CN1478886A - Chinese beimao spore liquid culture fermentationi technology - Google Patents
Chinese beimao spore liquid culture fermentationi technology Download PDFInfo
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Abstract
A liquid fermentation process for the Chinese pilous spore includes such steps as getting bacterial spawn, Erlenmeyer flask culture of bacterial spawns, liquid seed culture, and liquid fermenting. Its advantags are high successful rate, low cost and no environmental pollution. The resultant mycelia can be used for medical or health-care product. The fermented liquid can be used as oral liquir or beverage.
Description
Technical field
The present invention relates to the fermentation technology of the anamorph China pilose spore liquid spawn of Cordyceps sinensis, specifically use isolating China pilose spore (Hirsutella sinensis Liu, Guo, Yu ﹠amp from the Cordyceps sinensis; Zeng) the RCEF0273 bacterial strain carries out the liquid fungus seed processing method and the method for preserving of anamorph breeding.
Background technology
The pharmaceutical use of Cordyceps sinensis (Cordyceps sinensis (Berk.) Sacc.) is early on the books in Ancient Times in China, is traditional rare Chinese medicine, and is equally celebrated for their achievements with genseng, pilose antler, is one of three famous big tonics.Based on Cordyceps sinensis in the effect aspect the medicines and health protection, and the natural resource collection is excessive, be close to exhaustion, existing output is difficult to satisfy the demands, price also is doubled and redoubled, and the seventies, per 500 grams were only sold tens of units, but city's price has reached 10～33 yuan/gram at present, promptly 5000～16500 yuan/500 restrain, ordinary people is difficult to make inquiries.
The artificial culture Chinese caterpillar fungus becomes the important channel that solves this difficult problem.But the artificial culture difficulty of Cordyceps sinensis is very big, does not still have extensive example of succeeing.So the road of breeding-artificial fermentation's mycelium of Chinese caterpillar fungus anamorph has been gone in modern research one after another.A large amount of studies show that the main pharmacology medicine of artificial fermentation's Cordyceps mycelium changes into branch and is similar to the Chinese caterpillar fungus crude drug substantially, and this just provides theoretical foundation for the artificial fermentation of Cordyceps sinensis.
The primary difficulty of artificial fermentation's Cordyceps sinensis is exactly the anamorph problem of Cordyceps sinensis.Along with (2000) such as Zhao Jin etc. (1999) and Li Zengzhi adopt molecular biology method, the anamorph of having verified Cordyceps sinensis once more is China pilose spore; Simultaneously Lee increases application RAPD technology such as intelligence and has got rid of the corresponding relation of other several relevant fungies and Cordyceps sinensis, thereby has finished the anamorph battle of Cordyceps sinensis.
Because the anamorph China pilose spore of Cordyceps sinensis separates comparatively difficulty, people successively from the Cordyceps sinensis isolating relevant bacterial strain about 10 belong to totally 11 kinds, several bacterial strains wherein are taken as Cordyceps fungus and have been developed to medicine and protective foods, make capsule etc. as JINSHUIBAO, intelligence curing capsule, hundred.So nearly all document (comprising patented technology), the bacterial strain that institute uses is the extremely rare of China pilose spore or allied species; And strains tested is not a China pilose spore, but is called very common (the comprising that external some pass so-called Cordyceps strain in the past from China) of Cordyceps fungus, and has done the research of a large amount of pharmacology and medicine aspect.Hence one can see that, and the rare reference of the research of the submerged fermentation of most so-called Cordyceps sinensis at present is worth.
Because Cordyceps sinensis often is grown in high mountain snow line above (3000～4000 meters of height above sea level), suitable temperature is lower.The growth of its anamorph-China pilose spore needs lower temperature equally, and growth is very slow, and the artificial fermentation is difficulty very, and often fermentation period reaches 20～40 days, and this has improved industrial cost undoubtedly greatly.So artificial fermentation's problem of China pilose spore is perplexing the scientific research personnel always.
Two Chinese patents more approaching with this patent are respectively CN85101971 and CN1197843, but difference is more obvious.Mainly be: (1) used bacterial strain is not same bacterial strain; (2) culture medium prescription difference; (3) production technique difference; (4) the preservation problem of liquid spawn is not mentioned.
Summary of the invention
In order to address the above problem, keeping under the stable prerequisite of China pilose spore bacterial classification, the invention provides a kind of physical method and change culture medium prescription way of adopting and realize the batch production producing and manufacturing technique of China pilose spore and the method for preserving of bacterial classification.
Realize that the concrete technical solution of above-mentioned purpose is as follows:
1, a kind of China pilose spore liquid fungus seed processing method, described bacterial classification is isolated RCEF0273 bacterial strain (the Hirsutella sinensisLiu of Cordyceps sinensis (Cordyceps sinensis (Berk) Sacc.) stroma that gathers from Qinghai, Guo, Yu ﹠amp; Zeng); Its liquid fungus seed processing method comprises feedstock capture, strains separation, spawn culture, preservation and multistage fermentation, it is characterized in that:
(1) described spawn culture was divided into for two steps:
A, one level solid flask strain culture
The bacterial classification inoculation of separation and purification in 50ml triangular flask substratum, is put into 19 ℃ of incubators, and incubation time is 20 days, treats that mycelia is covered with triangular flask and produces spore to get final product;
B, secondary liquid shaking bottle seed culture
The 200ml substratum of in the 500ml triangular flask, packing into, with 1.5pa/kg autoclaving 30 minutes, to be cooled during to room temperature, will be to 500ml triangular flask substratum through the bacterial classification inoculation of 2 bottles of solid triangular flasks after the high speed dispersion, and place the constant-temperature shaking culture case, 19 ± 1 ℃ of temperature are cultivated under 180 rev/mins of conditions, and incubation time is 10 days～12 days;
(2) fermentation of described bacterial classification is three grades of liquid fungus seeds:
A, first class seed pot production
The 21L liquid nutrient medium of in the 50L airlift fermentor, packing into, the substratum temperature is 6.0～7.0 greater than 95 ℃, pH value, and adds edible defoaming agent, add-on is 0.03%, 14.7 * 10
4Under the Pa pressure, feed steam heating to 121 ℃, and pressurize 30～45 minutes; After the sterilization, when being cooled to 20 ℃ 4 bottles of above-mentioned cultured 500ml shake-flask seeds are inserted cultivation, culture temperature is 19-20 ℃, and air flow is 1: 0.5V/Vmin, pressurize 4.903～7.0845 * 10
4Pa through 7 days aerated culture, reaches the logarithmic growth after date and gets final product, and final volume of culture is 30L;
B, secondary seed jar are produced
The 300L liquid nutrient medium of in the 500L seeding tank, packing into, the pH value is 6.0～7.0, and adds edible defoaming agent, and add-on is 0.03%, and original position is heated to 121 ℃, 14.7 * 10
4Under the Pa pressure, pressurize 30～45 minutes; After the sterilization, when being cooled to 20 ℃, the 30L liquid seeds of first class seed pot all being inserted the secondary seed jar cultivate, culture temperature 19-20 ℃, air flow is 1: 0.5V/Vmin, pressurize 4.903～7.0845 * 10
4Pa through 7 days aerated culture, reaches the logarithmic growth after date and gets final product, and final volume of culture is 300L;
C, fermentor tank production
The 3T liquid nutrient medium of in the 5T fermentor tank, packing into, the pH value is 6.0～7.0, and the adding edible defoaming agent, add-on is 0.03%, original position is heated to 121 ℃, pressurize 40 minutes, be cooled to 20 ℃, cultured liquid seeds 300L all inserts fermentor cultivation with the secondary seed jar, and culture temperature is 19-20 ℃, air flow is 1: 0.4～0.5V/Vmin, tank pressure 4.903～7.0845 * 10
4Pa, incubation time 5 days;
A large amount of little bacterium balls occur and find a large amount of mycelia through sediments microscope inspection in fermented liquid, residual sugar is below 1%, and fermented liquid is than thickness, and color is obviously deepened, and can go out jar.
2, the raw material weight proportioning of described one-level solid triangular flask substratum is: potato 200g liquor, glucose 20g, maltose 10g, wort 20ml, peptone 10g, yeast powder 10g, MgSO
41.5g, KH
2PO
43g, ammonium citrate 0.4g, agar 20g, add water to 1L, pH6.5.
3, described secondary liquid shaking bottle seed culture medium raw material weight proportioning is: contain glucose 20g, milk powder 30g, dried silkworm chrysalis meal 30g, yeast powder 20g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and is 6.5 with saturated NaOH adjust pH.
4, described seeding tank production liquid medium starting material weight proportion is: contain white sugar 20g, milk powder 30g, dried silkworm chrysalis meal 30g, yeast powder 30g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and transfers pH to 6.5 with saturated NaOH.
5, described fermentor tank production liquid nutrient medium comprises: contain white sugar 20g, milk powder 20g, dried silkworm chrysalis meal 20g, yeast powder 20g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and transfers pH to 6.5 with saturated NaOH.
6, described defoamer is a polyoxypropylene ethylene oxide glyceryl ether, and consumption is 0.03% of a nutrient solution total amount.
7, in the access procedure of described secondary seed jar, can use high-speed dispersion equipment that seed is carried out online dispersion;
Described high-speed dispersion equipment or be interior cut high speed dispersion device, or be online high-speed dispersion equipment, its processing parameter be rotating speed at 200-5000 rev/min, intermittently or not intermittently pulverize, non-online homogeneous dispersion stirring-head diameter is 20-30mm.
8, the liquid shaking bottle seed can carry out preservation, and concrete grammar is to place refrigerator to preserve cultured liquid spawn, and the short term storage temperature is 4 ℃, and the preservation time mostly is 14 days most, and long-term preservation temperature is below-40 ℃.
When 9, the liquid seeds after the preservation is transferred, need to activate 12-24 hour in the constant-temperature shaking culture case, processing condition are 19 ± 1 ℃ of temperature, 180 rev/mins; Available afterwards high speed dispersion device carries out interval type to be disperseed, and each jitter time is no more than 5 seconds; The switchable liquid or solid substratum of liquid seeds after the dispersion.
Processing method of the present invention is fit to suitability for industrialized production, is not subjected to the limitation in time place, can be according to the increase in demand or the reduction industrial scale in market.Production technique of the present invention is simple, process stabilizing, easy-regulating, success ratio height; Adopt the inventive method production less investment, take up an area of little, no three wastes problem, be particularly suited for medium-sized and small enterprises and adopt.
The present invention has optimized zymotechnique.Fungi morphs easily and degenerates for animals and plants.Bacterial strain after the variation often growth velocity is accelerated rapidly, the angle that this institute once was separated to a strain China pilose spore becomes strain, well-grown under 25 ℃ of conditions, 3 days can expire ware, and biological activity then is significantly less than only could well-grown bacterial strain (as RCEF0273) about 19 ℃.Therefore, this institute pays special attention to the variation degenerate problem of China pilose spore bacterial strain.On this basis, study its zymotechnique and obtained success, do not influencing under the active condition, its fermentation period shortens greatly than domestic (as Taiwan), makes product have more market competitiveness.
The inventive method can adopt the different method for preserving continuity preservation time and shorten soak time with reality as required, for tissue production in time provides good quality strain.
The mycelium of explained hereafter of the present invention has unique advance: one, chromaticness is lighter, is canescence after the mycelium drying, is easy to accept into the client; Two, its cultural characteristic and original strain no significant difference behind the mycelium tieback solid of Sheng Chaning, promptly obvious variation does not take place in bacterial strain; Three, the mycelium of explained hereafter of the present invention is main if it were not for by the conidium amplification under liquid state, but be new vegetative point with the mycelia fragment of fracture, cultivates on this basis to obtain a large amount of mycelia; Four, the mycelium of Sheng Chaning can be used for Medicines and Health Product, and fermented liquid can be made oral liquid or beverage, thereby maximally utilises resource, pursues bigger economic and social benefit.
Embodiment
In conjunction with the embodiments the present invention is done to describe further.
Embodiment
This China pilose spore liquid fungus seed processing method comprises following production process:
Obtaining of A, bacterial classification
RCEF0273 bacterial strain (Hirsutella sinensis Liu, Guo, Yu ﹠amp; Zeng), separate Cordyceps sinensis (Cordyceps sinensis (Berk) Sacc.) stroma of gathering from Qinghai.
B, one level solid flask strain culture:
The bacterial classification inoculation of separation and purification in the triangular flask substratum, is put into 19 ℃ of incubators, cultivated 20 days, when treating that mycelia is covered with triangular flask and produces spore, change shake-flask culture over to; The raw material weight proportioning of one-level solid triangular flask substratum is: potato 200g liquor, glucose 20g, maltose 10g, wort 20ml, peptone 10g, yeast powder 10g, MgSO
41.5g, KH
2PO
43g, ammonium citrate 0.4g, agar 20g, add water to 1L, pH6.5.
C, liquid shaking bottle seed culture:
In the 500ml triangular flask, pack into the liquid seed culture medium of 200ml, 1.5pa/kg autoclaving 30 minutes, to be cooled during to room temperature, bacterial classification inoculation that will be after high speed dispersion is to triangular flask, and inoculum size is the little triangular flask bacterial classification of one-level (the little triangular flask of 50ml) that every 500ml triangle bottle graft 2 flask culture are good in the liquid seed culture medium, and place the constant-temperature shaking culture case, 19 ± 1 ℃, under 180 rev/mins of conditions, cultivate and got final product in 10 days; Secondary liquid shaking bottle seed culture medium raw material weight proportioning is: contain glucose 20g, milk powder 30g, dried silkworm chrysalis meal 30g, yeast powder 20g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and is 6.5 with saturated NaOH adjust pH.
D, liquid fermenting production: adopt three grade fermemtation
(1), first class seed pot production
The 21L liquid nutrient medium of in the 50L airlift fermentor, packing into, the substratum temperature is greater than 95 ℃, pH6.0～7.0, and the adding edible defoaming agent, i.e. polyoxypropylene ethylene oxide glyceryl ether, add-on is 0.03%, under 14.7 * 104Pa pressure, feed steam heating to 121 ℃, and pressurize 30～45 minutes, final volume of culture is 30L.After the sterilization, when being cooled to 20 ℃ 4 bottles of above-mentioned cultured 500ml shake-flask seeds are inserted cultivation, cultivate air flow 1: 0.5, pressurize, 4.903～7.0845 * 10 down for 19 ℃
4Pa through 7 days aerated culture, reaches the logarithmic growth after date, inserts the secondary seed jar.This grade seeding tank is produced the liquid medium starting material weight proportion: contain white sugar 20g, milk powder 30g, dried silkworm chrysalis meal 30g, yeast powder 30g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and transfers pH to 6.5 with saturated NaOH.
(2), the secondary seed jar is produced
At the 500L seeding tank, the 300L liquid nutrient medium of packing into, pH6.0～7.0, and add edible defoaming agent, i.e. polyoxypropylene ethylene oxide glyceryl ether, add-on is 0.03%, original position is heated to 121 ℃, under 14.7 * 104Pa pressure, pressurize 30～45min.After the sterilization, when being cooled to 20 ℃ first class seed pot seed 30L is all inserted the secondary seed jar and cultivate, in the access procedure, in pipeline, use high-speed dispersion equipment that seed is carried out online dispersion, 19 ℃ of culture temperature, air flow 1: 0.5V/Vmin, pressurize 4.903～7.0845 * 10
4Pa through 7 days aerated culture, reaches the logarithmic growth after date, inserts fermentor tank; This grade seeding tank is produced the liquid medium starting material weight proportion: contain white sugar 20g, milk powder 30g, dried silkworm chrysalis meal 30g, yeast powder 30g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and transfers pH to 6.5 with saturated NaOH.
(3), fermentor tank production
At the 5T fermentor tank, the 3T liquid nutrient medium liquid of packing into, pH6.0～7.0, add edible defoaming agent, be polyoxypropylene ethylene oxide glyceryl ether, add-on is 0.03%, and original position is heated to 121 ℃, pressurize 40 minutes, be cooled to 20 ℃, cultured liquid seeds 300L all inserts with the secondary seed jar, 19 ℃ of following aerated culture, 1: 0.4～0.5V/Vmin of air flow, tank pressure 4.903～7.0845 * 10
4Pa, incubation time 5 days.This grade fermentor tank is produced liquid nutrient medium and is comprised: contain white sugar 20g, milk powder 20g, dried silkworm chrysalis meal 20g, yeast powder 20g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and transfers pH to 6.5 with saturated NaOH.
A large amount of little bacterium balls occur and find that through sediments microscope inspection a large amount of mycelia are arranged in fermented liquid, residual sugar is below 1%, and fermented liquid is than thickness, and color is obviously deepened, and can go out jar.
Behind continuously centrifuged, obtain mycelium 50 ℃ of aseptic forced air dryings down.Afterwards, through the canned one-tenth capsule of micronizing (or pulverizing 300 mesh sieves).Fermented liquid the results showed and need not to concentrate, and makes functional oral liquid after changing local flavor, as improving sleep.
But the liquid spawn stored refrigerated, concrete operation method is: in 4 ℃ of stored refrigerated, the preservation time is 14 days at most with cultured liquid spawn.Before this, all can be used for inoculation, available high speed dispersion device disperses or does not disperse before the inoculation.If need long-time preservation liquid spawn can be placed under-40 ℃ or the lower temperature.
Concrete operation method when needing the liquid spawn of switching preservation is: by the inoculation of 5% inoculum size, place the constant-temperature shaking culture case by liquid spawn, 19 ± 1 ℃, under 180 rev/mins of conditions, cultivate and got final product in 7 days.
Claims (9)
1, a kind of China pilose spore liquid fungus seed processing method, described bacterial classification is isolated RCEF0273 bacterial strain (the Hirsutella sinensisLiu of Cordyceps sinensis (Cordyceps sinensis (Berk) Sacc.) stroma that gathers from Qinghai, Guo, Yu ﹠amp; Zeng); Its liquid fungus seed processing method comprises feedstock capture, strains separation, spawn culture, preservation and multistage fermentation, it is characterized in that:
(1) described spawn culture was divided into for two steps:
A, one level solid flask strain culture
The bacterial classification inoculation of separation and purification in 50ml triangular flask substratum, is put into 19 ℃ of incubators, and incubation time is 20 days, treats that mycelia is covered with triangular flask and produces spore to get final product;
B, secondary liquid shaking bottle seed culture
The 200ml substratum of in the 500ml triangular flask, packing into, with 1.5pa/kg autoclaving 30 minutes, to be cooled during to room temperature, will be to 500ml triangular flask substratum through the bacterial classification inoculation of 2 bottles of solid triangular flasks after the high speed dispersion, and place the constant-temperature shaking culture case, 19 ± 1 ℃ of temperature are cultivated under 180 rev/mins of conditions, and incubation time is 10 days～12 days;
(2) fermentation of described bacterial classification is three grades of liquid fungus seeds:
A, first class seed pot production
The 21L liquid nutrient medium of in the 50L airlift fermentor, packing into, the substratum temperature is 6.0～7.0 greater than 95 ℃, pH value, and adds edible defoaming agent, add-on is 0.03%, 14.7 * 10
4Under the Pa pressure, feed steam heating to 121 ℃, and pressurize 30～45 minutes; After the sterilization, when being cooled to 20 ℃ 4 bottles of above-mentioned cultured 500ml shake-flask seeds are inserted cultivation, culture temperature is 19-20 ℃, and air flow is 1: 0.5V/Vmin, pressurize 4.903～7.0845 * 10
4Pa through 7 days aerated culture, reaches the logarithmic growth after date and gets final product, and final volume of culture is 30L;
B, secondary seed jar are produced
The 300L liquid nutrient medium of in the 500L seeding tank, packing into, the pH value is 6.0～7.0, and adds edible defoaming agent, and add-on is 0.03%, and original position is heated to 121 ℃, 14.7 * 10
4Under the Pa pressure, pressurize 30～45 minutes; After the sterilization, when being cooled to 20 ℃, the 30L liquid seeds of first class seed pot all being inserted the secondary seed jar cultivate, culture temperature 19-20 ℃, air flow is 1: 0.5V/Vmin, pressurize 4.903～7.0845 * 10
4Pa through 7 days aerated culture, reaches the logarithmic growth after date and gets final product, and final volume of culture is 300L;
C, fermentor tank production
The 3T liquid nutrient medium of in 5T fermentor tank 3, packing into, the pH value is 6.0～7.0, and the adding edible defoaming agent, add-on is 0.03%, original position is heated to 121 ℃, pressurize 40 minutes, be cooled to 20 ℃, cultured liquid seeds 300L all inserts fermentor cultivation with the secondary seed jar, and culture temperature is 19-20 ℃, air flow is 1: 0.4～0.5V/Vmin, tank pressure 4.903～7.0845 * 10
4Pa, incubation time 5 days;
A large amount of little bacterium balls occur and find a large amount of mycelia through sediments microscope inspection in fermented liquid, residual sugar is below 1%, and fermented liquid is than thickness, and color is obviously deepened, and can go out jar.
2, China pilose spore liquid fungus seed processing method according to claim 1 is characterized in that: the raw material weight proportioning of described one-level solid triangular flask substratum is: potato 200g liquor, glucose 20g, maltose 10g, wort 20ml, peptone 10g, yeast powder 10g, MgSO
41.5g, KH
2PO
43g, ammonium citrate 0.4g, agar 20g, add water to 1L, pH6.5.
3, China pilose spore liquid fungus seed processing method according to claim 1 is characterized in that: described secondary liquid shaking bottle seed culture medium raw material weight proportioning is: contain glucose 20g, milk powder 30g, dried silkworm chrysalis meal 30g, yeast powder 20g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and is 6.5 with saturated NaOH adjust pH.
4, China pilose spore liquid fungus seed processing method according to claim 1 is characterized in that: described seeding tank is produced the liquid medium starting material weight proportion and is: contain white sugar 20g, milk powder 30g, dried silkworm chrysalis meal 30g, yeast powder 30g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and transfers pH to 6.5 with saturated NaOH.
5, China pilose spore liquid fungus seed processing method according to claim 1 is characterized in that: described fermentor tank is produced liquid nutrient medium and is comprised: contain white sugar 20g, milk powder 20g, dried silkworm chrysalis meal 20g, yeast powder 20g, MgSO in the 1000ml substratum
40.5g, KH
2PO
41g adds water after each component is mixed and mends to 1000ml, and uses saturated N
aOH transfers pH to 6.5.
6, China pilose spore liquid fungus seed processing method according to claim 1 is characterized in that: described defoamer is a polyoxypropylene ethylene oxide glyceryl ether, and consumption is 0.03% of a nutrient solution total amount.
7, China pilose spore liquid fungus seed processing method according to claim 1 is characterized in that: in the access procedure of described secondary seed jar, can use high-speed dispersion equipment that seed is carried out online dispersion;
Described high-speed dispersion equipment or be interior cut high speed dispersion device, or be online high-speed dispersion equipment, its processing parameter be rotating speed at 200-5000 rev/min, intermittently or not intermittently pulverize, non-online homogeneous dispersion stirring-head diameter is 20-30mm.
8, China pilose spore liquid fungus seed processing method according to claim 1, it is characterized in that: the liquid shaking bottle seed can carry out preservation, concrete grammar is to place refrigerator to preserve cultured liquid spawn, the short term storage temperature is 4 ℃, the preservation time mostly is 14 days most, and long-term preservation temperature is below-40 ℃.
9, China pilose spore liquid fungus seed processing method according to claim 1 is characterized in that: when the liquid seeds after the preservation is transferred, need to activate 12-24 hour in the constant-temperature shaking culture case, processing condition are 19 ± 1 ℃ of temperature, 180 rev/mins; Available afterwards high speed dispersion device carries out interval type to be disperseed, and each jitter time is no more than 5 seconds; The switchable liquid or solid substratum of liquid seeds after the dispersion.
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1323157C (en) * | 2005-09-23 | 2007-06-27 | 田向荣 | Chinese caterpillar fungus bionic culturing method |
| CN100448456C (en) * | 2006-05-22 | 2009-01-07 | 安徽农业大学 | Chinese pilose spore concentrated oral liquid |
| CN101134940B (en) * | 2007-07-22 | 2011-04-20 | 樊立 | Fermentation production method for Chinese caterpillar fungus fungus-bat moth hirsutella sinensis |
| CN101407765B (en) * | 2007-10-10 | 2011-07-06 | 青海久实虫草生物科技有限公司 | Method for producing Chinese piliferous mycelium powder |
| CN102329837A (en) * | 2011-09-01 | 2012-01-25 | 安徽农业大学 | Preparation method of Beauveria sp. extract with functions of tyrosinase inhibitor and antioxidant |
| CN103031285A (en) * | 2012-12-10 | 2013-04-10 | 浙江工业大学 | Cordyceps Chinese Hirsutella uridine-cytidine kinase, coding gene and application thereof |
| CN103045574A (en) * | 2012-12-10 | 2013-04-17 | 浙江工业大学 | Cordyceps sinensis hirsutella sinensis uridylic acid synthetase, as well as coding gene and application thereof |
| CN103431368A (en) * | 2013-09-05 | 2013-12-11 | 东方中科生命科学有限责任公司 | Preparation method and comprehensive utilization of submerged fermentation product of cordyceps sinensis liquid |
| CN103881912A (en) * | 2013-12-20 | 2014-06-25 | 云南大学 | Method for the long-term preservation of Hirsutella sinensis |
| CN104082034A (en) * | 2014-06-25 | 2014-10-08 | 广东省昆虫研究所 | Ophiocordyceps sinensis sporocarp artificial cultivation method |
| CN105624200A (en) * | 2016-02-05 | 2016-06-01 | 浙江工业大学 | Recycling method of hirsutella sinensis fermentation filtrate |
| CN106754401A (en) * | 2016-11-22 | 2017-05-31 | 青海珠峰冬虫夏草工程技术研究有限公司 | A kind of production method of hirsutella sinensis fungal |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1323157C (en) * | 2005-09-23 | 2007-06-27 | 田向荣 | Chinese caterpillar fungus bionic culturing method |
| CN100448456C (en) * | 2006-05-22 | 2009-01-07 | 安徽农业大学 | Chinese pilose spore concentrated oral liquid |
| CN101134940B (en) * | 2007-07-22 | 2011-04-20 | 樊立 | Fermentation production method for Chinese caterpillar fungus fungus-bat moth hirsutella sinensis |
| CN101407765B (en) * | 2007-10-10 | 2011-07-06 | 青海久实虫草生物科技有限公司 | Method for producing Chinese piliferous mycelium powder |
| CN102329837A (en) * | 2011-09-01 | 2012-01-25 | 安徽农业大学 | Preparation method of Beauveria sp. extract with functions of tyrosinase inhibitor and antioxidant |
| CN103031285B (en) * | 2012-12-10 | 2014-12-17 | 浙江工业大学 | Cordyceps Chinese Hirsutella uridine-cytidine kinase, coding gene and application thereof |
| CN103031285A (en) * | 2012-12-10 | 2013-04-10 | 浙江工业大学 | Cordyceps Chinese Hirsutella uridine-cytidine kinase, coding gene and application thereof |
| CN103045574A (en) * | 2012-12-10 | 2013-04-17 | 浙江工业大学 | Cordyceps sinensis hirsutella sinensis uridylic acid synthetase, as well as coding gene and application thereof |
| CN103431368A (en) * | 2013-09-05 | 2013-12-11 | 东方中科生命科学有限责任公司 | Preparation method and comprehensive utilization of submerged fermentation product of cordyceps sinensis liquid |
| CN103881912A (en) * | 2013-12-20 | 2014-06-25 | 云南大学 | Method for the long-term preservation of Hirsutella sinensis |
| CN103881912B (en) * | 2013-12-20 | 2016-05-04 | 云南大学 | A kind of long term storage method of Hirsutella sinensis |
| CN104082034A (en) * | 2014-06-25 | 2014-10-08 | 广东省昆虫研究所 | Ophiocordyceps sinensis sporocarp artificial cultivation method |
| CN104082034B (en) * | 2014-06-25 | 2015-05-20 | 广东省昆虫研究所 | Ophiocordyceps sinensis sporocarp artificial cultivation method |
| CN105624200A (en) * | 2016-02-05 | 2016-06-01 | 浙江工业大学 | Recycling method of hirsutella sinensis fermentation filtrate |
| CN105624200B (en) * | 2016-02-05 | 2019-06-14 | 浙江工业大学 | A kind of recoverying and utilizing method of Hirsutella sinensis ferment filtrate |
| CN106754401A (en) * | 2016-11-22 | 2017-05-31 | 青海珠峰冬虫夏草工程技术研究有限公司 | A kind of production method of hirsutella sinensis fungal |
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