CN110710669A - Preparation method of sparassis crispa mycelium oral liquid - Google Patents
Preparation method of sparassis crispa mycelium oral liquid Download PDFInfo
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- CN110710669A CN110710669A CN201910937436.6A CN201910937436A CN110710669A CN 110710669 A CN110710669 A CN 110710669A CN 201910937436 A CN201910937436 A CN 201910937436A CN 110710669 A CN110710669 A CN 110710669A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/60—Cultivation rooms; Equipment therefor
- A01G18/69—Arrangements for managing the environment, e.g. sprinklers
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Nutrition Science (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the technical field of food processing, and discloses a preparation method of a sparassis crispa mycelium oral liquid, which is prepared by converting various macromolecular active nutrient components in sparassis crispa mycelium into small molecular nutrients which are easier to absorb by a human body through fermentation and enzymolysis methods. The invention avoids the defect of short storage time, so that the final product is easier to store, and the safety of the product is ensured. Can also be applied to the fields of beauty products, medical nutrition foods and nutrition supplements and the production and processing of the beauty products and the medical nutrition foods and the nutrition supplements.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a preparation method of an Sparassis crispa mycelium oral liquid.
Background
Sparassis crispa, also known as Sparassis crispa, Latin is named as: sparassis crispa is named as giant hydrangea, which is the medium to large fleshy meat of the sporocarp of the genus Sparassis belonging to the order Aphyllophorales, the family Sparassaceae, the genus Sparassis, the meat is branched from a thick handle, and numerous zigzag lobes are formed at the branch ends and are shaped like giant hydrangea. The Sparassis crispa has relatively rare resource reserves, and is known as the king of 'Wan mushroom' as a rare dual-purpose edible and medicinal mushroom which is very rare in the world.
According to the analysis of the Japanese food analysis center, each 100g of sparassis crispa contains beta-glucan which is up to 43.6g and is 3 ~ 4 times higher than that of lucid ganoderma and agaricus blazei murill, so that the beta-glucan contained in the sparassis crispa is the most important of mushrooms, the glucan is polysaccharide formed by polymerizing glucose monomers and is divided into alpha-type and beta-type, the alpha-glucan is a main source of body energy, and the beta-glucan has no biological activity, and medical researches prove that the polysaccharide has various functions of immunoregulation, anti-tumor, anti-inflammation, antivirus, antioxidation, antiradiation, blood sugar reduction, blood fat reduction, liver protection and the like, and the majority of the polysaccharide with the anti-tumor activity is beta (1-3) D glucan with beta (1-6) glycosidic bond branches, and the researches prove that the beta (1-3) D from fungi generally has the tumor inhibiting effect, and the beta (1-3) D) glucan has the good tumor regulating effect, and the beta (1-3) D) glucan has the good hematopoietic function.
Secondly, since Sparassis crispa contains a large amount of antioxidant substances, free radicals in human bodies can cause oxidative damage to biological cells to cause aging of the organisms, destroy the disease resistance and defense capability of the organisms, and cause chronic diseases such as cancers, cardiovascular diseases and the like. Nowadays, the antioxidant capacity of natural food has been generally regarded by people, wherein superoxide dismutase SOD can effectively remove free radicals in vivo, prevent active oxygen from damaging the body, and has antioxidant and anti-aging effects. According to the determination, the content of superoxide dismutase in the sparassis crispa is at the head of various edible fungi.
And thirdly, the sparassis crispa contains a large amount of vitamins and minerals, the sparassis crispa contains vitamin C and vitamin E, the content of the vitamin E is in the front of the food of the resident bacteria algae, and the vitamins have the antioxidation effect. Sparassis crispa also contains ergosterol, can be converted into vitamin D under sunlight and ultraviolet irradiation, can promote calcium and phosphorus absorption, is beneficial for bone formation, and can prevent rickets in children, osteoporosis in adults and osteomalacia. The Sparassis crispa contains quite high potassium element and low sodium content, and the high-potassium low-sodium food has diuretic effect and is very beneficial to hypertension patients.
Based on the fact that the content of superoxide dismutase and the content of vitamin E in sparassis crispa are all in front of the food of the resident bacteria algae, the sparassis crispa seaweed food has obvious effects on preventing active oxygen from damaging organisms, resisting oxidation, resisting aging and the like. Therefore, the production process of an antioxidant and anti-aging sparassis crispa oral product is urgently needed.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a preparation method of an oral liquid of sparassis crispa mycelium. Can also be used in the fields of production and processing of cosmetic products, medical nutritional foods and nutritional supplements
In order to achieve the purpose, the invention adopts the following technical scheme
A preparation method of sparassis crispa mycelium oral liquid is prepared by converting various macromolecular active nutrient components in sparassis crispa mycelium into micromolecular nutrients which are easier to absorb by a human body through fermentation and enzymolysis methods, and comprises the following specific steps:
1) preparing a mother strain of sparassis crispa, namely taking a part of sparassis crispa fruiting body which is thick in meat and not fully stretched, washing the part with sterile water, and picking a small tissue block to be transplanted on a PDA inclined plane by using a scalpel in a sterile box; culturing in a constant temperature incubator at 25 deg.C for 11-13 days until the new mycelia grow vigorously, inoculating the new mycelia on PDA plate culture medium, performing purification culture at 25 deg.C, inoculating the obtained purified mycelia in a PDA slant test tube under aseptic condition, culturing in a constant temperature incubator at 25 deg.C in dark, and storing in a refrigerator at 4 deg.C after mycelia are fully grown to obtain Sparassis crispa stock;
2) and preparing a sparassis crispa liquid strain:
a. weighing potatoes, peeling 200g, 20g of glucose and 2g of peptone;
b. slicing potato, adding 1L water, boiling for 20 min, and filtering with gauze to obtain juice;
c. adding the glucose and peptone into potato juice, and diluting to a constant volume of 1L with purified water to obtain a liquid strain culture medium;
d. subpackaging the liquid strain culture medium into 250ml of suction bottles, wherein the loading amount is 100ml, plugging the bottle mouth with a cotton plug, and sterilizing for 30 minutes at the temperature of 121 ℃ under the pressure of 0.1 Mpa;
e. inoculating 6-8 sparassis crispa mother strain blocks into a liquid culture medium naturally cooled to room temperature under the aseptic condition, and statically culturing for 24 hours at the temperature of 24-26 ℃; culturing on electromagnetic bed at 24-26 deg.C for 7 days after mycelia germinate; when the culture solution is clear and transparent, suspending a large amount of mycelium pellets to obtain the Sparassis crispa liquid strain;
3) and preparing Sparassis crispa mycelium fermentation liquor:
a. preparation of sparassis crispa liquid culture medium: weighing 750g of starch, 500g of corn flour, 120g of peptone, 1200g of glucose and 50L of purified water to prepare a hydrangea sparassis liquid culture medium;
b. sterilizing a sparassis crispa liquid culture medium: placing the liquid culture medium in a liquid strain fermentation tank of the edible fungi, opening a starting switch, slightly opening an exhaust valve when the temperature reaches 100 ℃, and automatically timing for 40 minutes when the temperature reaches 121 ℃; wherein the air filter element is sterilized in advance, and then the air filter element is mounted, and the air pump is opened to blow the filter element; in the sterilization process, in order to avoid the valve from becoming a sterilization dead angle, the materials are discharged once every 15 minutes in the pressure maintaining process, three times in total, and 3 minutes are discharged each time;
c. cooling the liquid culture medium: opening a water inlet valve of the interlayer after sterilization, and cooling by introducing water; connecting a filter air inlet pipe to a fermentation tank air inlet valve under the protection of flame, and opening the air inlet valve to supply air into the tank when the pressure of the fermentation tank is reduced to be below 0.1MPa and the air pressure of an air inlet system is higher than the pressure in the tank by more than 0.02 MPa; when the air inlet valve is opened, the material liquid is prevented from flowing backwards slowly, so that pollution is caused; when the temperature in the tank is reduced to below 28 ℃, the inoculation operation can be carried out;
d. inoculating the Sparassis crispa liquid strain: two persons are required to cooperate during inoculation; firstly, making a circle by using thick iron wires according to the size of an inoculation port of a fermentation tank, wrapping the iron wire circle with absorbent cotton, and soaking the iron wire circle with 95% alcohol for later use; preparing a wet towel, placing the wet towel beside the inoculation port, and using the wet towel after extinguishing fire; the inoculation method comprises the following steps:
(1) wiping the outer cover and the periphery of the inoculation port twice by using 75% alcohol;
(2) opening an exhaust valve, and when the pressure in the tank is reduced to be close to zero, quickly closing the exhaust valve and igniting a fire ring;
(3) under the protection of flame, opening an inoculation port cover, quickly pouring the sparassis crispa liquid strain in the shake flask into the tank, and screwing the inoculation port cover;
(4) after the pressure in the tank is increased, a wet towel is used for extinguishing the fire ring, an exhaust valve is opened, and the pressure in the tank is adjusted to be 0.02 ~ 0.04.04 MPa and enters a culture state;
e. culturing Sparassis crispa hypha fermentation liquor at the constant temperature of 25 ℃ for 24 hours, sampling from an inoculation port every two hours, observing the germination and growth conditions of strains, and culturing in a fermentation tank for 5 days, wherein the bacterial liquid has no mixed bacteria pollution, pure color and no peculiar smell, only has the fragrance of one hypha, the quantity of the bacterial balls accounts for about 80% of ~ 90% of the total volume, and the Sparassis crispa hypha fermentation liquor can be fermented;
4) and preparing the sparassis crispa oral liquid:
a. an enzymolysis process: adding cellulase, protease and pectinase into Sparassis crispa mycelium fermentation broth for enzymolysis, controlling the temperature at 45-55 deg.C, pH6.5-7, and the enzymolysis time for 1.5 hr;
b. and (3) homogenizing and concentrating: treating the zymolyzed Sparassis crispa mycelium fermentation liquor by colloid mill with a homogenizer, adjusting the clearance between stator and rotor of the colloid mill to 10-50 μm, and crushing mycelium in the fermentation liquor, wherein the colloid mill flow is 1-5 ton/hr;
c. treating with a high-pressure homogenizer, adjusting the pressure of the high-pressure homogenizer to 20-60MPa, the high-pressure homogenizing flow rate to 0.5-3 ton/h, and the wall breaking rate of the mycelium detected by a microscope is 100%; after homogenizing, concentrating to make the solid content reach 30-55%;
d. filling and sterilizing the sparassis crispa oral liquid: filling the treated Sparassis crispa mycelium fermentation liquid, and sterilizing at 135 deg.C for 20 s to obtain oral liquid.
Due to the adoption of the technical scheme, the invention has the following advantages:
the method solves the market blank of the Sparassis crispa nutritional oral liquid, obtains a novel oral liquid product suitable for the public nutritional supplement through the optimization of the fermentation technology and the updating of the enzymolysis technology, and continuously innovates the production process, thereby providing a healthier nutritional supplement which is easy to absorb for consumers.
The invention belongs to pure fermentation type, avoids the addition of various food additives, simultaneously converts various macromolecular active nutrient components in the Sparassis crispa mycelium into micromolecular nutrients which are easier to absorb by human bodies through enzymolysis at the later stage of submerged fermentation, and the Sparassis crispa oral liquid is prepared according to aseptic operation rules strictly in the preparation process and through instantaneous high-temperature sterilization at last, so that the product is safer and healthier. The invention has the following advantages:
1. according to the invention, the natural fermentation type nutrient oral liquid is prepared by researching the biological fermentation process of the sparassis crispa and recording various physical and biological indexes in the fermentation process, and the safe and healthy edible fungus oral liquid is obtained according to the aseptic operation requirement strictly without adding any chemical in the preparation process.
2. The invention adopts enzymolysis purification process in the preparation process, thereby fully releasing the nutrient substances in the edible fungus cells, being easier for human body to absorb, and having certain nutrition supplement and health care effects for specific people.
3. The invention adopts the instant high-temperature inactivation method in the later stage of the manufacturing process, thereby avoiding the defect of short storage time, leading the final product to be easier to store and ensuring the safety of the product.
The invention can also be applied to beauty products, medical nutrition food and nutrition supplements, and the production and processing fields thereof, etc.
Detailed Description
A preparation method of sparassis crispa mycelium oral liquid is prepared by converting various macromolecular active nutrient components in sparassis crispa mycelium into micromolecular nutrients which are easier to absorb by a human body through fermentation and enzymolysis methods, and comprises the following specific steps:
1) and preparing a sparassis crispa mother strain:
taking the thick and incompletely stretched positions of Sparassis crispa fruiting body, washing with sterile water, and picking a small piece of tissue with a scalpel in a sterile box to be transplanted on the inclined surface of PDA. Culturing in a constant-temperature incubator at 25 ℃ for about 12 days until the new mycelia grow vigorously, inoculating the new mycelia on a PDA (PDA dextrose agar) plate culture medium, performing purification culture at 25 ℃, inoculating the obtained purified mycelia in a PDA slant test tube under aseptic conditions, performing constant-temperature dark culture at 25 ℃, and storing in a refrigerator at 4 ℃ after the mycelia grow to full length to obtain a Sparassis crispa stock seed;
2) and preparing a sparassis crispa liquid strain:
weighing potatoes, peeling 200g, glucose 20g and peptone 2 g;
b, slicing the potatoes, adding 1L of water, boiling for 20 minutes, and filtering with gauze to obtain juice for later use;
c, adding the glucose and the peptone into the potato juice, and fixing the volume to 1L by using purified water to obtain a liquid strain culture medium;
d, subpackaging the liquid strain culture medium into a 250ml suction flask, wherein the loading amount is 100ml, the bottle mouth is plugged by a cotton plug, and the sterilization is carried out for 30 minutes under the condition of high pressure steam of 0.1Mpa and 121 ℃;
e, under the aseptic condition, naturally cooling to room temperature in a liquid culture medium, inoculating 6-8 sparassis crispa mother strain blocks, and statically culturing for 24 hours at the temperature of 24-26 ℃; culturing on electromagnetic bed at 24-26 deg.C for 7 days after mycelia germinate; when the culture solution is clear and transparent, suspending a large amount of mycelium pellets to obtain the Sparassis crispa liquid strain;
3) and preparing Sparassis crispa mycelium fermentation liquor:
a, preparation of a sparassis crispa liquid culture medium: 750g of starch, 500g of corn flour, 120g of peptone, 1200g of glucose and 50L of purified water are weighed to prepare the sparassis crispa liquid culture medium.
b, sterilizing a sparassis crispa liquid culture medium: and (3) placing the liquid culture medium in an edible fungus liquid strain fermentation tank, opening a starting switch, slightly opening an exhaust valve when the temperature reaches 100 ℃, and automatically timing for 40 minutes when the temperature reaches 121 ℃. The air filter element is sterilized in advance, and then the air pump is opened to blow the air filter element. In the sterilization process, in order to avoid the valve from becoming a sterilization dead angle, the materials are discharged once every 15 minutes in the pressure maintaining process, three times in total, and 3 minutes are discharged every time.
c, cooling the liquid culture medium: and opening the interlayer water inlet valve to be filled with water for cooling after the sterilization is finished. Connecting a filter air inlet pipe to a fermentation tank air inlet valve under the protection of flame, and opening the air inlet valve to supply air to the tank when the pressure of the fermentation tank is reduced to be below 0.1MPa and the air pressure of an air inlet system is higher than the pressure in the tank by more than 0.02 MPa. When the air inlet valve is opened, the material liquid is slowly prevented from flowing backwards to cause pollution. The inoculation operation can be carried out when the temperature in the tank is reduced to below 28 ℃.
Firstly, making a circle by using thick iron wires according to the size of an inoculation port of a fermentation tank, wrapping absorbent cotton on the iron wire, soaking the iron wire by using 95% alcohol for standby, preparing a wet towel, placing the wet towel beside the inoculation port (for extinguishing fire), and inoculating, namely, 1, wiping the outer cover and the periphery of the inoculation port twice by using 75% alcohol, 2, opening an exhaust valve, quickly closing the exhaust valve and igniting a fire ring when the pressure in the tank is reduced to be close to zero, 3, opening an inoculation port cover under the protection of flame, quickly pouring the liquid strain of the sparassis crispa in a shake flask into the tank, screwing the inoculation cover 4, extinguishing the fire ring by using the wet towel after the pressure in the tank is increased, opening the exhaust valve, and adjusting the pressure in the tank to be 0.02 MPa 0.02 ~ 0.04.04 to enter a culture state.
And e, culturing the Sparassis crispa hypha fermentation liquor at the constant temperature of 25 ℃ for 24 hours, sampling from the inoculation port every two hours, observing the germination and growth conditions of the strains, and culturing in a fermentation tank for 5 days, wherein the bacterial liquid has no mixed bacteria pollution, pure color and no peculiar smell, the bacterial liquid has the fragrance of only one hypha, the quantity of the bacterial balls accounts for about 80% of ~ 90% of the total volume, and the Sparassis crispa hypha fermentation liquor can be fermented.
4) And preparing the sparassis crispa oral liquid:
a. enzymolysis of the sparassis crispa oral liquid: adding cellulase, protease and pectinase into Sparassis crispa mycelium fermentation liquid for enzymolysis together, controlling the temperature at 45-55 deg.C, pH6.5-7, and the enzymolysis time for 1.5 hr.
b. Homogenizing and concentrating the sparassis crispa oral liquid: treating the zymolyzed Sparassis crispa mycelium fermentation liquor by colloid mill with a homogenizer, adjusting the clearance between stator and rotor of the colloid mill to 10-50 μm, and crushing mycelium in the fermentation liquor, wherein the colloid mill flow is 1-5 ton/hr; c. treating with high pressure homogenizer, adjusting pressure of the high pressure homogenizer to 20-60MPa, high pressure homogenizing flow rate to 0.5-3 ton/hr, and microscopic wall breaking rate of mycelium to 100%; after homogenizing, concentrating to make the solid content reach 30-55%.
d. Filling and sterilizing the sparassis crispa oral liquid: filling the treated Sparassis crispa mycelium fermentation liquid, and sterilizing at 135 deg.C for 20 s to obtain oral liquid.
The invention belongs to a pure fermentation type, which is a method for converting various macromolecular active nutrient components in sparassis crispa mycelium into micromolecular nutrients which are easier to be absorbed by human bodies, the sparassis crispa mycelium is subjected to enzymolysis and crushing of cell walls under the action of cellulase and pectinase so as to fully release nutrient substances in cells, meanwhile, various macromolecular nutrient substances in cells are subjected to enzymolysis into micromolecular nutrient substances under the multiple actions of the cellulase, the pectinase and the protease, for example, macromolecular proteins are subjected to enzymolysis into micromolecular polypeptides and amino acids by protease, and later, the feed liquid is more refined under the triple actions of squeezing, strong impact and decompression expansion of the oral liquid through the use of a high-pressure homogenizer so as to be more beneficial to the absorption of human bodies and improve the absorption rate of human bodies, meanwhile, the layering of the oral liquid is effectively reduced, the appearance of the oral liquid is improved, the color is clearer, the, the mouthfeel is more mellow. The invention avoids the addition of various food additives, simultaneously converts various macromolecular active nutrient components in the sparassis crispa mycelium into micromolecular nutrients which are easier to absorb by human bodies through enzymolysis at the later stage of submerged fermentation, and the sparassis crispa oral liquid strictly follows the aseptic operation rules in the preparation process, and is finally sterilized at instantaneous high temperature through the preparation process, so that the product is safer and healthier.
The invention can also be applied to beauty products, medical nutrition food and nutrition supplements, and the production and processing fields thereof, etc.
Claims (1)
1. A preparation method of sparassis crispa mycelium oral liquid is characterized by comprising the following steps: the preparation method is characterized in that various macromolecular active nutrient components in the sparassis crispa mycelium are converted into micromolecular nutrients which are easier to absorb by a human body through fermentation and enzymolysis methods, and the preparation method comprises the following specific steps:
1) preparing a mother strain of sparassis crispa, namely taking a part of sparassis crispa fruiting body which is thick in meat and not fully stretched, washing the part with sterile water, and picking a small tissue block to be transplanted on a PDA inclined plane by using a scalpel in a sterile box; culturing in a constant temperature incubator at 25 deg.C for 11-13 days until the new mycelia grow vigorously, inoculating the new mycelia on PDA plate culture medium, performing purification culture at 25 deg.C, inoculating the obtained purified mycelia in a PDA slant test tube under aseptic condition, culturing in a constant temperature incubator at 25 deg.C in dark, and storing in a refrigerator at 4 deg.C after mycelia are fully grown to obtain Sparassis crispa stock;
2) and preparing a sparassis crispa liquid strain:
a. weighing potatoes, peeling 200g, 20g of glucose and 2g of peptone;
b. slicing potato, adding 1L water, boiling for 20 min, and filtering with gauze to obtain juice;
c. adding the glucose and peptone into potato juice, and diluting to a constant volume of 1L with purified water to obtain a liquid strain culture medium;
d. subpackaging the liquid strain culture medium into 250ml of suction bottles, wherein the loading amount is 100ml, plugging the bottle mouth with a cotton plug, and sterilizing for 30 minutes at the temperature of 121 ℃ under the pressure of 0.1 Mpa;
e. inoculating 6-8 sparassis crispa mother strain blocks into a liquid culture medium naturally cooled to room temperature under the aseptic condition, and statically culturing for 24 hours at the temperature of 24-26 ℃; culturing on electromagnetic bed at 24-26 deg.C for 7 days after mycelia germinate; when the culture solution is clear and transparent, suspending a large amount of mycelium pellets to obtain the Sparassis crispa liquid strain;
3) and preparing Sparassis crispa mycelium fermentation liquor:
a. preparation of sparassis crispa liquid culture medium: weighing 750g of starch, 500g of corn flour, 120g of peptone, 1200g of glucose and 50L of purified water to prepare a hydrangea sparassis liquid culture medium;
b. sterilizing a sparassis crispa liquid culture medium: placing the liquid culture medium in a liquid strain fermentation tank of the edible fungi, opening a starting switch, slightly opening an exhaust valve when the temperature reaches 100 ℃, and automatically timing for 40 minutes when the temperature reaches 121 ℃; wherein the air filter element is sterilized in advance, and then the air filter element is mounted, and the air pump is opened to blow the filter element; in the sterilization process, in order to avoid the valve from becoming a sterilization dead angle, the materials are discharged once every 15 minutes in the pressure maintaining process, three times in total, and 3 minutes are discharged each time;
c. cooling the liquid culture medium: opening a water inlet valve of the interlayer after sterilization, and cooling by introducing water; connecting a filter air inlet pipe to a fermentation tank air inlet valve under the protection of flame, and opening the air inlet valve to supply air into the tank when the pressure of the fermentation tank is reduced to be below 0.1MPa and the air pressure of an air inlet system is higher than the pressure in the tank by more than 0.02 MPa; when the air inlet valve is opened, the material liquid is prevented from flowing backwards slowly, so that pollution is caused; when the temperature in the tank is reduced to below 28 ℃, the inoculation operation can be carried out;
d. inoculating the Sparassis crispa liquid strain: two persons are required to cooperate during inoculation; firstly, making a circle by using thick iron wires according to the size of an inoculation port of a fermentation tank, wrapping the iron wire circle with absorbent cotton, and soaking the iron wire circle with 95% alcohol for later use; preparing a wet towel, placing the wet towel beside the inoculation port, and using the wet towel after extinguishing fire; the inoculation method comprises the following steps:
(1) wiping the outer cover and the periphery of the inoculation port twice by using 75% alcohol;
(2) opening an exhaust valve, and when the pressure in the tank is reduced to be close to zero, quickly closing the exhaust valve and igniting a fire ring;
(3) under the protection of flame, opening an inoculation port cover, quickly pouring the sparassis crispa liquid strain in the shake flask into the tank, and screwing the inoculation port cover;
(4) after the pressure in the tank is increased, a wet towel is used for extinguishing the fire ring, an exhaust valve is opened, and the pressure in the tank is adjusted to be 0.02 ~ 0.04.04 MPa and enters a culture state;
e. culturing Sparassis crispa hypha fermentation liquor at the constant temperature of 25 ℃ for 24 hours, sampling from an inoculation port every two hours, observing the germination and growth conditions of strains, and culturing in a fermentation tank for 5 days, wherein the bacterial liquid has no mixed bacteria pollution, pure color and no peculiar smell, only has the fragrance of one hypha, the quantity of the bacterial balls accounts for about 80% of ~ 90% of the total volume, and the Sparassis crispa hypha fermentation liquor can be fermented;
4) and preparing the sparassis crispa oral liquid:
a. an enzymolysis process: adding cellulase, protease and pectinase into Sparassis crispa mycelium fermentation broth for enzymolysis, controlling the temperature at 45-55 deg.C, pH6.5-7, and the enzymolysis time for 1.5 hr;
b. and (3) homogenizing and concentrating: treating the zymolyzed Sparassis crispa mycelium fermentation liquor by colloid mill with a homogenizer, adjusting the clearance between stator and rotor of the colloid mill to 10-50 μm, and crushing mycelium in the fermentation liquor, wherein the colloid mill flow is 1-5 ton/hr;
c. treating with a high-pressure homogenizer, adjusting the pressure of the high-pressure homogenizer to 20-60MPa, the high-pressure homogenizing flow rate to 0.5-3 ton/h, and the wall breaking rate of the mycelium detected by a microscope is 100%; after homogenizing, concentrating to make the solid content reach 30-55%;
d. filling and sterilizing the sparassis crispa oral liquid: filling the treated Sparassis crispa mycelium fermentation liquid, and sterilizing at 135 deg.C for 20 s to obtain oral liquid.
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| CN114540202A (en) * | 2020-07-10 | 2022-05-27 | 融和梦(福建)生物科技有限公司 | Preparation method of sparassis crispa dry powder, physiological function activator and application thereof |
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