CN106544277B - A kind of selenium-rich cordyceps culturing method - Google Patents
A kind of selenium-rich cordyceps culturing method Download PDFInfo
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- CN106544277B CN106544277B CN201610889346.0A CN201610889346A CN106544277B CN 106544277 B CN106544277 B CN 106544277B CN 201610889346 A CN201610889346 A CN 201610889346A CN 106544277 B CN106544277 B CN 106544277B
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Abstract
The invention discloses a kind of selenium-rich cordyceps culturing methods.The present invention provides a kind of method for preparing fruiting bodies of cordyceps militaris product, include the following steps: that (a1) using seed enriched medium culture cordyceps militaris link bacterial strain, obtains seed hot housing object；(a2) the seed hot housing object that step (a1) obtains is seeded in liquid seed culture medium and is cultivated, obtain liquid seeds；(a3) liquid seeds that step (a2) obtains are seeded in fructification selenium-rich culturing base and are cultivated, obtain fruiting bodies of cordyceps militaris.The present invention improves the utilization rate and absorptivity of selenium, greatly enhances the Se content in fructification using Se-enriched yeast powder as selenium source is provided while cordyceps militaris link bacterial strain fructification culture organic nitrogen source.Moreover, the selenium in Se-enriched yeast powder exists mostly in the form of organic selenium, it is the selenium source of a kind of safely and effectively selenium-rich fructification, can be promoted in the whole industry, have good economic and social benefit.
Description
Technical field
The present invention relates to a kind of selenium-rich cordyceps culturing methods.
Background technique
Cordyceps militaris (Cordyceps militaris) also known as northern Chinese caterpillar Fungus, belong to mycota, belong on taxology true
Bacterium door, Ascomycotina, gang pyrenomycetes, Spheeriales, Clavicipitaceae, Cordyceps, Cordyceps militaris distribution is wide, growth is fast, easily cultivates, contains
There are many active constituents, have different physiological roles to human body.
Modern research shows that the substance that Cordyceps militaris drug effect works typically belongs to ucleosides, glycitols, polysaccharide, amino
Acids, enzyme and mineral element etc..Fixed function factor mainly has cordycepin, adenosine, mannitol, Cordyceps sinensis polysaccharide, surpasses
Superoxide dismutase (SOD) etc..Wherein, cordycepin is most important one kind, is played in clinical medicine and health care important
Effect.In recent years, the effective component as Cordyceps militaris class product marking, the physiological function point of cordycepin and pharmacological action
Research, caused the highest attention of biological educational circles and medicine sector scientific worker, scientific worker is to cordycepin product
Developmental research is also paid attention to and specific further.With the active function to cordycepin gradually recognize and mechanism of action disclose, institute
Antibacterial, anti-inflammatory, the significant function that is antitumor, adjusting human endocrine and enhancing immune function of human body etc. of performance, can all obtain
It is further to pay attention to.It is new resource food that the 2009 Nian Yuan Ministry of Public Health, which ratify Cordyceps militaris, assert that it, as new health food, is winter worm
The favorable substitutes of summer grass.National health State Family Planning Commission approval Cordyceps militaris eliminates as new raw-food material to Cordyceps militaris within 2014
Amount and the limitation such as use scope, further embody safety and feasibility of the Cordyceps militaris as healthy food.
In China, the shortage of selenium is a common problem.And selenium is the necessary Elements of limit quantity of human body, is had anti-
Cancer, cardioprotection, treating liver necrosis, prevention and treatment myopia and cataract, removing toxic substances improve immunity, anti-aging and enhancing reproductive function
Etc. multiple pharmacological effects pass through the inorganic selenium source of supplement for selenium deficiency area --- sodium selenite, absorption rate is low, peace
Full property is poor, and selenium-supply should also be mainly preferably rich in organic selenium, because organic selenium is better than inorganic selenium in terms of utilization rate, and by micro-
Bio-absorbable, conversion, enrichment selenium be exactly a kind of very high selenium source of bioavailability, thus the exploitation of selenium-rich fungi causes
The attention of people.In general, edible bacterial type microorganism all has stronger selenium rich ability and higher yield.Moreover,
Cordyceps contain various bioactive components, and the same of the nutritive and medicinal value of Cordyceps militaris then can be improved in culture selenium-enriched cordceps militaris
When achieve the purpose that people's selenium-supply health care.Currently, cultivating the more of the source use of selenium in the technical solution of selenium-enriched cordceps militaris mostly
It is fertilizer nutrition liquid containing selenium, and selenium fertilizer nutrition liquid mostly uses sodium selenite as the source of selenium element, using sodium selenite conduct
The source Cordyceps militaris of selenium element is difficult to absorb, and leads to the Se content extremely pettiness in the fruiting bodies of cordyceps militaris cultivated, it is difficult to mention
The nutritive and medicinal value of high Cordyceps militaris, and the residual of excessive inorganic selenium brings unfavorable shadow to the utilization of Chinese caterpillar fungus culture medium
It rings.How the Cordyceps militaris product for many physiological functions that Efficient Development integrates Active Components in Various Cordyceps militaris Strains and selenium becomes one newly
Research topic.
Summary of the invention
The object of the present invention is to provide a kind of selenium-rich cordyceps culturing methods.
The present invention provides a kind of methods for preparing fruiting bodies of cordyceps militaris product, include the following steps:
(a1) seed enriched medium culture cordyceps militaris link bacterial strain is used, seed hot housing object is obtained；
(a2) the seed hot housing object that step (a1) obtains is seeded in liquid seed culture medium and is cultivated, obtained
Liquid seeds；
(a3) liquid seeds that step (a2) obtains are seeded in fructification selenium-rich culturing base and are cultivated, obtain pupa worm
Grass seed entity.
The raw material proportioning of the seed enriched medium are as follows: 1L nutrient solution first: 0.5-1.5kg millet；The nutrient solution first
In containing concentration be 1.5mg/100ml or more selenium.
Specifically containing the selenium of 1.5mg/100ml-20mg/100ml in the nutrient solution first.
Specifically containing the selenium of 1.96mg/100ml in the nutrient solution first.
Specifically containing the selenium of 3.92mg/100ml in the nutrient solution first.
Specifically containing the selenium of 5.88mg/100ml in the nutrient solution first.
Specifically containing the selenium of 19.76mg/100ml in the nutrient solution first.
The selenium for being 0.9mg/100ml or more containing concentration in the liquid seed culture medium.
The selenium for being specifically 0.9mg/100ml-10mg/100ml containing concentration in the liquid seed culture medium.
Specifically containing the selenium of 0.98mg/100ml in the nutrient solution.
Specifically containing the selenium of 1.96mg/100ml in the nutrient solution.
Specifically containing the selenium of 2.94mg/100ml in the nutrient solution.
Specifically containing the selenium of 9.88mg/100ml in the nutrient solution.
The raw material proportioning of the fructification selenium-rich culturing base are as follows: 100-150ml nutrient solution second: 100g wheat seed；It is described
The selenium for being 1.5mg/100ml or more containing concentration in nutrient solution second.
The selenium for being 1.5mg/100ml-20mg/100ml containing concentration in the nutrient solution second.
Specifically containing the selenium of 1.96mg/100ml in the nutrient solution second.
Specifically containing the selenium of 3.92mg/100ml in the nutrient solution second.
Specifically containing the selenium of 5.88mg/100ml in the nutrient solution second.
Specifically containing the selenium of 19.76mg/100ml in the nutrient solution second.
Any description above selenium is organic selenium.
Any description above selenium can be added in culture medium in the form of Se-enriched yeast powder.
Any description above selenium can also be added in culture medium in the form of Se-enriched yeast product.
Se content is 2000-2400mg/kg, organic selenium content >=98% in the Se-enriched yeast powder.
Organic selenium content is 1960-2400mg/kg in the Se-enriched yeast powder.
The organic selenium content of the Se-enriched yeast product is 9880mg/kg.
The preparation method of the Se-enriched yeast product be patent of invention (application No. is: 201210258191.2, grant date is
2014.09.10) in Se-enriched yeast product preparation method as described in example 2.
The raw material proportioning of the seed enriched medium is concretely: 1L nutrient solution: 0.5kg millet.
The raw material proportioning of the seed enriched medium is concretely: 1L nutrient solution: 1.0kg millet.
The raw material proportioning of the seed enriched medium is concretely: 1L nutrient solution: 1.5kg millet.
Any description above nutrient solution first is made of solute and solvent；The solute and its dense in the nutrient solution first
It spends as follows: Se-enriched yeast powder (or described Se-enriched yeast product) 1-3g/100ml, glucose 0.5-1.5g/100ml, peptone
0.5-0.7g/100ml, potassium dihydrogen phosphate 0.05-0.15g/100ml, magnesium sulfate 0.04-0.06g/100ml, vitamin B1 1-
3mg/100ml, vitamin B20.5-1.5mg/100ml, glycine 0.2-0.4mg/100ml, threonine 0.2-0.3mg/
100ml, valine 0.1-0.2mg/100ml, leucine 0.04-0.06mg/100ml, isoleucine 0.05-0.15mg/
100ml, phenylalanine 0.2-0.3mg/100ml, serine 0.05-0.15mg/100ml, proline 0.04-0.06mg/
100ml, hydroxyproline 0.02-0.04mg/100ml, cysteine 0.01-0.03mg/100ml, tryptophan 0.02-0.03mg/
100ml, methionine 0.04-0.06mg/100ml, lysine 0.05-0.15mg/100ml, glutamic acid 0.04-0.06mg/
100ml；The solvent is water.
The solute and its concentration in the nutrient solution first can are as follows: Se-enriched yeast powder (or described Se-enriched yeast product)
1-2g/100ml, glucose 0.5-1.0g/100ml, peptone 0.5-0.6g/100ml, potassium dihydrogen phosphate 0.05-0.10g/
100ml, magnesium sulfate 0.04-0.05g/100ml, vitamin B11-2mg/100ml, vitamin B2It is 0.5-1.0mg/100ml, sweet
Propylhomoserin 0.2-0.3mg/100ml, threonine 0.2-0.25mg/100ml, valine 0.1-0.15mg/100ml, leucine 0.04-
0.05mg/100ml, isoleucine 0.05-0.10mg/100ml, phenylalanine 0.2-0.25mg/100ml, serine 0.05-
0.10mg/100ml, proline 0.04-0.05mg/100ml, hydroxyproline 0.02-0.03mg/100ml, cysteine 0.01-
0.02mg/100ml, tryptophan 0.02-0.025mg/100ml, methionine 0.04-0.05mg/100ml, lysine 0.05-
0.10mg/100ml, glutamic acid 0.04-0.05mg/100ml.
The solute and its concentration in the nutrient solution first can are as follows: Se-enriched yeast powder (or described Se-enriched yeast product)
2-3g/100ml, glucose 1.0-1.5g/100ml, peptone 1.0-0.7g/100ml, potassium dihydrogen phosphate 0.10-0.15g/
100ml, magnesium sulfate 0.05-0.06g/100ml, vitamin B12-3mg/100ml, vitamin B2It is 1.0-1.5mg/100ml, sweet
Propylhomoserin 0.3-0.4mg/100ml, threonine 0.25-0.3mg/100ml, valine 0.15-0.2mg/100ml, leucine 0.05-
0.06mg/100ml, isoleucine 0.10-0.15mg/100ml, phenylalanine 0.25-0.3mg/100ml, serine 0.10-
0.15mg/100ml, proline 0.05-0.06mg/100ml, hydroxyproline 0.03-0.04mg/100ml, cysteine 0.02-
0.03mg/100ml, tryptophan 0.025-0.03mg/100ml, methionine 0.05-0.06mg/100ml, lysine 0.10-
0.15mg/100ml, glutamic acid 0.05-0.06mg/100ml.
The solute and its concentration in the nutrient solution first is concretely: Se-enriched yeast powder (or the Se-enriched yeast
Product) 2g/100ml, glucose 1g/100ml, peptone 0.6g/100ml, potassium dihydrogen phosphate 0.1g/100ml, magnesium sulfate
0.05g/100ml, vitamin B12mg/100ml, vitamin B21mg/100ml, glycine 0.3mg/100ml, threonine
0.25mg/100ml, valine 0.15mg/100ml, leucine 0.05mg/100ml, isoleucine 0.1mg/100ml, phenylpropyl alcohol ammonia
Sour 0.25mg/100ml, serine 0.1mg/100ml, proline 0.05mg/100ml, hydroxyproline 0.03mg/100ml, half Guang
Propylhomoserin 0.02mg/100ml, tryptophan 0.025mg/100ml, methionine 0.05mg/100ml, lysine 0.1mg/100ml,
Glutamic acid 0.05mg/100ml.
The solute and its concentration in the nutrient solution first is concretely: Se-enriched yeast powder (or the Se-enriched yeast
Product) 3g/100ml, glucose 1.5g/100ml, peptone 0.7g/100ml, potassium dihydrogen phosphate 0.15g/100ml, magnesium sulfate
0.06g/100ml, vitamin B13mg/100ml, vitamin B21.5mg/100ml, glycine 0.4mg/100ml, threonine
0.3mg/100ml, valine 0.2mg/100ml, leucine 0.06mg/100ml, isoleucine 0.15mg/100ml, phenylpropyl alcohol ammonia
Sour 0.3mg/100ml, serine 0.15mg/100ml, proline 0.06mg/100ml, hydroxyproline 0.04mg/100ml, half Guang
Propylhomoserin 0.03mg/100ml, tryptophan 0.03mg/100ml, methionine 0.06mg/100ml, lysine 0.15mg/100ml,
Glutamic acid 0.06mg/100ml.
The solute and its concentration in the nutrient solution first is concretely: Se-enriched yeast powder (or the Se-enriched yeast
Product) 1g/100ml, glucose 0.5g/100ml, peptone 0.5g/100ml, potassium dihydrogen phosphate 0.05g/100ml, magnesium sulfate
0.04g/100ml, vitamin B11mg/100ml, vitamin B20.5mg/100ml, glycine 0.2mg/100ml, threonine
0.2mg/100ml, valine 0.1mg/100ml, leucine 0.04mg/100ml, isoleucine 0.05mg/100ml, phenylpropyl alcohol ammonia
Sour 0.2mg/100ml, serine 0.05mg/100ml, proline 0.04mg/100ml, hydroxyproline 0.02mg/100ml, half Guang
Propylhomoserin 0.01mg/100ml, tryptophan 0.02mg/100ml, methionine 0.04mg/100ml, lysine 0.05mg/100ml,
Glutamic acid 0.04mg/100ml.
Any description above liquid seed culture medium is made of solute and solvent；The solute and its in the liquid seeds
Concentration in culture medium is as follows: potato leaches powder 0.5-1.5g/100ml, glucose 0.5-1.5g/100ml, Se-enriched yeast powder
(or described Se-enriched yeast product) 0.5-1.5g/100ml, peptone 0.4-0.6g/100ml, sodium chloride 0.05-0.15g/
100ml, potassium dihydrogen phosphate 0.02-0.04g/100ml, dipotassium hydrogen phosphate 0.04-0.06g/100ml, magnesium sulfate 0.04-0.06g/
100ml, vitamin B11-3mg/100ml, vitamin B20.5-1.5mg/100ml, glycine 0.2-0.4mg/100ml, Soviet Union's ammonia
Sour 0.2-0.3mg/100ml, valine 0.1-0.2mg/100ml, leucine 0.04-0.06mg/100ml, isoleucine 0.05-
0.15mg/100ml, phenylalanine 0.2-0.3mg/100ml, serine 0.05-0.15mg/100ml, proline 0.04-
0.06mg/100ml, hydroxyproline 0.02-0.04mg/100ml, cysteine 0.01-0.03mg/100ml, tryptophan 0.02-
0.03mg/100ml, methionine 0.04-0.06mg/100ml, lysine 0.05-0.15mg/100ml, glutamic acid 0.04-
0.06mg/100ml；The solvent is water.
The solute and its concentration in the liquid seed culture medium can are as follows: potato leaches powder 0.5-1.0g/
100ml, glucose 0.5-1.0g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 0.5-1.0g/100ml, albumen
Peptone 0.4-0.5g/100ml, sodium chloride 0.05-0.10g/100ml, potassium dihydrogen phosphate 0.02-0.03g/100ml, dipotassium hydrogen phosphate
0.04-0.05g/100ml, magnesium sulfate 0.04-0.05g/100ml, vitamin B11-2mg/100ml, vitamin B2 0.5-
1.0mg/100ml, glycine 0.2-0.3mg/100ml, threonine 0.2-0.25mg/100ml, valine 0.1-0.15mg/
100ml, leucine 0.04-0.05mg/100ml, isoleucine 0.05-0.10mg/100ml, phenylalanine 0.2-0.25mg/
100ml, serine 0.05-0.10mg/100ml, proline 0.04-0.05mg/100ml, hydroxyproline 0.02-0.03mg/
100ml, cysteine 0.01-0.02mg/100ml, tryptophan 0.02-0.025mg/100ml, methionine 0.04-0.05mg/
100ml, lysine 0.05-0.10mg/100ml, glutamic acid 0.04-0.05mg/100ml.
The solute and its concentration in the liquid seed culture medium can are as follows: potato leaches powder 1.0-1.5g/
100ml, glucose 1.0-1.5g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 1.0-1.5g/100ml, albumen
Peptone 0.5-0.6g/100ml, sodium chloride 0.10-0.15g/100ml, potassium dihydrogen phosphate 0.03-0.04g/100ml, dipotassium hydrogen phosphate
0.05-0.06g/100ml, magnesium sulfate 0.05-0.06g/100ml, vitamin B12-3mg/100ml, vitamin B2 1.0-
1.5mg/100ml, glycine 0.3-0.4mg/100ml, threonine 0.25-0.3mg/100ml, valine 0.15-0.2mg/
100ml, leucine 0.05-0.06mg/100ml, isoleucine 0.06-0.15mg/100ml, phenylalanine 0.25-0.3mg/
100ml, serine 0.10-0.15mg/100ml, proline 0.05-0.06mg/100ml, hydroxyproline 0.03-0.04mg/
100ml, cysteine 0.02-0.03mg/100ml, tryptophan 0.025-0.03mg/100ml, methionine 0.05-0.06mg/
100ml, lysine 0.10-0.15mg/100ml, glutamic acid 0.05-0.06mg/100ml.
The solute and its concentration in the liquid seed culture medium is concretely: potato leaches powder 1g/
100ml, glucose 1g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 1g/100ml, peptone 0.5g/100ml,
Sodium chloride 0.1g/100ml, potassium dihydrogen phosphate 0.03g/100ml, dipotassium hydrogen phosphate 0.05g/100ml, magnesium sulfate 0.05g/
100ml, vitamin B12mg/100ml, vitamin B21mg/100ml, glycine 0.3mg/100ml, threonine 0.25mg/
100ml, valine 0.15mg/100ml, leucine 0.05mg/100ml, isoleucine 0.1mg/100ml, phenylalanine
0.25mg/100ml, serine 0.1mg/100ml, proline 0.05mg/100ml, hydroxyproline 0.03mg/100ml, half Guang ammonia
Sour 0.02mg/100ml, tryptophan 0.025mg/100ml, methionine 0.05mg/100ml, lysine 0.1mg/100ml, paddy
Propylhomoserin 0.05mg/100ml.
The solute and its concentration in the liquid seed culture medium is concretely: potato leaches powder 1.5g/
100ml, glucose 1.5g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 1.5g/100ml, peptone 0.6g/
100ml, sodium chloride 0.15g/100ml, potassium dihydrogen phosphate 0.04g/100ml, dipotassium hydrogen phosphate 0.06g/100ml, magnesium sulfate
0.06g/100ml, vitamin B13mg/100ml, vitamin B21.5mg/100ml, glycine 0.4mg/100ml, threonine
0.3mg/100ml, valine 0.2mg/100ml, leucine 0.06mg/100ml, isoleucine 0.15mg/100ml, phenylpropyl alcohol ammonia
Sour 0.3mg/100ml, serine 0.15mg/100ml, proline 0.06mg/100ml, hydroxyproline 0.04mg/100ml, half Guang
Propylhomoserin 0.03mg/100ml, tryptophan 0.03mg/100ml, methionine 0.06mg/100ml, lysine 0.15mg/100ml,
Glutamic acid 0.06mg/100ml.
The solute and its concentration in the liquid seed culture medium is concretely: potato leaches powder 0.5g/
100ml, glucose 0.5g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 0.5g/100ml, peptone 0.4g/
100ml, sodium chloride 0.05g/100ml, potassium dihydrogen phosphate 0.02g/100ml, dipotassium hydrogen phosphate 0.04g/100ml, magnesium sulfate
0.04g/100ml, vitamin B11mg/100ml, vitamin B20.5mg/100ml, glycine 0.2mg/100ml, threonine
0.2mg/100ml, valine 0.1mg/100ml, leucine 0.04mg/100ml, isoleucine 0.05mg/100ml, phenylpropyl alcohol ammonia
Sour 0.2mg/100ml, serine 0.05mg/100ml, proline 0.04mg/100ml, hydroxyproline 0.02mg/100ml, half Guang
Propylhomoserin 0.01mg/100ml, tryptophan 0.02mg/100ml, methionine 0.04mg/100ml, lysine 0.0gmg/100ml,
Glutamic acid 0.04mg/100ml.
The raw material of the fructification selenium-rich culturing base is with concretely: 100g millet: 100ml nutrient solution.
The raw material of the fructification selenium-rich culturing base is with concretely: 100g millet: 120ml nutrient solution.
The raw material of the fructification selenium-rich culturing base is with concretely: 100g millet: 150ml nutrient solution.
Any description above nutrient solution second is made of solute and solvent；The solute and its dense in the nutrient solution second
It spends as follows: glucose 0.5-1.5g/100ml, sucrose 0.4-0.6g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product)
1-3g/100ml, peptone 0.5-0.7g/100ml, potassium dihydrogen phosphate 0.02-0.04g/100ml, dipotassium hydrogen phosphate 0.04-
0.06g/100ml, ferric citrate 0.02-0.04g/100ml, zinc chloride 0.02-0.04g/100ml, magnesium sulfate 0.04-
0.06g/100ml, vitamin B11-3mg/100ml, vitamin B20.5-1.5mg/100ml, vitamin B110.5-1.5mg/
100ml；The solvent is water.
The solute and its concentration in the nutrient solution second can are as follows: glucose 0.5-1.0g/100ml, sucrose 0.4-
0.5g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 1-2g/100ml, peptone 0.5-0.6g/100ml, phosphoric acid
Potassium dihydrogen 0.02-0.03g/100ml, dipotassium hydrogen phosphate 0.04-0.05g/100ml, ferric citrate 0.02-0.03g/100ml,
Zinc chloride 0.02-0.03g/100ml, magnesium sulfate 0.04-0.05g/100ml, vitamin B11-2mg/100ml, vitamin B2O.5-
1.0mg/100ml, vitamin B110.5-1.0mg/100ml。
The solute and its concentration in the nutrient solution second can are as follows: glucose 1.0-1.5g/100ml, sucrose 0.5-
0.6g/100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 2-3g/100ml, peptone 0.6-0.7g/100ml, phosphoric acid
Potassium dihydrogen 0.03-0.04g/100ml, dipotassium hydrogen phosphate 0.05-0.06g/100ml, ferric citrate 0.03-0.04g/100ml,
Zinc chloride 0.03-0.04g/100ml, magnesium sulfate 0.05-0.06g/100ml, vitamin B12-3mg/100ml, vitamin B21.0-
1.5mg/100ml, vitamin B111.0-1.5mg/100ml。
The solute and its concentration in the nutrient solution second is concretely: glucose 1g/100ml, sucrose 0.5g/
100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 2g/100ml, peptone 0.6g/100ml, potassium dihydrogen phosphate
0.03g/100ml, dipotassium hydrogen phosphate 0.05g/100ml, ferric citrate 0.03g/100ml, zinc chloride 0.03g/100ml, sulphur
Sour magnesium 0.05g/100ml, vitamin B12mg/100ml, vitamin B21mg/100ml, vitamin B11 1mg/100ml。
The solute and its concentration in the nutrient solution second is concretely: glucose 1.5g/100ml, sucrose 0.6g/
100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 3g/100ml, peptone 0.7g/100ml, potassium dihydrogen phosphate
0.04g/100ml, dipotassium hydrogen phosphate 0.06g/100ml, ferric citrate 0.04g/100ml, zinc chloride 0.04g/100ml, sulphur
Sour magnesium 0.06g/100ml, vitamin B13mg/100ml, vitamin B21.5mg/100ml, vitamin B11 1.5mg/100ml。
The solute and its concentration in the nutrient solution second is concretely: glucose 0.5g/100ml, sucrose 0.4g/
100ml, Se-enriched yeast powder (or described Se-enriched yeast product) 1g/100ml, peptone 0.5g/100ml, potassium dihydrogen phosphate
0.02g/100ml, dipotassium hydrogen phosphate 0.04g/100ml, ferric citrate 0.02g/100ml, zinc chloride 0.02g/100ml, sulphur
Sour magnesium 0.04g/100ml, vitamin B11mg/100ml, vitamin B20.5mg/100ml, vitamin B11 0.5mg/100ml。
The step (a1) is concretely: cordyceps militaris link bacterial strain is seeded to seed enriched medium, and (cordyceps militaris link bacterial strain is being trained
Initial concentration in the system of supporting is 104A/ml), 26 DEG C are cultivated 4 days, and the inoculum that strengthened (cultivating by cordyceps militaris link bacterial strain
Spore concentration in system is 1012A/gram culture).
The step (a2) is concretely: by the product of 10g step (a1) be seeded in 100ml liquid seed culture medium into
Row culture；It is protected from light culture 4 days under the conditions of concretely 25 DEG C of the culture, 220rpm, obtains cordyceps militaris link bacterial strain liquid seeds, liquid
The biomass of body seed is 50% (Cordyceps militaris seed weight in wet base is 50g in every 100ml culture medium).
The step (a3) is concretely: the product of 100ml step (a2) is seeded to 1000g fructification selenium-rich culturing base
In cultivated；
The incubation of the culture are as follows: bacterium germination dark culture stage (7-10 days): dark culturing, 23-25 DEG C of temperature, air
Relative humidity 60-70%, carbon dioxide volume fraction is less than 0.3%；Veraison cultivation stage (15-20 days): 14-26 DEG C of temperature
(day and night temperature is at 10 DEG C or so), relative air humidity 70-80%, carbon dioxide volume fraction is less than 0.2%, intensity of illumination
350lux, 9.5 hour/day of light application time, light source type is fluorescent lamp；The sporophore growth management stage (15 days): temperature 16-20
DEG C, relative air humidity 80%-85%, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, light application time 8.5
Hour/day, light source type are fluorescent lamp；Blue light cultivation stage (5-10 days): 16-20 DEG C of temperature, relative air humidity 80%-
85%, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, 8.5 hour/day of light application time, and light source type is indigo plant
Light.
The incubation of the culture is concretely: the bacterium germination dark culture stage (8 days): dark culturing, and 25 DEG C of temperature, air
Relative humidity 65%, carbon dioxide volume fraction is less than 0.3%；Veraison cultivation stage (20 days): 14-26 DEG C of temperature is (round the clock
The temperature difference is at 10 DEG C or so), relative air humidity 80%, carbon dioxide volume fraction is less than 0.2%, intensity of illumination 350lux, light
According to 9.5 hour/day of time, light source type is fluorescent lamp；The sporophore growth management stage (15 days): 118 DEG C of temperature, air is opposite
Humidity 80%-85%, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, 8.5 hour/day of light application time, light
Source Type is fluorescent lamp；Blue light cultivation stage (10 days): 18 DEG C of temperature, relative air humidity 80%-85%, carbon dioxide volume
Score is less than 0.1%, intensity of illumination 250Lux, and 8.5 hour/day of light application time, light source type is blue light.
The incubation of the culture is concretely: the bacterium germination dark culture stage (7 days): it is protected from light culture, 23 DEG C of temperature, and air
Relative humidity 70%, carbon dioxide volume fraction is less than 0.3%；Veraison cultivation stage (18 days): 14-26 DEG C of temperature, (round the clock
The temperature difference is at 10 DEG C or so), relative air humidity 70%, carbon dioxide volume fraction is less than 0.2%, intensity of illumination 350lux, light
According to 9.5 hour/day of time, light source type is fluorescent lamp；The sporophore growth management stage (15 days): 16 DEG C of temperature, air is opposite
Humidity 80%-85%, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, 8.5 hour/day of light application time, light
Source Type is fluorescent lamp；Blue light cultivation stage (5 days): 16 DEG C of temperature, relative air humidity 80%-85%, carbon dioxide volume
Score is less than 0.1%, intensity of illumination 250Lux, and 8.5 hour/day of light application time, light source type is blue light.
The incubation of the culture is concretely: the bacterium germination dark culture stage (10 days): it is protected from light culture, it is 24 DEG C of temperature, empty
Gas relative humidity 60%, carbon dioxide volume fraction is less than 0.3%；Veraison cultivation stage (15 days): 14-26 DEG C of (daytime of temperature
The night temperature difference is at 10 DEG C or so), relative air humidity 75%, carbon dioxide volume fraction is less than 0.2%, intensity of illumination 350lux,
9.5 hour/day of light application time, light source type are fluorescent lamp；The sporophore growth management stage (15 days): 20 DEG C of temperature, air phase
To humidity 80%-85%, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, light application time be 8.5 hours/
It, light source type is fluorescent lamp；Blue light cultivation stage (7 days): 20 DEG C of temperature, relative air humidity 80%-85%, carbon dioxide
Volume fraction is less than 0.1%, intensity of illumination 250Lux, and light application time is 8.5 hours/day, and light source type is blue light.
The product that the present invention also protects any description above method to be prepared.
It is 280mg/kg or more, even up to 360mg/kg that the product, which meets total Se content,.
The present invention also protects a kind of kit for cultivating or preparing fruiting bodies of cordyceps militaris product for Cordyceps militaris, including above
Any seed enriched medium, any description above liquid seed culture medium and any description above fructification selenium-rich culturing
Base.
Any description above cordyceps militaris link bacterial strain concretely cordyceps militaris link bacterial strain 50383: Chinese agriculture Microbiological Culture Collection
Administrative center (www.accc.org.cn, abbreviation ACCC), ACCC number: 50383.
Any description above Se-enriched yeast powder is concretely blue to match yeast selenium S200 (Shandong sage's fine jade biology Co., Ltd).
The present invention is experimentally confirmed, using Se-enriched yeast powder as the same of cordyceps militaris link bacterial strain fructification culture organic nitrogen source
When provide selenium source, especially selenoaminoacid form selenium source, guarantee fruiting bodies of cordyceps militaris growth simultaneously, promote fructification
Direct absorption to selenium improves the utilization rate and absorptivity of selenium, greatly enhances the Se content in fructification.Moreover, selenium-rich ferment
Selenium in female powder exists mostly in the form of organic selenium, there is no the residual of inorganic selenium and to the adaptability breeding problem of bacterial strain,
It is the selenium source of a kind of safely and effectively selenium-rich fructification, can be promoted in the whole industry, have good economic and social benefit.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even
Mean value.
Cordyceps militaris link bacterial strain 50383: Chinese agriculture Microbiological Culture Collection administrative center (www.accc.org.cn, referred to as
ACCC), ACCC is numbered: 50383.
Cordyceps FY1421 (Cordyceps militaris FY1421) is preserved in Chinese allusion quotation on April 21st, 2016
Type culture collection (abbreviation CCTCC；Address: Wuhan, China, Wuhan University；Postcode: 430072), deposit number is
CCTCC NO:M2016220.
Se-enriched yeast powder: Shandong sage's fine jade biology Co., Ltd, indigo plant match yeast selenium S200；Se content is in Se-enriched yeast powder
2000-2400mg/kg, organic selenium content >=98%.
Amino acid nutrient mother liquor the preparation method comprises the following steps: taking glycine, threonine, valine, leucine, isoleucine, benzene
Alanine, serine, proline, hydroxyproline, cysteine, tryptophan, methionine, lysine and glutamic acid add distillation
Water is settled to 1000ml；4 DEG C of storages, it is spare.
PDA solid medium: potato leaches powder 20g, glucose 20g, agar 20g, and distilled water is settled to 1000mL；
121 DEG C of high pressure steam sterilization 20min.
The preparation method of seed enriched medium: Se-enriched yeast powder, glucose, peptone, potassium dihydrogen phosphate, sulfuric acid are taken
Magnesium, amino acid nutrient mother liquor, vitamin B1And vitamin B2, add distilled water to be settled to 100ml, nutrient solution first be made；Nutrient solution first
It is uniformly mixed with millet, Steam by water bath 30min obtains seed enriched medium；121 DEG C of high pressure sterilization 25min.According to the method described above
Seed enriched medium first, seed enriched medium second and seed enriched medium third are prepared respectively.
Seed enriched medium first is uniformly mixed with 1000g millet by 1000ml nutrient solution first and is made；Each solute is in nutrition
Concentration in liquid first is as follows: Se-enriched yeast powder 2g/100ml, glucose 1g/100ml, peptone 0.6g/100ml, biphosphate
Potassium 0.1g/100ml, magnesium sulfate 0.05g/100ml, vitamin B12mg/100ml, vitamin B21mg/100ml, glycine
0.3mg/100ml, threonine 0.25mg/100ml, valine 0.15mg/100ml, leucine 0.05mg/100ml, isoleucine
0.1mg/100ml, phenylalanine 0.25mg/100ml, serine 0.1mg/100ml, proline 0.05mg/100ml, hydroxyl dried meat ammonia
Sour 0.03mg/100ml, cysteine 0.02mg/100ml, tryptophan 0.025mg/100ml, methionine 0.05mg/100ml,
Lysine 0.1mg/100ml, glutamic acid 0.05mg/100ml.
Seed enriched medium second is uniformly mixed with 1500g millet by 1000ml nutrient solution first and is made；Each solute is in nutrition
Concentration in liquid first is as follows: Se-enriched yeast powder 3g/100ml, glucose 1.5g/100ml, peptone 0.7g/100ml, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.15g/100ml, magnesium sulfate 0.06g/100ml, vitamin B13mg/100ml, vitamin B21.5mg/100ml, sweet ammonia
Sour 0.4mg/100ml, threonine 0.3mg/100ml, valine 0.2mg/100ml, leucine 0.06mg/100ml, isoleucine
0.15mg/100ml, phenylalanine 0.3mg/100ml, serine 0.15mg/100ml, proline 0.06mg/100ml, hydroxyl dried meat ammonia
Sour 0.04mg/100ml, cysteine 0.03mg/100ml, tryptophan 0.03mg/100ml, methionine 0.06mg/100ml,
Lysine 0.15mg/100ml, glutamic acid 0.06mg/100ml.
Seed enriched medium third is uniformly mixed with 500g millet by 1000ml nutrient solution first and is made；Each solute is in nutrient solution
Concentration in first is as follows: Se-enriched yeast powder 1g/100ml, glucose 0.5g/100ml, peptone 0.5g/100ml, biphosphate
Potassium 0.05g/100ml, magnesium sulfate 0.04g/100ml, vitamin B11mg/100ml, vitamin B20.5mg/100ml, glycine
0.2mg/100ml, threonine 0.2mg/100ml, valine 0.1mg/100ml, leucine 0.04mg/100ml, isoleucine
0.05mg/100ml, phenylalanine 0.2mg/100ml, serine 0.05mg/100ml, proline 0.04mg/100ml, hydroxyl dried meat ammonia
Sour 0.02mg/100ml, cysteine 0.01mg/100ml, tryptophan 0.02mg/100ml, methionine 0.04mg/100ml,
Lysine 0.05mg/100ml, glutamic acid 0.04mg/100ml.
The preparation method of liquid seed culture medium: potato is taken to leach powder, glucose, Se-enriched yeast powder, peptone, chlorination
Sodium, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, vitamin B1, vitamin B2And span amino acid mother liquor, adjusting pH is 6.8, adds distillation
Water is settled to 1000ml；121 DEG C of high pressure sterilization 20min.Liquid seed culture medium first, liquid is prepared respectively according to the method described above
Body seed culture medium second and liquid seed culture medium third.
In liquid seed culture medium first, the concentration of each solute is as follows: potato leaches powder 1g/100ml, glucose 1g/
100ml, Se-enriched yeast powder 1g/100ml, peptone 0.5g/100ml, sodium chloride 0.1g/100ml, potassium dihydrogen phosphate 0.03g/
100ml, dipotassium hydrogen phosphate 0.05g/100ml, magnesium sulfate 0.05g/100ml, vitamin B12mg/100ml, vitamin B2 1mg/
100ml, glycine 0.3mg/100ml, threonine 0.25mg/100ml, valine 0.15mg/100ml, leucine 0.05mg/
100ml, isoleucine 0.1mg/100ml, phenylalanine 0.25mg/100ml, serine 0.1mg/100ml, proline
0.05mg/100ml, hydroxyproline 0.03mg/100ml, cysteine 0.02mg/100ml, tryptophan 0.025mg/100ml, first
Methyllanthionine 0.05mg/100ml, lysine 0.1mg/100ml, glutamic acid 0.05mg/100ml.
In liquid seed culture medium second, the concentration of each solute is as follows: potato leaches powder 1.5g/100ml, glucose
1.5g/100ml, Se-enriched yeast powder 1.5g/100ml, peptone 0.6g/100ml, sodium chloride 0.15g/100ml, potassium dihydrogen phosphate
0.04g/100ml, dipotassium hydrogen phosphate 0.06g/100ml, magnesium sulfate 0.06g/100ml, vitamin B13mg/100ml, vitamin
B21.5mg/100ml, glycine 0.4mg/100ml, threonine 0.3mg/100ml, valine 0.2mg/100ml, leucine
0.06mg/100ml, isoleucine 0.15mg/100ml, phenylalanine 0.3mg/100ml, serine 0.15mg/100ml, dried meat ammonia
Sour 0.06mg/100ml, hydroxyproline 0.04mg/100ml, cysteine 0.03mg/100ml, tryptophan 0.03mg/100ml,
Methionine 0.06mg/100ml, lysine 0.15mg/100ml, glutamic acid 0.06mg/100ml.
In liquid seed culture medium third, the concentration of each solute is as follows: potato leaches powder 0.5g/100ml, glucose
0.5g/100ml, Se-enriched yeast powder 0.5g/100ml, peptone 0.4g/100ml, sodium chloride 0.05g/100ml, potassium dihydrogen phosphate
0.02g/100ml, dipotassium hydrogen phosphate 0.04g/100ml, magnesium sulfate 0.04g/100ml, vitamin B11mg/100ml, vitamin
B20.5mg/100ml, glycine 0.2mg/100ml, threonine 0.2mg/100ml, valine 0.1mg/100ml, leucine
0.04mg/100ml, isoleucine 0.05mg/100ml, phenylalanine 0.2mg/100ml, serine 0.05mg/100ml, dried meat ammonia
Sour 0.04mg/100ml, hydroxyproline 0.02mg/100ml, cysteine 0.01mg/100ml, tryptophan 0.02mg/100ml,
Methionine 0.04mg/100ml, lysine 0.05mg/100ml, glutamic acid 0.04mg/100ml.
The preparation method of fructification selenium-rich culturing base: glucose, sucrose, Se-enriched yeast powder, peptone, biphosphate are taken
Potassium, dipotassium hydrogen phosphate, ferric citrate, zinc chloride, magnesium sulfate, vitamin B1, vitamin B2And vitamin B11, pH to 7.5 is adjusted,
Add distilled water to be settled to 1000ml, nutrient solution second is made；Nutrient solution second is uniformly mixed with wheat seed, 121 DEG C of high pressure sterilizations
20min.It is rich that fructification selenium-rich culturing Ji Jia, fructification selenium-rich culturing base second and fructification is prepared respectively according to the method described above
Seleno culture medium third.
Fructification selenium-rich culturing Ji Jia is uniformly mixed with 100g wheat seed by 120ml nutrient solution and is made；Each solute is being sought
Concentration in nutrient solution second is as follows: glucose 1g/100ml, sucrose 0.5g/100ml, Se-enriched yeast powder 2g/100ml, peptone
0.6g/100ml, potassium dihydrogen phosphate 0.03g/100ml, dipotassium hydrogen phosphate 0.05g/100ml, ferric citrate 0.03g/100ml,
Zinc chloride 0.03g/100ml, magnesium sulfate 0.05g/100ml, vitamin B12mg/100ml, vitamin B21mg/100ml, dimension life
Plain B11 1mg/100ml。
Fructification selenium-rich culturing base second, which is uniformly mixed by 150ml nutrient solution with 100g wheat seed, to be made；Each solute is being sought
Concentration in nutrient solution second is as follows: glucose 1.5g/100ml, sucrose 0.6g/100ml, Se-enriched yeast powder 3g/100ml, peptone
0.7g/100ml, potassium dihydrogen phosphate 0.04g/100ml, dipotassium hydrogen phosphate 0.06g/100ml, ferric citrate 0.04g/100ml,
Zinc chloride 0.04g/100ml, magnesium sulfate 0.06g/100ml, vitamin B13mg/100ml, vitamin B21.5mg/100ml, dimension
Raw element B11 1.5mg/100ml。
Fructification selenium-rich culturing base third, which is uniformly mixed by 100ml nutrient solution with 100g wheat seed, to be made；Each solute is being sought
Concentration in nutrient solution second is as follows: glucose 0.5g/100ml, sucrose 0.4g/100ml, Se-enriched yeast powder 1g/100ml, peptone
0.5g/100ml, potassium dihydrogen phosphate 0.02g/100ml, dipotassium hydrogen phosphate 0.04g/100ml, ferric citrate 0.02g/100ml,
Zinc chloride 0.02g/100ml, magnesium sulfate 0.04g/100ml, vitamin B11mg/100ml, vitamin B20.5mg/100ml, dimension
Raw element B11 0.5mg/100ml。
Se-enriched yeast meal component in seed enriched medium first: being replaced with the common yeast powder of isoconcentration by culture medium A,
Remaining ingredient and preparation method are identical as seed enriched medium first.
Se-enriched yeast meal component in liquid seed culture medium first: being replaced with the common yeast powder of isoconcentration by culture medium B,
Remaining ingredient and preparation method are identical as seed culture medium first.
Culture medium C: the Se-enriched yeast meal component in fructification selenium-rich culturing base first is replaced with to the common yeast of isoconcentration
Powder, remaining ingredient and preparation method are identical as fructification selenium-rich culturing base first.
The method for detecting Se content: referring to bibliography, " Wang Ying, iron plum, Kangping benefit wait three kinds of measurement pupas such as .ICP-MS
Comparison [J] the spectroscopy of cordyceps sinensis Se content method and spectrum analysis, 2009,29 (3): 3, the 3- diamino connection in 815-818. "
Aniline spectrophotometry.
The method for detecting cordycepin: the measurement high-efficient liquid phase color of cordycepin and adenosine in NY/T 2116-2012 cordyceps product
Spectrometry.
The breeding of embodiment 1, cordyceps FY1421
It is starting strain with cordyceps militaris link bacterial strain 50383, by atmospheric pressure at room plasma (ARTP) mutagenic treatment, filters out
One plant of cordyceps militaris link bacterial strain is named as cordyceps FY1421 (Cordyceps militaris FY1421).Cordyceps
FY1421 (Cordyceps militaris FY1421) is preserved in China typical culture collection center on April 21st, 2016
(abbreviation CCTCC；Address: Wuhan, China, Wuhan University；Postcode: 430072), deposit number is CCTCC NO:M2016220.Pupa
Cordyceps Militaris FY1421 (Cordyceps militaris FY1421) CCTCC N0:M2016220 is referred to as cordyceps
FY1421。
The acquisition of embodiment 2, fruiting bodies of cordyceps militaris
1, cordyceps militaris link bacterial strain 50383 is seeded to PDA solid slope culture medium, 25 DEG C are protected from light culture 6 days.
2, with sterile physiological saline by the cultured PDA solid slope culture medium of step 1 spore elute, switching in
New PDA solid slope culture medium is protected from light culture 5 days, the cordyceps militaris link bacterial strain activated for 25 DEG C in incubator.
3, the spore elution for the cordyceps militaris link bacterial strain for being activated step 2 with 10ml sterile physiological saline, switching is in being equipped with
(initial concentration of the cordyceps militaris link bacterial strain in cultivating system is 10 in the eggplant bottle of 100g seed enriched medium4A/gram culture
Object), stationary culture is protected from light 5 days for 25 DEG C in incubator, the inoculum that strengthened (spore concentration 1012A/gram culture
Object).
The seed enriched medium that test group first uses is seed enriched medium first.
The seed enriched medium that test group second uses is seed enriched medium second.
The seed enriched medium that test group third uses is seed enriched medium third.
4, the reinforcing inoculum for taking 10g step 3 to prepare is inoculated in 100ml liquid seed culture medium, 25 DEG C,
It is protected from light culture 4 days under the conditions of 220rpm, obtains cordyceps militaris link bacterial strain liquid seeds, the biomass of liquid seeds is 50% (every 100ml
Cordyceps militaris seed weight in wet base is 50g in culture medium).
The liquid seed culture medium that test group first uses is liquid seed culture medium first.
The liquid seed culture medium that test group second uses is liquid seed culture medium second.
The liquid seed culture medium that test group third uses is liquid seed culture medium third.
5, the 100ml liquid seeds that step 4 obtains are inoculated in progress selenium-rich training in the fructification selenium-rich culturing base of 1000g
It supports.
The fructification selenium-rich culturing base that test group first uses is fructification selenium-rich culturing Ji Jia.
The fructification selenium-rich culturing base that test group second uses is fructification selenium-rich culturing base second.
The fructification selenium-rich culturing base that test group third uses is fructification selenium-rich culturing base third.
The incubation of test group first specifically: bacterium germination dark culture stage (8 days): dark culturing, 25 DEG C of temperature, air phase
To humidity 65%, carbon dioxide volume fraction is less than 0.3%；Veraison cultivation stage (20 days): 14-26 DEG C of temperature is (warm round the clock
Difference is at 10 DEG C or so), relative air humidity 80%, carbon dioxide volume fraction is less than 0.2%, intensity of illumination 350lux, illumination
9.5 hour/day of time, light source type are fluorescent lamp；The sporophore growth management stage (15 days): 118 DEG C of temperature, air is relatively wet
80%-85% is spent, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, 8.5 hour/day of light application time, light source
Type is fluorescent lamp；Blue light cultivation stage (10 days): 18 DEG C of temperature, relative air humidity 80%-85%, carbon dioxide volume point
Number is less than 0.1%, and intensity of illumination 250Lux, 8.5 hour/day of light application time, light source type is blue light.
The incubation of test group second specifically: bacterium germination dark culture stage (7 days): being protected from light culture, and 23 DEG C of temperature, air phase
To humidity 70%, carbon dioxide volume fraction is less than 0.3%；Veraison cultivation stage (18 days): 14-26 DEG C of temperature, (warm round the clock
Difference is at 10 DEG C or so), relative air humidity 70%, carbon dioxide volume fraction is less than 0.2%, intensity of illumination 350lux, illumination
9.5 hour/day of time, light source type are fluorescent lamp；The sporophore growth management stage (15 days): 16 DEG C of temperature, air is relatively wet
80%-85% is spent, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, 8.5 hour/day of light application time, light source
Type is fluorescent lamp；Blue light cultivation stage (5 days): 16 DEG C of temperature, relative air humidity 80%-85%, carbon dioxide volume point
Number is less than 0.1%, and intensity of illumination 250Lux, 8.5 hour/day of light application time, light source type is blue light.
The incubation of test group third specifically: bacterium germination dark culture stage (10 days): being protected from light culture, and 24 DEG C of temperature, air
Relative humidity 60%, carbon dioxide volume fraction is less than 0.3%；Veraison cultivation stage (15 days): 14-26 DEG C of temperature is (round the clock
The temperature difference is at 10 DEG C or so), relative air humidity 75%, carbon dioxide volume fraction is less than 0.2%, intensity of illumination 350lux, light
According to 9.5 hour/day of time, light source type is fluorescent lamp；The sporophore growth management stage (15 days): 20 DEG C of temperature, air is opposite
Humidity 80%-85%, carbon dioxide volume fraction is less than 0.1%, intensity of illumination 250Lux, and light application time is 8.5 hours/day,
Light source type is fluorescent lamp；Blue light cultivation stage (7 days): 20 DEG C of temperature, relative air humidity 80%-85%, carbon dioxide body
Fraction is less than 0.1%, intensity of illumination 250Lux, and light application time is 8.5 hours/day, and light source type is blue light.
Each test group fruiting bodies of cordyceps militaris average height reaches 8-12cm after culture, and about 65% fructification top is swollen
Greatly.Fructification is harvested after culture, it is 8% that 65 DEG C, which are dried to moisture, obtains fruiting bodies of cordyceps militaris dried product first (test group
First), fruiting bodies of cordyceps militaris dried product second (test group second) and fruiting bodies of cordyceps militaris dried product third (test group the third).
6, fruiting bodies of cordyceps militaris dried product first, the fruiting bodies of cordyceps militaris dried product second and Cordyceps militaris that prepared by detecting step 5
Se content and cordycepin content in entity dried product third.
Testing result are as follows: the Se content of fruiting bodies of cordyceps militaris dried product first is 360mg/kg, cordycepin content 0.21g/
kg；The Se content of fruiting bodies of cordyceps militaris dried product second is 355mg/kg, cordycepin content 0.18g/kg；Fruiting bodies of cordyceps militaris is dry
The Se content of product third is 350mg/kg, cordycepin content 0.15g/kg.Se content in fruiting bodies of cordyceps militaris dried product first
Highest.
7, the seed enriched medium first in step 3 is replaced with etc. to the culture medium A of quality, the liquid seeds in step 4
The culture medium for the quality such as culture medium first such as replaces at the culture medium B of quality, and the fructification selenium-rich culturing Ji Jia in step 5 is replaced with
C is operated according to the experiment condition of test group first in step 1-6, obtains control fruiting bodies of cordyceps militaris dried product I.Compare pupa
The Se content of cordyceps militaris sporocarp dried product I is 0mg/kg, cordycepin content 0.22g/kg.
8, the experiment of step 1-6 is carried out using cordyceps FY1421 substitution cordyceps militaris link bacterial strain 50383, uses examination in experiment
The culture medium and condition of culture for testing group first obtain fruiting bodies of cordyceps militaris dried product II；Pupa worm is substituted using cordyceps FY1421
Seed enriched medium first in step 3 is replaced with etc. the culture medium A of quality, the liquid strain in step 4 by careless bacterial strain 50383
The culture for the quality such as sub- culture medium first such as replaces at the culture medium B of quality, and the fructification selenium-rich culturing Ji Jia in step 5 is replaced with
Base C is operated according to the experiment condition of test group first in step 1-6, obtains control fruiting bodies of cordyceps militaris dried product II.Pupa worm
The Se content of grass seed entity dried product II is 375mg/kg, cordycepin content 23.3g/kg；It is drying to compare fruiting bodies of cordyceps militaris
The Se content of product II is 0mg/kg, cordycepin content 23.1g/kg.
9, using embodiment 2 institute in patent of invention (application No. is 201210258191.2, grant date 2014.09.10)
The Se-enriched yeast product preparation method stated prepares Se-enriched yeast product (organic selenium content 9880mg/kg).Pupa worm is used respectively
Careless bacterial strain 50383 and cordyceps FY1421 carry out culture medium and condition of culture that test group first is used in the experiment of step 1-6
(using the Se-enriched yeast products substitution Se-enriched yeast powder of the preparation of isoconcentration), obtains fruiting bodies of cordyceps militaris dried product III (pupa worm
Careless bacterial strain 50383) and fruiting bodies of cordyceps militaris dried product IV (cordyceps FY1421).The selenium of fruiting bodies of cordyceps militaris dried product III
Content is 280mg/kg, cordycepin content 0.15g/kg；The Se content of fruiting bodies of cordyceps militaris dried product IV is 290mg/kg, worm
Careless cellulose content is 22.4g/kg.The result shows that: the richness of fructification is carried out using selenium-rich culturing base of the present invention and selenium-rich culturing technique
Selenium culture can effectively obtain selenium-enriched cordceps militaris fructification, and it does not have the cordycepin synthesis capability of cordyceps militaris link bacterial strain itself
There is inhibition.Embodiment also illustrates that cordyceps FY1421 has the cordycepin synthesis capability of good fructification simultaneously.
Claims (3)
1. a kind of method for preparing fruiting bodies of cordyceps militaris product, includes the following steps:
(a1) seed enriched medium culture cordyceps militaris link bacterial strain is used, seed hot housing object is obtained；
(a2) the seed hot housing object that step (a1) obtains is seeded in liquid seed culture medium and is cultivated, obtain liquid
Seed；
(a3) liquid seeds that step (a2) obtains are seeded in fructification selenium-rich culturing base and are cultivated, obtain Cordyceps militaris
Entity；
The raw material proportioning of the seed enriched medium are as follows: 1L nutrient solution first: 0.5-1.5kg millet；Contain in the nutrient solution first
Having concentration is the selenium of 1.5mg/100ml-20mg/100ml；
The selenium for being 0.9mg/100ml-10mg/100ml containing concentration in the liquid seed culture medium；
The raw material proportioning of the fructification selenium-rich culturing base are as follows: 100-150ml nutrient solution second: 100g wheat seed；The nutrition
The selenium for being 1.5mg/100ml-20mg/100ml containing concentration in liquid second；
The cordyceps militaris link bacterial strain is cordyceps (Cordyceps militaris) 50383, ACCC number: 50383 or Cordyceps militaris
Bacterium (Cordyceps militaris) FY1421, deposit number are CCTCC NO:M2016220；
The selenium is organic selenium；
The selenium is added in culture medium in the form of Se-enriched yeast powder；
The nutrient solution first is made of solute and solvent；The solute and its concentration in the nutrient solution first are as follows: selenium-rich
Yeast powder 1-3g/100ml, glucose 0.5-1.5g/100ml, peptone 0.5-0.7g/100ml, potassium dihydrogen phosphate 0.05-
0.15g/100ml, magnesium sulfate 0.04-0.06g/100ml, vitamin B11-3mg/100ml, vitamin B2 0.5-1.5mg/
100ml, glycine 0.2-0.4mg/100ml, threonine 0.2-0.3mg/100ml, valine 0.1-0.2mg/100ml, bright ammonia
Sour 0.04-0.06mg/100ml, isoleucine 0.05-0.15mg/100ml, phenylalanine 0.2-0.3mg/100ml, serine
0.05-0.15mg/100ml, proline 0.04-0.06mg/100ml, hydroxyproline 0.02-0.04mg/100ml, cysteine
0.01-0.03mg/100ml, tryptophan 0.02-0.03mg/100ml, methionine 0.04-0.06mg/100ml, lysine
0.05-0.15mg/100ml, glutamic acid 0.04-0.06mg/100ml；The solvent is water；
The liquid seed culture medium is made of solute and solvent；The solute and its dense in the liquid seed culture medium
Spend as follows: potato leaches powder 0.5-1.5g/100ml, glucose 0.5-1.5g/100ml, Se-enriched yeast powder 0.5-1.5g/
100ml, peptone 0.4-0.6g/100ml, sodium chloride 0.05-0.15g/100ml, potassium dihydrogen phosphate 0.02-0.04g/100ml,
Dipotassium hydrogen phosphate 0.04-0.06 g/100ml, magnesium sulfate 0.04-0.06g/100ml, vitamin B11-3mg/100ml, dimension life
Plain B20.5-1.5mg/100ml, glycine 0.2-0.4mg/100ml, threonine 0.2-0.3mg/100ml, valine 0.1-
0.2mg/100ml, leucine 0.04-0.06mg/100ml, isoleucine 0.05-0.15mg/100ml, phenylalanine 0.2-
0.3mg/100ml, serine 0.05-0.15mg/100ml, proline 0.04-0.06mg/100ml, hydroxyproline 0.02-
0.04mg/100ml, cysteine 0.01-0.03mg/100ml, tryptophan 0.02-0.03mg/100ml, methionine 0.04-
0.06mg/100ml, lysine 0.05-0.15mg/100ml, glutamic acid 0.04-0.06mg/100ml；The solvent is water；
The nutrient solution second is made of solute and solvent；The solute and its concentration in the nutrient solution are as follows: glucose
0.5-1.5g/100ml, sucrose 0.4-0.6g/100ml, Se-enriched yeast powder 1-3g/100ml, peptone 0.5-0.7g/100ml,
Potassium dihydrogen phosphate 0.02-0.04g/100ml, dipotassium hydrogen phosphate 0.04-0.06g/100ml, ferric citrate 0.02-0.04g/
100ml, zinc chloride 0.02-0.04g/100ml, magnesium sulfate 0.04-0.06g/100ml, vitamin B11-3mg/100ml, dimension life
Plain B20.5-1.5mg/100ml, vitamin B110.5-1.5mg/100ml；The solvent is water.
2. the product that claim 1 the method is prepared.
3. product as claimed in claim 2, it is characterised in that: it is 280mg/kg or more that the product, which meets total Se content,.
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| CN108934781A (en) * | 2018-09-26 | 2018-12-07 | 张家港市藏联生物研究所有限公司 | A kind of high-yield culture medium of selenium-enriched cordceps militaris |
| CN109348990A (en) * | 2018-12-06 | 2019-02-19 | 四川农业大学 | A method of improving 808 polyoses content of mushroom kind |
| CN109937795B (en) * | 2019-04-01 | 2021-07-13 | 广西壮仁堂生物科技有限公司 | Method for cultivating cordyceps sinensis mycelia by taking jasmine as matrix |
| CN111657053B (en) * | 2020-06-01 | 2022-02-25 | 齐齐哈尔大学 | Simulated wild high-quality high-yield standardized cultivation method for cordyceps militaris |
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