myocardial function, regulating the immune system, regulating the central nervous system, anti-tumor effects, and anti-fatigue effects.

[0003] Wild Cordyceps sinensis mainly grows in high-altitude, cold regions, with extremely demanding and complex growing environments. Coupled with increasing demand and improper harvesting, natural Cordyceps sinensis resources are gradually decreasing. To address the shortage of natural Cordyceps sinensis, artificial cultivation is necessary to meet market demand. In recent years, research and practice on artificial Cordyceps sinensis cultivation have been extensive and in-depth both domestically and internationally. The technology of infecting host larvae with Cordyceps fungus, such as using silkworm pupae as infectants, and cultivating Cordyceps fruiting bodies is now mature. Cordyceps militaris (L.) Link, also known as North Cordyceps or North Cordyceps sinensis, has been increasingly studied, and its composition is quite similar to that of natural Cordyceps sinensis. Its main medicinal components are cordycepin, cordycepic acid, cordyceps polysaccharides, and SOD, etc. Cordyceps militaris has similar health benefits to Cordyceps sinensis, making it an ideal substitute for traditional Cordyceps sinensis.

[0004] Cordyceps militaris is one of the few usable species in the Cordyceps genus and is currently capable of large-scale industrial production. Artificially cultivated Cordyceps militaris, also known as North Cordyceps sinensis, has active ingredients close to those of natural Cordyceps sinensis, and its edible and medicinal value is comparable to traditional Cordyceps sinensis. Some components are even several times or dozens of times higher in content than in natural Cordyceps sinensis. Currently, there are two main types of artificially cultivated North Cordyceps sinensis: one type is produced by artificially raising silkworm pupae for mycelial growth. However, silkworm pupae are relatively expensive and cannot be supplied year-round, limiting year-round production. Furthermore, the inoculation process in cultivating Cordyceps sinensis from silkworm pupae is cumbersome, resulting in low production efficiency. The second type, the most common on the market, uses grains such as rice and wheat as substrates for Cordyceps sinensis cultivation. This method has relatively lower production costs compared to using silkworm pupae, and inoculation is convenient, quick, and efficient, significantly shortening the growth cycle of North Cordyceps sinensis and increasing yield in a short period. However, rice and wheat tend to stick together after steaming, preventing the inoculum from penetrating the substrate during inoculation and resulting in slow mycelial growth after germination. Furthermore, rice and wheat are prone to water loss, which in turn affects the later yield of cordyceps. Currently, most culture media used in artificial cordyceps cultivation prioritize increasing yield, neglecting to improve the quality of the cordyceps. This results in cordyceps with lower levels of nutrients (cordycepin, cordycepic acid, cordyceps polysaccharides, etc.) and less than ideal efficacy. Therefore, how to increase cordyceps yield and cultivate cordyceps with higher nutritional content is a key focus of current research.

[0005] Culture medium is a mixture of various nutrients formulated in specific proportions to provide the nutrients necessary for the growth, reproduction, and biosynthesis of microorganisms and their various metabolic products. The composition of the culture medium significantly impacts microbial growth, reproduction, biosynthesis, separation and purification, as well as the quality and yield of the products. Studies have shown that the quality and bioactive components of the fruiting bodies and mycelia of *Cordyceps militaris* cultivated in different culture media vary considerably. The types and proportions of carbon sources, nitrogen sources, minerals, and vitamins in the culture medium have a significant impact on the growth of *Cordyceps militaris* and the formation of cordycepin within it. Therefore, developing a *Cordyceps militaris* culture medium and cultivation method that is conducive to the growth of *Cordyceps militaris*, increases the content of cordycepin, cordyceps polysaccharides, and other nutrients, and improves the yield and quality of *Cordyceps militaris* is of great significance for the artificial cultivation of *Cordyceps militaris*. Summary of the Invention

[0006] The purpose of this invention is to provide a method for cultivating Cordyceps sinensis using Tremella fuciformis culture medium. This method uses Tremella fuciformis as the main material to prepare the culture medium for cultivating Cordyceps sinensis. The method is simple and low in cost. Cultivating Cordyceps sinensis using this method can greatly increase the content of cordycepin, cordycepic acid, cordyceps polysaccharide and adenosine in Cordyceps sinensis. The cultivated Cordyceps sinensis has high yield and good quality, contains rich polysaccharides, cordycepin and other substances, has high nutritional content, good medicinal effect and significant health care effect.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: A method for cultivating Cordyceps sinensis using Tremella fuciformis culture medium includes the following steps: (1) Soak dried white fungus in water, clean it, drain the water, and crush it into 1-2cm pieces for later use; take pest-free Job's tears, wash them, soak them in water for 2-2.5 hours, and drain them for later use. (2) Take 10-20 parts by weight of white fungus; 8-12 parts of coix seed; 5-6 parts of Chinese yam powder; 0.02-0.03 parts of glucose; 0.003-0.005 parts of lactose; 0.02-0.03 parts of peptone; 0.01-0.03 parts of potassium dihydrogen phosphate; 0.003-0.005 parts of glycine; 0.01-0.02 parts of magnesium sulfate and 32-50 parts of water, and set aside. (3) First, dissolve glucose, lactose, peptone, potassium dihydrogen phosphate, glycine, and magnesium sulfate in water to obtain a nutrient solution; then mix the nutrient solution with tremella, coix seed, and Chinese yam powder and stir evenly to obtain a mixture. The mixture was dispensed into culture containers and sterilized at 121°C for 30-35 minutes. After sterilization, it was cooled to room temperature to obtain the culture medium. (4) Inoculate the Cordyceps militaris spawn into the culture medium under sterile conditions, and then place it in a dark room at 18-20℃ and 60%-65% humidity for 8-10 days until the culture medium is covered with mycelium; (5) After the mycelium has completely absorbed the culture medium, carry out light color change culture at 20-22℃ and 60-70% humidity, with 10-12 hours of light per day until the mycelium on the surface of the medium completely changes color; (6) After the mycelium changes color, scratch the fungus and continue to cultivate until the fruiting bodies germinate. At a temperature of 20-24℃ and a humidity of 85-90%, ventilate for 1-2 hours and provide light for 7-9 hours every day. After the fruiting bodies have basically germinated on the surface of the substrate, harvest, dry and store to obtain Cordyceps militaris product.

[0008] The *Tremella fuciformis* Berk. used in the preparation of the culture medium in this invention is the fruiting body of the fungus *Tremella fuciformis*, also known as white fungus, snow fungus, and silver ear fungus. *Tremella fuciformis* is rich in nutrients, containing protein, fat, various amino acids, minerals, and glycogen. Its protein contains 17 amino acids, providing three-quarters of the essential amino acids required by the human body. It also contains various minerals such as calcium, phosphorus, iron, potassium, sodium, magnesium, and sulfur. Furthermore, it contains trehalose, pentosan, mannitol, and other glycogen, making it highly nutritious and possessing strengthening properties, making it a high-grade nourishing tonic. The acidic polysaccharides in *Tremella fuciformis* can enhance the body's immunity, mobilize lymphocytes, strengthen the phagocytic ability of white blood cells, and stimulate bone marrow hematopoiesis; *Tremella fuciformis* polysaccharides also have anti-tumor effects. White fungus is sweet in taste and neutral in nature, and enters the lung, stomach and kidney meridians; it has the effects of strengthening essence, tonifying the kidney, moistening the intestines, benefiting the stomach, replenishing qi, harmonizing blood, strengthening the heart, nourishing yin, moistening the lungs, promoting body fluid, strengthening the body, nourishing the brain, refreshing the mind, beautifying the skin, and prolonging life.

[0009] Job's tears (scientific name: *Coix chinensis* Tod.), also known as coix seed, is the seed of the *Coix lacryma-jobi* plant, a member of the Poaceae family. It is cool in nature, sweet and bland in taste, and enters the spleen, lung, and kidney meridians. It has the effects of promoting diuresis, strengthening the spleen, relieving numbness, and clearing heat and draining pus. Job's tears are rich in nutrients, containing 52%-80% carbohydrates, 13%-17% protein, and 4%-7% fat. The oil is mainly composed of unsaturated fatty acids, with linoleic acid accounting for 34%, and also contains a special coixol. Job's tears contain protein, amino acids, vitamins, minerals, dietary fiber, and other effective nutrients. When ground into flour and eaten as a staple food, it is a highly valuable health food. Job's tears can promote metabolism and reduce the burden on the gastrointestinal tract, making it a nourishing food for patients who are weak during or after illness.

[0010] Chinese yam powder, also known as yam powder, is made by drying and grinding Chinese yam (Latin name Dioscorea polystachya Turczaninow). Chinese yam is rich in mucoprotein, sugar, a large amount of starch, vitamins A1, B1, B2, C, and various amino acids and trace elements. Chinese yam powder generally has the effects of strengthening the spleen and stomach, tonifying the kidneys and replenishing essence. Consuming Chinese yam powder allows the absorption of rich plant protein and natural polysaccharides, as well as some choline and vitamins, nourishing the body and improving physical fitness. Chinese yam powder contains abundant natural saponins and various amino acids, as well as some vitamin E and vitamin C. These substances can regulate the body's endocrine system, nourish the skin, accelerate skin cell regeneration and metabolism, and promote beauty and prevent skin aging.

[0011] The beneficial effects of this invention are as follows: 1. The culture medium prepared by this invention, which is mainly composed of white fungus, supplemented with coix seed and Chinese yam powder, as well as glucose, lactose and other raw materials, is rich in nutrients. It promotes the growth of cordyceps and does not cause sticking after steaming like rice or wheat. The white fungus, coix seed and Chinese yam powder used contain abundant polysaccharides, amino acids, vitamins, minerals, starch and other substances. The culture medium prepared is conducive to promoting the growth of cordyceps and stimulating the production of cordycepin, cordyceps polysaccharides and other components in cordyceps.

[0012] 2. The raw materials for the Cordyceps culture medium of this invention are abundant and readily available. Using this medium for Cordyceps cultivation is simple, and under suitable conditions for dark culture, mycelial color change culture, and fruiting body culture, the Cordyceps exhibits good growth, a short cultivation cycle, and high production efficiency. The cultivated Cordyceps has high yield and good quality, containing abundant polysaccharides, cordycepin, cordycepic acid, and other substances. Cultivating Cordyceps using the method of this invention can significantly increase the content of cordycepin, cordycepic acid, and other components in Cordyceps. The cultivated Cordyceps contains over 7.5% Cordycepin polysaccharide, over 6.5 mg / g Cordycepin, over 9.2% Cordycepic acid, and over 810 mg / kg adenosine, exhibiting high nutritional content and excellent medicinal and health benefits. Detailed Implementation

[0013] Example 1 A method for cultivating Cordyceps sinensis using Tremella fuciformis culture medium includes the following steps: (1) Soak dried white fungus in water until it expands, clean it, drain the water, and crush it into 1-2cm pieces for later use; take pest-free Job's tears, wash them, soak them in water for 2 hours, and drain them for later use. (2) Take 10 parts of white fungus, 8 parts of coix seed, 5 parts of Chinese yam powder, 0.02 parts of glucose, 0.005 parts of lactose, 0.02 parts of peptone, 0.01 parts of potassium dihydrogen phosphate, 0.003 parts of glycine, 0.01 parts of magnesium sulfate and 32 parts of water according to the following weight proportions, and set aside; (3) First, dissolve glucose, lactose, peptone, potassium dihydrogen phosphate, glycine, and magnesium sulfate in water to obtain a nutrient solution; then mix the nutrient solution with tremella, coix seed, and Chinese yam powder and stir evenly to obtain a mixture. The mixture was dispensed into culture containers and sterilized at 121°C for 30 min. After sterilization, it was cooled to room temperature to obtain the culture medium. (4) Inoculate the Cordyceps militaris spawn into the culture medium under sterile conditions, and then place it in a dark room at 18-20℃ and 60%-65% humidity for 8 days in the dark until the culture medium is covered with mycelium; (5) After the mycelium has completely absorbed the culture medium, carry out light color change culture at 20-22℃ and 60-70% humidity, with 10 hours of light per day, until the mycelium on the surface of the medium completely changes color; (6) After the mycelium changes color, scratch the fungus and continue to cultivate until the fruiting bodies germinate. At a temperature of 20-24℃ and a humidity of 85-90%, ventilate for 1 hour and light for 7 hours a day. After the fruiting bodies have basically germinated on the surface of the substrate, harvest, dry and store to obtain Cordyceps militaris product.

[0014] Example 2 A method for cultivating Cordyceps sinensis using Tremella fuciformis culture medium includes the following steps: (1) Soak dried white fungus in water, clean it, drain the water, and crush it into 1-2cm pieces for later use; take pest-free Job's tears, wash them, soak them in water for 2.5 hours, and drain them for later use. (2) Take 15 parts of white fungus, 10 parts of coix seed, 5.5 parts of Chinese yam powder, 0.03 parts of glucose, 0.004 parts of lactose, 0.03 parts of peptone, 0.02 parts of potassium dihydrogen phosphate, 0.004 parts of glycine, 0.02 parts of magnesium sulfate and 45 parts of water according to the following weight proportions, and set aside. (3) First, dissolve glucose, lactose, peptone, potassium dihydrogen phosphate, glycine, and magnesium sulfate in water to obtain a nutrient solution; then mix the nutrient solution with tremella, coix seed, and Chinese yam powder and stir evenly to obtain a mixture. The mixture was dispensed into culture containers and sterilized at 121°C for 32 min. After sterilization, it was cooled to room temperature to obtain the culture medium. (4) Inoculate the Cordyceps militaris spawn into the culture medium under sterile conditions, and then place it in a dark room at 18-20℃ and 60%-65% humidity for 9 days in the dark until the culture medium is covered with mycelium; (5) After the mycelium has completely absorbed the culture medium, carry out light color change culture at 20-22℃ and 60-70% humidity, with 11 hours of light per day, until the mycelium on the surface of the medium completely changes color; (6) After the mycelium changes color, scratch the fungus and continue to cultivate until the fruiting bodies germinate. At a temperature of 20-24℃ and a humidity of 85-90%, ventilate for 1.5 hours and expose to light for 8 hours every day. After the fruiting bodies have basically all germinated on the substrate surface, harvest, dry and store to obtain Cordyceps militaris product.

[0015] Example 3 A method for cultivating Cordyceps sinensis using Tremella fuciformis culture medium includes the following steps: (1) Soak dried white fungus in water, clean it, drain the water, and crush it into 1-2cm pieces for later use; take pest-free Job's tears, wash them, soak them in water for 2.2 hours, and drain them for later use. (2) Take 20 parts of white fungus, 12 parts of coix seed, 6 parts of Chinese yam powder, 0.03 parts of glucose, 0.003 parts of lactose, 0.02 parts of peptone, 0.03 parts of potassium dihydrogen phosphate, 0.005 parts of glycine, 0.02 parts of magnesium sulfate and 50 parts of water according to the following weight proportions, and set aside. (3) First, dissolve glucose, lactose, peptone, potassium dihydrogen phosphate, glycine, and magnesium sulfate in water to obtain a nutrient solution; then mix the nutrient solution with tremella, coix seed, and Chinese yam powder and stir evenly to obtain a mixture. The mixture was dispensed into culture containers and sterilized at 121°C for 35 min. After sterilization, it was cooled to room temperature to obtain the culture medium. (4) Inoculate the Cordyceps militaris spawn into the culture medium under sterile conditions, and then place it in a dark room at 18-20℃ and 60%-65% humidity for 10 days in the dark until the culture medium is covered with mycelium; (5) After the mycelium has completely absorbed the culture medium, carry out light color change culture at 20-22℃ and 60-70% humidity, with 12 hours of light per day, until the mycelium on the surface of the medium completely changes color; (6) After the mycelium changes color, scratch the fungus and continue to cultivate until the fruiting bodies germinate. At a temperature of 20-24℃ and a humidity of 90%, ventilate for 2 hours and expose to light for 9 hours every day. After the fruiting bodies have basically germinated on the surface of the substrate, harvest, dry and store to obtain Cordyceps militaris product.

[0016] Quality Inspection The fruiting bodies of Cordyceps militaris cultivated according to the methods of Examples 1-3 of this invention were subjected to component content analysis, and the results are shown in Table 1 below: As can be seen from Table 1 above, the cordyceps cultivated using the method of this invention have high contents of cordyceps polysaccharide, cordycepin, cordycepic acid and adenosine, and the cordyceps have good quality and good medicinal effects.

Claims

1. A method for cultivating Cordyceps sinensis using a culture medium with Tremella fuciformis as the main ingredient, characterized in that, Includes the following steps: (1) Soak dried white fungus in water, clean it, drain the water, and crush it into 1-2cm pieces for later use; take pest-free Job's tears, wash them, soak them in water for 2-2.5 hours, and drain them for later use. (2) Take 10-20 parts by weight of white fungus; 8-12 parts of coix seed; 5-6 parts of Chinese yam powder; 0.02-0.03 parts of glucose; 0.003-0.005 parts of lactose; 0.02-0.03 parts of peptone; 0.01-0.03 parts of potassium dihydrogen phosphate; 0.003-0.005 parts of glycine; 0.01-0.02 parts of magnesium sulfate and 32-50 parts of water, and set aside. (3) First, dissolve glucose, lactose, peptone, potassium dihydrogen phosphate, glycine and magnesium sulfate in water to obtain nutrient solution; then mix the nutrient solution with tremella, coix seed and Chinese yam powder and stir evenly to obtain a mixture. Distribute the mixture into culture containers and sterilize at 121℃ for 30-35 minutes. After sterilization, cool to room temperature to obtain culture medium. (4) Inoculate the Cordyceps militaris spawn into the culture medium under sterile conditions, and then place it in a dark room at 18-20℃ and 60%-65% humidity for 8-10 days until the culture medium is covered with mycelium; (5) After the mycelium has completely absorbed the culture medium, carry out light color change culture at 20-22℃ and 60-70% humidity, with 10-12 hours of light per day until the mycelium on the surface of the medium completely changes color; (6) After the mycelium changes color, scratch the fungus and continue to cultivate until the fruiting bodies germinate. At a temperature of 20-24℃ and a humidity of 85-90%, ventilate for 1-2 hours a day and provide light for 7-9 hours. After the fruiting bodies have basically all germinated on the substrate surface, harvest, dry and store. The Cordyceps obtained from cultivation contains more than 7.5% Cordyceps polysaccharide, more than 6.5mg / g Cordycepin, more than 9.2% Cordycepic acid, and more than 810mg / kg adenosine.

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