CN115777444A - Method for cultivating cordyceps militaris by using wheat kernels as culture medium - Google Patents
Method for cultivating cordyceps militaris by using wheat kernels as culture medium Download PDFInfo
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- CN115777444A CN115777444A CN202211560020.5A CN202211560020A CN115777444A CN 115777444 A CN115777444 A CN 115777444A CN 202211560020 A CN202211560020 A CN 202211560020A CN 115777444 A CN115777444 A CN 115777444A
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Abstract
The invention relates to the technical field of cordyceps militaris culture, in particular to a method for cultivating cordyceps militaris by using wheat kernels as a culture medium. The method comprises the steps of carrying out peeling treatment on wheat, modifying by microwave irradiation, and then sterilizing at a lower temperature, so that the obtained solid culture medium is low in hardness and viscosity, and the yield per unit of cordyceps militaris is greatly increased. The method comprises the following steps: (1) Selecting wheat grains as a substrate, and adopting a peeling process, wherein the peeling rate of the wheat is 4-10%; (2) The wheat kernels are modified by microwave irradiation, the microwave irradiation temperature is 140-150 ℃, and the duration time is 5-15 min; (3) Sterilizing at lower temperature of 110-121 deg.c for 20-40 min. The other part is carried out according to the normal production flow of Cordyceps militaris. The wheat kernel solid culture medium treated by the method has moderate viscosity, can greatly improve the yield per unit of cordyceps militaris, and is convenient to harvest.
Description
Technical Field
The invention relates to the technical field of cordyceps militaris culture, in particular to a method for cultivating cordyceps militaris by using wheat kernels as a culture medium.
Background
With the continuous development of cordyceps militaris cultivation technology, most of the cordyceps militaris are cultivated by using grains such as rice and wheat as substrates, and particularly wheat is generally adopted for cultivation in northeast China. In the actual large-scale culture process, due to the direct application of wheat grains, the biotransformation rate of cordyceps militaris to wheat grains is not high.
The patent (publication No. CN 104641944A) discloses a method for cultivating cordyceps sinensis by using unhulled wheat as a culture medium, in the implementation process, the shell of steamed wheat grains is cracked and exposed out of the wheat, but the shell is wrapped by the wheat all the time, so that the gelatinization and adhesion of the wheat are effectively prevented. However, in the implementation process of the patent, the wheat is shelled, so that the culture medium is easy to scatter, and much inconvenience is brought to the automation links such as harvesting.
The patent (publication number: CN 109105148A) discloses a high-yield and high-quality cordyceps militaris cultivation medium and a cultivation method thereof, wherein the culture medium takes rice or wheat as a main cultivation substrate, corncobs, wheat bran, wood chips or rice husks accounting for 10% -40% of the mass of the rice or wheat are added, a nutrient solution with the mixture liquid ratio of 1.1-1.5 g/mL is added, and the cultivation medium is prepared after sterilization.
The patent (publication number: CN 108633624A) discloses a high-yield culture medium for fruiting bodies of cordyceps alpina, which is characterized by consisting of rice or wheat and nutrient solution according to a mass-volume ratio of 1.2-2, wherein each liter of the nutrient solution comprises 10-30 g of a carbon source, 5-10 g of a nitrogen source and 3-15 g of potassium salt, and finally water is used for carrying out volume fixing to 1L, pH 6.0-7.5, the carbon source is sucrose, glucose or soluble starch, the nitrogen source is peptone, beef extract, yeast extract or soybean, and the potassium salt is potassium chloride, potassium nitrate, potassium dihydrogen phosphate or dipotassium hydrogen phosphate and the like.
The patent (publication number: CN 105315043A) discloses a culture medium for cordyceps militaris, which is characterized in that: the raw materials comprise the following components in percentage by mass: 15-25 parts of potato, 1-2.5 parts of glucose, 0.05-0.15 part of monopotassium phosphate, 0.05-0.15 part of magnesium sulfate, 10.10-0.15 part of vitamin B, 5-10 parts of egg, 10-20 parts of silkworm chrysalis meal, 1-2 parts of calcium sulfate, 18-25 parts of rice and 20-35 parts of wheat, wherein the rice is rice grains and the wheat is wheat grains.
The patent uses unhulled wheat or wheat grain to cultivate, neglects another detail, through the cultivation of cordyceps militaris for many years, the structure composition and the nature of various cultivation media are deeply researched, it is found that the wheat not only has an outer wheat shell, but also contains wheat bran, because of the wrapping of the wheat bran, the wheat grain can not fully expand and gelatinize in the cooking process, the released nutrient substances are not enough, the aleurone layer is firmly wrapped by the cortex on the surface of the cooked wheat, the direct contact utilization of cordyceps mycelium is avoided, finally, the cordyceps mycelium can only utilize the nutrient components in the gap part of the wheat grain, the utilization rate of the medium is low, and the yield of the cordyceps militaris is low. In addition, researches show that microorganisms in the wheat are mainly distributed in the wheat cortex, the number of the microorganisms in the wheat is small, and the number of the microorganisms gradually decreases from the wheat cortex to the inner part. In order to realize substrate sterilization, a high-temperature and high-pressure sterilization process is generally adopted, and the reason that the temperature cannot be reduced is that the wheat bran layer microorganisms are higher in magnitude and reach 5lgCFU/g.
The wheat kernel is obtained by a simple method, the wheat kernels except the ventral sulcus are removed by friction and cutting, and batch production is realized at present. The culture medium is prepared by adopting the wheat kernels, and because the epidermis and the pericarp contain the alpha-amylase of the wheat grains, the viscosity of the culture medium is greatly increased due to the lack of the degradation of the alpha-amylase and the increase of the dissolved starch in the process of stewing and sterilization of the wheat kernels after the wheat hulls are removed, so that the cordyceps militaris hyphae are difficult to eat through, and the utilization rate of the matrix is still low.
Disclosure of Invention
The invention provides a method for cultivating cordyceps militaris by using wheat kernels as a culture medium. The invention creatively introduces the microwave irradiation modification technology and is matched with the wheat grain peeling technology, thereby not only reducing the temperature required by cooking sterilization and shortening the time required by starch gelatinization, but also degrading and degrading the starch in wheat kernels, particularly the molecular weight of amylopectin, and fundamentally reducing the viscosity of the wheat kernel culture medium due to the change of the rheological property of the dissolved wheat starch in the sterilization process, being beneficial to the infiltration of cordyceps militaris hyphae, and achieving the purpose of high yield.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for cultivating cordyceps militaris by using wheat kernels as a culture medium, which comprises the following steps:
(1) Performing microwave irradiation modification treatment on peeled wheat kernels to obtain a material 1;
(2) Mixing the material 1 obtained in the step (1) with water to obtain a culture medium, sterilizing the culture medium, and inoculating cordyceps militaris strains for culture.
Preferably, the peeling rate of the wheat kernels in the step (1) is 4-10%.
Preferably, the thickness of the wheat kernel paving material irradiated by the microwave in the step (1) is 1-2 cm.
Preferably, the microwave irradiation temperature in the step (1) is 140-150 ℃ and the time is 5-15 min.
Preferably, the temperature for sterilization in the step (2) is 110-121 ℃, and the time is 20-40 min.
Preferably, the mass ratio of the material 1 to the water in the step (2) is 1.8-2.0.
Preferably, the volume-to-mass ratio of the cordyceps militaris strain in the step (2) to the material 1 is 8-10 mL:100g.
Preferably, the culturing in step (2) is divided into dark culturing and light culturing.
Preferably, the dark culture time is 3-7 days, and the temperature is 16-19 ℃.
Preferably, the light culture time is 40-50 d, the illumination intensity is 50-1000 Lux, and the temperature is 16-22 ℃.
The technical names related to the invention are as follows:
wheat kernel: the wheat grain is used as a raw material, and the skin layers of the wheat except for the groin are removed by a peeling technology to obtain the wheat grain. The wheat kernel has six layers of peels, and the outer three layers of peels of the wheat kernel can be removed by peeling: epidermis, epicarp, and endocarp. In addition, the seed coat and the bead core layer are also detached and damaged to different degrees.
(structural description of wheat grain in FIG. 4)
Wheat groin: the back of the wheat grain is raised, the abdomen is depressed, and a groove is called as an abdominal furrow.
Storing fungi: in the grain just harvested, semiparasitic fungi such as Helminthosporium (Helminthosporium), alternaria (Alternaria), fusarium (Fusarium) can be cultured, and the semiparasitic fungi gradually decrease and are replaced by saprophytic fungi such as Penicillium, aspergillus, and Mycoplasma (Blastomyces), which are called storage fungi, in the processes of harvesting, dry-sun, warehousing and the like.
An aleurone layer: the aleurone layer is the innermost layer of the seed cortex of the cereal crop and is positioned between the seed coat and the endosperm, the aleurone layer accounts for 7-9% of the total mass of the seeds, the physiological active ingredients with high nutritive value (such as dietary fiber, mineral substances, beneficial lipids, vitamins, phenolic acids, lignans and the like) in the cereal crop are concentrated in the aleurone layer, and 20% of pure aleurone layer is equivalent to the nutrition of the whole grain. During the milling process, the aleurone layer of wheat and rice is usually peeled off as bran with the hull adhered together, and the aleurone layer is extremely difficult to physically peel off.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention utilizes peeled wheat kernels as culture medium to cultivate cordyceps, solves the problem that cordyceps militaris hypha can not utilize nutrient substances in wheat pericarp, and simultaneously is beneficial to the growth of cordyceps militaris hypha because storage fungi and possible toxins attached to the pericarp are removed and fermentation inhibiting molecules such as ferulic acid, sinapic acid and the like are also removed.
2. The invention provides a method for modifying wheat bran by using microwave, which effectively solves the problems of overlarge viscosity of a culture medium caused by removal of wheat bran and difficulty in infiltration and collection of hypha.
3. The final single yield of the cordyceps militaris (the original seed is from Fujian province) is remarkably improved, the improvement rate reaches more than 30%, and the economic benefit is remarkable.
In conclusion, the wheat kernels are used as the culture medium, the constraints of wheat husks and wheat husks are eliminated, the aleurone layer on the surface is exposed and is fully gelatinized, more nutrients such as starch, protein and the like are released, and the microwave irradiation modification treatment is carried out, so that the hardness and viscosity of the culture medium are reduced, the direct contact utilization of cordyceps militaris hyphae is facilitated, the single yield of cordyceps militaris is greatly improved, and the wheat kernels have important significance for the collection automation of cordyceps militaris solid fermentation and the improvement of production benefits.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is an enlarged view of wheat grain and kernel;
FIG. 2 shows the culture media of examples (left panel) and comparative examples (right panel);
FIG. 3 is a schematic diagram of medium hardness and viscosity tests (left: texture analyzer; right: field test chart);
figure 4 is an illustration of wheat grain structure.
Detailed Description
The invention provides a method for cultivating cordyceps militaris by using wheat kernels as a culture medium, which comprises the following steps:
(1) Performing microwave irradiation modification treatment on peeled wheat kernels to obtain a material 1;
(2) Mixing the material 1 obtained in the step (1) with water to obtain a culture medium, sterilizing the culture medium, and inoculating cordyceps militaris strains for culture.
In the invention, the peeling rate of the wheat kernels in the step (1) is 4-10%; preferably 5 to 9 percent; more preferably 6 to 8%; more preferably 7%.
In the invention, the thickness of the wheat kernel paving material irradiated by microwave in the step (1) is 1-2 cm; preferably 1.5cm.
In the invention, the microwave irradiation temperature in the step (1) is 140-150 ℃; preferably 142 to 148 ℃; further preferably 144 to 146 ℃; more preferably 145 deg.c.
In the invention, the microwave irradiation time in the step (1) is 5-15 min; preferably 7-13 min; more preferably 9-11 min; more preferably 10min.
In the invention, the microwave irradiation power in the step (1) is 25-35 kw, preferably 30kw.
In the invention, the microwave irradiation frequency in the step (1) is 2350-2550 MHz; preferably 2450MHz.
In the invention, the temperature for sterilizing in the step (1) is 110-121 ℃; preferably 113-119 ℃; more preferably 115 to 117 ℃; more preferably 116 deg.c.
In the invention, the sterilization time in the step (1) is 20-40 min; preferably 24-36 min; more preferably 28 to 32min; more preferably 30min.
In the invention, the mass ratio of the material 1 in the step (2) to water is 1.8-2.0; preferably 1.
In the invention, the volume-to-mass ratio of the cordyceps militaris strain in the step (2) to the material 1 is 8-10 mL:100g of the total weight of the mixture; preferably 9mL:100g.
In the present invention, the culturing in step (2) is divided into dark culturing and light culturing.
In the invention, the dark culture time is 3-7 d; preferably 4 to 6d; more preferably 5d.
In the invention, the temperature of dark culture is 16-19 ℃; preferably 17 to 18 ℃; further preferably 18 ℃.
In the invention, the time of the light culture is 40-50 d; preferably from 42 to 48d; more preferably 44 to 46d; more preferably 45d.
In the invention, the illumination intensity of the light culture is 50-1000 Lux; preferably 200 to 800Lux; more preferably 400 to 600Lux; more preferably 500Lux.
In the invention, the temperature of the light culture is 16-22 ℃; preferably 17 to 21 ℃; further preferably 18 to 20 ℃; more preferably 19 deg.c.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Tunnel microwave irradiation equipment was purchased from the fuhao intelligent equipment south beige ltd.
Example 1
A method for cultivating Cordyceps militaris by using wheat kernel as culture medium comprises the following steps:
(1) Wheat hulling: after the wheat grains are subjected to the first wheat wetting → screening → second wheat wetting → peeling process, the peeling rate is 4 percent, and the removal rate of microorganisms such as storage fungi and the like on the wheat grain cortex reaches 85 percent.
(2) Modification by microwave irradiation: the tunnel type microwave irradiation equipment is adopted, the power of the equipment is 30kw, the microwave frequency is 2450MHz, and the treatment capacity is 200kg/h. Starting the irradiation equipment and preheating. Conveying the wheat kernels to a conveyor belt through a vibrating feeder to perform microwave irradiation, and continuously irradiating for 5min at the temperature of 140 +/-5 ℃.
The thickness of the wheat kernel laying material is 1cm;
(3) Preparing a solid culture medium: and (3) mixing 100g of wheat kernels with water, and filling, wherein the ratio of materials to water is 1.8.
(4) Sterilizing at 121 deg.C for 30min, and cooling.
(5) Inoculation: the concentration of the cordyceps militaris original seed liquid is 10g/L, and the strain dilution ratio is 1; the diluted cordyceps militaris strain is inoculated into the solid culture medium, and the volume mass ratio of the diluted cordyceps militaris strain to the wheat kernel is 8mL:100g of the total weight of the mixture;
(6) Culturing: placing the inoculated cordyceps militaris culture medium into a culture room for dark culture, and culturing for 3 days at 16 ℃ until the surface of the cordyceps militaris culture medium is completely covered by cordyceps militaris mycelia; after completion of the dark culture, 50Lux light was applied and cultivation management was performed at 16 ℃.
(7) Harvesting: and harvesting at the right time 55 days after inoculation.
Example 2
A method for cultivating Cordyceps militaris by using wheat kernel as culture medium comprises the following steps:
(1) Wheat hulling: after the wheat grains are subjected to the first wheat wetting → screening → second wheat wetting → peeling process, the peeling rate is 10 percent, and the removal rate of microorganisms such as storage fungi and the like of the wheat grain cortex reaches 90 percent.
(2) Modification by microwave irradiation: the tunnel type microwave irradiation equipment is adopted, the power of the equipment is 30kw, the microwave frequency is 2450MHz, and the treatment capacity is 200kg/h. Starting the irradiation equipment and preheating. Conveying the wheat kernels to a conveyor belt through a vibrating feeder to perform microwave irradiation, and continuously irradiating for 6min at the temperature of 150 +/-5 ℃.
The thickness of the wheat kernel laying material is 2cm;
(3) Preparing a solid culture medium: and (3) mixing 100g of wheat kernels with water, and filling, wherein the ratio of materials to water is 1.
(4) And (3) sterilization and cooling: sterilizing at 110 deg.C for 30min; cooling and then standby.
(5) Inoculation: the concentration of the cordyceps militaris original seed liquid is 10g/L, and the strain dilution ratio is 1; the diluted cordyceps militaris strain is inoculated into the solid culture medium, and the volume mass ratio of the diluted cordyceps militaris strain to the wheat kernel is 10mL:100g of the total weight of the feed;
(6) Culturing: placing the inoculated cordyceps militaris culture medium into a culture room for dark culture, and culturing for 7 days at the temperature of 19 ℃ until the surface of the cordyceps militaris culture medium is completely covered by cordyceps militaris mycelia; after completion of the dark culture, the culture was managed at 22 ℃ by applying 1000Lux light.
(7) Harvesting: and harvesting at the right time 55 days after inoculation.
Example 3
A method for cultivating Cordyceps militaris by using wheat kernel as culture medium comprises the following steps:
(1) Wheat hulling: after the wheat grains are subjected to the first wheat wetting → screening → second wheat wetting → peeling process, the peeling rate is 6 percent, and the removal rate of microorganisms such as storage fungi and the like on the wheat grain cortex reaches 85 percent.
(2) Modification by microwave irradiation: the tunnel type microwave irradiation equipment is adopted, the power of the equipment is 30kw, the microwave frequency is 2450MHz, and the treatment capacity is 200kg/h. Starting the irradiation equipment and preheating. Conveying the wheat kernels to a conveyor belt through a vibrating feeder to perform microwave irradiation, and continuously irradiating for 11min at the temperature of 140 +/-5 ℃. The thickness of the wheat kernel laying material is 1.5cm;
(3) Preparing a solid culture medium: and (3) mixing 100g of wheat kernels with water, and filling, wherein the ratio of materials to water is 1.
(4) And (3) sterilization and cooling: sterilizing at 122 deg.C for 30min; cooling and then standby.
(5) Inoculation: the concentration of the cordyceps militaris original seed liquid is 10g/L, and the strain dilution ratio is 1; and (3) inoculating diluted cordyceps militaris strain into the solid culture medium, wherein the volume mass ratio of the diluted cordyceps militaris strain to the wheat kernel is 9mL:100g of the total weight of the mixture;
(6) Culturing: placing the inoculated cordyceps militaris culture medium into a culture room for dark culture, and culturing for 5 days at 18 ℃ until the surface of the cordyceps militaris culture medium is completely covered by cordyceps militaris mycelia; after completion of the dark culture, 500Lux light was applied thereto, and cultivation management was performed at a temperature of 19 ℃.
(7) Harvesting: and harvesting timely 55 days after inoculation.
Comparative example 1
The other method is the same as example 3, except that: the microwave irradiation temperature is 110 deg.C, and the time is 5min.
Comparative example 2
The other method is the same as example 3, except that: the microwave irradiation temperature is 120 deg.C, and the time is 3min.
Comparative example 3
The other method is the same as example 3, except that: the microwave irradiation temperature is 130 deg.C, and the time is 2min.
Experimental example 1
The hardness, viscosity and cordyceps militaris yield of the culture media of examples 1 to 3 and comparative examples 1 to 3 were measured, and the results are shown in tables 1 and 2.
Table 1: medium hardness and viscosity data for examples and comparative examples
Table 2: microwave irradiation parameters and Cordyceps militaris yield per unit of examples and comparative examples
As can be seen from the data in Table 1, the hardness and viscosity of the culture media prepared in examples 1-3 are moderate, which is beneficial to the direct utilization of Cordyceps, and the data in Table 2 can prove that the yield per unit of Cordyceps militaris in examples 1-3 is significantly higher than that in comparative examples 1-3. Therefore, the optimal technical effect can be achieved only by selecting the microwave irradiation parameter setting of the invention.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and amendments can be made without departing from the principle of the present invention, and these modifications and amendments should also be considered as the protection scope of the present invention.
Claims (10)
1. A method for cultivating cordyceps militaris by using wheat kernels as a culture medium is characterized by comprising the following steps:
(1) Performing microwave irradiation modification treatment on peeled wheat kernels to obtain a material 1;
(2) Mixing the material 1 obtained in the step (1) with water to obtain a culture medium, sterilizing the culture medium, and inoculating cordyceps militaris strains for culture.
2. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 1, wherein the peeling rate of the wheat kernels in the step (1) is 4-10%.
3. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 1, wherein the thickness of the wheat kernels subjected to microwave irradiation in the step (1) is 1-2 cm.
4. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 1, wherein the microwave irradiation temperature in the step (1) is 140-150 ℃ and the time is 5-15 min.
5. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 1, wherein the sterilization temperature in the step (2) is 110-121 ℃ and the time is 20-40 min.
6. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 1, wherein the mass ratio of the material 1 to water in the step (2) is 1.8-2.0.
7. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 1, wherein the volume-to-mass ratio of cordyceps militaris strains in the step (2) to the material 1 is 8-10 mL:100g.
8. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 1, wherein the cultivation in the step (2) is divided into dark cultivation and light cultivation.
9. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 8, wherein the dark cultivation time is 3-7 days and the temperature is 16-19 ℃.
10. The method for cultivating cordyceps militaris by using wheat kernels as a culture medium according to claim 8, wherein the light cultivation time is 40-50 days, the illumination intensity is 50-1000 Lux, and the temperature is 16-22 ℃.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101173221A (en) * | 2007-11-09 | 2008-05-07 | 乔善成 | Wheat cultivation Cordyceps militaris method |
| CN103843583A (en) * | 2014-03-06 | 2014-06-11 | 刘彦君 | Industrialized production method of green cordyceps militaris |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101173221A (en) * | 2007-11-09 | 2008-05-07 | 乔善成 | Wheat cultivation Cordyceps militaris method |
| CN103843583A (en) * | 2014-03-06 | 2014-06-11 | 刘彦君 | Industrialized production method of green cordyceps militaris |
Non-Patent Citations (1)
| Title |
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| 罗志刚等: "微波场对小麦淀粉性质的影响", 《化工学报》, vol. 58, no. 11, 15 November 2007 (2007-11-15), pages 2871 - 2875 * |
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