CN112042466A - Tremella cultivation method for extracting high-molecular tremella polysaccharides - Google Patents

Tremella cultivation method for extracting high-molecular tremella polysaccharides Download PDF

Info

Publication number
CN112042466A
CN112042466A CN202010965818.2A CN202010965818A CN112042466A CN 112042466 A CN112042466 A CN 112042466A CN 202010965818 A CN202010965818 A CN 202010965818A CN 112042466 A CN112042466 A CN 112042466A
Authority
CN
China
Prior art keywords
tremella
temperature
polysaccharide
days
cultivation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202010965818.2A
Other languages
Chinese (zh)
Other versions
CN112042466B (en
Inventor
王宇
杨建卫
冯兴成
孔文文
何勇
孔猛
杨守峰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shandong Tongkang Agricultural Development Co ltd
Original Assignee
Shandong Tongkang Agricultural Development Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shandong Tongkang Agricultural Development Co ltd filed Critical Shandong Tongkang Agricultural Development Co ltd
Priority to CN202010965818.2A priority Critical patent/CN112042466B/en
Publication of CN112042466A publication Critical patent/CN112042466A/en
Application granted granted Critical
Publication of CN112042466B publication Critical patent/CN112042466B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • A01G18/20Culture media, e.g. compost

Abstract

A tremella cultivation method for extracting high-molecular tremella polysaccharide relates to the technical field of tremella cultivation, solves the problem of low molecular weight of tremella polysaccharide obtained by the existing cultivation technology, and comprises the steps of obtaining tremella spores through low-temperature domestication, preparing a novel culture medium, dressing seeds, uniformly mixing lower-layer incense ash hypha and upper-layer tremella hypha in a clean area at a ratio of 1: 2, and then inoculating within 12 hours; optimizing the tremella cultivation process, including tremella strain domestication, novel culture medium preparation, sterilization, inoculation and optimization of culture conditions; after the tremella strain is domesticated at low temperature, the tremella strain is adaptive to the reduction of growth temperature, and cellulase, hemicellulase and polyphenol oxidase secreted by associated fungi cinerea are well controlled in the growth and development process of tremella; in order to optimize the initial growth of the tremella, important raw materials, namely tremella polysaccharide extraction waste residues and bagasse are added into a culture medium, so that the normal growth and development of the tremella at the initial stage are ensured; the tremella polysaccharide extraction waste residue in the culture medium is recycled, and the environmental protection problem of waste residue treatment is solved.

Description

Tremella cultivation method for extracting high-molecular tremella polysaccharides
Technical Field
The invention relates to a tremella cultivation method, in particular to a tremella cultivation method for extracting high-molecular tremella polysaccharide.
Background
Tremella, also known as Tremella fuciformis and Tremella fuciformis, has high nutritive value and medicinal efficacy, is a very precious edible fungus and is a name of 'crown in fungus'. The research and cultivation of white fungus in China is in the leading position all over the world, areas such as the Tongjiang Sichuan and the ancient field of Fujian are used as the genuine producing areas of white fungus, and cut-log (green periploca) cultivation and substitute (bottle cultivation and bag cultivation) cultivation are respectively used as representatives; the yield and quality of the tremella fuciformis planted in the Tongjiang section are the first in China, and 90% of the tremella fuciformis planted in China is used as a substitute for tremella fuciformis planted in a ancient field. The research on tremella mainly focuses on the aspects of biological characteristics, production and cultivation, pest control, processing technology, separation and purification, medicinal functions, economic value, genetic breeding and the like.
The tremella polysaccharide is a novel plant-derived high-efficiency humectant extracted and refined from tremella, is also considered as a plant-derived hyaluronic acid substance, has the molecular weight of 100-150 ten thousand, is the only plant-derived humectant with the molecular weight of more than 100 ten thousand in the cosmetic raw material world so far, and is particularly suitable for cosmetics such as essence, facial masks, cream, emulsion, skin lotion and the like.
In the cosmetic market, the demand of tremella polysaccharide for high molecules is gradually increased, and the situation that the molecular weight is high in innovation continuously appears in the market. At present, 160 ten thousand Da molecular weight tremella polysaccharide appears in the market, the demands of many customers are more than 180 ten thousand Da, and the high molecular weight tremella polysaccharide has a large application market space. However, when the existing tremella raw materials are extracted, the molecular weight of the extracted products is different from 90 to 160 ten thousand Da under the same conditions, and the fact that the raw materials play a very critical role in extracting tremella polysaccharides can be seen.
The analysis shows that the associated bacteria, namely cellulase, hemicellulase and polyphenol oxidase secreted by the cinerea virens in the tremella cultivation process have important influence on the molecular weight of tremella polysaccharides, so that the control of the content of secretion of the cinerea virens becomes a key factor for controlling the molecular weight of the tremella polysaccharides.
Disclosure of Invention
In order to solve the problems, the invention provides a tremella cultivation method for extracting high-molecular tremella polysaccharide, and provides a tremella cultivation method for cultivating high-molecular tremella polysaccharide. In order to achieve the purpose, the invention adopts the technical scheme that: a tremella cultivation method for extracting high-molecular tremella polysaccharide comprises the following steps:
1) domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 22 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving at the temperature of 18-20 ℃;
2) preparing a novel culture medium: 30-50% of wood chips, 10-20% of bran, 10-20% of corncobs, 10-20% of tremella polysaccharide extraction waste residues, 5-10% of bagasse, 1-5% of yeast peptone, 1-2% of gypsum and 1-2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) seed dressing, namely uniformly mixing the lower layer incense ash hypha and the upper layer tremella hypha at a ratio of 1: 2 in a clean area, and inoculating within 12 hours;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
Further, the temperature of the strain domesticated in the step 1) is lower than 3-5 ℃ of the growth environment of the strain before domestication, and the temperature is controlled in the cultivation stage to adjust the growth state of the incense ash hyphae, so that the secreted cellulase, hemicellulase, polyphenol oxidase and the like are controlled, and the molecular weight condition of polysaccharide in tremella is reduced.
Further, the domestication method in the step 1) adopts a constant-temperature incubator for cultivation.
Further, the growth cycle, the leaf diameter, the fresh weight and the dry weight of a single tremella and the molecular weight of extracted tremella polysaccharide are controlled by tremella polysaccharide extraction waste residues, bagasse and yeast peptone in the novel culture medium in the step 2).
Further, the step 3) is carried out by using a high-pressure high-temperature sterilization pot, so that the sterilization time is effectively shortened, and the production efficiency is improved.
Further, the tremella fuciformis strain is prepared in the step 4), seed dressing is carried out, tremella fuciformis is inoculated on a culture medium in a clean area, a small amount of incense ash hyphae are inoculated after the tremella fuciformis is cultured for 10 days, the tremella fuciformis is cultured for 7-10 days at the same temperature, and the tremella fuciformis is uniformly stirred when the incense ash hyphae spread in a whole test tube, so that the original seeds are obtained.
Further, fresh tremella can be directly extracted in the step 8), and drying is carried out at the negative pressure of 55-60 ℃ for ensuring the molecular weight due to the problem of difficult storage.
Compared with the prior art, the invention has the following beneficial effects: optimizing the tremella cultivation process, including tremella strain domestication, novel culture medium preparation, sterilization, inoculation and optimization of culture conditions; after the tremella strain is domesticated at low temperature, the tremella strain is adaptive to the reduction of growth temperature, and cellulase, hemicellulase and polyphenol oxidase secreted by associated fungi cinerea are well controlled in the growth and development process of tremella; in order to optimize the initial growth of the tremella, important raw materials, namely tremella polysaccharide extraction waste residues and bagasse are added into a culture medium, so that the normal growth and development of the tremella at the initial stage are ensured; the tremella polysaccharide extraction waste residues in the culture medium are recycled, so that the environmental protection problem of waste residue treatment is solved; the method can improve the molecular weight of mucopolysaccharide in the tremella and solve the environmental protection problem of tremella polysaccharide waste residue treatment, and is low in cost and suitable for industrial production.
Detailed Description
In order to make the technical solutions of the present invention better understood by those skilled in the art, the present invention is further described below with reference to specific examples.
Example one
1) Culturing Tremella fuciformis strains under aseptic conditions, culturing at the temperature of 23 ℃ accurately, and not performing any domestication treatment on original Tremella fuciformis strains;
2) preparing a novel culture medium: 40% of wood chips, 15% of bran, 15% of corncobs, 15% of tremella polysaccharide extraction waste residues, 8% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) seed dressing, namely uniformly mixing the lower layer incense ash hypha and the upper layer tremella hypha at a ratio of 1: 2 in a clean area, and inoculating within 12 hours;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
Example two
1) Domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 20 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving under the temperature of 15-17 ℃;
2) preparing a novel culture medium: 40% of wood chips, 15% of bran, 15% of corncobs, 15% of tremella polysaccharide extraction waste residues, 8% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) seed dressing, namely uniformly mixing the lower layer incense ash hypha and the upper layer tremella hypha at a ratio of 1: 2 in a clean area, and inoculating within 12 hours;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
EXAMPLE III
1) Domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 22 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving at the temperature of 18-20 ℃;
2) preparing a novel culture medium: 40% of wood chips, 15% of bran, 15% of corncobs, 15% of tremella polysaccharide extraction waste residues, 8% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) seed dressing, namely uniformly mixing the lower layer incense ash hypha and the upper layer tremella hypha at a ratio of 1: 2 in a clean area, and inoculating within 12 hours;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, measuring the molecular weight by a new method for extracting tremella polysaccharide, which is disclosed in patent application No. 201710243322.2;
example four
1) Domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 22 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving at the temperature of 18-20 ℃;
2) preparing a novel culture medium: 40% of wood chips, 15% of bran, 15% of corncobs, 23% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) seed dressing, namely uniformly mixing the lower layer incense ash hypha and the upper layer tremella hypha at a ratio of 1: 2 in a clean area, and inoculating within 12 hours;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
EXAMPLE five
1) Domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 22 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving at the temperature of 18-20 ℃;
2) preparing a novel culture medium: 40% of wood chips, 15% of bran, 15% of corncobs, 15% of tremella polysaccharide extraction waste residues, 8% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) dressing seeds, namely inoculating tremella on a culture medium in a clean area, culturing for 10 days, inoculating a little incense ash hypha, culturing for 7-10 days at the same temperature, and uniformly stirring when the incense ash hypha spreads the whole test tube to obtain stock seeds;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
EXAMPLE six
1) Domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 22 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving at the temperature of 18-20 ℃;
2) preparing a novel culture medium: 40% of wood chips, 15% of bran, 15% of corncobs, 15% of tremella polysaccharide extraction waste residues, 8% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) seed dressing, namely inoculating tremella on a culture medium in a clean area, culturing for 4-6 days, inoculating a proper amount of incense ash hypha, culturing for 7-10 days at the same temperature, and uniformly stirring the lower-layer incense ash hypha and the upper-layer tremella hypha in a ratio of 1: 1 to obtain stock seeds when the incense ash hypha spreads over a whole test tube;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
Results and analysis of the experiments
In each embodiment, 50 groups of tremella is picked randomly, the picked tremella is measured, and the average ear emergence rate, the average fresh weight of each tremella, the average dry weight of each tremella, the molecular weight and the growth condition of each tremella are obtained through statistics. Experimental results data and analysis were as follows:
table one: data on the results of the experiments in the examples
Group of Average ear growth (%) Average fresh weight per flower (g) Average single dry weight (g) Molecular weight (kDa) Problem of growth
Example one 98% 165 41 1657 Is free of
Example two 55% 125 35 1337 Low ear emergence rate
EXAMPLE III 98% 205 45 1964 Is free of
Example four 86% 170 41 1766 Low ear emergence rate
EXAMPLE five 90% 186 42 1923 The tremella hypha is unstable after seed dressing
EXAMPLE six 98% 152 48 1821 Is free of
Table two: influence of different domestication temperatures on molecular weight of tremella polysaccharide
Group of Incubation temperature (. degree.C.) Acclimation temperature (. degree.C.) Molecular weight (kDa)
Example one 23 No acclimation 1657
Example two 23 15-17℃ 1337
EXAMPLE III 23 18-20℃ 1964
And (4) analyzing results: the independent variables of the first, second and third examples are different domestication temperatures, under the condition that the culture medium condition and the seed dressing condition are kept the same, the molecular weight of the tremella polysaccharide obtained by the tremella mycelia domesticated at 18-20 ℃ is obviously increased after the tremella mycelia are domesticated at low temperature, the molecular weight of the tremella polysaccharide obtained by the tremella mycelia domesticated at 15-17 ℃ is lower, and the conditions that the tremella is not suitable for growth and the yield is low under the condition that the tremella mycelia domesticated at 15-17 ℃ are obtained by combining the average earning rate and the growth condition of the first table.
Table three: influence of different seed dressing modes on molecular weight of tremella polysaccharide
Group of Clip for fixing Seed dressing mode Average out Ear rate (%) Molecule Measurement of (kDa)
Fruit of Chinese wolfberry Applying (a) to Example (b) III In a clean area, the lower layer incense ash hypha and the upper layer tremella hypha are evenly mixed at a ratio of 1: 2 and inoculated within 12 hours. 98% 1964
Fruit of Chinese wolfberry Applying (a) to Example (b) Five of them Inoculating Tremella in clean region on culture medium, culturing for 10 days, inoculating small amount of incense ash mycelium, and culturing at the same temperature And (5) culturing for 7-10 days at the temperature, and obtaining the stock when the incense ash hyphae spread the whole test tube. 90% 1923
Fruit of Chinese wolfberry Applying (a) to Example (b) Six ingredients Inoculating Tremella in clean region on culture medium, culturing for 4-6 days, inoculating appropriate amount of incense ash mycelium, and culturing at the same temperature Culturing for 7-10 days, and allowing the lower layer of incense ash hyphae and the upper layer of tremella hyphae to enter at a ratio of 1: 1 when the incense ash hyphae spread over the whole test tube And uniformly mixing to obtain the original seed. 98% 1821
And (4) analyzing results: the independent variables of the third, fifth and sixth examples are different in seed dressing mode, and therefore, the molecular weight of the tremella polysaccharide obtained in the third and fifth examples is obviously higher than that of the tremella polysaccharide obtained in the sixth example, and the third and fifth examples can both produce and prepare tremella polysaccharide with high molecular weight. However, the tremella fuciformis obtained in the fifth embodiment has poor ear emergence rate and is concentrated, which shows that the seed dressing mode has poor stability and has certain influence on tremella fuciformis planting industrialization.
Table four: influence of different culture media on molecular weight of Tremella polysaccharides
Group of Media composition Molecular weight (kDa)
Practice of EXAMPLE III 40% of wood chips, 15% of bran, 15% of corncobs, 15% of tremella polysaccharide extraction waste residues, 8% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of phosphorus Dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and packaging. 1964
Practice of Example four 40% of wood chips, 15% of bran, 15% of corncobs, 23% of bagasse, 3% of yeast peptone, 2% of gypsum and 2% of dipotassium hydrogen phosphate; after mixing, the Pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and packaging. 1766
And (4) analyzing results: the difference between the example four and the example three is that the bagasse replaces the white fungus polysaccharide extraction waste residue, but the molecular weight of the white fungus polysaccharide in the example four is smaller than that in the example three, which indicates that the addition of the white fungus polysaccharide extraction waste residue has a positive effect on increasing the molecular weight of the white fungus polysaccharide.
The technical solutions of the present invention or similar technical solutions designed by those skilled in the art based on the teachings of the technical solutions of the present invention are all within the scope of the present invention.

Claims (7)

1. A tremella cultivation method for extracting high-molecular tremella polysaccharide is characterized by comprising the following steps:
1) domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 22 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving at the temperature of 18-20 ℃;
2) preparing a novel culture medium: 30-50% of wood chips, 10-20% of bran, 10-20% of corncobs, 10-20% of tremella polysaccharide extraction waste residues, 5-10% of bagasse, 1-5% of yeast peptone, 1-2% of gypsum and 1-2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) seed dressing, namely uniformly mixing the lower layer incense ash hypha and the upper layer tremella hypha at a ratio of 1: 2 in a clean area, and inoculating within 12 hours;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
2. The method for cultivating white fungus to extract high molecular weight tremella polysaccharide as claimed in claim 1, wherein the temperature of the domesticated strain in step 1) is lower than the growth environment of the pre-domesticated strain by 3-5 ℃, and the temperature is controlled in the cultivation stage to regulate the growth state of the cinerea mycelia, so that the secreted cellulase, hemicellulase, polyphenol oxidase and the like are controlled, and the molecular weight of the polysaccharide in the white fungus is reduced.
3. The method for cultivating white fungus by extracting high molecular weight white fungus polysaccharide according to claim 1, wherein the domestication method in the step 1) adopts a constant temperature incubator for cultivation.
4. The method for cultivating tremella fuciformis by extracting high-molecular tremella fuciformis polysaccharide as claimed in claim 1, wherein the growth cycle, leaf diameter, fresh weight of each tremella fuciformis, dry weight of each tremella fuciformis and molecular weight of the extracted tremella fuciformis polysaccharide are controlled by tremella fuciformis polysaccharide extraction waste residues, bagasse and yeast peptone in the novel culture medium in the step 2).
5. The method for cultivating tremella fuciformis from which high-molecular tremella polysaccharide is extracted according to claim 1, wherein the step 3) is performed by using a high-pressure high-temperature sterilization pot, so that the sterilization time is effectively shortened, and the production efficiency is improved.
6. The method for cultivating tremella fuciformis as claimed in claim 1, wherein fresh tremella fuciformis can be directly used for extraction in step 8), and drying is performed at a negative pressure of 55-60 ℃ to guarantee molecular weight due to difficulty in storage.
7. A tremella cultivation method for extracting high-molecular tremella polysaccharide is characterized by comprising the following steps:
1) domesticating Tremella fuciformis strains at low temperature under aseptic condition, culturing at the temperature of 23 ℃, performing spore ejection separation by adopting strong growth after eighty percent maturity, performing hypha culture and strain cultivation on the separated spores, changing the culture temperature to 22 ℃, and repeatedly screening Tremella fuciformis spores capable of thriving at the temperature of 18-20 ℃;
2) preparing a novel culture medium: 30-50% of wood chips, 10-20% of bran, 10-20% of corncobs, 10-20% of tremella polysaccharide extraction waste residues, 5-10% of bagasse, 1-5% of yeast peptone, 1-2% of gypsum and 1-2% of dipotassium hydrogen phosphate; mixing, pulverizing, adding water to make culture medium humidity reach 50-60%, mixing, and bagging;
3) performing high-temperature high-pressure sterilization, wherein the sterilization conditions are as follows: increasing pressure, sterilizing at 121 deg.C for 2-3 hr, and cooling to 22-26 deg.C in inoculation chamber for inoculation;
4) dressing seeds, namely inoculating tremella on a culture medium in a clean area, culturing for 10 days, inoculating a little incense ash hypha, culturing for 7-10 days at the same temperature, and uniformly stirring when the incense ash hypha spreads the whole test tube to obtain stock seeds;
5) inoculating 2-4g of tremella strain into the inoculating hole by using the inoculating spoon, and culturing the fungus bag in a sterile environment for 4-8 days at 22-26 ℃;
6) transferring to a fungus room for cultivation, sealing the fungus bag with small holes of 6 x 6cm to make the bacterial colony contact with air sufficiently, controlling the temperature at 20-24 deg.C and humidity at 80-90%, and culturing for 5-10 days; after the bud base differentiates the ear bud, controlling the temperature at 18-22 ℃ and the humidity at 90-95%, and cultivating for 10-15 days;
7) stopping spraying water when the diameter of the leaves is larger than 18cm, maintaining natural humidity, increasing ventilation, and picking after the fungus bags are obviously contracted, wrinkled and lightened at 18-22 deg.C for 5-10 days;
8) after vacuum low-pressure low-temperature drying, the molecular weight is measured by a new method for extracting tremella polysaccharide in patent application No. 201710243322.2.
CN202010965818.2A 2020-09-15 2020-09-15 Tremella cultivation method for extracting high-molecular tremella polysaccharides Active CN112042466B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010965818.2A CN112042466B (en) 2020-09-15 2020-09-15 Tremella cultivation method for extracting high-molecular tremella polysaccharides

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010965818.2A CN112042466B (en) 2020-09-15 2020-09-15 Tremella cultivation method for extracting high-molecular tremella polysaccharides

Publications (2)

Publication Number Publication Date
CN112042466A true CN112042466A (en) 2020-12-08
CN112042466B CN112042466B (en) 2021-08-03

Family

ID=73602818

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010965818.2A Active CN112042466B (en) 2020-09-15 2020-09-15 Tremella cultivation method for extracting high-molecular tremella polysaccharides

Country Status (1)

Country Link
CN (1) CN112042466B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115039639A (en) * 2022-08-17 2022-09-13 云南菌视界生物科技有限公司 Tremella liquid strain short-period production method and application of tremella liquid strain

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102174615A (en) * 2011-03-16 2011-09-07 浙江省农业科学院 Method for producing fungal polysaccharide with residues after extraction of polysaccharide using needle mushroom
US20120009241A1 (en) * 2010-07-12 2012-01-12 Agricultural Research Institute Skin wound dressing and preparing method thereof
CN102037849B (en) * 2009-10-12 2012-08-08 江寿根 Tremella fuciformis strain culture method
CN106749724A (en) * 2016-12-09 2017-05-31 昭平县科学技术指导站 A kind of method for extracting Auricularia polysaccharide
CN108359025A (en) * 2018-04-13 2018-08-03 苏州尚格生物科技有限公司 Tremella polysaccharides and the preparation method and application thereof
CN108522145A (en) * 2018-05-11 2018-09-14 福建省农业科学院农业质量标准与检测技术研究所 A kind of white fungus culture medium and cultivating white fungus method
CN110923281A (en) * 2019-12-09 2020-03-27 黑龙江菇友生物科技开发有限公司 Edible fungus polysaccharide extraction method, edible fungus polysaccharide and edible fungus beverage

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102037849B (en) * 2009-10-12 2012-08-08 江寿根 Tremella fuciformis strain culture method
US20120009241A1 (en) * 2010-07-12 2012-01-12 Agricultural Research Institute Skin wound dressing and preparing method thereof
CN102174615A (en) * 2011-03-16 2011-09-07 浙江省农业科学院 Method for producing fungal polysaccharide with residues after extraction of polysaccharide using needle mushroom
CN106749724A (en) * 2016-12-09 2017-05-31 昭平县科学技术指导站 A kind of method for extracting Auricularia polysaccharide
CN108359025A (en) * 2018-04-13 2018-08-03 苏州尚格生物科技有限公司 Tremella polysaccharides and the preparation method and application thereof
CN108522145A (en) * 2018-05-11 2018-09-14 福建省农业科学院农业质量标准与检测技术研究所 A kind of white fungus culture medium and cultivating white fungus method
CN110923281A (en) * 2019-12-09 2020-03-27 黑龙江菇友生物科技开发有限公司 Edible fungus polysaccharide extraction method, edible fungus polysaccharide and edible fungus beverage

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115039639A (en) * 2022-08-17 2022-09-13 云南菌视界生物科技有限公司 Tremella liquid strain short-period production method and application of tremella liquid strain

Also Published As

Publication number Publication date
CN112042466B (en) 2021-08-03

Similar Documents

Publication Publication Date Title
US10266695B2 (en) Cultivation of Xylaria species biomass as a binding agent in material production
US20070004022A1 (en) Industrial fermenting production process of Hirsutella hepiali Chen & Shen of anamorphic fungi related to Chinese Cordyceps Sinensis
CN103270887B (en) Silkworm chrysalis northern Chinese caterpillar Fungus industrial cultivation technique
CN107080008A (en) A kind of natural pile fermentation of golden flower Pu'er cooked teas loose tea is grown dim method
CN105961021B (en) White cordyceps militaris production process
CN112042466B (en) Tremella cultivation method for extracting high-molecular tremella polysaccharides
KR100823541B1 (en) Mushroom cultivation method
CN105493897A (en) Industrialized cultivation method of beech mushrooms
CN102550723A (en) Preparation of conventional Pu'er tea microbial colony seeds and application of microbial colony seeds in Pu'er tea fermentation
CN104403952A (en) New Lentinus tuber-regium strain, and culture method and application thereof
CN106867947A (en) A kind of method for preparing insecticide through microbial fermentation as raw material with China tree
CN112243798B (en) Energy-saving and efficient tremella strain breeding and planting method
CN1220768C (en) Variant strain of monoscus purpureus for generating lovastatin with high yield and its application
CN101736034A (en) Method for preparing low-citrinin monascus pigment through submerged fermentation
CN110079478B (en) Bacteriostatic agent of bacillus amyloliquefaciens, preparation method thereof and application of bacteriostatic agent in prevention and treatment of rhizoctonia solani flowers
CN109797107A (en) A kind of preparation method of hypha,hyphae fine powder
CN111149618A (en) Novel Wuzhi strain and artificial cultivation method
CN110100917A (en) A kind of loose tea fungus growing process
JP3277640B2 (en) Koji making method of puffing brewing raw material
KR20090090855A (en) Large-scale production of b-glucan through semi-continuous fermentation performed with sparassis crispa mycelia
CN107047057A (en) A kind of Antrodia camphorata inoculation method
CN100455654C (en) Heat tolerant asafetida mushroom mutant
CN108299045A (en) A kind of culture medium for cultivating schizophyllum commune
CN106967752A (en) A kind of method for preparing insecticide through microbial fermentation by raw material of Siberian cocklebur
CN113249232B (en) Large-scale high-yield preparation method of trichoderma harzianum

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant