CN106550763A - A kind of artificial culture method of Periostracum cicadae spore powder - Google Patents
A kind of artificial culture method of Periostracum cicadae spore powder Download PDFInfo
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Abstract
The present invention relates to a kind of artificial culture method of Periostracum cicadae spore powder, it is characterised in that methods described comprises the steps：Step 1, the strain seed liquor that bibulous carrier and liquid culture are obtained are fully contacted；The bibulous carrier of contact seed liquor is first carried out light culture, switchs to optical culture after mycelium is covered with by step 2；Step 3, harvests to spore powder., with hygroscopic material as carrier culture spore powder, process is simple, cycle is short, sporulation quantity are high for the method, and spore powder quality is high, has obvious advantage relative to existing solid culture.
Description
Technical field
The present invention relates to a kind of artificial culture method of Periostracum cicadae spore powder, and in particular to one kind is with hygroscopic material
Expect the cultural method of the culture Periostracum cicadae spore powder for carrier.
Background technology
Periostracum cicadae (Isaria cicadae Miquel) belongs to the Ascomycota of mycota (FUNGI)
(Ascomycot), cup fungi subphylum (Pezizomycotina), excrement shell Gammaproteobacteria (Sordariomycetes), meat
Seat Zoopagales (Hypocreales), Cordycepses section (Cordycipitaceae), Isaria category (Isaria), are China
Famous and precious Chinese crude drug, is to colonize in a kind of Cordyceps on Cicadae (being commonly called as " cicada ").Which medicinal existing 1000
History, is one of traditional rare medicinal herbss of China, with many medical values for many years.It is main into
Dividing has adenosine, Cordyceps polysaccharide, cordycepic acid (Mannitol), cordycepin, uracil, sterol, biology
Alkali, vitamin, inorganic salt, mineral element etc..Periostracum cicadae has unique and extensive effect, such as changes
Kind renal function, antitumor adjust immune system, and strengthening by means of tonics improves cell ability, resisting fatigue,
Blood pressure lowering, blood glucose, it is antipyretic, ease pain, improve the health care of sleep, improve lipid metabolism, promote hemopoietic, resist true
Bacterium, it is anti-ageing to wait for a long time.
Periostracum cicadae spore is the organ of multiplication of Periostracum cicadae, and spore has concentrated the complete of Periostracum cicadae culture in stage of incubation
Portion's nutrition, raw Periostracum cicadae spore powder is the high nutrition product of a kind of high-quality, specially good effect therefrom.Periostracum cicadae spore
In pure powder, up to 41.6 grams of protein content accounts for nearly the 70% of its nutrient values, is typical high egg
Food and drinks one gets without pay thing.Spore powder belongs to without cholesterol diet, only up to 7.04 grams of saturated fatty acid content, and unsaturated
Content of fatty acid is its 1.86 times.Up to 34.35 grams of dietary fiber, are the 1.37 of its nutrient values simultaneously
Times.This for for preventing and treating just secrete, beneficial to fat-reducing, prevention colon and rectal cancer, promote calcium absorption,
Reduction blood fat, prevention coronary heart disease play the role of important.In spore powder, the content of vitamin D is very high,
Up to 886 micrograms, are 177.2 times of its nutrient NRV contents, vitamin D also known as calciferol,
Antirachitic vitamin, is steroid derivatives, structure containing cyclopentanoperhy drophenanthrene.The work(of vitamin D
Can maintain normal Ca,P metabolism, thus the normal development to skeleton has epochmaking effect, lacks
Ricketss not only occur reactive volt-ampere hour, hinders growth, can also have a strong impact on reproductive capability.Spore powder chats
Material element calcium, potassium, magnesium, manganese, ferrum, zinc, manganese content are higher.In spore powder, total amino acid content reaches
23.8%, it is necessary to which aminoacid accounts for total amino acidss content 33.7%, in vegetable protein be it is very high, completely
The needs of people's life can be met.And in vegetable protein common several limiting amino acids, such as
Lys (lysine), Thr (threonine), Val (L-Valine), Met (methionine) content are higher.
Experimentation shows, Cordyceps cicadae Shing spore powder ethanol extract (Cordyceps cicadae Shing spore powder) and wild Cordyceps cicadae Shing
Crude polysaccharides (crude polysaccharides) to human gastric cancer HGC27 of In vitro culture, human leukaemia K562,
PBL985/GPD, human rhabdomyosarcoma RD, human cervical carcinoma Siha, Hela, human pancreas cancer SW1990,
The propagation of 9 plants of tumor cells such as people's glioma LN229 and human ileocecal carcinoma ccl-244 cells has necessarily
Inhibitory action.
With regard to the culture of spore powder, prior art be in Periostracum cicadae solid culture, by increase illumination,
Improve cultivation temperature, reduce the methods such as culture humidity to improve the yield of spore powder on culture.Culture
Base is all the culture medium with coremium culture.Cultural method difficulty is big and yield is low, and time-consuming.
The content of the invention
It is an object of the invention to provide a kind of artificial culture method of Periostracum cicadae spore powder, the method is inhaling
Wet stock is carrier culture spore powder, and process is simple, cycle is short, sporulation quantity are high, and spore powder quality
Height, has obvious advantage relative to existing solid culture.
The purpose of the present invention is achieved by the following technical solution：
A kind of artificial culture method of Periostracum cicadae spore powder, the method comprise the steps：
Step 1, the strain seed liquor that bibulous carrier and liquid culture are obtained are fully contacted；
The bibulous carrier of contact seed liquor is first carried out light culture, is turned after mycelium is covered with by step 2
For optical culture；
Step 3, harvests to spore powder.
Further, bibulous carrier described in step 1 is the material to seed liquor with adsorption function, such as yarn
Cloth, cotton, non-woven fabrics, grey cloth, handwoven cloth etc..
Further, the contact method described in step 1 can be in seed liquor by bibulous carrier direct invasion
In, or seed liquor is spread evenly across on carrier.
Further, Testa Tritici, analysis for soybean powder, Semen Maydis powder, wheat flour etc. are added in seed liquor described in step 1
The composition of culture medium viscosity can be increased；Further, its addition is 1～3% (w/v).
Further, bibulous carrier is placed in sealed environment in step 2 incubation.
Further, bibulous carrier is placed in sealed environment during step 2 light culture, optical culture mistake
Bibulous carrier is placed in open environment in journey.
Further, bibulous carrier is placed on culturing rack in step 2 incubation, culturing rack floor height
15～18cm.
Further, step 2 light culture condition be 20～24 DEG C of temperature, relative humidity 80～90%；Light is trained
Foster condition be 23～27 DEG C of temperature, relative humidity 90～95%.
Further, step 3 adjusts relative humidity before harvesting to spore powder for 55～60%.
Further, carrier is loaded when spore powder is harvested and shake off spore in plastic bag and collect by step 3,
To prevent spore diffusion from polluting then environment.
Further, liquid culture described in step 1 includes shake-flask culture, seed tank culture, fermentation tank training
Any in supporting or several method；The culture medium of the liquid culture is normal in Periostracum cicadae incubation
Fluid medium, such as Rhizoma Solani tuber osi-sucrose-agar culture medium (PSA), Potato-dextrose-fine jade
Fat culture medium (PDA), Potato-dextrose-water culture medium (PSB), yeast extract powder-compound
Aminoacid-white sand sugar culture-medium (SAAY), yeast extract powder-white sugar-soy bean protein hydrolysate culture medium
Deng；Preferred yeast leaches powder-aminoacids complex-white sand sugar culture-medium (SAAY).
Heretofore described Periostracum cicadae (Isaria cicadae Miquel) is dispersed species, is widely distributed in me
18 provinces, city, areas on the south the state Qinling Mountains-Huaihe River.Subtropical zones and torrid zone ground on the south China the Changjiang river
Area, and the Jinsha jiang River more on low lying areas and Yunnan-Tibet plateau, Nujiang, the Lancang River and the Yarlung Zangbo River etc.
River valley area.As long as in its season of growth, the annual 6-8 months, according to the collection side of general Chinese crude drug
Method can all be adopted, and be strain known in the state of the art, it is possible to which purchase from commercial channels is obtained.This
Bright embodiment adopts Paecilomyces cicadidae(Miquel)Samson bacterial strain [Paecilomyces cicadae disclosed in CN102851353A
(Miq.) Samson], on November 18th, 2009 in Chinese microorganism strain preservation conservator
Meeting common micro-organisms center (abbreviation CGMCC) registration preservation, preserving number is CGMCC No.3453.
Beneficial effects of the present invention：
1st, process is simple, energy-conservation, section material.With regard to the culture of spore powder, prior art adopts coremium
The solid medium of culture, improves spore powder on coremium by increasing illumination and raising cultivation temperature
Yield, the method needs to prepare solid medium, and needs to increase illumination and cultivation temperature, technique
Complexity, finished product are high, and difficulty is big.Bibulous carrier is directly contacted by the inventive method with seed liquor, need not
Solid medium is prepared, illumination and temperature need not be increased, process is simple greatly reduces energy consumption.
2nd, cultivation cycle is short.Prior art passes through solid culture method, 20 days after optical culture is switched to
Spore could be produced, and only 3～5 days spores can just be covered with moisture absorption load after the inventive method switchs to optical culture
Body, substantially reduces cultivation cycle.
3rd, sporulation quantity is high, and spore powder quality is good.Within unit interval and space, the product of the inventive method
Spore amount is apparently higher than existing solid culture method, and the inventive method does not affect spore powder quality, and which has
Effective component content is suitable with the spore powder of solid culture.
1 the inventive method of experimental example is compared with Conventional solid cultural method
The seed liquor that 1 liquid culture of Example is obtained, respectively by following three kinds of methods culture spore powder,
The spore powder quality that relatively sporulation quantity of three kinds of methods, cultivation cycle and three kinds of methods are obtained.
Method one：The seed liquor that 1 liquid culture of Example is obtained, adds 3% analysis for soybean powder, by enforcement
The method of example 8 is cultivated；
Method two：The seed liquor that 1 liquid culture of Example is obtained, adds 3% Testa Tritici liquor, by reality
The method for applying example 11 is cultivated；
Method three：The seed liquor that 1 liquid culture of Example is obtained, is entered by the method for comparative examples
Row culture.
The results are shown in Table 1.
1 two kinds of cultural method comparative results of table
| Cultural method | Method one | Method two | Method three |
| Sporulation quantity (g/m3)/20d | 448 | 301 | 238 |
| The solid culture cycle (my god) | 5 | 5 | 20 |
| Protein (g) | 45.33 | 43.20 | 41.6 |
| Dietary fiber (g) | 36.00 | 34.57 | 34.35 |
| Vitamin D (μ g) | 890 | 870 | 886 |
| Total amino acid content (%) | 24.0 | 23.9 | 23.8 |
Method one, two adopts gauze culture for the present invention, and method three is to be trained using solid medium
Support, by comparing as can be seen that in the case of strain source and condition of culture all same：
1st, from terms of cultivation cycle, method one, two is only 5 days, and method three is 20 days, the present invention
Method substantially shortens relative to existing solid culture method cultivation cycle；
2nd, from terms of sporulation quantity, in unit interval (20 days) and unit space (m3) in, method one,
Two are respectively 448g and 301g, and method three is 238g, and the inventive method trained apparently higher than existing solid
Foster method；
3rd, from terms of spore powder quality, the spore powder that distinct methods culture is obtained its protein, dietary fiber,
Vitamin D and amino acid content without significant difference, illustrate the spore powder quality of the inventive method culture
It is suitable with existing solid culture method.
In sum, present invention process is simple, cycle is short, sporulation quantity are high, and spore powder quality is high,
There is obvious advantage relative to existing solid culture.
Description of the drawings
Fig. 1 bibulous carriers
Fig. 2 culture nets
Fig. 3 light cultures under culturing rack upper sealing ring border
Fig. 4 optical cultures under open environment on culturing rack
Specific embodiment
In following instance, 25% bromo geramine of culturing room's ground spray, ozonization 30min.Experiment material
All sterilize, including aseptic clothes, gauze, dovetail clip, stainless steel cask, plastic bag, culture net etc..
1 liquid culture of embodiment
Culture medium prescription：Yeast powder leaches powder 0.5%, and aminoacids complex 1%, white sugar 3.5% are remaining
Moisturizing is measured to 100%；
Cultural method：Shake-flask culture；Condition of culture be 22 DEG C, 150 revs/min shaking tables of temperature in cultivate 72h；
It is in aubergine whne culture fluid or stops culture when there is the granular little mycelium pellet of Semen setariae, obtains seed liquor.
2 liquid culture of embodiment
With cultural method with embodiment 1, difference is to add in the seed liquor that culture is obtained to culture medium prescription
3% Testa Tritici liquor.
3 liquid culture of embodiment
With cultural method with embodiment 1, difference is to add in the seed liquor that culture is obtained to culture medium prescription
3% analysis for soybean powder.
4 liquid culture of embodiment
With cultural method with embodiment 1, difference is to add in the seed liquor that culture is obtained to culture medium prescription
2% Testa Tritici liquor.
5 liquid culture of embodiment
With cultural method with embodiment 1, difference is to add in the seed liquor that culture is obtained to culture medium prescription
1% Testa Tritici liquor, surplus moisturizing to 100%；
Cultural method：With embodiment 1.
6 liquid culture of embodiment
With cultural method with embodiment 1, difference is to add in the seed liquor that culture is obtained to culture medium prescription
1% analysis for soybean powder, surplus moisturizing to 100%；
Cultural method：With embodiment 1.
7 liquid culture of embodiment
Culture medium prescription：Yeast extract powder 3%, white sugar 3%, 0.5% surplus of soy bean protein hydrolysate are mended
Water is to 100%；
Cultural method：With embodiment 1,3% wheat flour is added in the seed liquor that culture is obtained.
8 spore powder cultural method of embodiment
Step 1, pours embodiment 1～7 arbitrary seed liquor into stainless steel cask, with three layers of gauze carrier
(such as Fig. 1) fully infiltrates, and slightly drains, and is placed in culture online (see Fig. 2)；
Step 2：Light culture：Culture net is positioned in plastic bag, dovetail clamp opening is used, is then put
On the culturing rack that floor height is 15cm or so (height per 1m can put 6 layers), enter in darkroom
Row light culture (see Fig. 3), 22 DEG C of cultivation temperature, humidity 85%；When mycelia is covered with gauze carrier
Terminate light culture (3-4 days)；
Optical culture：Culture net is removed from plastic bag, is placed directly within culturing rack that (i.e. carrier is upper and lower
Two sides and air contact), switch to optical culture (see Fig. 4), 25 DEG C of cultivation temperature, humidity 95%；
Terminate to cultivate (3-5 days) when spore is covered with culture.
Step 3, harvesting：Optical culture humidity 95% is adjusted to into 60% in harvesting the previous day, to allow which certainly
So it is dried；Spore powder is harvested with small brushes under hundred grades of laminar flow hoods.
9 spore powder cultural method of embodiment
Step 1,3 with embodiment 8, differ only in step 2：
Step 2：Light culture：Culture net is positioned in plastic bag, dovetail clamp opening is used, is placed in dark
Light culture, 22 DEG C of cultivation temperature, humidity 85% are carried out in room；Terminate when mycelia is covered with gauze carrier
Light culture (3-4 days)；
Optical culture：Will culture net removes from plastic bag and is placed in culture plate (i.e. only carrier upper surface and
Air contact), switch to optical culture, 25 DEG C of cultivation temperature, humidity 95%；Treat that spore is covered with culture
When terminate cultivate (3-5 days).
10 spore powder cultural method of embodiment
Step 1,3 with embodiment 8, differ only in step 2：
Step 2, light culture：Culture net is positioned in plastic bag, dovetail clamp opening is used, is placed in dark
Light culture, 22 DEG C of cultivation temperature, humidity 85% are carried out in room；Terminate when mycelia is covered with gauze carrier
Light culture (3-4 days)；
Optical culture：Switch to optical culture, 25 DEG C of cultivation temperature, humidity 95%；Treat that spore is covered with culture
When terminate cultivate (3-5 days).
11 spore powder cultural method of embodiment
Step 1,3 with embodiment 8, differ only in step 2：
Step 2, light culture：Culture net is placed in darkroom carries out light culture, 22 DEG C of cultivation temperature,
Humidity 85%；Terminate light culture (3-4 days) when mycelia is covered with gauze carrier；
Optical culture：Switch to optical culture, 25 DEG C of cultivation temperature, humidity 95%；Treat that spore is covered with culture
When terminate cultivate (3-5 days).
12 spore powder cultural method of embodiment
Step 1～3 are differed only in and for gauze to replace with cotton with embodiment 8.
13 spore powder cultural method of embodiment
Step 1～3 are differed only in and for gauze to replace with non-woven fabrics with embodiment 8.
14 spore powder cultural method of embodiment
Step 1～3 differ only in step 1 and seed liquor are coated on gauze with embodiment 8.
Comparative examples spore powder cultural method
Fluid medium and cultural method are with embodiment 1.Solid medium is Semen Tritici aestivi, according to different size
Container carries out subpackage (happy button box fills 0.33 kilogram, and square box fills 0.5 kilogram), and amount of water is 1：1.2,121 DEG C
Sterilizing 30 minutes, natural cooling.Inoculation (accessing the seed liquor of embodiment 1) after cooling, inoculum concentration is
10%, at 25 DEG C, optical culture 20 days under conditions of humidity 95% or so.It is dried after spore maturation and receives
Obtain.
The sporulation quantity of different cultural methods
| Liquid cultivating method | Spore powder cultural method | Sporulation quantity g/m3 |
| Embodiment 1 | Comparative examples | 238 |
| Embodiment 2 | Comparative examples | 217 |
| Embodiment 1 | Embodiment 11 | 245 |
| Embodiment 2 | Embodiment 11 | 301 |
| Embodiment 3 | Embodiment 11 | 302 |
| Embodiment 3 | Embodiment 10 | 326 |
| Embodiment 3 | Embodiment 9 | 374 |
| Embodiment 3 | Embodiment 8 | 448 |
| Embodiment 3 | Embodiment 12 | 439 |
| Embodiment 3 | Embodiment 13 | 447 |
| Embodiment 3 | Embodiment 14 | 464 |
| Embodiment 7 | Embodiment 14 | 425 |
Claims (10)
1. a kind of artificial culture method of Periostracum cicadae spore powder, it is characterised in that methods described includes as follows
Step：
Step 1, the strain seed liquor that bibulous carrier and liquid culture are obtained are fully contacted；
The bibulous carrier of contact seed liquor is first carried out light culture, is turned after mycelium is covered with by step 2
For optical culture；With
Step 3, harvests to spore powder.
2. the method for claim 1, it is characterised in that bibulous carrier described in step 1 is yarn
Any one in cloth, cotton, non-woven fabrics, grey cloth, handwoven cloth.
3. the method for claim 1, it is characterised in that the contact method described in step 1 is
By bibulous carrier direct invasion in seed liquor, or seed liquor is spread evenly across on carrier.
4. the method for claim 1, it is characterised in that add in seed liquor described in step 1
The composition of culture medium viscosity can be increased.
5. method as claimed in claim 4, it is characterised in that described to increase culture medium viscosity
Composition be Testa Tritici, analysis for soybean powder, Semen Maydis powder, any one or a few in wheat flour.
6. method as claimed in claim 5, it is characterised in that described to increase culture medium viscosity
Composition addition be 1～3%.
7. the method for claim 1, it is characterised in that by moisture absorption in step 2 incubation
Carrier is placed in sealed environment.
8. the method for claim 1, it is characterised in that will inhale during step 2 light culture
Wet carrier is placed in sealed environment, bibulous carrier is placed in open environment during optical culture.
9. the method for claim 1, it is characterised in that step 2 light culture condition is temperature
20～24 DEG C, relative humidity 80～90%；Optical culture condition be 23～27 DEG C of temperature, relative humidity 90～95%.
10. the method for claim 1, it is characterised in that step 3 is being carried out to spore powder
Before harvesting, adjustment relative humidity is 55～60%.
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| CN107699539A (en) * | 2017-10-18 | 2018-02-16 | 湖州新驰医药科技有限公司 | A kind of collection method of Periostracum cicadae conidia powder |
| CN112715276A (en) * | 2021-01-08 | 2021-04-30 | 浙江泛亚生物医药股份有限公司 | Culture method of cordyceps sobolifera spore powder |
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| CN104286032A (en) * | 2014-09-04 | 2015-01-21 | 青岛农业大学 | Preparation method of Talaromyces flavus spore powder, Talaromyces flavus wettable pulvis and preparation method of wettable pulvis |
| CN104855689A (en) * | 2015-05-28 | 2015-08-26 | 河南双成生物科技有限公司 | Method for preparing fatty acid-rich protein feed raw material from amino acid fermentation residual liquor |
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| CN112715276A (en) * | 2021-01-08 | 2021-04-30 | 浙江泛亚生物医药股份有限公司 | Culture method of cordyceps sobolifera spore powder |
| CN112715276B (en) * | 2021-01-08 | 2024-03-12 | 浙江泛亚生物医药股份有限公司 | Culture method of cordyceps sobolifera spore powder |
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