Use method of culture medium for culturing hyphae and preparation method of hyphae

Abstract

According to the use method of the culture medium for culturing the hyphae and the preparation method of the hyphae, the hyphae prepared by combining the edible fungus strains and the areca nuts can provide nutritional ingredients of pleurotus subsp. For a human body.

Description

[0001] This application is a divisional application with an application date of July 14, 2022, application number 202210823506.7, and invention name “A Mycelium Betel Nut”. Technical Field

[0002] The invention relates to the technical field of food processing, and in particular to a method for using a culture medium for culturing mycelium and a method for preparing mycelium. Background Art

[0003] It is well known that although betel nut is harmful to the oral cavity, it still contains a variety of nutrients and beneficial substances needed by the human body. At the same time, betel nut also contains more than 20 trace elements, 11 of which are essential trace elements for the human body, and people will have a certain refreshing effect when chewing betel nut.

[0004] The subpleural fungus, also known as winter mushroom, Yuan mushroom (Heilongjiang), yellow mushroom (Jilin), has a medium to slightly larger fruiting body, with a cap diameter of 9 to 12 cm, flat hemispherical to flat, semicircular or kidney-shaped, yellow-green, sticky with short villi, smooth edges, white flesh, slightly dense gills, white with light yellow, nearly extended, and a lateral stipe that is very short or almost absent. It grows on rotten wood of broad-leaved trees such as birch in autumn, growing in clusters in a shingle-like manner; it is distributed in Hebei, Heilongjiang, Jilin, Shanxi, Guangxi, Shaanxi, Sichuan, Yunnan, Tibet and other regions in my country, and is a wood decay fungus;

[0005] Among them, Pleurotus eryngii is the best specialty of Changbai Mountain area. It is the best among mushrooms, and its nutritional content is more than ten times that of ordinary vegetables. In particular, it is rich in fungal protein, fungal polysaccharides, polyols, amino acids, calcium, phosphorus and other nutrients. The protein content of every 100 grams of Pleurotus eryngii (dry product) is 8 grams. In terms of function, Pleurotus eryngii has the functions of increasing appetite, enhancing the body's resistance, alleviating abdominal distension, relieving liver pain, replenishing qi and not being hungry, regulating immune dysfunction, inhibiting tumors, and strengthening body immunity. Anti-cancer tests show that the inhibition rate of sarcoma 180 in mice is 70%, and the inhibition rate of Ehrlich cancer is 70%. In addition, it also has the functions of relaxing muscles and tendons, strengthening tendons and bones, and can treat waist and leg pain, numbness of hands and feet, and muscle discomfort.

[0006] In addition, edible fungi rich in nutrients, such as Cordyceps militaris, Ganoderma lucidum, Hydrangea, Antrodia cinnamomea and Morel, also have the effect of inhibiting tumors. Summary of the invention

[0007] In order to overcome the deficiencies in the background technology, the present invention discloses a method for using a culture medium for culturing mycelium and a method for preparing mycelium.

[0008] In order to achieve the above-mentioned invention object, the present invention adopts the following technical scheme:

[0009] The present invention provides a method for using a culture medium for culturing mycelium.

[0010] 1) Preparation of betel nut:

[0011] a. First, cut the clean betel nut in half, remove the core, and soak it in weak alkaline water for 24 to 36 hours;

[0012] b. Take out the betel nut and put it into a culture dish, and sterilize it at high temperature;

[0013] c. After sterilization, place the culture dish in a sterile inoculation box to cool naturally;

[0014] 2) Preparation of culture medium:

[0015] a. Take 200g peeled sweet potato slices, 20g glucose, 2g peptone and 1L water and boil for 20 minutes;

[0016] b. Then filter the culture solution through gauze and cool it to 18℃ for later use;

[0017] 3) Preparation of Pleurotus erythrorhizium:

[0018] a. Place the sterilized betel nut in a sterile inoculation box to cool naturally, inoculate the liquid strain of Pleurotus eryngii in a sterile environment, and add culture medium to complete the inoculation.

[0019] Preferably, when the betel nut is sterilized at high temperature, the culture dish containing the betel nut is first placed in a steamer, and then sterilized in high-pressure steam at 0.1 MPa and 120° C. for 40 minutes.

[0020] Preferably, in step a of 3), the weight ratio of culture solution to betel nut is 1:5.

[0021] Preferably, the Pleurotus ostreatus can be replaced by Cordyceps militaris, Ganoderma lucidum or Morchella.

[0022] The present invention provides a method for preparing mycelium.

[0023] 1) Preparation of liquid strains:

[0024] a. Inoculate 6 to 8 pieces of Pleurotus erythrocytes mother culture blocks into liquid culture medium under sterile conditions;

[0025] b. Then the liquid culture medium is placed at 18-22°C and cultured for 72 hours;

[0026] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 22° C., and the cultivation time is 14 days to obtain a liquid culture of Pleurotus eryngii;

[0027] 2) Preparation of Pleurotus erythrorhizium:

[0028] Ⅰ. Using the above-mentioned method to complete the inoculation of liquid strains of Pleurotus erythrorhizium;

[0029] Ⅱ. Place the inoculated betel nut in a sterile culture room at 21°C for constant temperature and light-proof culture. After 15 to 17 days of culture, you can see that the hyphae of Subpleurum have grown all over the surface and inside of the betel nut, indicating that the hyphae of Subpleurum have been obtained.

[0030] Preferably, the Pleurotus eryngii is replaced by Cordyceps militaris, Ganoderma lucidum or Morchella.

[0031] Preferably, when Cordyceps militaris is used as the inoculated mycelium,

[0032] 1) Preparation of liquid strains:

[0033] a. Inoculate 6 to 8 pieces of Cordyceps militaris mother culture blocks into the liquid culture medium under sterile conditions;

[0034] b. Then place the liquid culture medium at 20-21°C and culture it statically for 48-72 hours;

[0035] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 20 to 21°C for 8 to 12 days to obtain a liquid strain of Cordyceps militaris;

[0036] 2) Production of Cordyceps militaris mycelium:

[0037] Ⅰ. Use the above-mentioned method to complete the inoculation of liquid Cordyceps militaris fungus;

[0038] Ⅱ. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 4 to 7 days to obtain Cordyceps militaris mycelium.

[0039] Preferably, when using Ganoderma lucidum as inoculated mycelium,

[0040] 1) Preparation of liquid strains:

[0041] a. Inoculate 6 to 8 pieces of Ganoderma lucidum mother culture blocks into the liquid culture medium under sterile conditions;

[0042] b. Then place the liquid culture medium at 18-20°C and culture it statically for 48-72 hours;

[0043] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 20° C., and the cultivation time is 3 to 5 days to obtain a liquid strain of Ganoderma lucidum;

[0044] 2) Production of Ganoderma lucidum mycelium:

[0045] Ⅰ. Use the above-mentioned method to complete the inoculation of liquid strains of Ganoderma lucidum;

[0046] Ⅱ. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 4 days to obtain Ganoderma lucidum mycelium.

[0047] Preferably, when Morchella is used as the inoculated mycelium,

[0048] 1) Preparation of liquid strains:

[0049] a. Inoculate 6 to 8 pieces of Morchella mother culture blocks into the liquid culture medium under sterile conditions;

[0050] b. Then place the liquid culture medium at 18-20°C and culture it statically for 48-72 hours;

[0051] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 20° C., and the cultivation time is 7 to 10 days to obtain a liquid strain of Morchella;

[0052] 2) Production of Morel Mycelium:

[0053] Ⅰ. Use the above-mentioned method to complete the inoculation of liquid Morchella spp.;

[0054] Ⅱ. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 5 days to obtain Morchella mycelium.

[0055] Due to the adoption of the above-mentioned technical solution, the present invention has the following beneficial effects:

[0056] The present invention discloses a method for using a culture medium for culturing mycelium and a method for preparing mycelium.

[0057] 1. By growing edible fungus mycelium on betel nut-based culture medium, the types of culture medium for edible fungus mycelium have been expanded, and the process technology for mycelium cultivation has been innovated;

[0058] 2. The edible fungus mycelium betel nut not only retains the original product characteristics of betel nut, but also increases the nutrients of edible fungus mycelium;

[0059] 3. Due to the unique anti-tumor cell properties of the mycelium of Pleurotus eryngii, it can solve the disadvantage that betel nut itself is harmful to the human body. During the long fermentation process, the mycelium can break and decompose the crude fiber of betel nut, allowing arecoline to be more effectively leaked out. DETAILED DESCRIPTION

[0060] The technical solution of the present invention will be described below:

[0061] The mycelium described above needs to go through the following steps:

[0062] 1) Preparation of liquid culture of Pleurotus erythrorhizium:

[0063] a. In a liquid culture medium under sterile conditions, 6 to 8 pieces of subpleureus mother culture blocks are accessed, wherein the blocks are 8 to 10 mm wide cubes; and the sterile conditions are achieved by placing the culture dish where the liquid culture medium is located in a sterile inoculation box;

[0064] b. Then the liquid culture medium is placed at 18-22°C and cultured for 72 hours;

[0065] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18-22° C., and the cultivation time is 14 days, to obtain a liquid culture of Pleurotus eryngii; wherein the liquid culture of Pleurotus eryngii exhibits a state in which the culture solution is clear and transparent, and the mycelium is suspended in the culture solution in a spherical shape;

[0066] An embodiment of this step is:

[0067] a. Inoculate 6 pieces of Pleurotus erythrocytes mother culture blocks into liquid culture medium under sterile conditions;

[0068] b. Then the liquid culture medium was placed at 18°C ​​and incubated for 72 hours;

[0069] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18° C. and the cultivation time is 14 days to obtain a liquid culture of Pleurotus eryngii.

[0070] The second embodiment of this step is:

[0071] a. Inoculate 7 pieces of Pleurotus erythrocytes mother culture blocks into liquid culture medium under sterile conditions;

[0072] b. Then the liquid culture medium was placed at 19°C and incubated for 72 hours;

[0073] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 19° C. and the cultivation time is 14 days to obtain a liquid culture of Pleurotus eryngii.

[0074] The third embodiment of this step is:

[0075] a. Inoculate 8 pieces of Pleurotus erythrocytes mother culture blocks into liquid culture medium under sterile conditions;

[0076] b. Then the liquid culture medium was placed at 20°C and incubated for 72 hours;

[0077] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 20° C. and the cultivation time is 14 days to obtain a liquid culture of Pleurotus eryngii.

[0078] 2) Preparation of betel nut:

[0079] a. First, cut the clean betel nut in half, remove the core, and soak it in weakly alkaline water for 24 to 36 hours. The soaking time is selected according to the volume of the betel nut;

[0080] b. Take out the betel nuts and put them into a culture dish, and sterilize them at high temperature; in addition, the conditions for high temperature sterilization of betel nuts are as follows: first, place the culture dish containing betel nuts in a steamer, and then sterilize the soybean particles under 0.1Mpa pressure and 120℃ high pressure steam for 40 minutes.

[0081] c. After sterilization, place the culture dish in a sterile inoculation box to cool naturally;

[0082] 3) Preparation of culture medium:

[0083] a. Take 200g peeled sweet potato slices, 20g glucose, 2g peptone and 1L water and boil for 20 minutes;

[0084] b. Then filter the culture solution through gauze and cool it to 18℃ for later use;

[0085] 4) Preparation of Pleurotus erythrorhizium:

[0086] a. The sterilized betel nut is placed in a sterile inoculation box and cooled naturally, and the liquid strain of Pleurotus eryngii is accessed in a sterile environment, and the culture medium is added to complete the inoculation; wherein the weight ratio of the culture medium to the betel nut is 1:5;

[0087] b. Place the inoculated betel nut in a sterile culture room at 21°C for constant temperature and light-proof culture for 15 to 17 days to obtain the mycelium of Subpleurum. The manifestation of Subpleurum mycelium growing all over the betel nut is that the mycelium of Subpleurum grows all over the surface and inside of the betel nut. In addition, the culture time is also selected according to the volume of the betel nut.

[0088] In addition, the Pleurotus ostreatus in the mycelium can be replaced by Cordyceps militaris, Ganoderma lucidum or Morchella.

[0089] When using Cordyceps militaris as inoculated mycelium,

[0090] 1) Preparation of liquid strains:

[0091] a. Inoculate 6 to 8 pieces of Cordyceps militaris mother culture blocks into the liquid culture medium under sterile conditions;

[0092] b. Then place the liquid culture medium at 20-21°C and culture it statically for 48-72 hours;

[0093] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 20 to 21°C for 8 to 12 days to obtain a liquid strain of Cordyceps militaris;

[0094] When the number of mother inoculum blocks of Cordyceps militaris was 6, 7 and 8, the liquid culture medium was placed at 20°C, 20.5°C and 21°C, and left to stand for 72 hours, 60 hours and 48 hours, respectively. Then, the mycelium was cultured on an electromagnetic shaker at 20°C, 20.5°C and 21°C for 12 days, 10 days and 8 days, respectively.

[0095] 4) Production of Cordyceps militaris mycelium:

[0096] a. Place the sterilized betel nut in a sterile inoculation box and allow to cool naturally. Access the liquid strain of Cordyceps militaris in a sterile environment and add the culture medium to complete the inoculation.

[0097] b. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 4 to 7 days to obtain Cordyceps militaris mycelium; depending on the number of Cordyceps militaris mother culture blocks used, the culture time of betel nut is 7 days, 5.5 days and 7 days respectively.

[0098] When using Ganoderma lucidum as inoculated mycelium,

[0099] 1) Preparation of liquid strains:

[0100] a. Inoculate 6 to 8 pieces of Ganoderma lucidum mother culture blocks into the liquid culture medium under sterile conditions;

[0101] b. Then place the liquid culture medium at 18-20°C and culture it statically for 48-72 hours;

[0102] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 20° C., and the cultivation time is 3 to 5 days to obtain a liquid strain of Ganoderma lucidum;

[0103] When the number of Ganoderma lucidum mother inoculum blocks was 6, 7 and 8, the liquid culture medium was placed at 18°C, 19°C and 20°C, and left to stand for 72 hours, 60 hours and 48 hours, respectively. Then, the mycelium was cultured on an electromagnetic shaker at 18°C, 19°C and 20°C for 5 days, 4 days and 3 days, respectively.

[0104] 4) Production of Ganoderma lucidum mycelium:

[0105] a. Place the sterilized betel nut in a sterile inoculation box for natural cooling, access the liquid strain of Ganoderma lucidum in a sterile environment, and add the culture medium to complete the inoculation;

[0106] b. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light protection for 4 days to obtain Ganoderma lucidum mycelium.

[0107] Preferably, when Morchella is used as the inoculated mycelium,

[0108] 1) Preparation of liquid strains:

[0109] a. Inoculate 6 to 8 pieces of Morchella mother culture blocks into the liquid culture medium under sterile conditions;

[0110] b. Then place the liquid culture medium at 18-22°C and culture it statically for 48-72 hours;

[0111] c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 22° C., and the cultivation time is 7 to 10 days to obtain a liquid strain of Morchella;

[0112] When the number of mother inoculum blocks of Morchella was 6, 7 and 8 respectively, the liquid culture medium was placed at 18°C, 20°C and 22°C and left to stand for 72 hours, 60 hours and 48 hours respectively. Then, the mycelium was cultured on an electromagnetic shaker at 18°C, 20°C and 22°C for 7 days, 8.5 days and 10 days respectively.

[0113] 4) Production of Morel Mycelium:

[0114] a. Place the sterilized betel nut in a sterile inoculation box for natural cooling, access the liquid Morchella spp. in a sterile environment, and add the culture medium to complete the inoculation;

[0115] b. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 5 days to obtain Morchella mycelium.

[0116] The parts of the present invention that are not described in detail are prior art. It is obvious to those skilled in the art that the present invention is not limited to the details of the above-mentioned exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic characteristics of the present invention; therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is limited by the appended claims rather than the above description, and it is intended that all changes within the meaning and scope of the equivalent elements of the claims are included in the present invention.

Claims

1. A method for using a culture medium for culturing mycelium, characterized in that: 1) Preparation of betel nut: a. First, cut the clean betel nut in half, remove the core, and soak it in weak alkaline water for 24 to 36 hours; b. Take out the betel nut and put it into a culture dish, and sterilize it at high temperature; c. After sterilization, place the culture dish in a sterile inoculation box to cool naturally; 2) Preparation of culture medium: a. Take 200g peeled sweet potato slices, 20g glucose, 2g peptone and 1L water and boil for 20 minutes; b. Then filter the culture solution through gauze and cool it to 18℃ for later use; 3) Inoculation method of Pleurotus erythrocytes mycelium: a. Place the sterilized betel nut in a sterile inoculation box to cool naturally, inoculate the liquid strain of Pleurotus eryngii in a sterile environment, and add culture medium to complete the inoculation.

2. The method of use according to claim 1, characterized in that: When the betel nut is sterilized at high temperature, the culture dish containing the betel nut is firstly placed in a steamer, and then sterilized in high-pressure steam at 0.1 MPa and 120° C. for 40 minutes.

3. The method of use according to claim 1, characterized in that: In step a of 3), the weight ratio of culture solution to betel nut is 1:

5.

4. The method of use according to claim 1, characterized in that: The subpleureus fungus is replaced by Cordyceps militaris, Ganoderma lucidum or Morchella.

5. A method for producing mycelium, characterized in that: 1) Preparation of liquid strains: a. Inoculate 6 to 8 pieces of Pleurotus erythrocytes mother culture blocks into liquid culture medium under sterile conditions; b. Then the liquid culture medium is placed at 18-22°C and cultured for 72 hours; c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 22° C., and the cultivation time is 14 days to obtain a liquid culture of Pleurotus eryngii; 2) Preparation of Pleurotus erythrorhizium: Ⅰ. Using the method of any one of claims 1 to 4 to complete the inoculation of liquid strains of Pleurotus erythrorhizium; Ⅱ. Place the inoculated betel nut in a sterile culture room at 21°C for constant temperature and light-proof culture. After 15 to 17 days of culture, you can see that the mycelium of Subpleurum has grown all over the surface and inside of the betel nut, indicating that the mycelium of Subpleurum has been obtained.

6. The method of claim 5, wherein: The subpleureus fungus is replaced by Cordyceps militaris, Ganoderma lucidum or Morchella.

7. The method of claim 6, wherein: When using Cordyceps militaris as inoculated mycelium, 1) Preparation of liquid strains: a. Inoculate 6 to 8 pieces of Cordyceps militaris mother culture blocks into the liquid culture medium under sterile conditions; b. Then place the liquid culture medium at 20-21°C and culture it statically for 48-72 hours; c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 20 to 21°C for 8 to 12 days to obtain a liquid strain of Cordyceps militaris; 2) Production of Cordyceps militaris mycelium: Ⅰ. Using the method of any one of claims 1 to 4 to complete the inoculation of liquid Cordyceps militaris spawn; Ⅱ. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 4 to 7 days to obtain Cordyceps militaris mycelium.

8. The method of claim 6, wherein: When using Ganoderma lucidum as inoculated mycelium, 1) Preparation of liquid strains: a. Inoculate 6 to 8 pieces of Ganoderma lucidum mother culture blocks into the liquid culture medium under sterile conditions; b. Then place the liquid culture medium at 18-20°C and culture it statically for 48-72 hours; c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 20° C., and the cultivation time is 3 to 5 days to obtain a liquid strain of Ganoderma lucidum; 2) Production of Ganoderma lucidum mycelium: Ⅰ. Using the method of any one of claims 1 to 4 to complete the inoculation of liquid strains of Ganoderma lucidum; Ⅱ. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 4 days to obtain Ganoderma lucidum mycelium.

9. The method of claim 6, wherein: When using Morchella as inoculum, 1) Preparation of liquid strains: a. Inoculate 6 to 8 pieces of Morchella mother culture blocks into the liquid culture medium under sterile conditions; b. Then place the liquid culture medium at 18-20°C and culture it statically for 48-72 hours; c. After the mycelium germinates, the mycelium is placed on an electromagnetic shaker for cultivation, wherein the cultivation temperature is controlled at 18 to 20° C., and the cultivation time is 7 to 10 days to obtain a liquid strain of Morchella; 2) Production of Morel Mycelium: Ⅰ. Using the method of any one of claims 1 to 4 to complete the inoculation of liquid Morchella spp.; Ⅱ. Place the inoculated betel nut in a sterile culture room at 20°C for constant temperature and light-proof culture for 5 days to obtain Morchella mycelium.