CN105586265A - Solid fermentation method for cordyceps sinensis mycelia - Google Patents
Solid fermentation method for cordyceps sinensis mycelia Download PDFInfo
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Abstract
The invention discloses a solid fermentation method for cordyceps sinensis mycelia. The method comprises the steps of strain culture and solid fermentation culture, wherein Hirsutella sinensis is adopted as a strain, and a solid medium is inoculated with the strain for fermenting production. Effective ingredients of the product prepared according to the solid fermentation method are consistent with or approximate to those of wild cordyceps sinensis; besides, cost is low, process control is simple, pollution is avoided, and safety and effectiveness are achieved.
Description
Technical field
The invention belongs to fungal organism fermentation field, relate to a kind of solid fermentation method of aweto mycelium.
Background technology
Cordyceps sinensis Cordycepssinensis (Berk.) Sacc. is China's tradition rare Chinese medicine, with ginseng, pilose antlerEqually celebrated for their achievements, three is collectively referred to as " Chinese medicine Triratna ". It is many that Cordyceps sinensis contains cordycepin, Cordyceps sinensis polysaccharide, adenosine, cordycepic acid etc.Kind of active principle, have kidney tonifying benefit lung, hemostasis and phlegm, chatter cough, anticancer, improve the functions such as immunity, antifatigue. CauseWild cordyceps yield reduced year by year, the market price is constantly soaring in recent years, is called as " soft gold ".
For the present situation that solves that supply falls short of demand in market, artificially cultivating cordyceps becomes each scientific research institution and each large health products factory owner grindsDirection. On existing market, product is mainly gathered the asexual bacterium of Cordyceps sinensis as main taking liquid fermentation. Although but liquid fermentation is producedCycle is short, but has high input, and has the problems such as the outer loss of effective components of born of the same parents is serious, discharging of waste liquid amount is large. Solid-fermented techniqueCan effectively address the above problem.
Existing solid-fermented technique is not tight controlling to bacterial classification kind. 2005, Chinese fungus association by discussing fully andStrict demonstration, final unified identification Hirsutella sinensis (Hirsutellasinensis) is that Cordyceps sinensis is unique correctPhorozoon. (aweto-cephalosporin Cephalosporiumdongchongxiacae, Hirsutella hepiali Chen et Shen HirsutellaHepiali, Chinese synnema spore Synnematiumsinensis are all Hirsutella sinensis Hirsutellasinensis notThe qualified title of delivering is the synonym of Hirsutella sinensis). And some document or patent before experimental work or finish after,The bacterial classification that derives from wild cordyceps extraction not being identified, may not be in use though deliver with the name of Cordyceps sinensisState is formed by the fermentation of hair spore bacterium, exceedes more than 90 kind of bacterium because can extract to sort out from wild cordyceps, exceedes 10 kinds of bacteriumSimilar with aweto. For example application number is CN201410180618.0 and application number CN200510014467.2 twoIndividual patent application, has Cordyceps sinensis printed words, but according to its fermentation temperature, the former one-level mycelia cultivate temperature be 18 DEG C～37 DEG C, fermentation temperature is 18 DEG C～22 DEG C (and former Cordyceps sinensis is soaked 24 hours in 60 DEG C of hot water). The latter bacteriumPlanting cultivation temperature is 24 DEG C～27 DEG C, and fermentation temperature is 22 DEG C～26 DEG C, and the two bacterial classification used is Hirsutella sinensisPossibility is minimum, because hirsutella sinensis fungal is 12 DEG C～18 DEG C the most suitable growthes, reaches 21 DEG C and just stops growing, and reaches25 DEG C just start to have the phenomena of mortality.
Existing solid-fermented technique fermentation period is longer, and for example patent application of application number CN201410174424X is sent outThe ferment cycle needs 50～70 days, and in batching, contains oyster shell etc. and be not suitable as the batching of food or products and health products additive agent.
Summary of the invention
The object of the invention is, liquid fermentation production cordyceps sinensis mycelium powder cost expensive for wild cordycepsThe problem such as higher, provide a kind of simple, effectively, the preparation method of applicable suitability for industrialized production that cost is low.
In order to address the above problem, the invention provides a kind of solid fermentation preparation method of aweto mycelium.
Concrete steps are as follows:
One, bacterial classification is cultivated
(1) bacterial classification: bacterial classification is purchased from the Hirsutella sinensis of institute of microbiology of the Chinese Academy of Sciences (Hirsutellasinensis).
(2) liquid first class inoculum is cultivated:
Bacterial classification is cut into 5mm2Left and right small pieces, in the 1L conical flask of 150ml first class inoculum fluid nutrient medium, add 2 to filling～In the full temperature concussion culture medium that 3 is 150～180r/m at rotating speed, cultivate, cultivation temperature is 16～18 DEG C, cultivation 6～10d, can obtain Cordyceps sinensis first class inoculum;
First class inoculum fluid nutrient medium is configured to: mealy potato 1.5～3%w/v; Glucose 1.5～2.5%w/v;MgSO4·7H2O0.02～0.05％w/v，KH2PO40.05g～0.10%w/v, peptone 0.3～0.5%w/v, fermentFemale cream 0.4g～0.6%w/v, all the other are pure water, pH value nature;
(3) liquid two stage bacterial classification is cultivated:
By after real 30L strain fermentation ullage slake disappearing, cultured first class inoculum is put in spawn fermentation tank to inoculationAmount 5%～10%, in spawn fermentation tank, the amount of second class inoculum fluid nutrient medium is 21～24L, throughput is 0.15～0.35L/min cultivates 6～10d at 16～18 DEG C, can obtain liquid two stage bacterial classification;
Second class inoculum fluid nutrient medium is configured to: mealy potato 1.5～3%w/v; Glucose 1.5～2.5%w/v;MgSO4·7H2O0.02～0.05％w/v；KH2PO40.05%～0.10%w/v; Peptone 0.3～0.5%w/v; FermentFemale cream 0.4g～0.6%w/v; CaCO3Powder 0.05g～0.10%w/v; All the other are pure water, pH value nature;
Two, solid fermentation is cultivated:
(1) solid medium pre-treatment:
Solid medium is taking wheat or oat as main, put in the heating kettle that contains dextrose plus saccharose solution into boiling extremelyBe split up and bloom; Glucose sugar and sucrose ratio are 1:1, and sugared concentration is 1.5%～2.5%w/v; After draining the water, add oligomeric halfLactose 1%～3%w/w, dried silkworm chrysalis meal 2%～5%w/w, corn flour 2%～5%w/w, stir;
(2) sterilizing: above-mentioned solid medium is packed in fermentation tank, and 121 DEG C～126 DEG C sterilizing 30min, get after coolingGo out to wait to inoculate;
(3) inoculation: the second class inoculum of above-mentioned gained is inoculated in the solid medium after sterilization treatment with inoculating gun,Inoculum concentration is 10%～15%v/w of solid medium dry weight;
(4) fermented and cultured and gathering: fermenting cellar temperature is controlled to 16～18 DEG C, under half-light, cultivates through 30～40d,Culture medium entirety is pitchy, can take out;
(5) low-temp low-pressure drying and crushing: use low-temp low-pressure seasoning at 60～70 DEG C, 60～90Pa the culture taking outCondition under dry, time 8～12h, crushed after being dried.
Wherein, in sterilization steps, fermentation tank is special wide-mouth bottle can tank, and point tank body and lid two parts, in lidBetween leave the tubulose airway of the long diameter 12mm of 2cm, clog with tampon, be beneficial to through air; The interior diameter of fermentation tankShould not exceed 12cm, solid medium thickness should not exceed 5cm.
In inoculation step, described second class inoculum is inoculated into solid culture with the tampon that inoculating gun sees through on glass jar lidIn base.
In fermented and cultured and gathering in step, during solid fermentation, since the 15th day, every 24h, environment temperature is adjustedJoint to 8 DEG C～12 DEG C 2 hours, prevent that the heat production of bacterial classification high speed breeding from exceeding 20 DEG C, affects mycelial growth, thereby dragProlong fermentation period.
Whole solid fermentation carries out under half-light.
Only 30～40 days solid fermentation production cycle of the present invention, along with the prolongation of time, active ingredient is as Cordyceps sinensis polysaccharideDeng by mushroom as nutrient consumption, on the contrary reduce.
The feature of maximum of the present invention: 1) fermentation period is short, 30 days～40 days, and can not exceed 50 days, exceed moreActive ingredient is as more serious in losses such as Cordyceps sinensis polysaccharides. 2) active ingredient as adenosine, cordycepin, Cordyceps sinensis polysaccharide, cordycepic acid,Superoxide dismutases etc. reach or approach wild cordyceps level.
Detailed description of the invention
Embodiment 1
One, bacterial classification is cultivated
1. bacterial classification: bacterial classification is purchased from the Hirsutella sinensis of institute of microbiology of the Chinese Academy of Sciences (Hirsutellasinensis).
2. liquid first class inoculum is cultivated:
First class inoculum culture medium: mealy potato 15g; Glucose 15g; MgSO4·7H2O0.2g；KH2PO40.5g; EggWhite peptone 3g; Yeast extract 4g; Pure water constant volume 1000ml. Be divided in the conical flask of 8 1L. Sterilizing: 121 DEG C,30 minutes.
Bacterial classification is cut into 5mm2Left and right small pieces, add 2 to filling in the 1L conical flask of above-mentioned first class inoculum fluid nutrient mediumIn the full temperature concussion incubator that sheet is 150r/m at rotating speed, cultivate, cultivation temperature is 16 DEG C, cultivates 10d.
3. liquid two stage bacterial classification is cultivated:
Second class inoculum culture medium: mealy potato 315g; Glucose 315g; MgSO4·7H2O4.2g；KH2PO410.5g；Peptone 63g; Yeast extract 84g; CaCO3Powder 10.5g; All the other are pure water, altogether 21L.
Cultured first class inoculum 1.5L is put in spawn fermentation tank, and throughput is 0.15L/min, at 16 DEG CCultivate 10d, obtain liquid two stage bacterial classification, under aseptic condition, bacterial classification is divided in the conical flask of 1L.
Two, solid fermentation is cultivated:
1. solid fermentation formula and pre-treatment:
Formula: wheat 20Kg, glucose 300g, sucrose 300g, dried silkworm chrysalis meal 400g, corn flour 400g, oligomeric galaSugar 200g, water 40Kg.
Dextrose plus saccharose is added in water and boiled, add 20Kg wheat, boil to blooming, pull out and drain, add silkwormPupa powder, corn flour, galactooligosaccharide are mixed thoroughly.
2. sterilizing: above-mentioned solid fermentation culture medium is packed in fermentation glass jar, and every tank 150g, puts into subsequently high pressure and go out121 DEG C of sterilizing 30min in bacterium pot, wait to inoculate after cooling.
3. inoculation: above-mentioned second class inoculum is inoculated in solid medium with the tampon that inoculating gun sees through on glass jar lid,Inoculum concentration is every bottle of 7.5ml.
4. fermented and cultured and gathering: fermenting cellar temperature is controlled to 16 DEG C, and since the 15th day, every 24h was by environment temperatureDegree be adjusted to 8 DEG C～12 DEG C 2 hours. Under half-light, cultivate through 40d, in culture medium, the nutriment such as naked barley endosperm is completeTransformed by mycelia, entirety is pitchy, can take out.
5. low-temp low-pressure drying and crushing: by the culture taking out by low-temp low-pressure seasoning under 60 DEG C, the condition of 60PaDry 8h. Crushed after being dried to 120 order.
Embodiment 2
One, bacterial classification is cultivated
1. bacterial classification: bacterial classification is purchased from institute of microbiology of Chinese Academy of Sciences Hirsutella sinensis (Hirsutellasinensis).
2. liquid first class inoculum is cultivated:
First class inoculum culture medium: mealy potato 30g; Glucose 25g; MgSO4·7H2O0.5g；KH2PO41.0g; EggWhite peptone 5g; Yeast extract 6g; Pure water constant volume 1000ml. Be divided in the conical flask of 8 1L sterilizing: 121 DEG C,30 minutes.
Bacterial classification is cut into 5mm2Left and right small pieces, are added in the 1L conical flask that fills aforesaid liquid culture medium 3, at rotating speedIn full temperature concussion incubator for 180r/m, cultivate, cultivation temperature is 18 DEG C, cultivates 6d.
3. liquid two stage bacterial classification is cultivated:
Second class inoculum culture medium: mealy potato 720g; Glucose 600g; MgSO4·7H2O12g；KH2PO424g; EggWhite peptone 120g; Yeast extract 144g; CaCO3Powder 24g; All the other are pure water, altogether 24L.
Cultured first class inoculum 2.4L is put in spawn fermentation tank, and throughput is 0.35L/min, at 18 DEG CCultivate 6d, obtain liquid two stage bacterial classification, under aseptic condition, bacterial classification is divided in the conical flask of 1L.
Two, solid fermentation is cultivated:
1. solid fermentation formula and pre-treatment:
Formula: wheat 20Kg, glucose 500g, sucrose 500g, dried silkworm chrysalis meal 1000g, corn flour 1000g, oligomeric halfLactose 600g, water 40Kg.
Dextrose plus saccharose is added in water and boiled, add 20Kg wheat, boil to blooming, pull out and drain, add silkwormPupa powder, corn flour, galactooligosaccharide are mixed thoroughly.
2. sterilizing: above-mentioned solid fermentation culture medium is packed in fermentation glass jar, and every tank 150g, puts into subsequently high pressure and go out126 DEG C of sterilizing 30min in bacterium pot, wait to inoculate after cooling.
3. inoculation: above-mentioned second class inoculum is inoculated in solid medium with the tampon that inoculating gun sees through on glass jar lid,Inoculum concentration is every bottle of 15ml.
4. fermented and cultured and gathering: fermenting cellar temperature is controlled to 18 DEG C, and since the 15th day, every 24h was by environment temperatureDegree be adjusted to 8 DEG C～12 DEG C 2 hours. Under half-light through 30d cultivate, to naked barley endosperm completely by mycelium transform become blackBrown, can take out.
5. low-temp low-pressure drying and crushing: by the culture taking out by low-temp low-pressure seasoning under 70 DEG C, the condition of 90PaDry 12h. Crushed after being dried 120 orders.
The culture that embodiment 1 and embodiment 2 are obtained detects. Testing the each composition of detection through spectrum Buddhist nun is: adenosine222～225mg/kg; Cordycepin 117～175mg/kg; Cordyceps sinensis polysaccharide 2.6～4g/100g.
The active ingredient of product prepared by solid fermentation method according to the present invention is consistent with wild cordyceps or approaching, andCost is low, process control is simple, pollution-free, safe and effective.
Claims (4)
1. a solid fermentation method for aweto mycelium, is characterized in that, comprises the steps:
1), bacterial classification is cultivated:
(1) bacterial classification: bacterial classification is purchased from the Hirsutella sinensis of institute of microbiology of the Chinese Academy of Sciences (Hirsutellasinensis);
(2) liquid first class inoculum is cultivated:
Bacterial classification is cut into 5mm2Left and right small pieces, add 2～3 to filling in the conical flask of 150ml first class inoculum fluid nutrient medium, in the full temperature concussion culture medium that is 150～180r/m, cultivate at rotating speed, and cultivation temperature is 16～18 DEG C, cultivates 6～10d, can obtain Cordyceps sinensis first class inoculum;
Described first class inoculum fluid nutrient medium is configured to: mealy potato 1.5～3.0%w/v; Glucose 1.5～2.5%w/v; MgSO4·7H2O0.02～0.05％w/v，KH2PO40.05g～0.10%w/v, peptone 0.3～0.5%w/v, yeast extract 0.4g～0.6%w/v, all the other are pure water, pH value nature;
(3) liquid two stage bacterial classification is cultivated:
Cultured first class inoculum is put in spawn fermentation tank, and inoculum concentration is 5%～10% of second class inoculum fluid nutrient medium consumption, and throughput is 0.15～0.35L/min, at 16～18 DEG C, cultivates 6～10d, can obtain liquid two stage bacterial classification;
Described second class inoculum fluid nutrient medium is configured to: mealy potato 1.5～3%w/v; Glucose 1.5～2.5%w/v; MgSO4·7H2O0.02～0.05％w/v；KH2PO40.05g～0.10%w/v; Peptone 0.3～0.5%w/v; Yeast extract 0.4g～0.6%w/v; CaCO3Powder 0.05～0.10%w/v; All the other are pure water, pH value nature;
2), solid fermentation is cultivated:
(1) solid medium pre-treatment:
Solid medium, taking wheat or oat as main, is put into boiling in the heating kettle that contains dextrose plus saccharose solution and is bloomed to being split up; Glucose sugar and sucrose ratio are 1:1, and sugared concentration is 1.5%～2.5%w/v; After draining the water, add galactooligosaccharide 1%～3%w/w, dried silkworm chrysalis meal 2%～5%w/w, corn flour 2%～5%w/w, stir;
(2) sterilizing: above-mentioned solid medium is packed in fermentation tank, and 121 DEG C～126 DEG C sterilizing 30min, wait to inoculate after cooling;
(3) inoculation: the second class inoculum of above-mentioned gained is inoculated in the solid medium after sterilization treatment with inoculating gun, and inoculum concentration is 10%～15%v/w of solid medium dry weight;
(4) fermented and cultured and gathering: fermenting cellar temperature is controlled to 16～18 DEG C, cultivates through 30～40d under half-light, become pitchy to culture medium, can take out;
(5) low-temp low-pressure drying and crushing: the culture taking out is dry under 60～70 DEG C, the condition of 60～90Pa by low-temp low-pressure seasoning, time 8～12h, crushed after being dried.
2. solid fermentation method according to claim 1, wherein, in sterilization steps,
Tubulose airway is left in the centre of fermentation tank lid, clogs with tampon, is beneficial to through air.
3. solid fermentation method according to claim 2, wherein,
The interior diameter of fermentation tank is no more than 12cm, and solid medium thickness is no more than 5cm.
4. solid fermentation method according to claim 1, wherein, in fermented and cultured and gathering in step,
During solid fermentation since the 15th day, every 24h by environment temperature be adjusted to 8 DEG C～12 DEG C 2 hours.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107151632A (en) * | 2017-07-12 | 2017-09-12 | 西藏藏草宜生生物科技有限公司 | Hirsutella hepiali Chen et Shen filament and its fermentation technique |
| CN108739068A (en) * | 2018-06-08 | 2018-11-06 | 山西万海澳生物科技有限责任公司 | A kind of production technology of aweto mycelium and fructification |
| CN109082382A (en) * | 2018-06-15 | 2018-12-25 | 浙江工业大学 | Cordyceps sinensis Hirsutella sinensis ZJB18002 and its application |
| CN116970483A (en) * | 2023-08-02 | 2023-10-31 | 贵州贵旺生物科技有限公司 | Cordyceps sinensis flower production line and process based on strain fermentation optimization method |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107151632A (en) * | 2017-07-12 | 2017-09-12 | 西藏藏草宜生生物科技有限公司 | Hirsutella hepiali Chen et Shen filament and its fermentation technique |
| CN108739068A (en) * | 2018-06-08 | 2018-11-06 | 山西万海澳生物科技有限责任公司 | A kind of production technology of aweto mycelium and fructification |
| CN108739068B (en) * | 2018-06-08 | 2020-05-19 | 山西万海澳生物科技有限责任公司 | Production process of cordyceps sinensis mycelia and cordyceps sinensis sporophores |
| CN109082382A (en) * | 2018-06-15 | 2018-12-25 | 浙江工业大学 | Cordyceps sinensis Hirsutella sinensis ZJB18002 and its application |
| CN109082382B (en) * | 2018-06-15 | 2021-06-08 | 浙江工业大学 | Cordyceps sinensis hirsutella sinensis ZJB18002 and application thereof |
| CN116970483A (en) * | 2023-08-02 | 2023-10-31 | 贵州贵旺生物科技有限公司 | Cordyceps sinensis flower production line and process based on strain fermentation optimization method |
| CN116970483B (en) * | 2023-08-02 | 2024-03-15 | 贵州贵旺生物科技有限公司 | Cordyceps sinensis flower production line and process based on strain fermentation optimization method |
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Application publication date: 20160518 |