CN108812051A - A kind of cordyceps culturing method - Google Patents
A kind of cordyceps culturing method Download PDFInfo
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- CN108812051A CN108812051A CN201810499060.0A CN201810499060A CN108812051A CN 108812051 A CN108812051 A CN 108812051A CN 201810499060 A CN201810499060 A CN 201810499060A CN 108812051 A CN108812051 A CN 108812051A
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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
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Abstract
The invention discloses a kind of cordyceps culturing method, this method selects suitable culture medium and condition of culture, by mycelium to Cr3+Enrichment, by the Cr of mineralized3+Change organic chromium, effectively avoids inorganic states Cr3+Autoxidation is Cr6+, reduce human body and directly absorb metal Cr3+The injury that human body is generated.And by research, it has also been found that, the chromium in Cordyceps militaris also has facilitation, while organic chromium and Cordyceps sinensis polysaccharide collective effect to the conversion of polysaccharide in its mycelium, can improve the pancreas function of diabetic, to reduce the use of human insulin.
Description
Technical field
The present invention relates to Spawn incubation technical fields, and in particular to a kind of cordyceps culturing method.
Background technique
Cordyceps militaris Cordyceps militaris L.Link also known as northern Chinese caterpillar Fungus, Cordceps militaris, pupa grass, pupa grass bacterium etc.,
According to newest categorizing system, Cordyceps militaris belongs to Ascomycota Ascomycota, Hypocreales Hypocreales, cordyceps sinensis Cordycepps
Cordycipitaceae, Cordyceps Cordyceps (Sung et al., 2007).Cordyceps militaris has similar with cordyceps sinensis
Active constituent and pharmacological action (Kim andYun, 2005；Li et al., 2006；Wang Jianfang and Yang Chunqing, 2005), cordyceps sinensis hair
Ferment product has the various biologicals such as antibacterial, inoxidizability, anti-inflammatory, antitumor, Immune-enhancing effect activity.Wherein Cordyceps sinensis polysaccharide is to more
Kind diabetic animal models have significant hypoglycemic effect, and mechanism may inhibit hepatic glucose output with it, promote liver
The vigor of glucose metabolism enzyme and the content for reducing glucose transporter, have been increasingly becoming the ideal substitute of cordyceps sinensis.
Global diabetic was up to 300,000,000 by 2025 up to 1.5 hundred million.The existing diabetic about 30,000,000-of China
40000000, it can be more than 50,000,000 by 2025, be the third place in the world diabetes big country after India, the U.S..The harm of diabetes
It is various chronic complicating diseases, if diabetic nephropathy can seriously cause uremia, (cerebral infarction, coronary disease can occur for cardiovascular and cerebrovascular disease
Disease, myocardial infarction), eye disease (cataract, retinopathy, bleeding, glaucoma etc.), peripheral neuropathy (numb in every limb, whole body
Itch etc.), peripheral angiopathy (lower limb toe end necrosis, diabetes, exhaust walk lamely), it is various infection (gingivitis, decayed tooth,
Be easier to aggravate after urinary tract infections and infection, deteriorate it is not easy to control), once stress (such as severe pneumonia, operation, wound wound not
Easily healing).Above-mentioned final influence quality of life, brings considerable distress and inconvenience to patient, and it is heavy to bring to household and entire society
The burden of weight.The medical totle drilling cost of the annual city treatment diabetes B in China and its complication is 208.60 hundred million yuan at present, is accounted for complete
The 4.38% of state's medical treatment Health Expenditure, wherein the medical totle drilling cost for having the diabetes B of complication is 164.51 hundred million yuan.Now
Clinical treatment diabetes are that blood glucose is controlled by insulin injection, will cause the dependence of insulin using insulin for a long time,
Cause the irreversible injury of pancreas.It is badly in need of a kind of antihypelipidemic foodstuff that safety is without side-effects now, sugar is improved by diet
The pancreatic secretion function of urinating patient, mitigates the dependence of patients towards insulin.
Diabetes B (T2DM) is a kind of common incretion metabolism disease, and main pathological manifestations are supported for insulin
Anti- and islet function deficiency, and with serious hyperinsulinemia, wherein insulin resistance diabetes B generation,
It plays an important role in development.(PCS alleviates insulin resistance T2DM model mice hyperglycemia blood lipid symptom to Cordyceps sinensis polysaccharide, together
When reduce insulin resistance index, prompt Cordyceps sinensis polysaccharide that there is potential insulin-sensitizing effect.PCS promotes high sugar and insulin
The insulin-stimulated glucose intake of the fat cell of IR is induced, while showing that its hypoglycemic mechanism may be with promotion periphery
The glucose metabolism of tissue is related.PCS is that have protection beta Cell of islet simultaneously, promote impaired β cell repair act on and
The effect of PCS promotion peripheral tissues' glucose utilization.
Nature chromium is with Cr to Cr6+Various valence states exist, but with Cr, Cr2+、Cr3+、Cr6+It is most common, wherein Cr6+It is
It is toxic to human body.Cr in natural environment2+With Cr3+Stability under inorganic ions state is poor, holds by Cr very much2+With Cr3+Turn
Become Cr6+Inorganic ions, Cr3+It is the essential trace elements of the human body, the intracorporal chromium of people is almost Cr entirely3+, United States Department of Agriculture's nutrition
The authoritative institutions such as center current research reconfirms, Cr3+Inorganic chromium enhances insulin active very little, after being transformed into Organic Chromium then
Have the function of significantly reinforcing insulin active.Important component as glucose tolerance factor.Studies have shown that organic
Chromium stability is strong, as fat-soluble non-electrolyte, can pass through cell membrane and directly act on histocyte, enhance insulin
Activity improves human body glycometabolism, has important role to the treatment of diabetes, is suitable for diabetes patient and impaired glucose tolerance, sugar
Urinate sick people at highest risk and Healthy People.There are no cultivated by chromium-rich by mineralized in current cordyceps culturing method
Cr3+Change the report in terms of organic.
Summary of the invention
The purpose of the present invention is to provide a kind of new cordyceps culturing methods, and this method is by selecting suitable culture medium
And condition of culture, by Cr3+Enrichment, by the Cr of mineralized3+Change organic, effectively avoids inorganic states Cr3+From
So it is oxidized to Cr6+, while organic chromium and Cordyceps sinensis polysaccharide collective effect, the pancreas function of diabetic can be improved, thus
Reduce the use of human insulin.
To achieve the above object, the technical scheme is that：
A kind of cordyceps culturing method, described method includes following steps：
(1) Cordyceps militaris spawn of 0-4 DEG C of preservation is subjected to VB1Slant strains culture：
A, Cordyceps militaris spawn culture medium is prepared：200 portions of potatoes in parts by weight are cut into the fritter of 0.8-1.2cm,
It is boiled in boiling water 20 minutes, is then filtered, takes juice, rear 20 parts of glucose of addition, 0.5 part of magnesium sulfate, 1 part of potassium dihydrogen phosphate,
Vitamin B10.01 part, 20 parts of agar, after thawing, constant volume is at 1000mL, natural ph；
B, Cordyceps militaris spawn culture medium dispenses, sterilizing：
Prepared bacterium culture medium is sub-packed in 18mm × 180mm test tube, is sealed, under the conditions of 121 DEG C, sterilizing
20min；
C, cooling, inoculation：
It has gone out and has taken out Cordyceps militaris spawn culture medium after bacterium, naturally cooled to 20 DEG C -30 DEG C, aseptically, accessed pupa
Cordyceps species；
D, it cultivates：
The Cordyceps militaris spawn culture medium that will be connected kind is placed in constant incubator, 22-25 DEG C culture 4-8 days, mycelia is abundant
Growth；
E, it selects：
Growing way is got well to the Cordyceps militaris VB for degradation phenomena do not occur1Slant strains are selected；
(2) it is inoculated with：
By Cordyceps militaris VB1Slant strains are inoculated on Cordyceps militaris bacterium culture medium under sterile working, are in temperature
20 DEG C, revolving speed be 150-160r/min shaking table in cultivate 5d, the good strain of liquid fermentation is inoculated on chromium-rich culture medium, often
Bottle inoculum concentration is 20mL；
The Cordyceps militaris bacterium culture medium is exactly the bacterium culture medium for not adding agar, is formed according to parts by weight
It is calculated as：200 parts of potato, 20 parts of glucose, 0.5 part of magnesium sulfate, 1 part of potassium dihydrogen phosphate, vitamin B10.01 part, 1000 parts of water.
The composition of the chromium-rich culture medium is in parts by weight：30 parts of oat, CrCl30.4-0.6 parts, potassium dihydrogen phosphate
1 part, 50 parts of water.Its preparation method is：First by 1 part of potassium dihydrogen phosphate and CrCl30.4-0.6 parts are added in 50 parts of water and stir
Dissolution, load weighted oat is fitted into the tissue culture bottle of 500mL for 30 parts, and the potassium dihydrogen phosphate and CrCl dissolved is added3Mixing
Solution covers lid, then 121 DEG C of sterilizing 60min, takes out cooling, for use.
(3) chromium-rich culture：
It after Cordyceps militaris is accessed chromium-rich culture medium after cooling, is placed in constant incubator, 16-20 DEG C of culture 8-12
It, after mycelia sufficiently grows, carries out being transferred to 18-20 DEG C, relative humidity 85-95%, intensity of illumination 150-250 after machinery disturbs bacterium
Daily illumination 11-13 hours under the conditions of lux, cultivate 10-15 days；It is produced, is removed with tweezers when fructification grows to 8-12cm
The culture medium of fructification connection carries out the harvesting of fructification.
Preferably, the condition of chromium-rich culture is：After Cordyceps militaris is accessed chromium-rich culture medium after cooling, it is placed in constant temperature
In incubator, 20 DEG C are cultivated 10 days, after mycelia sufficiently grows, are carried out machinery and are disturbed bacterium, be then transferred into 18 DEG C, relative humidity
90%, illumination 12 hours daily under the conditions of 200 lux of intensity of illumination, it cultivates 15 days；, tweezers when fructification grows to 10cm
Extraction.
Wherein, the preparation method of step (2) described liquid spawn culture medium is：200 portions of potatoes in parts by weight are cut
At the fritter of 0.8-1.2cm, is boiled in boiling water 20 minutes, be then filtered, take juice, rear to be added 20 parts of glucose, magnesium sulfate
0.5 part, 1 part of potassium dihydrogen phosphate, vitamin B10.01 part, after thawing, add water 1000mL, natural ph.
More specifically, it is every bottle of 250mL that step (2) the Cordyceps militaris bacterium culture medium, which is packed into capacity, then 121 DEG C
Sterilize 30min, cooling inoculation, and every bottle meets liquid spawn 20mL.The chromium-rich culture medium is sub-packed in 500ml can after preparing
In bottle, the sealing of edible mushroom special sealing membrane sterilizes 20 minutes under the conditions of 121 DEG C；It has gone out to take out after bacterium and has held chromium-rich culture medium
Can, naturally cool to 20 DEG C -30 DEG C.
The invention has the advantages that：
This method is by selecting suitable culture medium and condition of culture, by Cr3+Enrichment, by mineralized
Cr3+Change organic chromium, effectively avoids inorganic states Cr3+Autoxidation is Cr6+, reduce human body and directly absorb metal Cr3+To people
The injury that body generates.
And by research, it has also been found that, the chromium in Cordyceps militaris also has facilitation to the conversion of polysaccharide in its mycelium, rises
Unexpected technical effect, while organic chromium and Cordyceps sinensis polysaccharide collective effect have been arrived, the pancreas of diabetic can be improved
Gland function, to reduce the use of human insulin.
Detailed description of the invention
Fig. 1 is CrCl in chromium-rich culture medium3Influence of the additive amount to fruiting body yield；
Fig. 2 is the CrCl of various concentration3Influence of the additive amount to chromium content in fructification；
Fig. 3 is influence of the chromium concn to polysaccharide in mycelium.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..It is of the present invention various
Reagent, test material (including Cordyceps militaris spawn) can be obtained commercially.
Embodiment 1
A kind of cordyceps culturing method, described method includes following steps：
(1) Cordyceps militaris spawn of 0-4 DEG C of preservation is subjected to VB1Slant strains culture：
A, Cordyceps militaris spawn culture medium is prepared：200 portions of potatoes in parts by weight are cut into the fritter of 0.8-1.2cm,
It is boiled in boiling water 20 minutes, is then filtered, takes juice, rear 20 parts of glucose of addition, 0.5 part of magnesium sulfate, 1 part of potassium dihydrogen phosphate,
Vitamin B10.01 part, 20 parts of agar, after thawing, constant volume is at 1000mL, natural ph；
B, Cordyceps militaris spawn culture medium dispenses, sterilizing：
Prepared bacterium culture medium is sub-packed in 18mm × 180mm test tube, is sealed, under the conditions of 121 DEG C, sterilizing
20min；
C, cooling, inoculation：
It has gone out and has taken out Cordyceps militaris spawn culture medium after bacterium, naturally cooled to 30 DEG C, aseptically, accessed cordyceps
Kind；
D, it cultivates：
The Cordyceps militaris spawn culture medium for connecting kind is placed in constant incubator, 25 DEG C are cultivated 4 days, and mycelia sufficiently grows；
E, it selects：
Growing way is got well to the Cordyceps militaris VB for degradation phenomena do not occur1Slant strains are selected；
(2) it is inoculated with：
Cordyceps militaris bacterium culture medium is prepared first：200 portions of potatoes in parts by weight are cut into the small of 0.8-1.2cm
Block boils 20 minutes in boiling water, is then filtered, takes juice, rear 20 parts of glucose of addition, and 0.5 part of magnesium sulfate, potassium dihydrogen phosphate
1 part, vitamin B10.01 part, after thawing, add water 1000mL, natural ph.
Prepare chromium-rich culture medium：First by 1 part of potassium dihydrogen phosphate and CrCl30.5 part is added to stirring and dissolving in 50 parts of water, will
30 parts of load weighted oat is fitted into the tissue culture bottle of 500mL, and the potassium dihydrogen phosphate and CrCl dissolved is added3Mixed solution covers
Lid, then 121 DEG C of sterilizing 60min take out cooling, for use.
Then Cordyceps militaris bacterium culture medium is packed into capacity is every bottle of 250mL, then 121 DEG C of sterilizings 30min, cooling
Inoculation, every bottle meets liquid spawn 20mL.The chromium-rich culture medium is sub-packed in 500ml can after preparing, and edible mushroom is special
It is sealed with sealed membrane, is sterilized 20 minutes under the conditions of 121 DEG C；Gone out the can for taking out after bacterium and holding chromium-rich culture medium, natural
It is cooled to 20 DEG C -30 DEG C.
Again by Cordyceps militaris VB1Slant strains are inoculated on Cordyceps militaris bacterium culture medium under sterile working, in temperature
It is to cultivate 5d in the shaking table of 160r/min for 20 DEG C, revolving speed, the good strain of liquid fermentation is inoculated on chromium-rich culture medium, every bottle
Inoculum concentration is 20mL；
(3) chromium-rich culture：
It after Cordyceps militaris is accessed chromium-rich culture medium after cooling, is placed in constant incubator, 18 DEG C are cultivated 8 days, bacterium
After silk is sufficiently grown, carry out machinery disturb after bacterium be transferred to 20 DEG C, it is relative humidity 95%, every under the conditions of 250 lux of intensity of illumination
Its illumination 11 hours is cultivated 10 days；It is produced when fructification grows to 12cm or so with tweezers, removes the culture medium of fructification connection
Carry out the harvesting of fructification.
Embodiment 2
A kind of cordyceps culturing method, described method includes following steps：
(1) Cordyceps militaris spawn of 0-4 DEG C of preservation is subjected to VB1Slant strains culture：
A, Cordyceps militaris spawn culture medium is prepared：200 portions of potatoes in parts by weight are cut into the fritter of 0.8-1.2cm,
It is boiled in boiling water 20 minutes, is then filtered, takes juice, rear 20 parts of glucose of addition, 0.5 part of magnesium sulfate, 1 part of potassium dihydrogen phosphate,
Vitamin B10.01 part, 20 parts of agar, after thawing, constant volume is at 1000mL, natural ph；
B, Cordyceps militaris spawn culture medium dispenses, sterilizing：
Prepared bacterium culture medium is sub-packed in 18mm × 180mm test tube, is sealed, under the conditions of 121 DEG C, sterilizing
20min；
C, cooling, inoculation：
It has gone out and has taken out Cordyceps militaris spawn culture medium after bacterium, naturally cooled to 25 DEG C, aseptically, accessed cordyceps
Kind；
D, it cultivates：
The Cordyceps militaris spawn culture medium for connecting kind is placed in constant incubator, 24 DEG C are cultivated 6 days, and mycelia sufficiently grows；
E, it selects：
Growing way is got well to the Cordyceps militaris VB for degradation phenomena do not occur1Slant strains are selected；
(2) it is inoculated with：
Cordyceps militaris bacterium culture medium is prepared first：200 portions of potatoes in parts by weight are cut into the small of 0.8-1.2cm
Block boils 20 minutes in boiling water, is then filtered, takes juice, rear 20 parts of glucose of addition, and 0.5 part of magnesium sulfate, potassium dihydrogen phosphate
1 part, vitamin B10.01 part, after thawing, add water 1000mL, natural ph.
Prepare chromium-rich culture medium：First by 1 part of potassium dihydrogen phosphate and CrCl30.6 part is added to stirring and dissolving in 50 parts of water, will
30 parts of load weighted oat is fitted into the tissue culture bottle of 500mL, and the potassium dihydrogen phosphate and CrCl dissolved is added3Mixed solution covers
Lid, then 121 DEG C of sterilizing 60min take out cooling, for use.
Then it is every bottle of 250mL that Cordyceps militaris bacterium culture medium, which is packed into capacity, and then 121 DEG C of sterilizing 30min, cooling connect
Kind, every bottle meets liquid spawn 20mL.The chromium-rich culture medium is sub-packed in 500ml can after preparing, and edible mushroom is dedicated
Sealed membrane sealing, sterilizes 20 minutes under the conditions of 121 DEG C；Gone out the can for taking out after bacterium and holding chromium-rich culture medium, naturally cold
But to 20 DEG C -30 DEG C.
Again by Cordyceps militaris VB1Slant strains are inoculated on Cordyceps militaris bacterium culture medium under sterile working, in temperature
It is to cultivate 5d in the shaking table of 160r/min for 20 DEG C, revolving speed, the good strain of liquid fermentation is inoculated on chromium-rich culture medium, every bottle
Inoculum concentration is 20mL；
(3) chromium-rich culture：
It after Cordyceps militaris is accessed chromium-rich culture medium after cooling, is placed in constant incubator, 20 DEG C are cultivated 10 days, bacterium
After silk is sufficiently grown, carry out machinery and disturb bacterium, be then transferred into 18 DEG C, relative humidity 90%, under the conditions of 200 lux of intensity of illumination
Daily illumination 12 hours is cultivated 15 days, is produced when fructification grows to 10cm with tweezers, remove the culture medium of fructification connection into
The harvesting of row fructification.
Embodiment 3
A kind of cordyceps culturing method, described method includes following steps：
(1) Cordyceps militaris spawn of 0-4 DEG C of preservation is subjected to VB1Slant strains culture：
A, Cordyceps militaris spawn culture medium is prepared：200 portions of potatoes in parts by weight are cut into the fritter of 0.8-1.2cm,
It is boiled in boiling water 20 minutes, is then filtered, takes juice, rear 20 parts of glucose of addition, 0.5 part of magnesium sulfate, 1 part of potassium dihydrogen phosphate,
Vitamin B10.01 part, 20 parts of agar, after thawing, constant volume is at 1000mL, natural ph；
B, Cordyceps militaris spawn culture medium dispenses, sterilizing：
Prepared bacterium culture medium is sub-packed in 18mm × 180mm test tube, is sealed, under the conditions of 121 DEG C, sterilizing
20min；
C, cooling, inoculation：
It has gone out and has taken out Cordyceps militaris spawn culture medium after bacterium, naturally cooled to 20 DEG C, aseptically, accessed cordyceps
Kind；
D, it cultivates：
The Cordyceps militaris spawn culture medium for connecting kind is placed in constant incubator, 22 DEG C are cultivated 8 days, and mycelia sufficiently grows；
E, it selects：
Growing way is got well to the Cordyceps militaris VB for degradation phenomena do not occur1Slant strains are selected；
(2) it is inoculated with：
Cordyceps militaris bacterium culture medium is prepared first：200 portions of potatoes in parts by weight are cut into the small of 0.8-1.2cm
Block boils 20 minutes in boiling water, is then filtered, takes juice, rear 20 parts of glucose of addition, and 0.5 part of magnesium sulfate, potassium dihydrogen phosphate
1 part, vitamin B10.01 part, after thawing, add water 1000mL, natural ph.
Prepare chromium-rich culture medium：First by 1 part of potassium dihydrogen phosphate and CrCl30.4 part is added to stirring and dissolving in 50 parts of water, will
30 parts of load weighted oat is fitted into the tissue culture bottle of 500mL, and the potassium dihydrogen phosphate and CrCl dissolved is added3Mixed solution lid
Lid, then 121 DEG C of sterilizing 60min take out cooling, for use；
Then it is every bottle of 250mL that Cordyceps militaris bacterium culture medium, which is packed into capacity, and then 121 DEG C of sterilizing 30min, cooling connect
Kind, every bottle meets liquid spawn 20mL.The chromium-rich culture medium is sub-packed in 500ml can after preparing, and edible mushroom is dedicated
Sealed membrane sealing, sterilizes 20 minutes under the conditions of 121 DEG C；Gone out the can for taking out after bacterium and holding chromium-rich culture medium, naturally cold
But to 25 DEG C.
Again by Cordyceps militaris VB1Slant strains are inoculated on Cordyceps militaris bacterium culture medium under sterile working, in temperature
It is to cultivate 5d in the shaking table of 150r/min for 20 DEG C, revolving speed, the good strain of liquid fermentation is inoculated on chromium-rich culture medium, every bottle
Inoculum concentration is 20mL；
(3) chromium-rich culture：
It after Cordyceps militaris is accessed chromium-rich culture medium after cooling, is placed in constant incubator, 16 DEG C are cultivated 12 days, bacterium
After silk is sufficiently grown, carry out machinery disturb after bacterium be transferred to 20 DEG C, it is relative humidity 85%, every under the conditions of 150 lux of intensity of illumination
Its illumination 13 hours is cultivated 15 days；It is produced when fructification grows to 8cm or so with tweezers, removes the culture medium of fructification connection
Carry out the harvesting of fructification.
Inventor also proves beneficial effects of the present invention by testing as follows, specific as follows：
Firstly, inventor is separated and measured to trivalent Organic Chromium by capillary electrophoresis, chromium in Cordyceps militaris is determined
Existence form be organic, and also carried out following test：
One, CrCl in chromium-rich culture medium3Influence of the additive amount to fruiting body yield
From figure 1 it appears that the biomass of fructification is with chromium concn when the concentration of chromium is 0.6 μ g/mL or less
Increase and increase, it is 21.3g/ bottle that fructification biomass, which reaches maximum, in 0.6 μ g/mL, and when 0.6 μ g/mL of concentration of chromium is sub in fact
Body yield highest, biological transformation ratio 71%, and test and existed using the Cordyceps militaris biological transformation ratio of present conventional cultivation method
60% or so.
Two, the CrCl of various concentration3Influence of the additive amount to chromium content in fructification
From figure 2 it can be seen that chromium concn be 0.6 μ g/mL hereinafter, in fructification chromium content with chromium concn increase
And increase, chromium content is positively correlated with chromium concn, reaches maximum in the 0.6 μ g/mL of concentration of chromium, and Organic Chromium is 314.33 μ g/
G, more than 0.6 μ g/mL, chromium content is begun to decline.
Three, influence of the chromium concn to polysaccharide in mycelium
From figure 3, it can be seen that chromium concn has facilitation to the conversion of polysaccharide in mycelium, in 0.4 μ g/mL hereinafter, bacterium
Polyoses content is increased with the raising of chromium concn in filament, does not add the culture medium of chromium ion, the content of Cordyceps sinensis polysaccharide is
0.23mg/g is 0.44mg/mL in 0.6 μ g/mL, is 2 times of conventional cultivation method, turns at this time for polyoses content in mycelium
The optium concentration of change, when more than 0.6 μ g/g, polyoses content starts to reduce.
Four, influence of the different cultural methods to Cordyceps militaris blood glucose
Induced using alloxan and urinate sick mouse model, the Cordyceps militaris of routine culture mode and chromium-rich training method according to
2 dosage groups (600mg/kg, 300mg/kg), positive controls (diabetic mice of diabetes pill 1000mg/kg), pyridine carboxylic acid
2 dosage groups of chromium (6 μ g/kg, 10 μ g/kg) and negative control group.0.20ml/10g volume gastric infusion is pressed, one time a day, even
Continuous 7d.7h fasting before last dose, administration 1h posterior orbit veniplex blood sampling, separation serum is by determination of glucose oxidase empty stomach
Blood glucose.It is found by experiment that chromium-rich Cordyceps militaris has preferably alloxan diabetes animal model in 600mg/kg gastric infusion
Therapeutic effect, have significant difference (P < 0.01, P < 0.05) compared with model control group, while chromium-rich Cordyceps militaris is in 600mg/kg
Intact animal blood glucose is had not significant impact under dosage, prompt chromium-rich Cordyceps militaris without significantly stimulation insulin releasing act on or
Insulin-like effects, and diabetes pill 1000mg/kg has influence to the blood glucose of intact animal.
Although above having used general explanation and specific embodiment, the present invention is described in detail, at this
On the basis of invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Therefore,
These modifications or improvements without departing from theon the basis of the spirit of the present invention are fallen within the scope of the claimed invention.
Claims (6)
1. a kind of cordyceps culturing method, which is characterized in that described method includes following steps：
(1) Cordyceps militaris spawn for saving 0-4 DEG C carries out VB1Slant strains culture：
A, Cordyceps militaris spawn culture medium is prepared：200 portions of potatoes in parts by weight are cut into the fritter of 0.8-1.2cm, are being boiled
It is boiled in water 20 minutes, is then filtered, takes juice, it is rear to be added 20 parts of glucose, it 0.5 part of magnesium sulfate, 1 part of potassium dihydrogen phosphate, ties up
Raw element B10.01 part, 20 parts of agar, after thawing, constant volume is at 1000mL to get natural ph；
B, Cordyceps militaris spawn culture medium dispenses, sterilizing：
Prepared Cordyceps militaris spawn culture medium is sub-packed in 18mm × 180mm test tube, is sealed, under the conditions of 121 DEG C, sterilizing
20min；
C, cooling, inoculation：
It has gone out and has taken out Cordyceps militaris spawn culture medium after bacterium, naturally cooled to 20 DEG C -30 DEG C, aseptically, accessed Cordyceps militaris
Strain；
D, it cultivates：
The Cordyceps militaris spawn culture medium that will be connected kind is placed in constant incubator, 22-25 DEG C culture 4-8 days, mycelia sufficiently grows；
E, it selects：
Growing way is got well to the Cordyceps militaris VB for degradation phenomena do not occur1Slant strains are selected；
(2) it is inoculated with：
By Cordyceps militaris VB1Slant strains are inoculated on Cordyceps militaris bacterium culture medium under sterile working, temperature be 20 DEG C,
5d is cultivated in the shaking table that revolving speed is 150-160r/min, the good strain of liquid fermentation is inoculated on chromium-rich culture medium, every bottle connects
Kind amount is 20mL；
(3) chromium-rich culture：
After Cordyceps militaris is accessed chromium-rich culture medium after cooling, be placed in constant incubator, 16-20 DEG C culture 8-12 days,
After mycelia sufficiently grows, carry out being transferred to 18-20 DEG C, relative humidity 85-95%, intensity of illumination 150-250 Le after machinery disturbs bacterium
Gram illumination daily 11-13 hour under the conditions of this, culture 10-15 days；It is produced when fructification grows to 8-12cm with tweezers, removes son
The culture medium of entity connection carries out the harvesting of fructification.
2. cordyceps culturing method according to claim 1, which is characterized in that step (2) described liquid spawn culture medium
Composition be in parts by weight：200 parts of potato, 20 parts of glucose, 0.5 part of magnesium sulfate, 1 part of potassium dihydrogen phosphate, vitamin
B10.01 part, 1000 parts of water.
3. cordyceps culturing method according to claim 1, which is characterized in that step (2) the Cordyceps militaris strain
It is every bottle of 250mL that culture medium, which is packed into capacity, then 121 DEG C of sterilizings 30min, cooling inoculation, and every bottle meets liquid spawn 20mL.
4. cordyceps culturing method according to claim 1, which is characterized in that the group of step (2) the chromium-rich culture medium
At being in parts by weight：30 parts of oat, CrCl30.4-0.6 parts, 1 part of potassium dihydrogen phosphate, 50 parts of water.
5. cordyceps culturing method according to claim 1, which is characterized in that step (2) the chromium-rich culture medium is being matched
It is sub-packed in after making in 500ml can, the sealing of edible mushroom special sealing membrane sterilizes 20 minutes under the conditions of 121 DEG C；It has gone out bacterium
The can for holding chromium-rich culture medium is taken out afterwards, naturally cools to 20 DEG C -30 DEG C.
6. cordyceps culturing method according to claim 1, which is characterized in that the condition of chromium-rich culture is：By Cordyceps militaris
After liquid accesses chromium-rich culture medium after cooling, it is placed in constant incubator, 20 DEG C are cultivated 10 days, after mycelia sufficiently grows, into
Row machinery disturbs bacterium, is then transferred into 18 DEG C, relative humidity 90%, illumination daily 12 hours under the conditions of 200 lux of intensity of illumination,
Culture 15 days, is produced when fructification grows to 10cm with tweezers.
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| CN1297995A (en) * | 2001-01-12 | 2001-06-06 | 杨忠才 | Production process of chromium-rich cordyceps mycelium |
| CN103125275A (en) * | 2013-03-15 | 2013-06-05 | 熊艳 | Cultivation method of cordyceps militaris |
| CN104604522A (en) * | 2015-01-19 | 2015-05-13 | 河南省民兴茧丝绸有限责任公司 | Method for producing cordyceps militaris |
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| CN1297995A (en) * | 2001-01-12 | 2001-06-06 | 杨忠才 | Production process of chromium-rich cordyceps mycelium |
| CN103125275A (en) * | 2013-03-15 | 2013-06-05 | 熊艳 | Cultivation method of cordyceps militaris |
| CN104604522A (en) * | 2015-01-19 | 2015-05-13 | 河南省民兴茧丝绸有限责任公司 | Method for producing cordyceps militaris |
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