CN102668880A - Method for culturing cordyceps militaris bacterium - Google Patents
Method for culturing cordyceps militaris bacterium Download PDFInfo
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Abstract
The invention discloses a method for culturing cordyceps militaris bacterium. The method is as follows: finished bacterium powder is obtained after natural cordyceps militaris is subjected to separation and purification, culturing on a test tube slant, culturing of liquid first-grade seeds and second-grade seeds in oscillators, culturing of small-size seeds in a seed fermenter, concentration and spray-drying. The method for culturing cordyceps militaris bacterium disclosed by the invention has the beneficial effects that: the yield of cordyceps militaris can be increased and the price of cordyceps militaris can be reduced fundamentally; and the method plays a role in relieving pain of all peoples suffering from diseases and makes a wonder for traditional Chinese medicine treasure.
Description
Technical field
The present invention relates to a kind of aweto strain cultural method.
Background technology
Cordyceps sinensis (formal name used at school: Cordyceps sinensis), have another name called Chinese Chinese caterpillar fungus, be called Cordyceps sinensis again, be called for short Chinese caterpillar fungus, Eumycota, Ascomycotina.It is the traditional rare traditional Chinese medicine of China; It is that aweto by Hypocreales Clavicipitaceae Cordyceps parasitizes the Hepialus larva in the alpine meadow soil; Larva is ossify; Under optimum conditions, taken out by the bombys batryticatus head end and bear long bar-shaped stroma and form summer, i.e. the complex of the fruit body of aweto and bombys batryticatus sclerotium (larva corpse) formation.It mainly originates in province and municipal severe cold areas and snow-capped mountains and marshlands such as Chinese Qinghai, Tibet, Xinjiang, Sichuan, Yunnan, Gansu, Guizhou.
Real Cordyceps sinensis is wild, is grown near the careless slope 3000 meters snow lines to the alpine meadow shrub zone of 5000m of height above sea level, natural environment is required high.In summer, the insect ovum originates in ground, becomes through hatching in about month to pierce moist soft soil layer behind the larva.A kind of fungus attack in the soil larva, in the larva body, grow.Through a winter, arrived to 1 year spring, fungal hyphae begins growth, grows ground during to summer, and outward appearance resembles a little grass, and like this, the body of larva and fungal hyphae have been formed a complete Cordyceps sinensis jointly.As nutriment, growth is rapid polypide for the bacterium spore, and polypide is generally four to five centimetres, and the bacterium spore can grow to the length of polypide within one day, and Chinese caterpillar fungus at this moment is called the head grass, and is best in quality; The bacterium spore grew to about the twice of polypide in second day, was called two grass, and quality is taken second place.Because can grow coring after ossifing, so be known as Cordyceps sinensis.
Among the pharmacology modern study result, it is about 7% that Qinghai Cordyceps sinensis contains cordycepic acid, carbohydrate 28.9%; Fat is about 8.4%, and protein is about 25%, and 82.2% is unsaturated fatty acid in the fat; In addition, still contain cobalamin, ergot lipidol, hexose alcohol, alkaloid etc.
Narrow, the natural parasitic rate in area is low, harsh to the living environment conditional request because wild cordyceps distributes, so resource own is more limited.Again because Cordyceps sinensis main product ground ecotope suffers that the people is heavy damage, a large amount of blindly unreasonable excavating cause resource to reduce day by day in recent years, and output descends year by year.
The Cordyceps sinensis of Qinghai-Tibet Platean is distributed in the Nagqu Diqu in Tibet, Nyingchi Prefecture, Qamdo Prefecture, the cajaput area in Qinghai, Golog state one band, the Rangtang, Aba state in Sichuan; Litang, state, Ganzi, Batang, Dege, Deqie, state, Deqen in Yunnan Province, a band to the north of the middle pasture; Ground such as to the west of the Maqu, state, Gannan, Gansu Province.
Cordyceps sinensis is the distinctive one type of valuable ingredient of traditional Chinese medicine of China; Have high value medical health care and add economic worth; At home and abroad enjoying high reputation, have the laudatory title of " the legendary jewellery in east ", is the traditional foreign exchange earning commodity of China; So Cordyceps sinensis is a wonderful work in China's traditional medicine treasure-house.
The value medical health care and the economic worth that have Cordyceps sinensis only are the highest, so at home and abroad mycota, the world of medicine, food circle, biosphere etc. are devoted to the emphasis researched and developed for many years.
Cordyceps sinensis has antifatigue, anoxia enduring, anti-ageing, antitumor and improve multiple medical care effect such as immune function of human body.To human body can play nourishing the lung and the kidney, hemostasis and phlegm, relieving asthma, expand tracheae, calm anti-each bacterioid, effect such as hypotensive.Contain cordycepin in the Cordyceps sinensis, towards the multiple essential amino acid of oxalic acid; The vitamin of healthy trace elements with household, Cordyceps sinensis polysaccharide, superoxide dismutase multiple nutrients material and anticancer antidotal active substances such as (SOD); One of Main Ingredients and Appearance cordycepin is a kind of have antibiosis and material that suppresses the cell division effect, the hyperplasia of ability anticancer.Both at home and abroad the isotope detection technology is to cordycepin in the Cordyceps sinensis research proof Cordyceps sinensis and the effect that not only has tonifying kidney and strengthening yang, beneficial smart Dingchuan towards oxalic acid, and can resist cell ageing significantly.
Cordyceps sinensis is grown in the high and cold area in China western part and the west and south, and growing environment is special, resource-constrained; Along with further investigation both domestic and external for many years, further recognize Cordyceps sinensis to the medical treatment of human body and nourishing research, further recognize the important value that Cordyceps sinensis is had the medical treatment of human body and nourishing; Demand constantly increases, because the natural cordyceps resource is originally just few, adds that worm grass resources quantity falls sharply; Output constantly descends, and disparities between supply and demand are increasingly sharpened, and causes the market supply and demand to disconnect; Cost an arm and a leg,, drop into artificial introduction and acclimatization Cordyceps sinensis to this situation; Among the research work of aspects such as cultivation heredity and application,, open up medicine source biofermentation technique and have great value and development prospect really as a kind of new technology new technology that development potentiality is arranged.
Summary of the invention
The technical issues that need to address of the present invention just are to overcome the defective of prior art, and a kind of aweto strain cultural method is provided.
For addressing the above problem, the present invention adopts following technical scheme:
The invention provides a kind of aweto strain cultural method; Said method for nature Cordyceps sinensis through separation and purification, test tube slant kind, oscillator liquid one-level kind, secondary kind, small-sized seed fermentation seeding tank, produce fermentation tank, concentrate and the drying of dusting, finished product bacterium powder.
The specific requirement of purification procedures is:
(1) material requirements: the piece of tissue partition method of carrying out Cordyceps sinensis; Parting material is annual by the end of October to November; The high and cold grassy marshland soil in Qinghai-Tibet Platean has just begun to freeze the material adopted period; Aweto just infected Hepialus larva to get into the bombys batryticatus phase not long-time in period this, and tender little stroma bud has just grown worm head 0.2-0.5cm, and other assorted bacterium are less in the polypide;
(2) separation operating method has following 3 steps:
A. the separation of bombys batryticatus body (being sclerotium): before the separation, water scrubs clean body surfaces, uses sterile water wash again 2 one 3 times; Mercuric chloride solution with 0.1% one 0.2% carries out the surface sterilization of about 3 one 5min to parting material; Use sterile water wash again.Choose with pereiopoda is the leading portion part on boundary.Cut epidermis with scalpel, avoid digestive tract, get the haemocoele mycelia and be cut into the sesame seed size, be pressed in the plating medium, 1 one 2 in every ware.Place 15 1 19 ℃ of cultivations, when treating that bacterium colony grows 0.2 1 0.5cm, select a small amount of mycelia and on plating medium, divide pure 2 one 4 times repeatedly, behind definite other assorted bacterium of nothing, move into test tube and preserve and cultivate;
B. the stroma bud separates: downcut the stroma bud of cleaning from the bombys batryticatus crown, insert sterilization 2 one 3min in the 0.1% mercuric chloride liquid, clean with sterile water, cut the middle part piece of tissue and be pressed in the medium; The same a of condition of culture;
C. ascospore separates: entangle the ripe stroma of ascospore with transparent paper bag, let the ascospore spring stick on the paper bag, then; In the Glucose Liquid that thecasporous paper bag immersion 25% is arranged; Wash spore, put into 15 1 19 ℃ of cultivations, every day microscopy; When robe is sprouted, inhale single spore with micro pipette and drip in flat board and cultivate; The same a of condition of culture;
Also can take back the Cordyceps sinensis of whole maturation indoorly, encase bombys batryticatus and the stroma that connects polypide, only reserve the part of being pregnant with cotton paper, in desinfection chamber, traverse, underlying a slice slide keeps main body humidity at any time; Microscopy when seeing that ascospore is bound to slide, was drawn onto with micro pipette in the medium of plate and cultivated every day; The same a of condition of culture.
(3) selecting for use of medium:
A. potato dextrose agar (PDA), this kind prescription is the collective media of all Chinese caterpillar fungus Pseudomonas fungies, can fill a prescription with this at the separation and Culture initial stage, but growth is very not vigorous, and be prone to aging, degenerate;
B. enriched medium (1): many property albumen navel 10g, glucose 50g, dipotassium hydrogen phosphate 1g, magnesium sulfate 0.5g, the silkworm chrysalis 30g that lives, growth hormone 0.5 μ g, agar 20g adds water 1000mL, pH:5.0;
Enriched medium (II): albumen dried meat 40g, glucose 40g, peeling fresh potato 100g, dipotassium hydrogen phosphate 1g; Magnesium sulfate 0.5g, beef extract 10g, growth hormone 0.5 μ g, bat larva (grinding) 30g alive; Agar 20g, high and cold meadow soil leachate 10OOmL, pH:5.0.
On enriched medium, bacterium colony is more vigorous and rapid than growth on the PDA medium; Add rich II and number be superior to I number again.
After the aweto separation and purification, promptly can be used as various experiments and enlarge production application;
(1) it enlarges the method for producing often has three kinds: solid leaves standstill cultivation, oscillating culture and big jar of aerobic fementation cultivation;
A. leave standstill cultivation: mainly be used in solid culture, cultivate or the like like test tube slant, triangular flask cultivation, rice.Can make the bacterium normal growth as long as in leaving standstill cultivation, grasp temperature and illumination.After the conidium maturation on the inclined-plane, can deposit in and preserve 8-14 month in 1-2 ℃ the refrigerator, also can be used as seed and directly be used for producing;
B. shaken cultivation: adopt the liquid culture bacterial classification or on a small scale breeding cultivate all available shaken cultivation; The medium of shaken cultivation deducts agar to solid culture medium and gets final product; Equipment is preferably selected the constant-temperature shaking culture machine for use, puts liquid nutrient medium with triangular flask, inserts the test tube solid spawn can cultivate.Make through continuous vibration that various compositions mix and unlikely deposition in the liquid nutrient medium, promote gas contact and exchange simultaneously, make in the oxygen entering liquid nutrient medium, be beneficial to the formation of mycelial growth and conidium with liquid;
C. big jar of aerobic fementation cultivated: when bacterium powder such as large-scale production mycelia and conidium, and must be with big jar of aerobic fementation cultivation; This method ventilation is to adopt methods such as air-breathing or vavuum pump decompression, through filter removal of impurities bacterium, sends into and supplies growing of aweto in the pot liquid medium.
Fermentation requires temperature at 20-25 ℃ in the aweto jar, and tank pressure 392.3-686.5kPa, throughput are 0.5-1.0VVm*; The liquid nutrient medium that injects fermentation tank is a jar 65%-75% who holds; Inoculum concentration is 10%, and mixing speed is 180r/min, and incubation time 72-96h can be put jar and to concentrate and dust;
Fermentation medium: fresh potato (peeling) 8%, sucrose 2%, corn starch 0.5%, dried silkworm chrysalis meal 1%, peptone 0.4%, ammonium sulfate 0.2%, pH value 5.5-6.0;
Fermented and cultured finished product standard: conidium is several parent sporophores that come off entirely, and sporophore quantity increases when not obvious, count check under mirror, and every milliliter contains conidium 1,800,000,000-2,500,000,000; Residual sugar is lower than 1%; Amino nitrogen is lower than 0.2mg/mL, can become finished product bacterium liquid.
Effect of the present invention is: through aweto strain cultural method of the present invention; Can increase the output of Cordyceps sinensis; Fundamentally reduce the price of Cordyceps sinensis, the misery that palliates a disease for extensive patients plays a role, for increasing another wonderful work again in China's traditional medicine treasure-house.
Embodiment
The invention provides a kind of aweto strain cultural method; Said method for nature Cordyceps sinensis through separation and purification, test tube slant kind, oscillator liquid one-level kind, secondary kind, small-sized seed fermentation seeding tank, produce fermentation tank, concentrate and the drying of dusting, finished product bacterium powder.
The specific requirement of purification procedures is:
(1) material requirements: the piece of tissue partition method of carrying out Cordyceps sinensis; Parting material is annual by the end of October to November; The high and cold grassy marshland soil in Qinghai-Tibet Platean has just begun to freeze the material adopted period; Aweto just infected Hepialus larva to get into the bombys batryticatus phase not long-time in period this, and tender little stroma bud has just grown worm head 0.2-0.5cm, and other assorted bacterium are less in the polypide;
(2) separation operating method has following 3 steps:
A. the separation of bombys batryticatus body (being sclerotium): before the separation, water scrubs clean body surfaces, uses sterile water wash again 2 one 3 times; Mercuric chloride solution with 0.1% one 0.2% carries out the surface sterilization of about 3 one 5min to parting material; Use sterile water wash again.Choose with pereiopoda is the leading portion part on boundary.Cut epidermis with scalpel, avoid digestive tract, get the haemocoele mycelia and be cut into the sesame seed size, be pressed in the plating medium, 1 one 2 in every ware.Place 15 1 19 ℃ of cultivations, when treating that bacterium colony grows 0.2 1 0.5cm, select a small amount of mycelia and on plating medium, divide pure 2 one 4 times repeatedly, behind definite other assorted bacterium of nothing, move into test tube and preserve and cultivate;
B. the stroma bud separates: downcut the stroma bud of cleaning from the bombys batryticatus crown, insert sterilization 2 one 3min in the 0.1% mercuric chloride liquid, clean with sterile water, cut the middle part piece of tissue and be pressed in the medium; The same a of condition of culture;
C. ascospore separates: entangle the ripe stroma of ascospore with transparent paper bag, let the ascospore spring stick on the paper bag, then; In the Glucose Liquid that thecasporous paper bag immersion 25% is arranged; Wash spore, put into 15 1 19 ℃ of cultivations, every day microscopy; When robe is sprouted, inhale single spore with micro pipette and drip in flat board and cultivate; The same a of condition of culture;
Also can take back the Cordyceps sinensis of whole maturation indoorly, encase bombys batryticatus and the stroma that connects polypide, only reserve the part of being pregnant with cotton paper, in desinfection chamber, traverse, underlying a slice slide keeps main body humidity at any time; Microscopy when seeing that ascospore is bound to slide, was drawn onto with micro pipette in the medium of plate and cultivated every day; The same a of condition of culture.
(3) selecting for use of medium:
A. potato dextrose agar (PDA), this kind prescription is the collective media of all Chinese caterpillar fungus Pseudomonas fungies, can fill a prescription with this at the separation and Culture initial stage, but growth is very not vigorous, and be prone to aging, degenerate;
B. enriched medium (1): many property albumen navel 10g, glucose 50g, dipotassium hydrogen phosphate 1g, magnesium sulfate 0.5g, the silkworm chrysalis 30g that lives, growth hormone 0.5 μ g, agar 20g adds water 1000mL, pH:5.0;
Enriched medium (II): albumen dried meat 40g, glucose 40g, peeling fresh potato 100g, dipotassium hydrogen phosphate 1g; Magnesium sulfate 0.5g, beef extract 10g, growth hormone 0.5 μ g, bat larva (grinding) 30g alive; Agar 20g, high and cold meadow soil leachate 10OOmL, pH:5.0.
On enriched medium, bacterium colony is more vigorous and rapid than growth on the PDA medium; Add rich II and number be superior to I number again.
After the aweto separation and purification, promptly can be used as various experiments and enlarge production application;
(1) it enlarges the method for producing often has three kinds: solid leaves standstill cultivation, oscillating culture and big jar of aerobic fementation cultivation;
A. leave standstill cultivation: mainly be used in solid culture, cultivate or the like like test tube slant, triangular flask cultivation, rice.Can make the bacterium normal growth as long as in leaving standstill cultivation, grasp temperature and illumination.After the conidium maturation on the inclined-plane, can deposit in and preserve 8-14 month in 1-2 ℃ the refrigerator, also can be used as seed and directly be used for producing;
B. shaken cultivation: adopt the liquid culture bacterial classification or on a small scale breeding cultivate all available shaken cultivation; The medium of shaken cultivation deducts agar to solid culture medium and gets final product; Equipment is preferably selected the constant-temperature shaking culture machine for use, puts liquid nutrient medium with triangular flask, inserts the test tube solid spawn can cultivate.Make through continuous vibration that various compositions mix and unlikely deposition in the liquid nutrient medium, promote gas contact and exchange simultaneously, make in the oxygen entering liquid nutrient medium, be beneficial to the formation of mycelial growth and conidium with liquid;
C. big jar of aerobic fementation cultivated: when bacterium powder such as large-scale production mycelia and conidium, and must be with big jar of aerobic fementation cultivation; This method ventilation is to adopt methods such as air-breathing or vavuum pump decompression, through filter removal of impurities bacterium, sends into and supplies growing of aweto in the pot liquid medium.
Fermentation requires temperature at 20-25 ℃ in the aweto jar, and tank pressure 392.3-686.5kPa, throughput are 0.5-1.0VVm*; The liquid nutrient medium that injects fermentation tank is a jar 65%-75% who holds; Inoculum concentration is 10%, and mixing speed is 180r/min, and incubation time 72-96h can be put jar and to concentrate and dust;
Fermentation medium: fresh potato (peeling) 8%, sucrose 2%, corn starch 0.5%, dried silkworm chrysalis meal 1%, peptone 0.4%, ammonium sulfate 0.2%, pH value 5.5-6.0;
Fermented and cultured finished product standard: conidium is several parent sporophores that come off entirely, and sporophore quantity increases when not obvious, count check under mirror, and every milliliter contains conidium 1,800,000,000-2,500,000,000; Residual sugar is lower than 1%; Amino nitrogen is lower than 0.2mg/mL, can become finished product bacterium liquid.
What should explain at last is: obviously, the foregoing description only be for clearly the present invention is described and is done for example, and be not qualification to embodiment.For the those of ordinary skill in affiliated field, on the basis of above-mentioned explanation, can also make other multi-form variation or change.Here need not also can't give exhaustive to all embodiments.And conspicuous variation of being amplified out thus or change still are among protection scope of the present invention.
Claims (4)
1. aweto strain cultural method; It is characterized in that: said method for nature Cordyceps sinensis through separation and purification, test tube slant kind, oscillator liquid one-level kind, secondary kind, small-sized seed fermentation seeding tank, produce fermentation tank, concentrate and the drying of dusting, finished product bacterium powder.
2. aweto strain cultural method as claimed in claim 1 is characterized in that, the specific requirement of purification procedures is:
(1) material requirements: the piece of tissue partition method of carrying out Cordyceps sinensis; Parting material is annual by the end of October to November; The high and cold grassy marshland soil in Qinghai-Tibet Platean has just begun to freeze the material adopted period; Aweto just infected Hepialus larva to get into the bombys batryticatus phase not long-time in period this, and tender little stroma bud has just grown worm head 0.2-0.5cm, and other assorted bacterium are less in the polypide;
(2) separation operating method has following 3 steps:
A. the separation of bombys batryticatus body (being sclerotium): before the separation, water scrubs clean body surfaces, uses sterile water wash again 2 one 3 times; Mercuric chloride solution with 0.1% one 0.2% carries out the surface sterilization of about 3 one 5min to parting material; Use sterile water wash again; Choose with pereiopoda is the leading portion part on boundary; Cut epidermis with scalpel, avoid digestive tract, get the haemocoele mycelia and be cut into the sesame seed size; Be pressed in the plating medium, 1 one 2 in every ware places 15 1 19 ℃ of cultivations; When treating that bacterium colony grows 0.2 1 0.5cm; Select a small amount of mycelia and on plating medium, divide pure 2 one 4 times repeatedly, behind definite other assorted bacterium of nothing, move into test tube and preserve and cultivate;
B. the stroma bud separates: downcut the stroma bud of cleaning from the bombys batryticatus crown, insert sterilization 2 one 3min in the 0.1% mercuric chloride liquid, clean with sterile water, cut the middle part piece of tissue and be pressed in the medium; The same a of condition of culture;
C. ascospore separates: entangle the ripe stroma of ascospore with transparent paper bag, let the ascospore spring stick on the paper bag, then; In the Glucose Liquid that thecasporous paper bag immersion 25% is arranged; Wash spore, put into 15 1 19 ℃ of cultivations, every day microscopy; When robe is sprouted, inhale single spore with micro pipette and drip in flat board and cultivate; The same a of condition of culture;
Also can take back the Cordyceps sinensis of whole maturation indoorly, encase bombys batryticatus and the stroma that connects polypide, only reserve the part of being pregnant with cotton paper, in desinfection chamber, traverse, underlying a slice slide keeps main body humidity at any time; Microscopy when seeing that ascospore is bound to slide, was drawn onto with micro pipette in the medium of plate and cultivated every day; The same a of condition of culture;
(3) selecting for use of medium:
A. potato dextrose agar (PDA), this kind prescription is the collective media of all Chinese caterpillar fungus Pseudomonas fungies, can fill a prescription with this at the separation and Culture initial stage, but growth is very not vigorous, and be prone to aging, degenerate;
B. enriched medium (1): many property albumen navel 10g, glucose 50g, dipotassium hydrogen phosphate 1g, magnesium sulfate 0.5g, the silkworm chrysalis 30g that lives, growth hormone 0.5 μ g, agar 20g adds water 1000mL, pH:5.0;
Enriched medium (II): albumen dried meat 40g, glucose 40g, peeling fresh potato 100g, dipotassium hydrogen phosphate 1g; Magnesium sulfate 0.5g, beef extract 10g, growth hormone 0.5 μ g, bat larva (grinding) 30g alive; Agar 20g, high and cold meadow soil leachate 10OOmL, pH:5.0;
On enriched medium, bacterium colony is more vigorous and rapid than growth on the PDA medium; Add rich II and number be superior to I number again.
3. aweto strain cultural method as claimed in claim 2 is characterized in that, after the aweto separation and purification, promptly can be used as various experiments and enlarges production application;
(1) it enlarges the method for producing often has three kinds: solid leaves standstill cultivation, oscillating culture and big jar of aerobic fementation cultivation;
A. leave standstill cultivation: mainly be used in solid culture, as test tube slant, triangular flask cultivate, rice cultivates or the like, as long as in leaving standstill cultivation, grasp temperature and illumination can make the bacterium normal growth; After the conidium maturation on the inclined-plane, can deposit in and preserve 8-14 month in 1-2 ℃ the refrigerator, also can be used as seed and directly be used for producing;
B. shaken cultivation: adopt the liquid culture bacterial classification or on a small scale breeding cultivate all available shaken cultivation; The medium of shaken cultivation deducts agar to solid culture medium and gets final product; Equipment is preferably selected the constant-temperature shaking culture machine for use, puts liquid nutrient medium with triangular flask, inserts the test tube solid spawn can cultivate; Make through continuous vibration that various compositions mix and unlikely deposition in the liquid nutrient medium, promote gas contact and exchange simultaneously, make in the oxygen entering liquid nutrient medium, be beneficial to the formation of mycelial growth and conidium with liquid;
C. big jar of aerobic fementation cultivated: when bacterium powder such as large-scale production mycelia and conidium, and must be with big jar of aerobic fementation cultivation; This method ventilation is to adopt methods such as air-breathing or vavuum pump decompression, through filter removal of impurities bacterium, sends into and supplies growing of aweto in the pot liquid medium.
4. aweto strain cultural method as claimed in claim 3 is characterized in that, fermentation requires temperature at 20-25 ℃ in the aweto jar, and tank pressure 392.3-686.5kPa, throughput are 0.5-1.0VVm*; The liquid nutrient medium that injects fermentation tank is a jar 65%-75% who holds; Inoculum concentration is 10%, and mixing speed is 180r/min, and incubation time 72-96h can be put jar and to concentrate and dust;
Fermentation medium: fresh potato (peeling) 8%, sucrose 2%, corn starch 0.5%, dried silkworm chrysalis meal 1%, peptone 0.4%, ammonium sulfate 0.2%, pH value 5.5-6.0;
Fermented and cultured finished product standard: conidium is several parent sporophores that come off entirely, and sporophore quantity increases when not obvious, count check under mirror, and every milliliter contains conidium 1,800,000,000-2,500,000,000; Residual sugar is lower than 1%; Amino nitrogen is lower than 0.2mg/mL, can become finished product bacterium liquid.
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| CN110731234A (en) * | 2019-10-22 | 2020-01-31 | 安发（福建）生物科技有限公司 | method for rapid spore production of cordyceps militaris |
| CN112772294A (en) * | 2021-01-05 | 2021-05-11 | 中国科学院动物研究所 | Separation method and application of cordyceps sinensis strain |
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