CN104774888A - Cordycepin fermentation solid medium and preparation method and application thereof - Google Patents
Cordycepin fermentation solid medium and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a cordycepin fermentation solid medium and a preparation method and application thereof. The medium is prepared through the following steps of firstly, fully stirring and dissolving glucose, yeast extract, peptone, monopotassium phosphate, dipotassium phosphate, magnesium sulfate and Tween 80 with water as solvent, and adjusting the pH value of the solution to be within the range of 5.5 to 6.0 to prepare a fermentation culture solution; secondly, evenly mixing the fermentation culture solution obtained in the first step with expanded perlite, filling a fermentation tank with the mixture after spray drying is conducted on the mixture, sterilizing the mixture through vapor under the temperature of 121 DEG C for 20 minutes, and obtaining the cordycepin fermentation solid medium. The cordycepin fermentation solid medium is large in sporocarp amount, high in synchronism and strong in viability; the growth of cordyceps militaris and the accumulation of cordycepin are promoted by compensating for cordycepin precursor adenine and glycerol trioleate through spraying, and the yield and quality of cordycepin can be improved.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of cordycepin fermentation solid substratum and preparation method thereof and application.
Background technology
Cordycepin is one of activeconstituents main in Chinese caterpillar fungus.Cordycepin has various biological activity, as antitumor, antiproliferative, anti-metastasis, antibacterial, antiviral, immunomodulatory and anti-inflammatory etc.The preparation of cordycepin mainly contains chemosynthesis and biosynthesizing two kinds of modes.Because current chemosynthesis cordycepin production cost is high, synthesis technique is complicated, and yield is low, and product purification is more difficult, so cordycepin is prepared primarily of biological synthesis process.Biological synthesis process is prepared Chinese caterpillar fungus and is have two kinds of approach: one is that solid fermentation obtains Cordyceps sporophore, more therefrom extracts; Two is by Chinese caterpillar fungus liquid fermenting, extracting directly from fermented liquid.Because liquid fermenting is than the advantage of solid fermentation in fermentation-scale, biomass growth rate, stand density and controllability, Chinese caterpillar fungus liquid fermenting extracts cordycepin becomes main cordycepin preparation method.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of cordycepin fermentation solid substratum producing a large amount of sporophore.
The technical problem that the present invention also will solve is to provide the preparation method of above-mentioned solid medium.
The technical problem that the present invention finally will solve is to provide the application of above-mentioned solid medium.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
A preparation method for cordycepin fermentation solid substratum, it comprises the steps:
1) by glucose 40 ~ 50g/L, yeast extract paste 3 ~ 10g/L, peptone 5 ~ 15g/L, potassium primary phosphate 0.2 ~ 1.0g/L, dipotassium hydrogen phosphate 0.2 ~ 1.0g/L, magnesium sulfate 0.2 ~ 1.0g/L, tween 80 0.5 ~ 5.0g/L, solvent is water, regulator solution pH value 5.5 ~ 6.0 after abundant stirring and dissolving, obtained fermentation culture;
2) by step 1) in fermentation culture mix with pearlstone, spray-dried rear loading fermentor tank, 121 DEG C of steam sterilizing 20min, obtain cordycepin fermentation solid substratum.
Step 1) in, by glucose 42g/L, yeast extract paste 6g/L, peptone 10g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, tween 80 2g/L, solvent is water, regulator solution pH value 5.8 after abundant stirring and dissolving, obtained fermentation culture.
Step 2) in, described pearlstone particle diameter is 4 ~ 8mm, preferred 6mm.
Step 2) in, fermentation culture is 30 ~ 50ml/g with the volume mass ratio of pearlstone, preferred 40ml/g.
Step 2) in, the material moisture after spraying dry controls at 2.5-5ml/g, preferred 4ml/g.
The cordycepin fermentation solid substratum that above-mentioned preparation method prepares is also within protection scope of the present invention.
The application of above-mentioned cordycepin fermentation solid substratum in fermentative production cordycepin is also within protection scope of the present invention.
Concrete application method is, under the Cordyceps militaris spawn 5ml stroke-physiological saline solution that PDA slant medium has been grown is washed, make bacteria suspension, be seeded to sterilizing good be equipped with in the fermentation culture shaking flask of 50ml, in 27 DEG C, 180rpm constant-temperature table cultivates 4 days, add sterilized water 450ml, after mixing, be seeded in the good cordycepin fermentation solid substratum of sterilizing with 1wt% again, abundant stirring makes bacterial classification be uniformly distributed in cordycepin fermentation solid substratum, relative humidity is maintained with the moisturizing of 1wt% ratio during every 24h, and stir stirring, pass into from fermenter base the sterile air that relative humidity is 80% simultaneously, ventilation ratio is 0.5min
-1, cultivate after 4 days for 27 DEG C, ventilation ratio reduces to 0.5h
-1, within every 5 days, spray with 5wt% the nutrient solution added containing 0.5g/L VITAMIN B4 and 2 ~ 6g/L (preferred 4g/L) triolein and make it produce a large amount of sporophores, all the other conditions are constant, then cultivate secondary fermentation in 21 days and terminate, add the water that 1.5 times are expected volume admittedly, 80 DEG C are stirred stirring 2h, collecting by filtration filtrate, poach process carries out three times, merging filtrate, is concentrated into solid material volume, obtains cordycepin extracting solution.
Above-mentioned Cordyceps militaris spawn is the Cordyceps militaris spawn arbitrarily with cordycepin throughput, such as, can be to buy from Chinese industrial Microbiological Culture Collection administrative center, is numbered the bacterial strain of CICC 14014.
This technology, by the absorption of nutrient solution on 20 order pearlstones, plays the advantage of solid fermentation, overcomes its defect, utilizes specific surface area abundant in 20 order pearlstones to reach a large amount of raw carpogenic object.As everyone knows, cordycepin is enriched in Cordyccps-militaris-(L.)-link. Sporophore top usually.
By literature survey and research, we find that the direct precursor of cordycepin is VITAMIN B4, and another structural unit of cordycepin is 3 '-ribodesose.In Cordyceps militaris (L.) Link. fermentation, we find that adding VITAMIN B4 in the fermentation medium can improve cordycepin output greatly.And the cordycepin content after we also find to add vegetables oil in Cordyceps militaris (L.) Link. fermented liquid has again further rising.
The fermention medium of Cordyceps militaris (L.) Link. classical culture protocols to be using glucose, peptone and yeast extract paste be main component.These compositions can only meet the basic g and D needs of Cordyceps militaris (L.) Link., and cordycepin output is lower.And after adding VITAMIN B4, provide certain cordycepin precursor, cordycepin output is increased.But 3 '-deoxyribose unit is by fatty acid metabolism approach, through acetyl-CoA, then through secondary metabolic pathways synthesis, 3 '-ribodesose of this and first feed-forward nets be synthesized by phosphopentose pathway have difference.Therefore, under the condition lacking acetyl-CoA supply, cordycepin content is difficult to improve again.And plant wet goods glyceryl ester can be decomposed into a large amount of acetyl-CoAs by fatty acid metabolism in Cordyceps militaris (L.) Link., and then the output of cordycepin can be improved further.
Beneficial effect of the present invention:
1,20 order pearlstones of the present invention are as supporting dielectric, and its specific surface area is 10m
2(internal diameter is the fermentor tank of 10 meters to/g, and under non-stirred condition, liquid-gas interface is only 78.5m
2, be only equivalent to 8g20 order pearlstone), porosity reaches 50-90%, and these are conducive to the generation of mass transfer and sporophore growth and spore, and synchronism is good.
2, the different oxygen supply conditions needed for Cordyceps militaris (L.) Link. fungus bulk-growth and cordycepin accumulation, utilize ventilation ratio to change, influence factor is few, more simple and convenient.
3, the present invention adds VITAMIN B4 and triolein, provides two kinds of precursors of cordycepin, is more conducive to the accumulation of cordycepin.
Accompanying drawing explanation
Fig. 1 is the effect diagram of different culture media to cordycepin content, and wherein X-coordinate 1 is embodiment 4, and 2 is embodiment 5, and 3 is embodiment 6, and 4 is comparative example 1, and 5 is comparative example 2, and ordinate zou represents cordycepin content in the fermented liquid or cordycepin extracting solution recorded.
Fig. 2 is the effect diagram of different culture media to sporophore concentration, and wherein X-coordinate 1 is embodiment 4, and 2 is embodiment 5, and 3 is embodiment 6, and 4 is comparative example 1, and 5 is comparative example 2, and ordinate zou represents the sporophore concentration recorded.
Embodiment
Following examples further illustrate content of the present invention, but should not be construed as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, the amendment do the inventive method, step or condition or replacement, all belong to scope of the present invention.
If do not specialize, the conventional means that technique means used in following examples is well known to those skilled in the art.
Embodiment 1: Cordyceps militaris (L.) Link. fermentation solid medium preparing.
1) glucose 42g/L, yeast extract paste 6g/L, peptone 10g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, tween 80 2g/L, solvent is water, regulator solution pH value 5.8 after abundant stirring and dissolving, obtained fermentation culture.
2) by step 1) in the pearlstone of fermentation culture and particle diameter 6mm according to volume mass than being 40ml/g mix and blend, after spray-dried, material moisture controls to load fermentor tank, 121 DEG C of steam sterilizing 20min at 4ml/g, obtains cordycepin fermentation solid substratum.
Embodiment 2: prepared by Cordyceps militaris (L.) Link. fermention medium.
1) glucose 42g/L, yeast extract paste 6g/L, peptone 10g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, tween 80 2g/L, solvent is water, regulator solution pH value 5.8 after abundant stirring and dissolving, obtained fermentation culture.
2) by step 1) in the pearlstone of fermentation culture and particle diameter 4mm according to volume mass than being 30ml/g mix and blend, after spray-dried, material moisture controls to load fermentor tank, 121 DEG C of steam sterilizing 20min at 5ml/g, obtains cordycepin fermentation solid substratum.
Embodiment 3: prepared by Cordyceps militaris (L.) Link. fermention medium.
1) glucose 42g/L, yeast extract paste 6g/L, peptone 10g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, tween 80 2g/L, solvent is water, regulator solution pH value 5.8 after abundant stirring and dissolving, obtained fermentation culture.
2) by step 1) in the pearlstone of fermentation culture and particle diameter 8mm according to volume mass than being 50ml/g mix and blend, after spray-dried, material moisture controls to load fermentor tank, 121 DEG C of steam sterilizing 20min at 2.5ml/g, obtains cordycepin fermentation solid substratum.
Embodiment 4: Cordyceps militaris (L.) Link. solid fermentation substratum is improving the application in Cordyceps militaris (L.) Link. cordycepin output.
1, the Cordyceps militaris spawn bought from Chinese Research for Industrial Microbial Germ preservation administrative center (is numbered: CICC14014) be deposited in ampoul tube, be in lyophilised state, is needed before the experiments to recover bacterial activity.In Bechtop, clean ampoul tube with the absorbent cotton of dipped 70% alcohol, ampoul tube top is heated on flame, drip several sterilized waters to heating place and make cracking glasses.Strike down the ampoul tube top of having ftractureed with tweezers, add the physiological saline of 0.5ml 0.9%, vibration makes freeze-drying thalline dissolve and is suspension.Getting 0.2ml thallus suspension liquid adds in slant medium, 25 DEG C of constant temperature culture 7d.
2, under the bacterial classification 5ml stroke-physiological saline solution that PDA slant medium has been grown is washed, make bacteria suspension, be seeded to sterilizing good be equipped with in the fermentation culture shaking flask of 50ml, in 27 DEG C, 180rpm constant-temperature table cultivates 4 days, add sterilized water 450ml, after mixing, be seeded to the good embodiment of sterilizing 1 with 1wt% more admittedly to expect in substratum, abundant stirring makes bacterial classification be uniformly distributed in substratum, suitable relative humidity is maintained with the moisturizing of 1wt% ratio during every 24h, and stir stirring, pass into from fermenter base the sterile air that relative humidity is 80% simultaneously, ventilation ratio is 0.5min
-1, cultivate after 4 days for 27 DEG C, ventilation ratio reduces to 0.5h
-1, within every 5 days, spray with 5wt% the nutrient solution added containing 0.5g/L VITAMIN B4 and 4g/L triolein and make it produce a large amount of sporophores, all the other conditions are constant, then cultivate secondary fermentation in 21 days and terminate.Add the water that 1.5 times are expected volume admittedly, 80 DEG C are stirred stirring 2h, collecting by filtration filtrate.Poach process carries out three times, merging filtrate, is concentrated into solid material volume, obtains cordycepin extracting solution.
Embodiment 5: Cordyceps militaris (L.) Link. fermention medium is improving the application in Cordyceps militaris (L.) Link. fermented liquid on cordycepin content
1, the Cordyceps militaris spawn bought from Chinese Research for Industrial Microbial Germ preservation administrative center (is numbered: CICC14014) be deposited in ampoul tube, be in lyophilised state, is needed before the experiments to recover bacterial activity.In Bechtop, clean ampoul tube with the absorbent cotton of dipped 70% alcohol, ampoul tube top is heated on flame, drip several sterilized waters to heating place and make cracking glasses.Strike down the ampoul tube top of having ftractureed with tweezers, add the physiological saline of 0.5ml 0.9%, vibration makes freeze-drying thalline dissolve and is suspension.Getting 0.2ml thallus suspension liquid adds in slant medium, 25 DEG C of constant temperature culture 7d.
2, under the bacterial classification 5ml stroke-physiological saline solution that PDA slant medium has been grown is washed, make bacteria suspension, be seeded to sterilizing good be equipped with in the fermentation culture shaking flask of 50ml, in 27 DEG C, 180rpm constant-temperature table cultivates 4 days, add sterilized water 450ml, after mixing, be seeded to the good embodiment of sterilizing 2 with 1wt% more admittedly to expect in substratum, abundant stirring makes bacterial classification be uniformly distributed in substratum, suitable relative humidity is maintained with the moisturizing of 1wt% ratio during every 24h, and stir stirring, pass into from fermenter base the sterile air that relative humidity is 80% simultaneously, ventilation ratio is 0.5min
-1, cultivate after 4 days for 27 DEG C, ventilation ratio reduces to 0.5h
-1, within every 5 days, spray with 5wt% the nutrient solution added containing 0.5g/L VITAMIN B4 and 2g/L triolein and make it produce a large amount of sporophores, all the other conditions are constant, then cultivate secondary fermentation in 21 days and terminate.Add the water that 1.5 times are expected volume admittedly, 80 DEG C are stirred stirring 2h, collecting by filtration filtrate.Poach process carries out three times, merging filtrate, is concentrated into solid material volume, obtains cordycepin extracting solution.
Embodiment 6: Cordyceps militaris (L.) Link. fermention medium is improving the application in Cordyceps militaris (L.) Link. fermented liquid on cordycepin content
1, the Cordyceps militaris spawn bought from Chinese Research for Industrial Microbial Germ preservation administrative center (is numbered: CICC14014) be deposited in ampoul tube, be in lyophilised state, is needed before the experiments to recover bacterial activity.In Bechtop, clean ampoul tube with the absorbent cotton of dipped 70% alcohol, ampoul tube top is heated on flame, drip several sterilized waters to heating place and make cracking glasses.Strike down the ampoul tube top of having ftractureed with tweezers, add the physiological saline of 0.5ml 0.9%, vibration makes freeze-drying thalline dissolve and is suspension.Getting 0.2ml thallus suspension liquid adds in slant medium, 25 DEG C of constant temperature culture 7d.
2, under the bacterial classification 5ml stroke-physiological saline solution that PDA slant medium has been grown is washed, make bacteria suspension, be seeded to sterilizing good be equipped with in the fermentation culture shaking flask of 50ml, in 27 DEG C, 180rpm constant-temperature table cultivates 4 days, add sterilized water 450ml, after mixing, be seeded to the good embodiment of sterilizing 3 with 1wt% more admittedly to expect in substratum, abundant stirring makes bacterial classification be uniformly distributed in substratum, suitable relative humidity is maintained with the moisturizing of 1wt% ratio during every 24h, and stir stirring, pass into from fermenter base the sterile air that relative humidity is 80% simultaneously, ventilation ratio is 0.5min
-1, cultivate after 4 days for 27 DEG C, ventilation ratio reduces to 0.5h
-1, within every 5 days, spray with 5wt% the nutrient solution added containing 0.5g/L VITAMIN B4 and 6g/L triolein and make it produce a large amount of sporophores, all the other conditions are constant, then cultivate secondary fermentation in 21 days and terminate.Add the water that 1.5 times are expected volume admittedly, 80 DEG C are stirred stirring 2h, collecting by filtration filtrate.Poach process carries out three times, merging filtrate, is concentrated into solid material volume, obtains cordycepin extracting solution.
Comparative example 1:
1, glucose 42g/L, yeast extract paste 6g/L, peptone 10g/L, potassium primary 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, regulator solution pH value 5.8 after abundant stirring and dissolving, obtained fermentation culture.
2, the Cordyceps militaris spawn bought from Chinese Research for Industrial Microbial Germ preservation administrative center (is numbered: CICC14014) be deposited in ampoul tube, be in lyophilised state, is needed before the experiments to recover bacterial activity.In Bechtop, clean ampoul tube with the absorbent cotton of dipped 70% alcohol, ampoul tube top is heated on flame, drip several sterilized waters to heating place and make cracking glasses.Strike down the ampoul tube top of having ftractureed with tweezers, add the physiological saline of 0.5ml 0.9%, vibration makes freeze-drying thalline dissolve and is suspension.Getting 0.2ml thallus suspension liquid adds in slant medium, 25 DEG C of constant temperature culture 7d.
3, under the bacterial classification 5ml stroke-physiological saline solution that PDA slant medium has been grown is washed, make bacteria suspension, be seeded to sterilizing good be equipped with in the fermentation culture shaking flask of 50ml, in 27 DEG C, 180rpm constant-temperature table cultivates 4 days, add sterilized water 450ml, after mixing, be seeded in the good fermentation culture of sterilizing with 1%w/w again, abundant stirring makes bacterial classification be uniformly distributed in substratum, suitable relative humidity is maintained with the moisturizing of 1%w/w ratio during every 24h, and stir stirring, pass into from fermenter base the sterile air that relative humidity is 80% simultaneously, ventilation ratio is 0.5min
-1, cultivate after 4 days for 27 DEG C, ventilation ratio reduces to 0.5h
-1, within every 5 days, spray with 5%w/w all the other conditions of nutrient solution added containing 0.5g/L VITAMIN B4 and 4g/L triolein constant, then cultivate secondary fermentation in 21 days and terminate.Collecting by filtration filtrate, obtains cordycepin extracting solution.
Comparative example 2:
1, glucose 42g/L, yeast extract paste 6g/L, peptone 10g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, tween 80 2g/L, solvent is water, regulator solution pH value 5.8 after abundant stirring and dissolving, obtained fermentation culture.
2, the fermentation culture in step 1 and the pearlstone of particle diameter 6mm are compared for 40ml/g mix and blend according to volume mass, after spray-dried, material moisture controls to load fermentor tank, 121 DEG C of steam sterilizing 20min at 4ml/g, obtains cordycepin fermentation solid substratum.
3, the Cordyceps militaris spawn bought from Chinese Research for Industrial Microbial Germ preservation administrative center (is numbered: CICC14014) be deposited in ampoul tube, be in lyophilised state, is needed before the experiments to recover bacterial activity.In Bechtop, clean ampoul tube with the absorbent cotton of dipped 70% alcohol, ampoul tube top is heated on flame, drip several sterilized waters to heating place and make cracking glasses.Strike down the ampoul tube top of having ftractureed with tweezers, add the physiological saline of 0.5ml 0.9%, vibration makes freeze-drying thalline dissolve and is suspension.Getting 0.2ml thallus suspension liquid adds in slant medium, 25 DEG C of constant temperature culture 7d.
4, under the bacterial classification 5ml stroke-physiological saline solution that PDA slant medium has been grown is washed, make bacteria suspension, be seeded to sterilizing good be equipped with in the fermentation culture shaking flask of 50ml, in 27 DEG C, 180rpm constant-temperature table cultivates 4 days, add sterilized water 450ml, after mixing, be seeded in the good cordycepin fermentation solid substratum of sterilizing with 1wt% again, abundant stirring makes bacterial classification be uniformly distributed in substratum, suitable relative humidity is maintained with the moisturizing of 1wt% ratio during every 24h, and stir stirring, pass into from fermenter base the sterile air that relative humidity is 80% simultaneously, ventilation ratio is 0.5min
-1, cultivate after 4 days for 27 DEG C, ventilation ratio reduces to 0.5h
-1, all the other conditions are constant, then cultivate secondary fermentation in 21 days and terminate.Add the water that 1.5 times are expected volume admittedly, 80 DEG C are stirred stirring 2h, collecting by filtration filtrate.Poach process carries out three times, merging filtrate, is concentrated into solid material volume, obtains cordycepin extracting solution.
Fermented liquid cordycepin content measures
In the fermented liquid of embodiment 4-7, the content of cordycepin passes through high effective liquid chromatography for measuring.Fermented liquid centrifuging and taking supernatant liquor pure water dilutes 6 times, and vibration mixing, detects cordycepin.Chromatographic condition: chromatographic column: Ultimate AQ-C18 (4.6mm × 250mm, 5 μm), moving phase: methyl alcohol: phosphate solution (10mmol/L KH
2pO
4solution)=15:85, column temperature 30 DEG C, flow velocity 1ml/min, sample size 20 μ L, determined wavelength is 260nm.
Result: the substratum that embodiment 4 obtains is about 200% and 243% than the cordycepin content increase of comparative example 1-2 respectively.
Sporophore Concentration Testing:
Before preparing solid medium, weigh perlitic quality, be designated as M
0, liquid nutrient medium is designated as 0; After fermentation ends, after filtration drying, weigh fermentation solid total mass, be designated as M
always; Fermentation cumulative volume is V, and sporophore concentration is designated as with (M
always-M
0)/V.
Result: the substratum that embodiment 4 obtains is about 118% and 140% than the sporophore concentration increase of comparative example 1-2 respectively.
Claims (8)
1. a preparation method for cordycepin fermentation solid substratum, is characterized in that, it comprises the steps:
1) by glucose 40 ~ 50g/L, yeast extract paste 3 ~ 10g/L, peptone 5 ~ 15g/L, potassium primary phosphate 0.2 ~ 1.0g/L, dipotassium hydrogen phosphate 0.2 ~ 1.0g/L, magnesium sulfate 0.2 ~ 1.0g/L, tween 80 0.5 ~ 5.0g/L, solvent is water, regulator solution pH value 5.5 ~ 6.0 after abundant stirring and dissolving, obtained fermentation culture;
2) by step 1) in fermentation culture mix with pearlstone, spray-dried rear loading fermentor tank, 121 DEG C of steam sterilizing 20min, obtain cordycepin fermentation solid substratum.
2. the preparation method of cordycepin fermentation solid substratum according to claim 1, is characterized in that, step 1) in, by glucose 42g/L, yeast extract paste 6g/L, peptone 10g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, tween 80 2g/L, solvent is water, regulator solution pH value 5.8 after abundant stirring and dissolving, obtained fermentation culture.
3. the preparation method of cordycepin fermentation solid substratum according to claim 1, is characterized in that, step 2) in, described pearlstone particle diameter is 4 ~ 8mm.
4. the preparation method of cordycepin fermentation solid substratum according to claim 1, is characterized in that, step 2) in, fermentation culture is 30 ~ 50ml/g with the volume mass ratio of pearlstone.
5. the preparation method of cordycepin fermentation solid substratum according to claim 1, is characterized in that, step 2) in, the material moisture after spraying dry controls at 2.5 ~ 5ml/g.
6. the cordycepin fermentation solid substratum that the preparation method in Claims 1 to 5 described in any one prepares.
7. the application of cordycepin fermentation solid substratum according to claim 6 in fermentative production cordycepin.
8. application according to claim 7, it is characterized in that, under the Cordyceps militaris spawn 5ml stroke-physiological saline solution that PDA slant medium has been grown is washed, make bacteria suspension, be seeded to sterilizing good be equipped with in the fermentation culture shaking flask of 50ml, in 27 DEG C, 180rpm constant-temperature table cultivates 4 days, add sterilized water 450ml, after mixing, be seeded in the good cordycepin fermentation solid substratum of sterilizing with 1wt% again, abundant stirring makes bacterial classification be uniformly distributed in cordycepin fermentation solid substratum, relative humidity is maintained with the moisturizing of 1wt% ratio during every 24h, and stir stirring, pass into from fermenter base the sterile air that relative humidity is 80% simultaneously, ventilation ratio is 0.5min
-1, cultivate after 4 days for 27 DEG C, ventilation ratio reduces to 0.5h
-1, within every 5 days, spray with 5wt% the nutrient solution added containing 0.5g/L VITAMIN B4 and 2 ~ 6g/L triolein and make it produce a large amount of sporophores, all the other conditions are constant, then cultivate secondary fermentation in 21 days and terminate, add the water that 1.5 times are expected volume admittedly, 80 DEG C are stirred stirring 2h, collecting by filtration filtrate, poach process carries out three times, merging filtrate, is concentrated into solid material volume, obtains cordycepin extracting solution.
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| 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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| 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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