CN109122050A - A kind of cultural method of Cordyceps militaris - Google Patents
A kind of cultural method of Cordyceps militaris Download PDFInfo
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- CN109122050A CN109122050A CN201810988558.3A CN201810988558A CN109122050A CN 109122050 A CN109122050 A CN 109122050A CN 201810988558 A CN201810988558 A CN 201810988558A CN 109122050 A CN109122050 A CN 109122050A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
Abstract
The present invention provides a kind of cultural methods of Cordyceps militaris, it is related to technical field of bioengineering, more specifically a kind of raising cordycepin, the Cordyceps militaris artificial culture method of cordycepic acid and Cordyceps sinensis polysaccharide content, Cordyceps militaris spore suspension is inoculated in Cordyceps militaris mycelia culture medium and first carries out logical hyperoxia air dark culture, lead to hyperoxia air jet flow in progress illumination in last three days of cultural hypha, obtain mycelia, the mycelium inoculation is carried out to logical hyperoxia airlight in Cordyceps militaris growth medium according to fructification culture, obtain fruiting bodies of cordyceps militaris, when cultivating mycelia and fructification, it is 14~18 DEG C that gas temperature is adjusted when ventilation, it is 18~22 DEG C that temperature is adjusted when stuffiness, the cordycepin of the fruiting bodies of cordyceps militaris, cordycepic acid and Cordyceps sinensis polysaccharide content are compared with traditional shaken cultivation or medicine bottle culture, there is apparent increase, and significantly Shorten the artificial cultivation cycle of Cordyceps militaris.
Description
Technical field
The present invention relates to technical field of bioengineering, it is more to relate more specifically to a kind of raising cordycepin, cordycepic acid and cordyceps sinensis
The Cordyceps militaris artificial culture method of sugared content.
Background technique
Cordyceps militaris (Cordyceps militaris), is called northern Chinese caterpillar Fungus, Cordceps militaris, and abbreviation pupa grass belongs to ascus Asia
Door, gang pyrenomycetes, Spheeriales, Clavicipitaceae, Cordyceps are to parasitize a Lepidopterous Vespertilionidae insect Hepialus larva
On stroma or other species Vespertilionidae insect larvae on complex, have it is antitumor, anti-inflammatory, antiviral, enhancing body
Immunity, enhancing macrophage activity and other effects, are one of China's rare traditional Chinese medicines.
Contain the nutrition such as cordycepin, cordycepic acid, Cordyceps sinensis polysaccharide, protein, amino acid, vitamin, minerals in Cordyceps militaris
Ingredient, wherein cordycepin, cordycepic acid and Cordyceps sinensis polysaccharide are the distinctive substances of Cordyceps militaris, are referring mainly to for Cordyceps militaris special efficacy
Mark.Cordycepin (cordycepin) also known as 3 '-desoxyadenossine are one of main effective component in Cordyceps militaris, clinically mainly
For treating cancer, leukaemia, expansion bronchus, significantly increase feritin, anti-aging the effects of.The height of cordycepic acid content is weighing apparatus
Measure one of the main standard of cordyceps sinensis quality, it is considered that the high Cordyceps militaris medical value of cordycepic acid content is high.Cordycepic acid can prevent
Treat cerebral thrombosis, cerebral hemorrhage, myocardial infarction, long-term decline, anti-liver tissue fibrosis, anti peroxidation of lipid.Cordyceps sinensis polysaccharide can mention
High immune function of human body, increasing leukocyte improve respiratory system, have and adrenal effect is promoted to inhibit tumour growth, and have
The pharmacological actions such as antitumor, anti-radiation, hypoglycemic and lipoprotein, cough-relieving, resolving sputum, moistening lung and anti-aging, clinic have been used for controlling
Treat malignant tumour.
The artificial culture of Cordyceps militaris has the culture techniques such as artificial solid culture technology and liquid fermentation and culture.Currently, improving
The method of Cordyceps cordycepin content has two classes, and one kind is the mutation breeding mode by bacterial strain, improves the cordyceps sinensis of original strain
Cellulose content is horizontal, and another kind of contained by adding the method raising cordycepin of inducer in training systern and incubation
Amount, Chinese invention patent CN102630490B induce Cordyceps militaris to improve cordycepin content by the manganese ion of addition higher concentration,
The cordycepin for adopting this technical solution the Cordyceps militaris of acquisition increases 2.3 compared with the cordycepin content for being not added with manganese ion induction
Times, but also still in reduced levels.The method of the cordycepic acid and Cordyceps sinensis polysaccharide content that improve Cordyceps militaris mainly passes through liquid
Submerged fermentation improves, still, submerged fermentation obtain be Cordyceps militaris mycelium, the content of the cordycepic acid generated generally exists
3~4.5% or so, and the obtained culture that ferments not only includes Cordyceps militaris itself, also contains some culture based draffs, is influenced
The purity of cordycepic acid and Cordyceps sinensis polysaccharide.
Summary of the invention
For this reason, it may be necessary to a kind of cultural method of Cordyceps militaris be provided, to solve current Tissue Culture of Cordyceps militaris cordycepin, cordyceps sinensis
Acid and the lower problem of Cordyceps sinensis polysaccharide content.
To achieve the above object, a kind of cultural method of Cordyceps militaris is inventor provided, comprising the following steps:
Culture mycelia: Cordyceps militaris spore suspension is inoculated in Cordyceps militaris mycelia culture medium, and height is first led under the conditions of being protected from light
Oxygen 7~11d of air jet flow, then continue logical hyperoxia air jet flow 3d under illumination condition, obtain Cordyceps militaris mycelia；
Culture fructification: the Cordyceps militaris mycelia is inoculated in Cordyceps militaris growth medium, and height is led under illumination condition
60~80d of oxygen air jet flow, the Cordyceps militaris growth medium are calculated as 800~1800 portions of rices by weight, obtain pupa worm
Grass seed entity.
Cordyceps militaris spore suspension of the present invention are as follows: will buy or be protected purchased from Chinese industrial microorganism fungus kind on the market
The Cordyceps militaris spawn (number: CICC 14013) for hiding administrative center is inoculated in Cordyceps militaris seed culture medium, conventionally
Culture, obtains cordyceps militaris link bacterial strain, then separate list cystospore according to a conventional method from above-mentioned cordyceps militaris link bacterial strain, i.e., is placed in strain
Bacterial strain is turned out in conventional medium, is aseptically placed in PD fluid nutrient medium with a small amount of pink colour spore of sterilizing toothpick picking
The conidium that the middle cordyceps militaris link bacterial strain for preparing spore suspension or freely matching is obtained by culture is placed in PD fluid nutrient medium
Be prepared, above-mentioned list cystospore or conidium be conventionally subjected to activation culture, be made content be 1.0 ×
107The Cordyceps militaris spore suspension of CFU/ml, then by the Cordyceps militaris spore suspension by the culture mycelia and culture fructification
Step is cultivated.Spore germination and vegetative stage, do not need illumination, in favor of generating stroma.Spore suspension is being inhaled
When receiving the nutritional ingredient in the Cordyceps militaris mycelia culture medium, need suitably to fill into fresh air to prevent carbon dioxide buildup mistake
It is more, hinder mycelia to generate stroma.Inventor is had found by a large amount of comparative tests, is passed directly into rich oxygen content air and is filled into than oscillation
Fresh air is more able to satisfy mycelial growth the growth needs of mycelia.Especially low, the present invention has when cultivating mycelia
It is not protected from light shaking table shaken cultivation in traditional, Cordyceps militaris spore suspension is first inoculated in Cordyceps militaris mycelia culture medium by the present invention,
First lead to 7~11d of hyperoxia air jet flow under the conditions of being protected from light, then continue logical hyperoxia air jet flow 3d under illumination condition, does so
Benefit first is that in order to activate mycelium, second is that being saved in order to which the nutrition for preferably absorbing mycelium converts active components
The step of preparation seed liquor, due to being passed through high oxygen-containing air, 7~11d just obtains a large amount of Cordyceps militaris mycelia, with conventional method
Need 18~25d just and can obtain the period of mycelia to compare, the time greatly shortens, then the Cordyceps militaris mycelia of the activation is inoculated in
Illumination is carried out in Cordyceps militaris growth medium leads to high oxygen-containing air fructification culture, obtained fruiting bodies of cordyceps militaris cordycepin,
The content of cordycepic acid and Cordyceps sinensis polysaccharide is all larger than the product of traditional shaking table shaken cultivation.
The generative growth phase of fructification needs bright illumination, in order to obtain fruiting bodies of cordyceps militaris best in quality, this
Invention use intensity of illumination for 200~240lux, is being passed through air of the oxygen content more than or equal to 80%, gas flow for 100~
Under conditions of 180vvm, by mycelium inoculation in rice, mycelia is allowed to maximally utilise the nutritional ingredient in rice, grown
Rate and active component content all obtain larger raising.Gap between rice is making full use of for hyperoxia air and rice nutrient
Good condition is created, the growth cycle of fructification is shortened, also makes the cordycepin, cordycepic acid and cordyceps sinensis of fruiting bodies of cordyceps militaris more
Sugared purity is higher.
Further, in the culture mycelia step, condition of culture are as follows: when be passed through oxygen content be more than or equal to 80% air
When, gas temperature is adjusted to 14~18 DEG C, adjustment temperature is 18~22 DEG C when stuffiness.
Further, be passed through oxygen content more than or equal to 80% air process conditions it is as follows: gas flow be 100~
180vvm, duration of ventilation are 1.5~2.5h/ times, are ventilated 4~6 times daily.
Further, when hyperoxia air jet flow is led in illumination, intensity of illumination is 200~500lux.
The yield and quality of Artificial Cultivation of Cordyceps militaris Link depends on strain and cultivation condition, and the hereditary effect of strain can also generate
Certain restrictions effect, light and cultivation temperature are also key factor.The life that culture is conducive to Cordyceps militaris mycelia is protected from light before culture
Long and cordycepin, cordycepic acid and Cordyceps sinensis polysaccharide conversion is formed, and the later period must light-exposed just be conducive to fruit-body formation, cultivate in
Proper temperature is conducive to the growth for inhibiting contaminated bacteria, and improves yield, but temperature is too low, extends cultivation period, by anti-
Multiple verification experimental verification, when being passed through hyperoxia air, adjustment gas temperature is 14~18 DEG C, can sufficiently activate Cordyceps militaris cell,
When not being passed through hyperoxia air, need for temperature to be adjusted to 18~22 DEG C, such temperature difference can make its product under hyperoxia air environment
Tire out more active materials.Illumination is not needed the early period of cordyceps mycelium growth phase, intense light irradiation has inhibition to mycelia growth
Effect, fruiting body differentiation stage then need lighting process appropriate, and in the growth course of fructification, illuminance, temperature and son are real
Cordycepin content in body is positively correlated.
Further, when being passed through air of the oxygen content more than or equal to 80%, gas flow is 100~180vvm, ventilation
Time is 1.5~2.5h/ times, is ventilated 4~6 times daily.
Duration of ventilation need to be held according to hypha growth condition when being passed through rich oxygen content air, otherwise, if nutrition in culture medium
Ingredient is depleted, continues to be passed through oxygen for mycelia growth without positive facilitation, and waste ventilation cost, through trying
It tests observation to learn, a duration of ventilation is that 1.5~2.5h effect is best.
Further, in the culture fructification step, condition of culture is as follows: being more than or equal to 80% when being passed through oxygen content
When air, adjustment gas temperature is 14~18 DEG C, and intensity of illumination is under 200~1200lux, and when stuffiness, adjustment temperature is 18
~22 DEG C.
Similar culture mycelia step, it is 14~18 DEG C that gas temperature is adjusted in logical hyperoxia air, and is adjusted when stuffiness
Temperature is 18~22 DEG C, and the active material of fructification can be stimulated to accumulate and accelerate growth to the maximum extent.
Further, the Cordyceps militaris mycelia culture medium, by weight, including following component: dehydrated potato powder 7~14
Part, 600~1500 parts of water purification, 7~14 parts of glucose, 2~5 parts of peptone, 0.01~0.05 part of magnesium sulfate, potassium dihydrogen phosphate
0.01~0.05 part and vitamin B10.01~0.10 part.
Cordyceps militaris cultural hypha of the present invention uses fluid nutrient medium, has the advantages that first is that mycelial growth rate is fast, bacterium
Filament yield is high, second is that the yield of the metabolite cordycepin of Cordyceps militaris, cordycepic acid and Cordyceps sinensis polysaccharide can be improved, and liquid
Culture overcomes extracts the disadvantages of metabolite amount of labour is big, time-consuming is more, low output directly from fruiting bodies of cordyceps militaris, third is that just
In industrialized production.
Carbon source is the basis of Cordyceps militaris synthetic carbohydrate and amino acid, is also important energy source.Artificial cultivation
When, the available carbon source of Cordyceps militaris has glucose, sucrose, maltose, starch, pectin etc., wherein especially small with glucose, sucrose etc.
The utilizing status of molecule carbohydrate is best.Carbon of the present invention using glucose and dehydrated potato powder as Cordyceps militaris activation and mycelia growth
Source, using rice as the carbon source of sporophore growth, energy and carbon utilization efficiency highest, and in fructification light passing according to logical height
When oxygen air jet flow, gap between rice is that hyperoxia air and rice nutrient make full use of the good condition that creates, contracting
The growth cycle of short fructification also makes the cordycepin, cordycepic acid and Cordyceps sinensis polysaccharide purity of fruiting bodies of cordyceps militaris higher.
Nitrogen is the important member of the inorganic nitrogens such as organic nitrogens and ammonium salt such as protein, the nucleic acid of Cordyceps militaris itself synthesis
Element, wherein organic nitrogen source is better than inorganic nitrogen-sourced, and there are many organic nitrogen that can be utilized, such as amino acid, peptone, beancake powder, dried silkworm chrysalis meal
It is compared Deng, the present invention by screening, still using conventional peptone as organic nitrogen donor.
Cordyceps militaris is slant acidity fungi, and mycelia growth and development optimal pH is 5.2~6.8, but in sterilizing and incubation
Middle pH value can decline.So answering heightening pH value 1~1.5 when preparing culture medium, suitable phosphoric acid can be added when preparing culture medium
The buffer substances such as potassium dihydrogen, pH value in regulation culture base simultaneously provide minerals.
Chinese caterpillar fungus hypha cannot synthesize necessary vitamin, be suitably added vitamin B1Be conducive to the growth and development of mycelia.
K in mineral matter element+、Mg+、Ca2+Deng and some biotins can promote the life of Cordyceps militaris mycelia and fructification
It is long.
Further, the Cordyceps militaris spore suspension content is 1.0 × 107CFU/ml。
It being different from the prior art, Cordyceps militaris spore suspension is inoculated in Cordyceps militaris mycelia culture medium by above-mentioned technical proposal,
High oxygen-containing air is passed through first to carry out dark culture, carried out illumination cultivation in same culture medium at last three days of cultural hypha and obtain
Mycelia eliminates and spore spreads cultivation the step of preparing seed liquor, due to being passed through high oxygen-containing air, just obtains within 10~14 days a large amount of
Mycelia, compared with conventional method needs 18~25 days can just obtain the period of mycelia, the time is greatly shortened, then the mycelia is connect
Kind carries out illumination in Cordyceps militaris growth medium and leads to high oxygen-containing air fructification culture, obtained fruiting bodies of cordyceps militaris cordyceps sinensis
Cellulose content is 9.6mg/g, cordycepic acid content is 2.4% and Cordyceps sinensis polysaccharide soma weight is 5.8, and the son that conventional surge culture obtains
Entity cordycepin content is 5.0mg/g, cordycepic acid content is 2.0% and Cordyceps sinensis polysaccharide soma weight is 4.4.
Specific embodiment
Technology contents, construction feature, the objects and the effects for detailed description technical solution, below in conjunction with specific reality
Example is applied to be explained in detail.
Embodiment one
The present embodiment provides a kind of method for being passed through high oxygen-containing air culture Cordyceps militaris, specific steps are as follows:
Cordyceps militaris spawn is purchased from Chinese industrial Microbiological Culture Collection administrative center, number CICC 14013.
Separation list cystospore: above-mentioned Cordyceps militaris spawn is inoculated in Cordyceps militaris seed culture medium, the Cordyceps militaris kind
Sub- culture medium by weight, including following component: 10 parts of dehydrated potato powder, 1000 parts of water purification, 7 parts of glucose, 3 parts of peptone,
10 parts of agar .05 parts of vitaminB10,0.03 part of vitamin B2,0.02 part of potassium dihydrogen phosphate and 0.03 part of magnesium sulfate, are protected from light training
It supports 7 days, obtains cordyceps militaris link bacterial strain, aseptically choose list cystospore from advantage cordyceps militaris link bacterial strain with sterilizing toothpick；
It prepares spore suspension: the list cystospore is placed in sterile water, it is 1.0 × 10 that content, which is made,7CFU/ml's
Spore suspension；
Culture mycelia: the spore suspension is inoculated in Cordyceps militaris mycelia culture medium to carry out the ventilation of high oxygen-containing air advanced
Row dark culture 8 days, then cultivated 3 days at illumination 400lux, mycelial color is observed, when it becomes Chinese red mycelium pellet,
Stop ventilation, obtains mycelia, the Cordyceps militaris mycelia culture medium, by weight, including following component: 7 parts of dehydrated potato powder, net
700 parts of water, 8 parts of glucose, 3 parts of peptone, 0.02 part of magnesium sulfate, 0.01 part of potassium dihydrogen phosphate and vitamin B10.05 part,
At 16 DEG C, oxygen content being passed through with the gas flow of 100vvm and is cultivated equal to 80% air, when ventilation, is 1.5h/ times a length of,
Gas temperature is adjusted to 21 DEG C after stopping ventilation by ventilation 5 times daily；
Culture fructification: the mycelium inoculation is carried out to logical hyperoxia airlight in Cordyceps militaris growth medium according to fructification
Culture, the Cordyceps militaris growth medium are calculated as 1000 portions of rices by weight, and adjustment gas temperature is 14 DEG C, with 120vvm
Gas flow be passed through the air that oxygen content is 85% and cultivated, when ventilation, is 2h/ time a length of, and when stuffiness, adjusting temperature is
19 DEG C, it is 1000lux in intensity of illumination, cultivates 60 days, be 8~10cm to fruiting bodies of cordyceps militaris length, can gather in.
Embodiment two
The present embodiment provides a kind of normal airs that is passed through to carry out the method that Cordyceps militaris is manually cultivated.
The present embodiment cultivates 11d what is different from the first embodiment is that the culture all dark cultures of mycelia step.Remaining condition
It is identical.
Embodiment three
The present embodiment is what is different from the first embodiment is that culture mycelia step and cultivate that fructification step is passed through is common sky
Gas, remaining condition are identical.
Example IV
For the present embodiment unlike embodiment three, culture mycelia step is passed through normal air under the conditions of being protected from light, remaining
Condition is identical.
Embodiment five
The present embodiment provides a kind of method of shaken cultivation Cordyceps militaris, specific steps are as follows:
Cordyceps militaris spawn be purchased from Chinese industrial Microbiological Culture Collection administrative center, number CICC 14013,
The step of preparing spore suspension is the same as embodiment one.
Unlike, the present embodiment uses traditional shaking table succusion culture seed liquor: the spore suspension is inoculated in
In conventional PDA liquid medium, it is protected from light culture 7 days under the conditions of 23 DEG C, obtains seed liquor；
Culture mycelia: the seed liquor is inoculated in Cordyceps militaris mycelia culture medium and carries out oscillation training illumination 400lux culture
11 days, at 26 DEG C, with 250rpm/min shaken cultivation, mycelial color is observed, when it becomes Chinese red mycelium pellet, is stopped
It only vibrates, obtains mycelia, the Cordyceps militaris mycelia culture medium, by weight, including following component: 7 parts of dehydrated potato powder, water purification
700 parts, 8 parts of glucose, 3 parts of peptone, 0.02 part of magnesium sulfate, 0.01 part of potassium dihydrogen phosphate and vitamin B10.05 part；
Culture fructification: carrying out illumination fructification culture for the mycelium inoculation in Cordyceps militaris growth medium, described
Cordyceps militaris growth medium is calculated as 1000 portions of rices by weight, and divulge information 3h daily, imposes intensity of illumination 1000lux illumination
8h is cultivated 60 days, until fructification length is 8~12cm, can harvest fruiting bodies of cordyceps militaris.
Embodiment six
The present embodiment provides a kind of method of shaken cultivation Cordyceps militaris mycelia, specific steps it is different from embodiment five
In cultured mycelia step is that oscillation is protected from light CMC model, is not passed through hyperoxia air or normal air, remaining step and embodiment
Two is identical.
Contain according to the cordycepin of the obtained Cordyceps militaris mycelia of the technical solution of embodiment one to embodiment six, cordycepic acid
Amount as shown in Table 1 and Table 2, leads to hyperoxia air respectively, the mycelia soma that normal air and obstructed air surge shaking table culture obtain
Weight and polysaccharide body dry weight content are as shown in Table 3 and Table 4.
1 embodiment one of table obtained cordyceps silk cordycepin content (mg/g) into embodiment six
| Number of days | 1 | 3 | 5 | 7 | 9 | 11 |
| Embodiment one | 0.42 | 1.96 | 6.86 | 9.64 | 12.8 | 12.5 |
| Embodiment two | 0.23 | 0.68 | 1.40 | 3.33 | 4.24 | 4.19 |
| Embodiment three | 0.31 | 1.30 | 3.25 | 5.04 | 5.57 | 5.43 |
| Example IV | 0.17 | 0.40 | 0.79 | 1.96 | 3.28 | 3.12 |
| Embodiment five | 0 | 0.95 | 1.10 | 1.29 | 1.50 | 1.23 |
| Embodiment six | 0 | 0.13 | 0.77 | 1.15 | 1.00 | 1.00 |
2 embodiment one of table obtained cordyceps filaria oxalic acid content (g/100g) into embodiment six
| Number of days | 1 | 3 | 5 | 7 | 9 | 11 |
| Embodiment one | 0.1 | 1.6 | 1.8 | 2.4 | 3.1 | 3.1 |
| Embodiment two | 0.1 | 1.2 | 1.5 | 1.4 | 1.6 | 1.5 |
| Embodiment three | 0.1 | 1.5 | 1.4 | 2.0 | 2.5 | 2.4 |
| Example IV | 0.1 | 1.5 | 1.6 | 1.6 | 1.7 | 1.6 |
| Embodiment five | 0 | 1.0 | 1.3 | 1.3 | 1.6 | 1.5 |
| Embodiment six | 0 | 0.7 | 1.4 | 1.3 | 1.6 | 1.5 |
Table 3 leads to hyperoxia air, normal air and the obtained cordyceps mycelium dry weight of obstructed air surge shaking table culture
(%)
Table 4 leads to hyperoxia air, normal air and the obtained polysaccharide body dry weight (%) of obstructed air surge shaking table culture
By Tables 1 and 2 result it is found that generally, leading to the content of cordycepin and cordycepic acid in the mycelia of hyperoxia air jet flow
Highest, taking second place for logical normal air culture, vibrates the minimum of shaking table culture, is equally logical in illumination and the comparison of dark culture
Hyperoxia air gives illumination in last three days in culture, and the content of cordycepin and cordycepic acid is also higher in obtained mycelia, together
The rule of sample also appears in the scheme for being passed through normal air and oscillation shaking table culture.By the result of table 3 and table 4 it is found that logical height
Oxygen air, normal air and the obtained cordyceps mycelium dry weight of obstructed air surge shaking table culture and polysaccharide body dry weight are presented
High to Low rule out as the increase cordyceps mycelium dry weight and polysaccharide body dry weight of cultivated days are with increase, but arrives
It does not continue to increase after 9th day, cordycepin and cordycepic acid content also have this rule to occur, using the technical side that the present invention is optimal
Case, the Cordyceps militaris turned out have the advantages that the content of cordycepin in every gram of Cordyceps militaris mycelia is 12.8mg, every 100g pupa worm
The content of cordycepic acid is 3.1g in careless mycelia, and the dry weight of Cordyceps militaris mycelia is 8.4%, and polysaccharide body dry weight is 5.7%.
Embodiment seven
What is different from the first embodiment is that adjustment gas temperature is 18 DEG C when carrying out being passed through hyperoxia air every time, with
The gas flow of 180vvm is passed through oxygen content and is cultivated equal to 99% air, and duration of ventilation is 2.5h/ times, tune when stuffiness
Whole temperature is 22 DEG C, and when cultivating the illumination cultivation in mycelia, intensity of illumination is adjusted to 200lux, the light in culture fructification
When according to culture, intensity of illumination is adjusted to 1100lux.
Similarly, it in different other embodiments, cultivates in mycelia step, gas temperature can be when leading to hyperoxia air
The adjustment of 14~18 DEG C of ranges, temperature can be adjusted in 18~22 DEG C of ranges when stuffiness, and intensity of illumination can be in 200~500lux
Range adjustment, gas flow can adjust in 100~180vvm range, and when cultivating fructification, temperature can be with when logical hyperoxia air
Adjusted in 14~18 DEG C of ranges, temperature can be adjusted in 18~22 DEG C of ranges when stuffiness, intensity of illumination can 200~
The adjustment of 1200lux range, gas flow can be adjusted in 100~180vvm range.The obtained pupa worm of above optimal case
Grass, the content of cordycepin is 12.8mg in every gram of Cordyceps militaris mycelia, and the content of cordycepic acid is 3.1g in every 100g Cordyceps militaris mycelia,
The dry weight of Cordyceps militaris mycelia is 8.4%, and polysaccharide body dry weight is 5.7%.
It should be noted that being not intended to limit although the various embodiments described above have been described herein
Scope of patent protection of the invention.Therefore, it based on innovative idea of the invention, change that embodiment described herein is carried out and is repaired
Change or the equivalent structure or equivalent process transformation made by using the contents of the present specification, directly or indirectly by the above technology
Scheme is used in other related technical areas, is included within scope of patent protection of the invention.
Claims (8)
1. a kind of cultural method of Cordyceps militaris, which comprises the following steps:
Culture mycelia: Cordyceps militaris spore suspension is inoculated in Cordyceps militaris mycelia culture medium, and it is empty that hyperoxia is first led under the conditions of being protected from light
7~11d is supported in air culture, then continues logical hyperoxia air jet flow 3d under illumination condition, obtains Cordyceps militaris mycelia；
Culture fructification: the Cordyceps militaris mycelia is inoculated in Cordyceps militaris growth medium, and it is empty that hyperoxia is led under illumination condition
60~80d is supported in air culture, and the Cordyceps militaris growth medium is calculated as 800~1800 portions of rices by weight, obtains Cordyceps militaris
Entity.
2. cultural method according to claim 1, which is characterized in that in the culture mycelia step, condition of culture are as follows: when
When being passed through oxygen content and being more than or equal to 80% air, gas temperature is adjusted to 14~18 DEG C, adjustment temperature is 18 when stuffiness
~22 DEG C.
3. cultural method according to claim 2, which is characterized in that be passed through the technique that oxygen content is more than or equal to 80% air
Condition is as follows: gas flow is 100~180vvm, and duration of ventilation is 1.5~2.5h/ times, is ventilated 4~6 times daily.
4. cultural method according to claim 2, which is characterized in that when hyperoxia air jet flow is led in illumination, intensity of illumination is
200~500lux.
5. cultural method according to claim 4, which is characterized in that when being passed through air of the oxygen content more than or equal to 80%,
Gas flow is 100~180vvm, and duration of ventilation is 1.5~2.5h/ times, is ventilated 4~6 times daily.
6. cultural method according to claim 1, which is characterized in that in the culture fructification step, condition of culture is such as
Under: when be passed through oxygen content be more than or equal to 80% air when, adjustment gas temperature be 14~18 DEG C, gas flow be 100~
180vvm, intensity of illumination are under 200~1200lux, and when stuffiness, adjustment temperature is 18~22 DEG C.
7. cultural method according to claim 1, which is characterized in that the Cordyceps militaris mycelia culture medium, by weight,
Including following component: 7~14 parts of dehydrated potato powder, 600~1500 parts of water purification, 7~14 parts of glucose, 2~5 parts of peptone, sulfuric acid
0.01~0.05 part of magnesium, 0.01~0.05 part of potassium dihydrogen phosphate and vitamin B10.01~0.10 part.
8. cultural method according to claim 1, which is characterized in that the Cordyceps militaris spore suspension content be 1.0 ×
107CFU/ml。
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