CN102498926A - Production method of liquid lyophilized strain of agaric - Google Patents
Production method of liquid lyophilized strain of agaric Download PDFInfo
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- CN102498926A CN102498926A CN2011102959067A CN201110295906A CN102498926A CN 102498926 A CN102498926 A CN 102498926A CN 2011102959067 A CN2011102959067 A CN 2011102959067A CN 201110295906 A CN201110295906 A CN 201110295906A CN 102498926 A CN102498926 A CN 102498926A
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Abstract
The invention discloses a production method of a liquid lyophilized strain of agaric, and belongs to the field of strain culture of edible mushrooms. The production method comprises the steps of: culturing a mother strain of the agaric in a bevel culture medium in an aseptic condition till mycelia are distributed on the bevel completely; grafting the mycelia on the bevel to a liquid culture medium of a shake flask to stand and performing shake culture amplification; grafting the mature strain cultured in the shake flask to a fermenting tank for culture amplification; performing centrifugation or filtration on strain liquid cultured mature in the fermenting tank and crushing the mycelium pellets; adding a cryoprotectant to crushed mycelia and pre-freezing the mycelia in a refrigerator; and drying the pre-frozen mycelia in a freezing and drying machine and cutting the mycelia into pieces to obtain the lyophilized strain. The mycelium pellets formed by the liquid cultured part are uniform in size, great in quantity, and have multiple germinating points, and are fast to germinate and short in mushroom production period. The liquid lyophilized strain produced by the invention is still active after half-year storage and is convenient to graft.
Description
Technical field
The present invention relates to edible fungus species and cultivate field, specifically a kind of production method of auricularia auriculajudae liquid freeze-drying lactobacillus.
Background technology
The employed bacterial classification of auricularia auriculajudae artificial cultivation adopts the solid fermentation method preparation more; But this method production cycle is long, labour intensity is big, efficiency is low, be subject to living contaminants; Compare with solid spawn; Liquid spawn has the cycle weak point, cell age is neatly sent out advantages such as bacterium is fast, convenient, the difficult pollution of inoculation, is the developing direction of following edible mashroom cultivating.But liquid spawn also has the shortcoming that is difficult for storage and transportation, has limited the application of liquid seeding technique in field of edible fungus culture.
Summary of the invention
Technical assignment of the present invention provides a kind of production method of auricularia auriculajudae liquid freeze-drying lactobacillus.
Technical assignment of the present invention realizes that by following mode this production method step is following:
1) under aseptic condition, the female kind of auricularia auriculajudae to be cultivated in slant medium, condition of culture is: 24～32 ℃ of temperature, 5～8 days time, treat that mycelia is covered with till the inclined-plane;
2) inclined-plane mycelia culture transferring is left standstill to cultivate and carries out the shaken cultivation amplification again after 1～2 day to shaking a bottle liquid nutrient medium;
3) will cultivate the ripe bottle bacterial classification that shakes is inoculated into fermentation tank by 5～10% inoculum concentration and cultivates amplification;
4) fermentation tank culture is ripe bacterium liquid carries out centrifugal or filtration treatment collection mycelium pellet, and mycelium pellet is carried out break process;
5) add freeze drying protectant to the good mycelium of fragmentation, and in refrigerator pre-freeze 0.5～2 hour;
6) mycelium that pre-freeze is good is divided into piece at the freeze drier inner drying after 18～30 hours and is freeze-drying lactobacillus.
Described slant medium is processed by following raw materials in weight portion:
10～25 parts of potatos, 1～5 part of sucrose, 1～2.5 part in agar.
Described liquid nutrient medium preparation method is following:
An amount of potato, wheat bran are put into the water extractive, in aforesaid liquid, add an amount of glucose, magnesium sulfate, potassium dihydrogen sulfate and Cobastab then
1, mix the back and obtain liquid nutrient medium;
The weight portion proportioning of described each raw material is following:
10～20 parts of potatos, 1～5 part of glucose, 0.01～0.08 part in magnesium sulfate, 0.05～0.2 part of potassium dihydrogen sulfate, Cobastab
10.01～0.1 part, 1～10 part in wheat bran.
The described bottle liquid seeds condition of culture that shakes is: 25～32 ℃ of temperature, shaking speed 100～200r/min, liquid amount are 100～180ml/500ml triangular flask, and cultivation cycle is 4～7 days.
Described fermentation tank liquid seeds condition of culture is: 25～32 ℃ of temperature, rotating speed 100～180r/min, and throughput 0.02～0.1Mpa, cultivation cycle is 4～7 days.
Described freeze drying protectant is one or more the mixture in skimmed milk, sodium glutamate, D-sorbierite, vitamin C, sucrose, glucose and the trehalose.
The production method of auricularia auriculajudae liquid freeze-drying lactobacillus of the present invention is compared with prior art; Mycelium pellet that liquid culture of the present invention partly forms size homogeneous, biomass is big and the mycelium pellet number is many; Can normally sprout behind the bacterium powder inoculation solid culture medium after the freeze-drying; And sprout point many, send out that bacterium is fast, the fruiting cycle is short, inoculate the 24h mycelia and be covered with charge level, covered with the bacterium bag in 3-15 days; In addition, can preserve the half a year vigor under the liquid freeze-dried vaccine room temperature that the present invention produces and not fail, inoculation is convenient; The freeze-dried vaccine powder is dissolved in can spraying in an amount of sterile water shines inoculation; Suitability for industrialized production is convenient in the stable performance of long-term maintenance bacterial classification, has higher popularization and application values.
Embodiment
Embodiment 1:
Under aseptic condition, the female kind of auricularia auriculajudae to be cultivated in slant medium, condition of culture is: 24 ℃ of temperature, 5 days time, treat that mycelia is covered with till the inclined-plane; Inclined-plane mycelia culture transferring is left standstill to cultivate and carries out the shaken cultivation amplification again after 1 day to shaking a bottle liquid nutrient medium, and condition of culture is: 25 ℃ of temperature, shaking speed 100r/min, liquid amount are the 100ml/500ml triangular flask, and cultivation cycle is 4 days; Be inoculated into fermentation tank by 5% inoculum concentration and cultivate amplification cultivating the ripe bottle bacterial classification that shakes, condition of culture is: 25 ℃ of temperature, rotating speed 100r/min, and throughput 0.02Mpa, cultivation cycle is 4 days; The bacterium liquid that fermentation tank culture is ripe carries out centrifugal or filtration treatment is collected mycelium pellet, and mycelium pellet is carried out break process; Add freeze drying protectant to the good mycelium of fragmentation, and in refrigerator pre-freeze 0.5 hour; The mycelium that pre-freeze is good is divided into piece at the freeze drier inner drying after 18 hours and is freeze-drying lactobacillus.
The raw material weight proportioning of above-mentioned slant medium is following:
Potato 10kg, sucrose 1kg, agar 1kg.
Aforesaid liquid medium preparation method is following:
10kg potato, 1kg wheat bran are put into the boiling water extractive, in the above-mentioned juice of enduring, add 1kg glucose, 0.01kg magnesium sulfate, 0.05kg potassium dihydrogen sulfate and 0.01kg Cobastab then
1, mix the back and obtain liquid nutrient medium.
Embodiment 2:
Under aseptic condition, the female kind of auricularia auriculajudae to be cultivated in slant medium, condition of culture is: 32 ℃ of temperature, 8 days time, treat that mycelia is covered with till the inclined-plane; Inclined-plane mycelia culture transferring is left standstill to cultivate and carries out the shaken cultivation amplification again after 2 days to shaking a bottle liquid nutrient medium, and condition of culture is: 32 ℃ of temperature, shaking speed 200r/min, liquid amount are the 180ml/500ml triangular flask, and cultivation cycle is 7 days; Be inoculated into fermentation tank by 10% inoculum concentration and cultivate amplification cultivating the ripe bottle bacterial classification that shakes, condition of culture is: 32 ℃ of temperature, rotating speed 180r/min, and throughput 0.1Mpa, cultivation cycle is 7 days; The bacterium liquid that fermentation tank culture is ripe carries out centrifugal or filtration treatment is collected mycelium pellet, and mycelium pellet is carried out break process; Add freeze drying protectant to the good mycelium of fragmentation, and in refrigerator pre-freeze 2 hours; The mycelium that pre-freeze is good is divided into piece at the freeze drier inner drying after 30 hours and is freeze-drying lactobacillus.
The raw material weight proportioning of above-mentioned slant medium is following:
Potato 25kg, sucrose 5kg, agar 2.5kg.
Aforesaid liquid medium preparation method is following:
20kg potato, 10kg wheat bran are put into the boiling water extractive, in the above-mentioned juice of enduring, add 5kg glucose, 0.08kg magnesium sulfate, 0.2kg potassium dihydrogen sulfate and 0.1kg Cobastab then
1, mix the back and obtain liquid nutrient medium.
Embodiment 3:
Under aseptic condition, the female kind of auricularia auriculajudae to be cultivated in slant medium, condition of culture is: 28 ℃ of temperature, 7 days time, treat that mycelia is covered with till the inclined-plane; Inclined-plane mycelia culture transferring is left standstill to cultivate and carries out the shaken cultivation amplification again after 1.5 days to shaking a bottle liquid nutrient medium, and condition of culture is: 29 ℃ of temperature, shaking speed 150r/min, liquid amount are the 140ml/500ml triangular flask, and cultivation cycle is 5 days; Be inoculated into fermentation tank by 8% inoculum concentration and cultivate amplification cultivating the ripe bottle bacterial classification that shakes, condition of culture is: 29 ℃ of temperature, rotating speed 140r/min, and throughput 0.06Mpa, cultivation cycle is 5 days; The bacterium liquid that fermentation tank culture is ripe carries out centrifugal or filtration treatment is collected mycelium pellet, and mycelium pellet is carried out break process; Add freeze drying protectant to the good mycelium of fragmentation, and in refrigerator pre-freeze 1.2 hours; The mycelium that pre-freeze is good is divided into piece at the freeze drier inner drying after 19 hours and is freeze-drying lactobacillus.
The raw material weight proportioning of above-mentioned slant medium is following:
Potato 12kg, sucrose 3kg, agar 1.5kg.
Aforesaid liquid medium preparation method is following:
15kg potato, 6kg wheat bran are put into the boiling water extractive, in the above-mentioned juice of enduring, add 3kg glucose, 0.04kg magnesium sulfate, 0.12kg potassium dihydrogen sulfate and 0.05kg Cobastab then
1, mix the back and obtain liquid nutrient medium.
When the freeze-drying lactobacillus among above-mentioned three embodiment is cultivated, be dissolved in sterile water earlier, be inoculated in the fungus cultivation pocket then, cultivate 24 hours mycelia and be covered with charge level, covered with the bacterium bag in 15 days, the fruiting amount improves 15% than solid spawn.
Claims (6)
1. the production method of an auricularia auriculajudae liquid freeze-drying lactobacillus belongs to edible fungus species and cultivates the field, it is characterized in that this production method step is following:
1) under aseptic condition, the female kind of auricularia auriculajudae to be cultivated in slant medium, condition of culture is: 24～32 ℃ of temperature, 5～8 days time, treat that mycelia is covered with till the inclined-plane;
2) inclined-plane mycelia culture transferring is left standstill to cultivate and carries out the shaken cultivation amplification again after 1～2 day to shaking a bottle liquid nutrient medium;
3) will cultivate the ripe bottle bacterial classification that shakes is inoculated into fermentation tank by 5～10% inoculum concentration and cultivates amplification;
4) fermentation tank culture is ripe bacterium liquid carries out centrifugal or filtration treatment collection mycelium pellet, and mycelium pellet is carried out break process;
5) add freeze drying protectant to the good mycelium of fragmentation, and in refrigerator pre-freeze 0.5～2 hour;
6) mycelium that pre-freeze is good is divided into piece at the freeze drier inner drying after 18～30 hours and is freeze-drying lactobacillus.
2. the production method of auricularia auriculajudae liquid freeze-drying lactobacillus according to claim 1 is characterized in that described slant medium processed by following raw materials in weight portion:
10～25 parts of potatos, 1～5 part of sucrose, 1～2.5 part in agar.
3. the production method of auricularia auriculajudae liquid freeze-drying lactobacillus according to claim 1 is characterized in that described liquid nutrient medium preparation method is following:
An amount of potato, wheat bran are put into the water extractive, in aforesaid liquid, add an amount of glucose, magnesium sulfate, potassium dihydrogen sulfate and Cobastab then
1, mix the back and obtain liquid nutrient medium;
The weight portion proportioning of described each raw material is following:
10～20 parts of potatos, 1～5 part of glucose, 0.01～0.08 part in magnesium sulfate, 0.05～0.2 part of potassium dihydrogen sulfate, Cobastab
10.01～0.1 part, 1～10 part in wheat bran.
4. the production method of auricularia auriculajudae liquid freeze-drying lactobacillus according to claim 1; It is characterized in that the described bottle liquid seeds condition of culture that shakes is: 25～32 ℃ of temperature, shaking speed 100～200r/min; Liquid amount is 100～180ml/500ml triangular flask, and cultivation cycle is 4～7 days.
5. the production method of auricularia auriculajudae liquid freeze-drying lactobacillus according to claim 1 is characterized in that described fermentation tank liquid seeds condition of culture is: 25～32 ℃ of temperature, rotating speed 100～180r/min, and throughput 0.02～0.1Mpa, cultivation cycle is 4～7 days.
6. the production method of auricularia auriculajudae liquid freeze-drying lactobacillus according to claim 1 is characterized in that described freeze drying protectant is one or more the mixture in skimmed milk, sodium glutamate, D-sorbierite, vitamin C, sucrose, glucose and the trehalose.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103449914A (en) * | 2013-08-27 | 2013-12-18 | 广西壮族自治区农业科学院植物保护研究所 | Application of trehalose to prolonging storage life of pleurotus ostrcatus strain, and culture mediums and method for prolonging storage life of pleurotus ostrcatus strain |
| CN106578325A (en) * | 2016-12-12 | 2017-04-26 | 吉林农业大学 | Auricularia auricula-judae mycelium pellet and mung bean shaved ice drink and making method thereof |
| CN109089729A (en) * | 2018-09-07 | 2018-12-28 | 贵州高山百益食用菌发展有限公司 | A kind of making and use method of external source corncob nutrition packet |
| CN114503873A (en) * | 2020-11-16 | 2022-05-17 | 上海国森生物科技有限公司 | Agaricus bisporus liquid strain reduction strain and cultivation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103449914A (en) * | 2013-08-27 | 2013-12-18 | 广西壮族自治区农业科学院植物保护研究所 | Application of trehalose to prolonging storage life of pleurotus ostrcatus strain, and culture mediums and method for prolonging storage life of pleurotus ostrcatus strain |
| CN103449914B (en) * | 2013-08-27 | 2015-07-01 | 广西壮族自治区农业科学院植物保护研究所 | Application of trehalose to prolonging storage life of pleurotus ostrcatus strain, and culture mediums and method for prolonging storage life of pleurotus ostrcatus strain |
| CN106578325A (en) * | 2016-12-12 | 2017-04-26 | 吉林农业大学 | Auricularia auricula-judae mycelium pellet and mung bean shaved ice drink and making method thereof |
| CN109089729A (en) * | 2018-09-07 | 2018-12-28 | 贵州高山百益食用菌发展有限公司 | A kind of making and use method of external source corncob nutrition packet |
| CN114503873A (en) * | 2020-11-16 | 2022-05-17 | 上海国森生物科技有限公司 | Agaricus bisporus liquid strain reduction strain and cultivation method thereof |
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Application publication date: 20120620 |