CN112210500B - Method for culturing mushroom fungus spherical mycelium - Google Patents

Method for culturing mushroom fungus spherical mycelium Download PDF

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CN112210500B
CN112210500B CN202011107293.5A CN202011107293A CN112210500B CN 112210500 B CN112210500 B CN 112210500B CN 202011107293 A CN202011107293 A CN 202011107293A CN 112210500 B CN112210500 B CN 112210500B
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mycelium
spherical
culture
culture medium
strain
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CN112210500A (en
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程汉强
张明军
张方方
邹天宝
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Wuhan Diao Pharmaceutical Co ltd
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor

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Abstract

The invention discloses a liquid strain culture medium for mushroom fungus spherical mycelium, which contains arginine salt or arginine;the liquid strain culture medium is prepared from the following raw materials in percentage by weight: wherein the water-soluble starch is 2-3%, the yeast powder is 0.5-1.5%, the arginine salt is 0.2-2.0%, the potassium dihydrogen phosphate is 0.05-0.35%, the magnesium sulfate is 0.05-0.20%, the biotin is 0.0001-0.0005%, the vitamin B is 0.05-0.0005%10.005-0.020% of water, and the balance being 100%. The invention can form seeds with large quantity of spherical mycelium, high ratio of spherical mycelium to the ratio of spherical mycelium and high toughness and compactness of spheres by selectively adding arginine salt or arginine, the fermentation inoculation amount can be large, the spherical mycelium has strong mechanical shearing resistance and is not easy to break, the invention can adapt to looser fermentation hardware conditions, and the small and medium spherical mycelium in the seeds can directly continue to grow in a spherical shape without worrying about the control of the balling condition of the filamentous mycelium, thereby greatly simplifying the production and shortening the fermentation time.

Description

Method for culturing mushroom fungus spherical mycelium
Technical Field
The invention relates to the technical field of biological fermentation, in particular to a culture method of mushroom fungus spherical mycelium.
Background
The liquid fermentation culture of fungi (including mycelial fungi and large edible fungi) mycelium is the application embodiment of modern biological fermentation technology in fungus culture, and has been widely used in the fields of pharmacy, food, cosmetics industry and the like as widely applied industrial microorganisms. Liquid fermentation cultures typically include slant seed culture, liquid seed culture, and fermentation culture stages. Currently, most studies are made on the fermentation culture stage of mushroom (large edible fungi) mycelium, but the attention on seed preparation is relatively less, and the studies on the formulation of seed culture medium specifically for spherical mycelium fermentation are more rare.
At present, in the process of liquid fermentation culture of fungal mycelia, different mycelium morphologies, such as filamentous, silky, spherical and the like, can be generated due to different conditions during culture. The amount of certain metabolites varies greatly for different forms of mycelium, and in addition, the conditions for formation of spherical mycelium, including the medium formulation, culture conditions and fermentation process control parameters, vary greatly depending on the species of the fungus. Therefore, conditions and control methods suitable for culturing certain strains are researched and found in a targeted manner, and the method has very important practical significance and application value.
In the actual research and production process, liquid seeds for fermentation are used for obtaining a large amount of activated mycelia with strong multiplication capacity, and at the moment, if the liquid seeds are not subjected to targeted regulation and control, the mycelia are mostly filamentous in shape, and after the liquid seeds are inoculated into a fermentation tank, the mycelia can quickly grow in the fermentation tank. In the seed stage, the hyphae are generally very thin and weak, the hypha cell wall is thin, the hyphae are in a shape of multiple filaments and clusters, the ball is not formed or the ball forming proportion is low, even if the ball is formed, the ball compactness (density) is low, and the ball spike is soft and not firm. At this time, when fermentation culture is performed after inoculation, if the inoculation amount is small, the culture time is long, the balling is less, and the efficiency is low; if the inoculation amount is large, the mycelium is difficult to form balls, spherical aberration or silky clusters; in addition, the influence of the high aeration flow rate, the aeration mode, the stirring mode and the stirring speed on the factors of shearing and crushing of the mycelium and the mycelium pellets and the like easily causes difficult mycelium balling and influences the final fermentation effect. Therefore, based on the differences of various properties such as compactness, toughness and shear resistance of seed hyphae, the ventilation quantity, the ventilation mode, the stirring speed and the like need to be strictly controlled and timely adjusted according to the fermentation condition during large-scale fermentation. Moreover, mycelium is difficult to form balls, spherical aberration or silky clusters due to other reasons during fermentation, which affects final fermentation. Often, the reason is that the fermentation process is not controlled in place and is regulated more tightly, so that the control cost is greatly increased and is doubled and half done.
Disclosure of Invention
The invention provides a culture method of mushroom fungus spherical mycelium, which aims to solve the technical problems.
In order to achieve the purpose, the technical scheme of the invention is realized as follows:
the invention also relates to a culture method of the mushroom fungus spherical mycelium, which comprises the following steps:
step 1, filling the culture components into a container according to a formula proportion, stirring and dissolving completely, and subpackaging into 1000ml of triangular flasks with the filling amount of 400 ml;
step 2, sterilizing for 30 minutes by moist heat steam at 121 ℃;
step 3, cooling to normal temperature to inoculate the solid strain in the eggplant-shaped bottle;
step 4, smashing the solid strain in the eggplant-shaped bottle into bacterial mud in advance by using an aseptic smashing device, and then inoculating the bacterial mud into a triangular flask of a liquid strain culture medium;
and 5, placing the inoculated triangular flask in a constant-temperature shaking table for culturing at the culture temperature: 25 +/-1 ℃; culture rotation speed: 100-; culturing for 10-12 days;
the liquid strain culture medium contains arginine salt or arginine; the liquid strain culture medium is prepared from the following raw materials in percentage by weight: wherein the water-soluble starch is 2-3%, the yeast powder is 0.5-1.5%, the arginine salt is 0.2-2.0%, the potassium dihydrogen phosphate is 0.05-0.35%, the magnesium sulfate is 0.05-0.20%, the biotin is 0.0001-0.0005%, the vitamin B is 0.05-0.0005%10.005-0.020% of water, and the balance being 100%.
The step 1 to the step 3 are methods for preparing a strain culture medium; the step 4 to the step 5 are the culture method of liquid strains.
Compared with the prior art, the invention has the advantages that:
the invention can form seeds with large quantity of spherical hyphae per unit volume, high occupation ratio of the spherical hyphae and high toughness and compactness of spheres by selectively adding arginine salt or arginine. The fermentation inoculation amount of the spherical mycelium seeds can be large, the spherical mycelium is strong in mechanical shearing resistance and not easy to break, and can adapt to more loose fermentation hardware conditions, such as: high ventilation rate, violent ventilation mode and/or stirring mode, higher stirring speed and the like. The small spherical mycelium in the seed can grow continuously in spherical form without worrying about the control of the balling condition of the filamentous mycelium, thereby greatly simplifying production and shortening fermentation time.
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Detailed Description
The technical solutions of the embodiments of the present invention will be described clearly and completely below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention relates to a method for culturing mushroom fungus spherical mycelium,
the invention also discloses a culture method for the spherical mycelium of the mushroom, which comprises a preparation method of a strain culture medium and a culture method of liquid strains;
the preparation method of the strain culture medium comprises the following steps:
step 1, filling the culture components into a container according to a formula proportion, stirring and dissolving completely, and subpackaging into 1000ml of triangular flasks with the filling amount of 400 ml;
step 2, sterilizing for 30 minutes by moist heat steam at 121 ℃;
step 3, cooling to normal temperature to inoculate the solid strain in the eggplant-shaped bottle;
the culture method of the liquid strain comprises the following steps:
step 4, smashing the solid strain in the eggplant-shaped bottle into bacterial mud in advance by using an aseptic smashing device, and then inoculating the bacterial mud into a triangular flask of a liquid strain culture medium;
and 5, placing the inoculated triangular flask in a constant-temperature shaking table for culturing at the culture temperature: 25 +/-1 ℃; culture rotation speed: 100-; culturing for 10-12 days;
the liquid strain culture medium contains arginine salt or arginine; the liquid strain culture medium is prepared from the following raw materials in percentage by weight: wherein the water-soluble starch is 2-3%, the yeast powder is 0.5-1.5%, the arginine salt is 0.2-2.0%, the potassium dihydrogen phosphate is 0.05-0.35%, the magnesium sulfate is 0.05-0.20%, the biotin is 0.0001-0.0005%, the vitamin B is 0.05-0.0005%10.005-0.020% of water, and the balance being 100%.
The specific embodiment of the invention is as follows:
the technical solution of the present invention is further described with reference to the following examples.
EXAMPLE 1
The strain culture medium for culturing spherical mycelium by liquid fermentation of shiitake mushroom comprises the following components in percentage by weight:
2% of water-soluble starch, 0.5% of yeast powder, 0.20% of arginine salt, 0.05% of potassium dihydrogen phosphate, 0.05% of magnesium sulfate, 0.0001% of biotin and vitamin B10.005% and water to 100%;
comprises a preparation method of a strain culture medium and a culture method of liquid strains;
the preparation method of the strain culture medium comprises the following steps:
step 1, filling the culture components into a container according to a formula proportion, stirring and dissolving completely, and subpackaging into 1000ml of triangular flasks with the filling amount of 400 ml;
step 2, sterilizing for 30 minutes by moist heat steam at 121 ℃;
step 3, cooling to normal temperature to inoculate the solid strain in the eggplant-shaped bottle;
the culture method of the liquid strain comprises the following steps:
step 4, smashing the solid strain in the eggplant-shaped bottle into bacterial mud in advance by using an aseptic smashing device, and then inoculating the bacterial mud into a triangular flask of a liquid strain culture medium;
and 5, placing the inoculated triangular flask in a constant-temperature shaking table for culturing at the culture temperature: 25 +/-1 ℃; culture rotation speed: 100-; culturing for 10-12 days.
EXAMPLE 2
The strain culture medium for culturing spherical mycelium by liquid fermentation of shiitake mushroom comprises the following components in percentage by weight:
2.5 percent of water-soluble starch, 1.0 percent of yeast powder, 0.5 percent of arginine salt, 0.25 percent of monopotassium phosphate, 0.15 percent of magnesium sulfate, 0.0002 percent of biotin and vitamin B10.005% and water to 100%.
Comprises a preparation method of a strain culture medium and a culture method of liquid strains;
the preparation method of the strain culture medium comprises the following steps:
step 1, filling the culture components into a container according to a formula proportion, stirring and dissolving completely, and subpackaging into 1000ml of triangular flasks with the filling amount of 400 ml;
step 2, sterilizing for 30 minutes by moist heat steam at 121 ℃;
step 3, cooling to normal temperature to inoculate the solid strain in the eggplant-shaped bottle;
the culture method of the liquid strain comprises the following steps:
step 4, smashing the solid strain in the eggplant-shaped bottle into bacterial mud in advance by using an aseptic smashing device, and then inoculating the bacterial mud into a triangular flask of a liquid strain culture medium;
and 5, placing the inoculated triangular flask in a constant-temperature shaking table for culturing at the culture temperature: 25 +/-1 ℃; culture rotation speed: 100-; culturing for 10-12 days.
EXAMPLE 3
The strain culture medium for culturing spherical mycelium by liquid fermentation of shiitake mushroom comprises the following components in percentage by weight:
2.5 percent of water-soluble starch, 1.0 percent of yeast powder, 1.0 percent of arginine salt, 0.25 percent of monopotassium phosphate, 0.25 percent of magnesium sulfate, 0.0003 percent of biotin and vitamin B10.01 percent and water to 100 percent.
Comprises a preparation method of a strain culture medium and a culture method of liquid strains;
the preparation method of the strain culture medium comprises the following steps:
step 1, filling the culture components into a container according to a formula proportion, stirring and dissolving completely, and subpackaging into 1000ml of triangular flasks with the filling amount of 400 ml;
step 2, sterilizing for 30 minutes by moist heat steam at 121 ℃;
step 3, cooling to normal temperature to inoculate the solid strain in the eggplant-shaped bottle;
the culture method of the liquid strain comprises the following steps:
step 4, smashing the solid strain in the eggplant-shaped bottle into bacterial mud in advance by using an aseptic smashing device, and then inoculating the bacterial mud into a triangular flask of a liquid strain culture medium;
and 5, placing the inoculated triangular flask in a constant-temperature shaking table for culturing at the culture temperature: 25 +/-1 ℃; culture rotation speed: 100-; culturing for 10-12 days.
EXAMPLE 4
The strain culture medium for culturing spherical mycelium by liquid fermentation of shiitake mushroom comprises the following components in percentage by weight:
3% of water-soluble starch, 1.5% of yeast powder, 2.0% of arginine salt, 0.35% of potassium dihydrogen phosphate, 0.20% of magnesium sulfate, 0.0005% of biotin and vitamin B10.02% and water to 100%.
Comprises a preparation method of a strain culture medium and a culture method of liquid strains;
the preparation method of the strain culture medium comprises the following steps:
step 1, filling the culture components into a container according to a formula proportion, stirring and dissolving completely, and subpackaging into 1000ml of triangular flasks with the filling amount of 400 ml;
step 2, sterilizing for 30 minutes by moist heat steam at 121 ℃;
step 3, cooling to normal temperature to inoculate the solid strain in the eggplant-shaped bottle;
the culture method of the liquid strain comprises the following steps:
step 4, smashing the solid strain in the eggplant-shaped bottle into bacterial mud in advance by using an aseptic smashing device, and then inoculating the bacterial mud into a triangular flask of a liquid strain culture medium;
and 5, placing the inoculated triangular flask in a constant-temperature shaking table for culturing at the culture temperature: 25 +/-1 ℃; culture rotation speed: 100-; culturing for 10-12 days.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention in any way, and any person skilled in the art can make many changes or modifications to the equivalent embodiments without departing from the scope of the present invention.

Claims (2)

1. A method for culturing mushroom fungus spherical mycelium is characterized by comprising the following steps: the preparation method of the strain culture medium comprises the following steps:
step 1, filling the culture components into a container according to a formula proportion, stirring and dissolving completely, and subpackaging into 1000ml of triangular flasks with the filling amount of 400 ml;
step 2, sterilizing for 30 minutes by moist heat steam at 121 ℃;
step 3, cooling to normal temperature to inoculate the solid strain in the eggplant-shaped bottle;
step 4, smashing the solid strain in the eggplant-shaped bottle into bacterial mud in advance by using an aseptic smashing device, and then inoculating the bacterial mud into a triangular flask of a liquid strain culture medium;
and 5, placing the inoculated triangular flask in a constant-temperature shaking table for culturing at the culture temperature: 25 +/-1 ℃; culture rotation speed: 100-; culturing for 10-12 days;
the liquid strain culture medium contains arginine salt or arginine; the liquid strain culture medium is prepared from the following raw materials in percentage by weight: wherein the water-soluble starch is 2-3%, the yeast powder is 0.5-1.5%, the arginine salt is 0.2-2.0%, the potassium dihydrogen phosphate is 0.05-0.35%, the magnesium sulfate is 0.05-0.20%, the biotin is 0.0001-0.0005%, the vitamin B is 0.05-0.0005%10.005-0.020% of water, and the balance being 100%.
2. The method for culturing spherical mycelium of Lentinus edodes according to claim 1, wherein: the step 1 to the step 3 are preparation methods of strain culture media; the steps 4 to 5 are the culture method of liquid strains.
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