CN105985150A - Isaria cicadae coremium liquid culture medium - Google Patents
Isaria cicadae coremium liquid culture medium Download PDFInfo
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Abstract
The invention provides a liquid culture medium used for artificially culturing isaria cicadae coremium, wherein the culture medium mainly includes 0.2-3% of yeast extract powder, 0.1-2% of soy protein hydrolysate and the balance being water to 100%, and also can includes 1-8% of saccharose, 0.005-0.1% of calcium chloride and 0.01-0.5% of potassium dihydrogen phosphate. The liquid culture medium is significantly increased in yield of the isaria cicadae coremium and the content of active components therein, has stable product quality and is suitable for large-scale industrial production.
Description
Technical field
The present invention relates to fluid medium in Periostracum cicadae artificial culture coremium, specifically, relate to Periostracum cicadae and manually train
The new technology supported.
Background technology
Periostracum cicadae (Isaria cicadae Miquel) belong to mycota (FUNGI) Ascomycota (Ascomycota),
Cup fungi subphylum (Pezizomycotina), excrement shell Gammaproteobacteria (Sordariomycetes), Hypocreales (Hypocreales),
Cordyceps section (Cordycipitaceae), Isaria belong to (Isaria).
Periostracum cicadae (Isaria cicadae Miquel) is dispersed species, its strain can control oneself collection separation can also purchase
Buy.The Periostracum cicadae 18 provinces on the south China Qinling Mountains-Huaihe River, city, district have widely distributed.In China the Changjiang river with
The subtropical zone in south and torrid areas, and many in low lying areas and the Jinsha jiang River on plateau, Yunnan-Tibet, Nujiang, the Lancang River
With areas, river valley such as the Yarlung Zangbo River.As long as in its season of growth, the annual 6-8 month, according to general Chinese medicine
The acquisition method of material all can be adopted.
Periostracum cicadae is the Chinese crude drug that China is famous and precious, is to colonize in a kind of Cordyceps on Cicadae (being commonly called as " cicada ").Its
Medicinal existing more than 1000 year history, is one of traditional rare medicinal herbs of China, has many medical values.
Main component has: adenosine, Cordyceps polysaccharide, cordycepic acid (mannitol), cordycepin, uracil, sterol,
Alkaloid, vitamin, inorganic salt, mineral element etc..
Periostracum cicadae has the function of regulation human immunity, improves appetite, hypnotic, strengthens self disease-resistant ability.
In the middle of these compositions, Cordyceps polysaccharide has the biological activity of uniqueness, have antitumor, antibacterial, antiviral,
Radioprotective, anti-ageing effect of waiting for a long time, immunoregulation effect becomes the focus of research and development.Adenosine can suppress maincenter
The irritability of neuron, can expand crown and peripheral vessels, increase coronary artery blood flow, reduce blood pressure,
Decreased heart rate etc. act on.Adenosine also has the effects such as antiplatelet aggregation, radioprotective and antitumor.The most such as
This, the active component ISP-1 in Periostracum cicadae has two-way immunoregulatory activity, can be greatly improved organ and move
Planting the success rate of operation, the rehabilitation to patient also has obvious good effect.
From Mr. Chen Zhuan in 1993 have studied the artificial culture of Periostracum cicadae, Periostracum cicadae artificial culture coremium
Seed liquor and fermentation liquid have been used up 20% Rhizoma Solani tuber osi liquor, 3% white sugar, surplus moisturizing to 100%.
Prior art there is no the relevant report using identical culture medium prescription for Periostracum cicadae artificial culture coremium
And application.
Summary of the invention
It is an object of the invention to provide a kind of fluid medium for artificial culture Periostracum cicadae coremium.
The present invention further provides a kind of method for artificial culture Periostracum cicadae coremium.
A kind of fluid medium for artificial culture Periostracum cicadae coremium, the raw material of this culture medium specifically includes that
Yeast extract powder, soy bean protein hydrolysate, water.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.2～3%,
Soy bean protein hydrolysate 0.1～2%, surplus moisturizing to 100%.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.3～1%,
Soy bean protein hydrolysate 0.2～1%, surplus moisturizing to 100%.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.4～0.8%,
Soy bean protein hydrolysate 0.3～0.8%, surplus moisturizing to 100%.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.4%,
Soy bean protein hydrolysate 0.3%, surplus moisturizing to 100%.
Further, the raw material composition of described culture medium can also add sucrose 1～8%；Further, sugarcane
Sugar addition is 2～5%；Further, sucrose addition is 3～4%；Further, sucrose addition
It is 3.5%；Further, described sucrose is white sugar.
Further, the raw material composition of described culture medium can also add calcium chloride 0.005～0.1%, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.01～0.5%；Further, the addition of calcium chloride and potassium dihydrogen phosphate be calcium chloride 0.008～
0.05%, potassium dihydrogen phosphate 0.02～0.2%；Further, the addition of calcium chloride and potassium dihydrogen phosphate is chlorine
Change calcium 0.01～0.02%, potassium dihydrogen phosphate 0.05～0.1%；Further, calcium chloride and potassium dihydrogen phosphate
Addition is calcium chloride 0.01%, potassium dihydrogen phosphate 0.05%.
Culture medium of the present invention is used for Cordyceps cicadae strain through slant strains cultivation, liquid submerged culture, seed
Tank liquid culture, fermentor liquid fermentation culture and solid go out grass cultivation, obtain spore by above-mentioned incubation step
Stalk bundle, spore powder and mycoplasma；Wherein slant strains cultivation, liquid submerged culture, seed tank liquid culture,
Fermentor liquid fermentation culture and solid go out grass cultivation all can use conventional method.
A kind of method that the present invention further provides artificial culture Periostracum cicadae coremium, the method includes slant strains
Cultivation, liquid submerged culture, seed tank liquid culture, fermentor liquid fermentation culture and solid go out grass cultivation
Step, wherein said liquid submerged culture, seed tank liquid culture, fermentor liquid fermentation culture and solid
Go out liquid medium starting material in grass incubation step and specifically include that yeast extract powder, soy bean protein hydrolysate, water.
Described cultural method step also includes seed tank amplification culture.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.2～3%,
Soy bean protein hydrolysate 0.1～2%, surplus moisturizing to 100%.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.3～1%,
Soy bean protein hydrolysate 0.2～1%, surplus moisturizing to 100%.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.4～0.8%,
Soy bean protein hydrolysate 0.3～0.8%, surplus moisturizing to 100%.
Further, the proportioning raw materials of described culture medium is (percentage by weight): yeast extract powder 0.4%,
Soy bean protein hydrolysate 0.3%, surplus moisturizing to 100%.
Further, the raw material composition of described culture medium can also add sucrose 1～8%；Further, sugarcane
Sugar addition is 2～5%；Further, sucrose addition is 3～4%；Further, sucrose addition
It is 3.5%；Further, described sucrose is white sugar.
Further, the raw material composition of described culture medium can also add calcium chloride 0.005～0.1%, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.01～0.5%；Further, the addition of calcium chloride and potassium dihydrogen phosphate be calcium chloride 0.008～
0.05%, potassium dihydrogen phosphate 0.02～0.2%；Further, the addition of calcium chloride and potassium dihydrogen phosphate is chlorine
Change calcium 0.01～0.02%, potassium dihydrogen phosphate 0.05～0.1%；Further, calcium chloride and potassium dihydrogen phosphate
Addition is calcium chloride 0.01%, potassium dihydrogen phosphate 0.05%.
In the present invention, Periostracum cicadae (Isaria cicadae Miquel) used is dispersed species, can make by oneself and can also purchase
Buy.The Periostracum cicadae 18 provinces on the south China Qinling Mountains-Huaihe River, city, district have widely distributed.In China the Changjiang river
On the south subtropical zone and torrid areas, and many in low lying areas and the Jinsha jiang River on plateau, Yunnan-Tibet, Nujiang, billows deep blue
Area, the river valley such as river and the Yarlung Zangbo River.As long as in its season of growth, the annual 6-8 month, in general
The acquisition method of medical material all can be adopted.The Periostracum cicadae adopted can be separated to Cordyceps cicadae strain, concrete steps according to purgation
As follows:
The separation method of bacterial strain is: on super mirror workbench, aseptically, the fresh Cordyceps that will adopt
Clean with aseptic water washing, it is placed in the culture dish of sterilized a diameter of 15cm, the interior sclerotium portion of Cordyceps
Divide (polypide) absorbent cotton parcel with drenching, with moisturizing.The load of a sterilizing is placed below the sporophore of Cordyceps
Slide, aweto sclerotium part paves with little slide so that pregnant part microscope slide can directly not contacted, its away from
From about about 0.5cm.The illumination box that then culture dish is positioned over 20 DEG C ± 0.5 DEG C is cultivated, and treats Cicadae
When flower spore launches on microscope slide, around spore, drip the PDA culture medium just dissolved a little, removal
Slide, is placed in moisturizing in the culture dish of another sterilizing and cultivates, with a small amount of bacterium of Inoculating needle picking after growing mycelia
Silk turns another plate (SDAY culture medium) and cultivates (ibid), until the bacterium colony growing single purification is
Only.And verify through morphology or molecular biology method, this bacterial strain is Periostracum cicadae (Isaria cicadae
Miquel) phorozoon.
Paecilomyces cicadidae(Miquel)Samson bacterial strain [Paecilomyces cicadae disclosed in the preferred CN102851353A of the present invention
(Miq.) Samson] general in China Committee for Culture Collection of Microorganisms on November 18th, 2009
Logical microorganism center (being called for short CGMCC) registration preservation, preserving number is CGMCC No.3453.
The present invention has following advantage compared with original culture medium technology: eliminate the loaded down with trivial details of former use Rhizoma Solani tuber osi liquor
Technique, has saved labour force, improves labor productivity；Solve Rhizoma Solani tuber osi different cultivars and Various Seasonal
Quality differs problem；The present invention provides soy bean protein hydrolysate with yeast extract powder proportioning first for artificial Cicadae
In flower coremium liquid culture, Periostracum cicadae coremium yield and active constituent content thereof significantly improve, it is ensured that product
Steady quality, improves production environment, it is adaptable to large-scale industrial production.
Detailed description of the invention:
Below in conjunction with detailed description of the invention, the present invention is further illustrated:
Embodiment 1
Culture medium prescription: soy bean protein hydrolysate 0.3%, yeast extract powder 0.4%, white sugar 3.5%, chlorination
Calcium 0.01%, potassium dihydrogen phosphate 0.05%, surplus distilled water supplies 100%.
Embodiment 2
Culture medium prescription: soy bean protein hydrolysate 0.3%, yeast extract powder 1%, white sugar 2%, calcium chloride
0.05%, potassium dihydrogen phosphate 0.02%, surplus distilled water supplies 100%.
Embodiment 3
Culture medium prescription: soy bean protein hydrolysate 1%, yeast extract powder 0.2%, white sugar 5%, calcium chloride
0.008%, potassium dihydrogen phosphate 0.2%, surplus distilled water supplies 100%.
Embodiment 4
Culture medium prescription: soy bean protein hydrolysate 0.1%, yeast extract powder 3%, white sugar 1%, calcium chloride
0.1%, potassium dihydrogen phosphate 0.01%, surplus distilled water supplies 100%.
Embodiment 5
Culture medium prescription: soy bean protein hydrolysate 2%, yeast extract powder 0.2%, white sugar 6%, calcium chloride
0.005%, potassium dihydrogen phosphate 0.5%, surplus distilled water supplies 100%.
Embodiment 6
Culture medium prescription: soy bean protein hydrolysate 0.6%, yeast extract powder 0.4%, white sugar 4%.
Embodiment 7
Culture medium prescription: soy bean protein hydrolysate 0.5%, yeast extract powder 0.4%, white sugar 3.5%.
Embodiment 8
Culture medium prescription: soy bean protein hydrolysate 1.5%, yeast extract powder 0.3%, white sugar 3.5%.
Embodiment 9 screening of medium formula is tested
1, primary inclined plane spawn culture
Culture medium prescription: 20% Rhizoma Solani tuber osi liquor+2% sucrose+2% agar, remaining moisturizing to 100%；pH6.5.
Specific practice is: boiled by potato decortication, after filter and remove residue, according to the ratio of 200 grams of preparations 1 liter
Preparation, adds sucrose, the constant volume of 2%, adds 2% agar and melts, and subpackage enters test tube, at 0.1Mpa
Under pressure, 121 DEG C, sterilizing 30min, exits complete, puts into inclined-plane natural cooling.Under aseptic condition
Cordyceps cicadae strain CGMCCNo.3453 is received on inclined-plane, 24 DEG C, cultivates 15 days, it is thus achieved that produce mother and plant
(Paecilomyces cicadidae(Miquel)Samson bacterial strain [Paecilomyces cicadae (Miq.) Samson] involved in the present invention in
On November 18th, 2009 (is called for short at China Committee for Culture Collection of Microorganisms's common micro-organisms center
CGMCC) registration preservation, preserving number is CGMCC No.3453).
2, liquid shaking bottle seed culture
Culture medium prescription: formula 1-15, refers to table 1.
Specific practice: 500ml triangular flask liquid amount 1/3, takes well-grown Periostracum cicadae (Isaria cicadae
Miquel) inclined-plane seed, homogenization, it is inoculated in shaking flask by the inoculum concentration of 2%, at water bath with thermostatic control shaking table
Upper 25 DEG C, 140r/min cultivate 3 days.Cultured seed is grey black, inside has substantial amounts of small rice grain big
Little fungus ball, close, bacterium solution has mushroom tone sweet taste；
Table 1 different liquids culture medium artificial culture Periostracum cicadae goes out grass situation and Main Ingredients and Appearance contrast table
3, seed tank amplification culture
The same liquid seeds of culture medium used by seed tank.
Specific practice: loading 20L fluid medium in 30L airlift fermentor, culture medium temperature is more than
95 DEG C, pH value be 6.0～7.0, under 14.7 × 104Pa pressure, be passed through steam and be heated to 121 DEG C, and
Pressurize 30～45 minutes；After sterilizing, by 6 bottles of above-mentioned cultured 500ml shaking flask kinds when being cooled to 20 DEG C
Son access cultivate, cultivation temperature is 25～26 DEG C, and ventilation is 1:0.5V/V min, pressurize 4.5～
7.0 × 104Pa, through 20-48h aerobic culture, reaches logarithmic growth after date, and final volume of culture is 20L.
When bubbles volume is more, can add edible defoaming agent, addition is 0.03%,；
In seed tank 1-18 hour is lag phase, within 18-38 hour, is fast growing period (exponential phase),
It within 38-48 hour, it is the fast-growth later stage.Incubation time was advisable with 48 hours.
4, ferment tank is cultivated
The same liquid seeds of culture medium used by fermentation tank.
Specific practice: loading 200L fluid medium in 300L fermentation tank, pH value is 6.0～7.0,
And adding edible defoaming agent, addition is 0.03%, logical steam and In Situ Heating to 121 DEG C,
Under 14.7 × 104Pa pressure, pressurize 30～45 minutes；After sterilizing, when being cooled to 26 DEG C, by seed tank
50L liquid seeds all accesses fermentor cultivation, cultivation temperature 25～26 DEG C, and ventilation is 1:0.5
V/V min, pressurize 4.903～7.0845 × 104Pa, through 36-48h aerobic culture, reach logarithmic growth
After date, final volume of culture is 200L.When bubbles volume is more, edible defoaming agent can be added, add
Entering amount is 0.03%；
In fermentation tank 0-12 hour is lag phase, within 12-38 hour, is fast growing period, for growth after 38 hours
Stable phase, reaches peak in 48 hours, enters phase of decline afterwards.With 48 hours eventually to fermentation, proceed to solid culture.
When larger-scale produces, can be using fermentation tank as secondary seed tank, the fermentation liquid that then will ferment
Proceed to the fermentation tank of bigger volume, to obtaining more culture fluid.
5, solid goes out grass cultivation
Cleaning up in Semen Tritici aestivi loads rinse bath, the Semen Tritici aestivi drainage after cleaning, subpackage is to cultivating box, often
330 grams of box-packed material, adds 470ml pure water, seals.Put high-pressure steam sterilizing pan, 121 DEG C, steam
Vapour sterilizing 50min, natural cooling.With inoculator, the seed liquor in the fermentation tank in above-mentioned 4 is compared by 5%
Example is injected in the culture medium of the aforementioned table 1 after sterilizing, fully shakes mixing.Postvaccinal cultivation box is moved
Cultivate to culturing room.Using stereoscopic culture frame to cultivate, at the initial stage, culturing room's temperature is 22 DEG C～24 DEG C, secretly
Cultivate 3～5 days, until mycelium grows until covering with charge level to compost；Change full spectrum light afterwards into shine
Cultivating, intensity of illumination 100lux～200lux, keeping culturing room's temperature is 20 DEG C～22 DEG C, timing ventilation
Aerofluxus, keeps culturing room's air fresh.(cultivate 23-25 days, during Biomass maximum) when coremium maturation,
Gather in time.Sporophore is taken out from cultivating box, mycoplasma is separated with sporophore, dehumidifying, 50 DEG C of drying,
Packaging.
After cultivation terminates, determine coremium output capacity (i.e. going out carelessness) and main effective ingredient gland simultaneously
Glycosides, polysaccharide, protein content.The results are shown in Table 2.
Table 2 different liquids culture medium artificial culture Periostracum cicadae goes out grass situation and Main Ingredients and Appearance contrast table
| Numbering | Go out careless % | Adenosine content % | Polyoses content % | Protein content % |
| 1 | 12.6 | 0.104 | 4.32 | 32.5 |
| 2 | 12.5 | 0.100 | 4.00 | 32.1 |
| 3 | 12.8 | 0.102 | 4.33 | 34.5 |
| 4 | 10.1 | 0.083 | 3.69 | 30.2 |
| 5 | 11.2 | 0.094 | 4.02 | 32.0 |
| 6 | 10.7 | 0.097 | 3.92 | 31.9 |
| 7 | 12.1 | 0.101 | 3.75 | 32.0 |
| 8 | 11.2 | 0.096 | 3.24 | 31.5 |
| 9 | 11.8 | 0.098 | 3.92 | 32.7 |
| 10 | 10.5 | 0.087 | 3.28 | 30.8 |
| 11 | 10.7 | 0.086 | 3.65 | 30.2 |
| 12 | 10.7 | 0.086 | 3.54 | 30.8 |
| 13 | 8.1 | 0.087 | 3.20 | 28.5 |
| 14 | 7.6 | 0.056 | 2.74 | 29.9 |
| 15 | 7.9 | 0.068 | 3.00 | 30.8 |
Experiment conclusion:
From upper table data it can be seen that in the formula adding soy bean protein hydrolysate (formula 1-12), produced
Periostracum cicadae coremium quantity is above not adding the formula (formula 13-15) of soy bean protein hydrolysate, and Semen sojae atricolor water is described
Solve albumen Periostracum cicadae cultivate in it is critical that；In addition to yield, the main active adenosine of Periostracum cicadae is with many
Sugar is above not adding the formula of soy bean protein hydrolysate.So the liquid that soy bean protein hydrolysate is applied to Periostracum cicadae is sent out
It ferment culture medium prescription is a quantum jump in Chinese caterpillar fungus fermentation technique.
From the result of testing inspection it can also be seen that only add yeast powder culture fluid (formula 4,10,11,
12,14), active component is significantly lower than the culture fluid (formula 1-3) adding yeast extract powder.
Embodiment 10 study on the stability
Cultivate by embodiment 9 method, investigate different batches fluid medium produce Periostracum cicadae yield and
Quality, the results are shown in Table 3.(culture medium prescription is the formula of embodiment 1, and Cordyceps cicadae strain is CGMCCNo.
3453)。
The change of effective ingredient and stability in table 3 fluid present invention culture medium coremium
| Batch number | Go out carelessness (%) | Polysaccharide (%) | Adenosine (%) |
| 20140516 | 12.3 | 5.173 | 0.106 |
| 20140520 | 12.1 | 5.200 | 0.101 |
| 20140523 | 11.8 | 5.205 | 0.098 |
| 20140527 | 12.0 | 5.179 | 0.099 |
| 20140530 | 12.3 | 5.132 | 0.091 |
Result shows, the Periostracum cicadae coremium that fluid present invention culture medium produces goes out careless more than 10%, polysaccharide
Content is higher than 5%, and adenosine content is higher than 0.09%, and constant product quality is good.
Embodiment 11 investigates yield and the quality of the liquid culture based formulas production Periostracum cicadae of different strains
Bacterial strain obtain: according to " research of Entomocenous Fungus, paecilomyces Cicadae " (Chen Zhuan, Acta Fungus Sinica, 1991
Year), " artificial culture of Periostracum cicadae and pharmacological research thereof " (Chen Zhuan etc., Acta Fungus Sinica, 1993
Year, 12 (2)) method disclosed in document.The separation method of bacterial strain is: on super mirror workbench,
Under aseptic condition, by clean for the fresh Cordyceps aseptic water washing adopted, it is placed in sterilized a diameter of 15cm
Culture dish in, the interior sclerotium part (polypide) of Cordyceps is with the absorbent cotton parcel drenched, with moisturizing.Cordyceps
Sporophore below place the microscope slide of a sterilizing, aweto sclerotium part paves with little slide, so that can pregnant portion
Divide and directly do not contact microscope slide, its distance about about 0.5cm.Then culture dish is positioned over 20 DEG C ±
The illumination box of 0.5 DEG C is cultivated, and when Periostracum cicadae spore launches on microscope slide, around spore, dropping is just
The PDA culture medium dissolved is a little, removes slide, is placed in moisturizing in the culture dish of another sterilizing and cultivates, treats
Turn another plate (SDAY culture medium) with a small amount of mycelia of Inoculating needle picking after growing mycelia and cultivate (ibid),
Obtain five bacterial strains: BACC0019, BACC0033, BACC0036, BACC0041
BACC0056.And verify through morphology and molecular biology method, the bacterial strain of separation is Periostracum cicadae worm
Grass (Isaria cicadae Miquel) phorozoon.
Cultural method: cultivate by embodiment 9 method.
Culture medium prescription: with the formula implementing 1.
The change of the coremium that table 4 fluid present invention culture medium different strains is cultivated and stability
| Bacterial strain number | Go out carelessness (%) | Polysaccharide (%) | Adenosine (%) |
| BACC0019 | 9.85 | 4.52 | 0.083 |
| BACC0033 | 9.47 | 5.10 | 0.082 |
| BACC0036 | 9.20 | 4.35 | 0.083 |
| BACC0041 | 9.43 | 4.08 | 0.075 |
| BACC0056 | 8.75 | 5.05 | 0.076 |
Result shows, the different Periostracum cicadae bacterial strain coremiums that fluid present invention culture medium produces go out careless 8.75%
Above, polyoses content is higher than 4.08%, and adenosine content is higher than 0.075%, and constant product quality is good.
Embodiment 12 study on the stability
Fluid present invention culture medium, is applied not only to the specific slant medium of embodiment 9 and Mycelium culture base
It is obtained in that good result, and when slant medium changes, or in Periostracum cicadae Mycelia body, make
Good result can be obtained equally by this culture medium.
The change of slant medium: change the culture medium in embodiment 9 into SDAY culture medium, i.e. peptone
10g/L, yeast leaches cream 10g/L, glucose 40g/L, agar 20g/L.Seed liquor and solid culture
With embodiment 9, cultivation results is: go out carelessness 12.6%, polyoses content 5.23%, adenosine content 0.105%.
Fermentor cultivation mycelium: cultivating identical by 1 in embodiment 9,2,3,4 steps, fermentation tank is trained
The mycelium supported, is dried after centrifugal, obtains mycelium product, survey its main chemical compositions content, with
Former cultural method compares, and the results are shown in Table 5:
Table 5 uses the change of effective ingredient in mycelium after fluid medium
| Batch number | Mycelia yield % | Adenosine % | Polyoses content % |
| 20140423(CK) | 4.2 | 0.099 | 7.52 |
| 20140430 | 5.0 | 0.105 | 8.82 |
| 20140428 | 5.4 | 0.112 | 9.24 |
These results suggest that, fluid present invention culture medium and different slant mediums or Mycelium culture group
Close, cultivate the product quality obtained and quantity is all significantly improved, and constant product quality.
Embodiment 13
Fluid medium: embodiment 2-8；
Cultural method: with embodiment 9；
The results are shown in Table 6:
| Embodiment | Go out carelessness (%) | Polysaccharide (%) | Adenosine (%) |
| 2 | 10.0 | 4.75 | 0.085 |
| 3 | 12.0 | 5.04 | 0.093 |
| 4 | 10.1 | 4.50 | 0.078 |
| 5 | 10.8 | 4.82 | 0.087 |
| 6 | 11.7 | 4.95 | 0.090 |
| 7 | 10.5 | 4.73 | 0.093 |
| 8 | 11.0 | 4.83 | 0.093 |
Claims (9)
1. the fluid medium for artificial culture Periostracum cicadae coremium, it is characterised in that described culture medium
Raw material specifically include that yeast extract powder, soy bean protein hydrolysate, water.
2. fluid medium as claimed in claim 1, it is characterised in that the proportioning raw materials of described culture medium
For (percentage by weight): yeast extract powder 0.2～3%, soy bean protein hydrolysate 0.1～2%, surplus moisturizing is extremely
100%.
3. fluid medium as claimed in claim 2, it is characterised in that the proportioning raw materials of described culture medium
For (percentage by weight): yeast extract powder 0.3～1%, soy bean protein hydrolysate 0.2～1%, surplus moisturizing is extremely
100%.
4. fluid medium as claimed in claim 3, it is characterised in that the proportioning raw materials of described culture medium
For (percentage by weight): yeast extract powder 0.4～0.8%, soy bean protein hydrolysate 0.3～0.8%, surplus is mended
Water is to 100%.
5. the fluid medium as described in any one of claim 1-4, it is characterised in that described culture medium
Raw material composition adds sucrose 1～8%.
6. fluid medium as claimed in claim 5, it is characterised in that the raw material composition of described culture medium
Middle addition calcium chloride 0.005～0.1%, potassium dihydrogen phosphate 0.01～0.5%.
7. fluid medium as claimed in claim 8, it is characterised in that described calcium chloride and biphosphate
The addition of potassium is calcium chloride 0.008～0.05%, potassium dihydrogen phosphate 0.02～0.2%.
8. a method for artificial culture Periostracum cicadae coremium, this cultural method includes slant strains cultivation, liquid
Shake-flask culture, seed tank liquid culture, fermentor liquid fermentation culture and solid go out grass incubation step, wherein
Described liquid submerged culture, seed tank liquid culture, fermentor liquid fermentation culture and solid go out grass and cultivate step
The arbitrary described fluid medium of claim 1-7 is used in rapid.
9. method as claimed in claim 8, wherein said cultural method also includes seed tank amplification culture
Step.
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