CN104774888B - A kind of cordycepin fermentation solid culture medium and preparation method and application - Google Patents
A kind of cordycepin fermentation solid culture medium and preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of cordycepin fermentation solid culture medium and its preparation method and application, the culture medium is prepared by following steps：1) by glucose, yeast extract, peptone, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, Tween 80, solvent is water, is sufficiently stirred for after dissolving adjusting solution ph 5.5~6.0, fermentation culture is made；2) by step 1) in fermentation culture be well mixed with expanded perlite, it is spray-dried after load fermentation tank, 121 DEG C of steam sterilizing 20min, obtain cordycepin fermentation solid culture medium.Fructification amount that this method is provided is big, and synchronism is good, energetic, and adds cordycepin precursor adenine by spray and olein promotes the growth of Cordyceps militaris and the accumulation of cordycepin, it is possible to increase the yield and quality of cordycepin.
Description
Technical field
The invention belongs to microbial technology field, and in particular to a kind of cordycepin fermentation solid culture medium and preparation method thereof
With application.
Background technology
Cordycepin is one of main active component in cordyceps sinensis.Cordycepin has various biological activity, such as antitumor, anti-
Propagation, anti-rotation shifting, antibacterial, antiviral, immunological regulation and anti-inflammatory etc..The preparation of cordycepin mainly has chemical synthesis and biosynthesis
Two ways.Because current chemical synthesis cordycepin production cost is high, synthesis technique is complicated, and yield is low, and product purification is more difficult,
So cordycepin is mainly prepared by biological synthesis process.Biological synthesis process prepares cordyceps sinensis and have two kinds of approach：One is that solid fermentation is obtained
Cordyceps militaris sporocarp is obtained, then is therefrom extracted；Two be, by cordyceps sinensis liquid fermentation, directly to be extracted from zymotic fluid.Due to liquid fermentation
Than advantage of the solid fermentation in fermentation-scale, biomass growth rate, stand density and controllability, cordyceps sinensis liquid fermentation is extracted
Cordycepin turns into main cordycepin preparation method.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of cordycepin fermentation solid training for producing a large amount of fructifications
Support base.
The technical problem of the invention also to be solved is to provide the preparation method of above-mentioned solid medium.
The technical problem of the invention finally to be solved is to provide the application of above-mentioned solid medium.
In order to solve the above technical problems, the technical solution adopted by the present invention is as follows：
A kind of preparation method of cordycepin fermentation solid culture medium, it comprises the following steps：
1) by 40~50g/L of glucose, 3~10g/L of yeast extract, 5~15g/L of peptone, potassium dihydrogen phosphate 0.2~
1.0g/L, 0.2~1.0g/L of dipotassium hydrogen phosphate, 0.2~1.0g/L of magnesium sulfate, 0.5~5.0g/L of Tween 80, solvent is water, is filled
Divide and solution ph 5.5~6.0 is adjusted after stirring and dissolving, fermentation culture is made；
2) by step 1) in fermentation culture be well mixed with expanded perlite, it is spray-dried after load fermentation tank,
121 DEG C of steam sterilizing 20min, obtain cordycepin fermentation solid culture medium.
Step 1) in, by glucose 42g/L, yeast extract 6g/L, peptone 10g/L, potassium dihydrogen phosphate 0.5g/L, phosphoric acid hydrogen
Dipotassium 0.5g/L, magnesium sulfate 0.5g/L, Tween 80 2g/L, solvent is water, is sufficiently stirred for after dissolving adjusting solution ph 5.8, system
Obtain fermentation culture.
Step 2) in, described expanded perlite particle diameter is 4~8mm, preferably 6mm.
Step 2) in, the volume mass ratio of fermentation culture and expanded perlite is 30~50ml/g, preferably 40ml/g.
Step 2) in, the material moisture after spray drying is controlled in 2.5-5ml/g, preferably 4ml/g.
The cordycepin fermentation solid culture medium that above-mentioned preparation method is prepared is also within protection scope of the present invention.
Application of the above-mentioned cordycepin fermentation solid culture medium in fermenting and producing cordycepin is also in protection scope of the present invention
Within.
Specific application process is, by the Cordyceps militaris spawn grown on PDA slant mediums 5ml sterile salines
Wash down, bacteria suspension is made, be seeded in the fermentation culture shaking flask equipped with 50ml sterilized, in 27 DEG C, 180rpm constant temperature shakes
Bed culture 4 days, adds sterilized water 450ml, after being well mixed, then the cordycepin fermentation solid training sterilized is seeded to 1wt%
Support in base, be sufficiently stirred for making strain be uniformly distributed in cordycepin fermentation solid culture medium, with 1wt% ratio moisturizings during per 24h
Relative humidity is maintained, and stirs stirring, while being passed through the filtrated air that relative humidity is 80% from fermenter base, ventilation ratio is
0.5min-1, after 27 DEG C are cultivated 4 days, ventilation ratio is reduced to 0.5h-1, added with 5wt% sprays containing 0.5g/L adenines within every 5 days and
The nutrient solution of 2~6g/L (preferably 4g/L) olein makes it produce substantial amounts of fructification, and remaining condition is constant, is further cultured for
After fermentation in 21 days terminates；1.5 times of water for expecting volume admittedly are added, filtrate is collected by filtration in 80 DEG C of stirring stirring 2h；Water boils process progress
Three times, merging filtrate is concentrated into solid material volume, obtains cordycepin extract solution.
Above-mentioned Cordyceps militaris spawn is any Cordyceps militaris spawn with cordycepin production capacity, for example, bought in
State's Research for Industrial Microbial Germ preservation administrative center, numbering is CICC 14014 bacterial strain.
Absorption of this technology by nutrient solution on 20 mesh expanded perlites, plays the advantage of solid fermentation, overcomes it to lack
Fall into, reached using the specific surface area enriched in 20 mesh expanded perlites and largely give birth to carpogenic purpose.It is well known that cordycepin
Generally it is enriched at the top of fruiting bodies of cordyceps militaris.
We have found that the direct precursor of cordycepin is adenine by literature survey and research, and another knot of cordycepin
Structure unit is 3 '-deoxyribose.In Cordyceps militaris is fermented, it has been found that adding adenine in the fermentation medium can greatly improve
Cordycepin output.And, it has been found that add the cordycepin content after vegetable oil in Cordyceps militaris zymotic fluid has further liter again
It is high.
Cordyceps militaris classical culture protocols are using the fermentation medium that glucose, peptone and yeast extract are main component.
These compositions can only meet the basic growth and development needs of Cordyceps militaris, and cordycepin output is relatively low.And add after adenine and provide
Certain cordycepin precursor so that cordycepin output increases.But 3 '-deoxyribose unit is by fatty acid metabolism
Approach, through acetyl-CoA, then is synthesized by secondary metabolic pathways, and 3 '-deoxyribose of this and first feed-forward nets is by phosphoric acid penta
What sugared approach was synthesized has difference.Therefore, under conditions of acetyl-CoA supply is lacked, cordycepin content is difficult to improve again.And plant
The glyceride such as oil can be decomposed into substantial amounts of acetyl-CoA in Cordyceps militaris by fatty acid metabolism, and then can further improve
The yield of cordycepin.
Beneficial effects of the present invention：
1st, 20 mesh expanded perlite of the present invention is as supporting dielectric, and its specific surface area is 10m2(internal diameter is /g
10 meters of fermentation tank, gas-liquid interface is only 78.5m under the conditions of non-stirred2, just correspond to 8g20 mesh expanded perlite), hole
Rate reaches 50-90%, and these are conducive to the generation of mass transfer and sporophore growth and spore, and synchronism is good.
2nd, the different oxygen supply conditions according to needed for being accumulated Cordyceps militaris thalli growth and cordycepin, are changed using ventilation ratio,
Influence factor is few, more simple and convenient.
3rd, the present invention adds adenine and olein there is provided the two of cordycepin kind precursor, is more beneficial for cordycepin
Accumulation.
Brief description of the drawings
Fig. 1 is influence figure of the different culture media to cordycepin content, and wherein abscissa 1 is that embodiment 4,2 is embodiment 5,3
It is that comparative example 1,5 is comparative example 2 for embodiment 6,4, ordinate represents cordycepin in the zymotic fluid measured or cordycepin extract solution
Content.
Fig. 2 is influence figure of the different culture media to fructification concentration, and wherein abscissa 1 is that embodiment 4,2 is embodiment 5,3
It is that comparative example 1,5 is comparative example 2 for embodiment 6,4, ordinate represents the fructification concentration measured.
Embodiment
Following examples further illustrate present disclosure, but should not be construed as limiting the invention.Without departing substantially from
In the case of spirit and essence of the invention, the modifications or substitutions made to the inventive method, step or condition belong to the present invention
Scope.
Unless otherwise specified, the conventional hand that technological means used in following examples is well known to those skilled in the art
Section.
Embodiment 1：It is prepared by Cordyceps militaris fermentation solid culture medium.
1) glucose 42g/L, yeast extract 6g/L, peptone 10g/L, potassium dihydrogen phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/
L, magnesium sulfate 0.5g/L, Tween 80 2g/L, solvent is water, is sufficiently stirred for after dissolving adjusting solution ph 5.8, and fermentation training is made
Nutrient solution.
2) by step 1) in fermentation culture and particle diameter 6mm expanded perlite it is mixed for 40ml/g according to volume mass ratio
Close stirring, it is spray-dried after, material moisture control 4ml/g load fermentation tank, 121 DEG C of steam sterilizing 20min, obtain worm
Careless element fermentation solid culture medium.
Embodiment 2：It is prepared by Cordyceps militaris fermentation medium.
1) glucose 42g/L, yeast extract 6g/L, peptone 10g/L, potassium dihydrogen phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/
L, magnesium sulfate 0.5g/L, Tween 80 2g/L, solvent is water, is sufficiently stirred for after dissolving adjusting solution ph 5.8, and fermentation training is made
Nutrient solution.
2) by step 1) in fermentation culture and particle diameter 4mm expanded perlite it is mixed for 30ml/g according to volume mass ratio
Close stirring, it is spray-dried after, material moisture control 5ml/g load fermentation tank, 121 DEG C of steam sterilizing 20min, obtain worm
Careless element fermentation solid culture medium.
Embodiment 3：It is prepared by Cordyceps militaris fermentation medium.
1) glucose 42g/L, yeast extract 6g/L, peptone 10g/L, potassium dihydrogen phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/
L, magnesium sulfate 0.5g/L, Tween 80 2g/L, solvent is water, is sufficiently stirred for after dissolving adjusting solution ph 5.8, and fermentation training is made
Nutrient solution.
2) by step 1) in fermentation culture and particle diameter 8mm expanded perlite it is mixed for 50ml/g according to volume mass ratio
Close stirring, it is spray-dried after, material moisture control 2.5ml/g load fermentation tank, 121 DEG C of steam sterilizing 20min, obtain
Cordycepin fermentation solid culture medium.
Embodiment 4：Application of the Cordyceps militaris solid fermentation culture medium on Cordyceps militaris cordycepin output is improved.
1st, the Cordyceps militaris spawn (numbering bought from Chinese industrial Microbiological Culture Collection administrative center：CICC14014) protect
Ensconce in ampoul tube, in lyophilised state, need to recover bacterial activity before the experiments.In superclean bench, with dipped
The absorbent cotton of 70% alcohol cleans ampoul tube, ampoul tube top is heated on flame, to heating at drop it is a few drop sterilized waters make glass
Glass ftractures.The ampoul tube top ftractureed is struck down with tweezers, 0.5ml 0.9% physiological saline is added, vibration makes lyophilized thalline
Dissolve and be in suspension.0.2ml thallus suspension liquids are taken to add in slant medium, 25 DEG C of incubated 7d.
2nd, under the strain grown on PDA slant mediums is washed with 5ml sterile salines, bacteria suspension is made, is inoculated with
Into the fermentation culture shaking flask equipped with 50ml sterilized, in 27 DEG C, 180rpm constant-temperature tables culture 4 days adds sterilized water
450ml, after being well mixed, then is seeded to the embodiment 1 sterilized with 1wt% and expected admittedly in culture medium, be sufficiently stirred for making strain equal
It is even to be distributed in culture medium, appropriate relative humidity is maintained with the moisturizing of 1wt% ratios during per 24h, and stir stirring, while from hair
Fermentation tank bottom is passed through the filtrated air that relative humidity is 80%, and ventilation ratio is 0.5min-1, after 27 DEG C are cultivated 4 days, ventilation ratio is reduced to
0.5h-1, adding the nutrient solution containing 0.5g/L adenines and 4g/L oleins with 5wt% sprays within every 5 days produces it
Substantial amounts of fructification, remaining condition is constant, is further cultured for after fermentation in 21 days and terminates.Add 1.5 times of water for expecting volume admittedly, 80 DEG C of stirrings
Stirring 2h, is collected by filtration filtrate.Water boils process and carried out three times, and merging filtrate is concentrated into solid material volume, obtains cordycepin extraction
Liquid.
Embodiment 5：Application of the Cordyceps militaris fermentation medium in Cordyceps militaris zymotic fluid is improved on cordycepin content
1st, the Cordyceps militaris spawn (numbering bought from Chinese industrial Microbiological Culture Collection administrative center：CICC14014) protect
Ensconce in ampoul tube, in lyophilised state, need to recover bacterial activity before the experiments.In superclean bench, with dipped
The absorbent cotton of 70% alcohol cleans ampoul tube, ampoul tube top is heated on flame, to heating at drop it is a few drop sterilized waters make glass
Glass ftractures.The ampoul tube top ftractureed is struck down with tweezers, 0.5ml 0.9% physiological saline is added, vibration makes lyophilized thalline
Dissolve and be in suspension.0.2ml thallus suspension liquids are taken to add in slant medium, 25 DEG C of incubated 7d.
2nd, under the strain grown on PDA slant mediums is washed with 5ml sterile salines, bacteria suspension is made, is inoculated with
Into the fermentation culture shaking flask equipped with 50ml sterilized, in 27 DEG C, 180rpm constant-temperature tables culture 4 days adds sterilized water
450ml, after being well mixed, then is seeded to the embodiment 2 sterilized with 1wt% and expected admittedly in culture medium, be sufficiently stirred for making strain equal
It is even to be distributed in culture medium, appropriate relative humidity is maintained with the moisturizing of 1wt% ratios during per 24h, and stir stirring, while from hair
Fermentation tank bottom is passed through the filtrated air that relative humidity is 80%, and ventilation ratio is 0.5min-1, after 27 DEG C are cultivated 4 days, ventilation ratio is reduced to
0.5h-1, adding the nutrient solution containing 0.5g/L adenines and 2g/L oleins with 5wt% sprays within every 5 days produces it
Substantial amounts of fructification, remaining condition is constant, is further cultured for after fermentation in 21 days and terminates.Add 1.5 times of water for expecting volume admittedly, 80 DEG C of stirrings
Stirring 2h, is collected by filtration filtrate.Water boils process and carried out three times, and merging filtrate is concentrated into solid material volume, obtains cordycepin extraction
Liquid.
Embodiment 6：Application of the Cordyceps militaris fermentation medium in Cordyceps militaris zymotic fluid is improved on cordycepin content
1st, the Cordyceps militaris spawn (numbering bought from Chinese industrial Microbiological Culture Collection administrative center：CICC14014) protect
Ensconce in ampoul tube, in lyophilised state, need to recover bacterial activity before the experiments.In superclean bench, with dipped
The absorbent cotton of 70% alcohol cleans ampoul tube, ampoul tube top is heated on flame, to heating at drop it is a few drop sterilized waters make glass
Glass ftractures.The ampoul tube top ftractureed is struck down with tweezers, 0.5ml 0.9% physiological saline is added, vibration makes lyophilized thalline
Dissolve and be in suspension.0.2ml thallus suspension liquids are taken to add in slant medium, 25 DEG C of incubated 7d.
2nd, under the strain grown on PDA slant mediums is washed with 5ml sterile salines, bacteria suspension is made, is inoculated with
Into the fermentation culture shaking flask equipped with 50ml sterilized, in 27 DEG C, 180rpm constant-temperature tables culture 4 days adds sterilized water
450ml, after being well mixed, then is seeded to the embodiment 3 sterilized with 1wt% and expected admittedly in culture medium, be sufficiently stirred for making strain equal
It is even to be distributed in culture medium, appropriate relative humidity is maintained with the moisturizing of 1wt% ratios during per 24h, and stir stirring, while from hair
Fermentation tank bottom is passed through the filtrated air that relative humidity is 80%, and ventilation ratio is 0.5min-1, after 27 DEG C are cultivated 4 days, ventilation ratio is reduced to
0.5h-1, adding the nutrient solution containing 0.5g/L adenines and 6g/L oleins with 5wt% sprays within every 5 days produces it
Substantial amounts of fructification, remaining condition is constant, is further cultured for after fermentation in 21 days and terminates.Add 1.5 times of water for expecting volume admittedly, 80 DEG C of stirrings
Stirring 2h, is collected by filtration filtrate.Water boils process and carried out three times, and merging filtrate is concentrated into solid material volume, obtains cordycepin extraction
Liquid.
Comparative example 1：
1st, glucose 42g/L, yeast extract 6g/L, peptone 10g/L, potassium dihydrogen phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/
L, magnesium sulfate 0.5g/L, Tween 80 0.5-5.0g/L, solvent is water, is sufficiently stirred for after dissolving adjusting solution ph 5.8, is made
Fermentation culture.
2nd, the Cordyceps militaris spawn (numbering bought from Chinese industrial Microbiological Culture Collection administrative center：CICC14014) protect
Ensconce in ampoul tube, in lyophilised state, need to recover bacterial activity before the experiments.In superclean bench, with dipped
The absorbent cotton of 70% alcohol cleans ampoul tube, ampoul tube top is heated on flame, to heating at drop it is a few drop sterilized waters make glass
Glass ftractures.The ampoul tube top ftractureed is struck down with tweezers, 0.5ml 0.9% physiological saline is added, vibration makes lyophilized thalline
Dissolve and be in suspension.0.2ml thallus suspension liquids are taken to add in slant medium, 25 DEG C of incubated 7d.
3rd, under the strain grown on PDA slant mediums is washed with 5ml sterile salines, bacteria suspension is made, is inoculated with
Into the fermentation culture shaking flask equipped with 50ml sterilized, in 27 DEG C, 180rpm constant-temperature tables culture 4 days adds sterilized water
450ml, after being well mixed, then is seeded in the fermentation culture sterilized with 1%w/w, is sufficiently stirred for being uniformly distributed strain
In culture medium, appropriate relative humidity is maintained with the moisturizing of 1%w/w ratios during per 24h, and stirs stirring, while from fermentation tank
Bottom is passed through the filtrated air that relative humidity is 80%, and ventilation ratio is 0.5min-1, after 27 DEG C are cultivated 4 days, ventilation ratio is reduced to
0.5h-1, every 5 days with remaining condition of 5%w/w spray nutrient solutions of the addition containing 0.5g/L adenines and 4g/L oleins
It is constant, it is further cultured for after fermentation in 21 days and terminates.Filtrate is collected by filtration, cordycepin extract solution is obtained.
Comparative example 2：
1st, glucose 42g/L, yeast extract 6g/L, peptone 10g/L, potassium dihydrogen phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/
L, magnesium sulfate 0.5g/L, Tween 80 2g/L, solvent is water, is sufficiently stirred for after dissolving adjusting solution ph 5.8, and fermentation training is made
Nutrient solution.
2nd, the fermentation culture in step 1 and particle diameter 6mm expanded perlite are mixed according to volume mass ratio for 40ml/g
Close stirring, it is spray-dried after, material moisture control 4ml/g load fermentation tank, 121 DEG C of steam sterilizing 20min, obtain worm
Careless element fermentation solid culture medium.
3rd, the Cordyceps militaris spawn (numbering bought from Chinese industrial Microbiological Culture Collection administrative center：CICC14014) protect
Ensconce in ampoul tube, in lyophilised state, need to recover bacterial activity before the experiments.In superclean bench, with dipped
The absorbent cotton of 70% alcohol cleans ampoul tube, ampoul tube top is heated on flame, to heating at drop it is a few drop sterilized waters make glass
Glass ftractures.The ampoul tube top ftractureed is struck down with tweezers, 0.5ml 0.9% physiological saline is added, vibration makes lyophilized thalline
Dissolve and be in suspension.0.2ml thallus suspension liquids are taken to add in slant medium, 25 DEG C of incubated 7d.
4th, under the strain grown on PDA slant mediums is washed with 5ml sterile salines, bacteria suspension is made, is inoculated with
Into the fermentation culture shaking flask equipped with 50ml sterilized, in 27 DEG C, 180rpm constant-temperature tables culture 4 days adds sterilized water
450ml, after being well mixed, then is seeded in the cordycepin fermentation solid culture medium sterilized with 1wt%, is sufficiently stirred for making strain
It is uniformly distributed in culture medium, appropriate relative humidity is maintained with the moisturizing of 1wt% ratios during per 24h, and stirs stirring, while from
Fermenter base is passed through the filtrated air that relative humidity is 80%, and ventilation ratio is 0.5min-1, after 27 DEG C are cultivated 4 days, ventilation ratio drop
For 0.5h-1, remaining condition is constant, is further cultured for after fermentation in 21 days and terminates.Add 1.5 times of water for expecting volume admittedly, 80 DEG C of stirring stirrings
2h, is collected by filtration filtrate.Water boils process and carried out three times, and merging filtrate is concentrated into solid material volume, obtains cordycepin extract solution.
Zymotic fluid cordycepin content is determined
The content of cordycepin passes through high effective liquid chromatography for measuring in embodiment 4-7 zymotic fluid.In zymotic fluid centrifuging and taking
Clear liquid pure water dilutes 6 times, and vibration is mixed, and detects cordycepin.Chromatographic condition：Chromatographic column：Ultimate AQ-C18(4.6mm×
250mm, 5 μm), mobile phase：Methanol：Phosphate solution (10mmol/L KH2PO4Solution)=15：85,30 DEG C of column temperature, flow velocity
1ml/min, the μ L of sample size 20, Detection wavelength is 260nm.
As a result：Cordycepin content increase of the culture medium made from embodiment 4 respectively than comparative example 1-2 is about 200% He
243%.
Fructification Concentration Testing：
The quality of perlite is weighed before solid medium is prepared, M is designated as0, fluid nutrient medium is designated as 0；Fermentation ends
Afterwards, after filtration drying, fermentation solid gross mass is weighed, M is designated asAlways；Fermentation cumulative volume is V, and fructification concentration is designated as with (MAlways-
M0)/V。
As a result：Fructification concentration increase of the culture medium made from embodiment 4 respectively than comparative example 1-2 is about 118% He
140%.
Claims (5)
1. application of a kind of cordycepin fermentation solid culture medium in fermenting and producing cordycepin, it is characterised in that will be on PDA inclined-planes
The Cordyceps militaris spawn grown on culture medium is washed down with 5ml sterile salines, and bacteria suspension is made, and is seeded to being equipped with of having sterilized
In 50ml fermentation culture shaking flask, in 27 DEG C, 180rpm constant-temperature tables culture 4 days adds sterilized water 450ml, is well mixed
Afterwards, then it is seeded to 1wt% in the cordycepin fermentation solid culture medium sterilized, is sufficiently stirred for making strain be uniformly distributed in cordyceps sinensis
In plain fermentation solid culture medium, relative humidity is maintained with the moisturizing of 1wt% ratios during per 24h, and stirs stirring, while from fermentation tank
Bottom is passed through the filtrated air that relative humidity is 80%, and ventilation ratio is 0.5min-1, after 27 DEG C are cultivated 4 days, ventilation ratio is reduced to
0.5h-1, adding the nutrient solution containing 0.5g/L adenines and 2~6g/L oleins with 5wt% sprays within every 5 days produces it
Raw substantial amounts of fructification, remaining condition is constant, is further cultured for after fermentation in 21 days and terminates；1.5 times of water for expecting volume admittedly are added, 80 DEG C are stirred
Stirring 2h is mixed, filtrate is collected by filtration；Water boils process and carried out three times, and merging filtrate is concentrated into solid material volume, obtains cordycepin extraction
Liquid；
Wherein, the preparation method of described cordycepin fermentation solid culture medium, comprises the following steps：
1) by 40~50g/L of glucose, 3~10g/L of yeast extract, 5~15g/L of peptone, 0.2~1.0g/L of potassium dihydrogen phosphate,
0.2~1.0g/L of dipotassium hydrogen phosphate, 0.2~1.0g/L of magnesium sulfate, 0.5~5.0g/L of Tween 80, solvent is water, is sufficiently stirred for
Solution ph 5.5~6.0 is adjusted after dissolving, fermentation culture is made；
2) by step 1) in fermentation culture be well mixed with expanded perlite, it is spray-dried after load fermentation tank, 121 DEG C
Steam sterilizing 20min, obtains cordycepin fermentation solid culture medium.
2. application according to claim 1, it is characterised in that step 1) in, by glucose 42g/L, yeast extract 6g/L, egg
White peptone 10g/L, potassium dihydrogen phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, Tween 80 2g/L, solvent is
Water, is sufficiently stirred for after dissolving adjusting solution ph 5.8, fermentation culture is made.
3. application according to claim 1, it is characterised in that step 2) in, described expanded perlite particle diameter for 4~
8mm。
4. application according to claim 1, it is characterised in that step 2) in, the body of fermentation culture and expanded perlite
Product mass ratio is 30~50ml/g.
5. application according to claim 1, it is characterised in that step 2) in, the material moisture control after spray drying
In 2.5~5ml/g.
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| CN105969653B (en) * | 2016-05-13 | 2018-01-09 | 南通大学 | A kind of bioreactor and its fermentation process of Cordyceps militaris standing for fermentation |
| CN107582604A (en) * | 2017-10-13 | 2018-01-16 | 贵阳中医学院 | A kind of Cordyceps militaris conversion bark of eucommia solid fermentation product and its preparation method and application |
| CN107586726A (en) * | 2017-10-13 | 2018-01-16 | 贵阳中医学院 | Improve Cordyceps militaris rhizoma Gastrodiae liquid fermentation method and the application of cordycepin content |
| CN111826410A (en) * | 2020-08-10 | 2020-10-27 | 辽东学院 | Method for obtaining high-yield cordycepin by using solid culture medium |
| CN113308505A (en) * | 2021-05-31 | 2021-08-27 | 东莞理工学院 | Method for improving fermentation yield of cordycepin |
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| Title |
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| 利用膨胀珍珠岩生产平菇菌种技术研究;申进文等;《食用菌》;20090723(第4期);第1.2节和第3节 * |
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