CN105087401A - Preparation method of artificial ophiocordyceps sobolifera - Google Patents
Preparation method of artificial ophiocordyceps sobolifera Download PDFInfo
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- CN105087401A CN105087401A CN201510595005.8A CN201510595005A CN105087401A CN 105087401 A CN105087401 A CN 105087401A CN 201510595005 A CN201510595005 A CN 201510595005A CN 105087401 A CN105087401 A CN 105087401A
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Abstract
The invention discloses a culture method of artificial ophiocordyceps sobolifera. The method comprises the following steps of first-stage culture: sterilizing a first-stage culture medium, performing slant inoculation, smashing hyphae, collecting a fungus liquid into another conical flask containing a culture medium, and performing shaking culture by a shaker for six days; second-stage culture (using a cow milk culture medium): performing sterilizing and cooling, adding seed fermentation broth, carrying out shaking to uniform, and performing shaking culture by a shaker for six days; third-stage enlarged culture (using a culture medium component of pure cow milk containing glucose): performing culture fermentation, conducting decompressed suction filtration to separate ophiocordyceps sobolifera fermentation culture liquid from a mycelium mixture, drying fermentation filtrate by a spray dryer to obtain dry filtrate powder, drying mycelium filter cakes by a dryer, grinding the cakes, and sieving the ground cakes through an 80-mesh sieve, thereby obtaining dry mycelium powder.
Description
Technical background
Cicada fungus is a kind of traditional Chinese medicine, and pharmacological research shows that cicada fungus has analgesia, calmness, antipyretic, the antitumor and anti-ageing effect of waiting for a long time.Natural cicada fungus is as a kind of traditional Chinese medicine material, and output is very rare, and quality is difficult to control.From cicada fungus separation and purification go out Paecilomyces cicadae bacterial strain carry out cicada fungus artificial culture can the application of Some Drugs and field of health care products.Due to active substance a large amount of in cicada fungus artificial culture process by strain secretes in substratum, therefore time by artificial cicada fungus application and health care of food product field, need the separation and Extraction of carrying out active substance in substratum, thus significantly improve production cost, or only use thalline and waste the substratum being rich in active substance.
In addition, the myriocin that artificial cicada fungus bacterial strain produces is important activity of fighting against senium material.It can strengthen the mitochondrial function of cell, promotes cellular respiration, promotes the generation of Antioxidative Factors in cell, reduces the oxidative damage of cell.But there are some researches show, the myriocin concentration that artificial cicada fungus produces is lower, when not having better means to improve myriocin concentration, needs to take certain methods to strengthen the drug effect of myriocin to reach health-care effect.
Summary of the invention
The object of this invention is to provide the preparation method of a kind of artificial cicada fungus, comprise the effective constituent adopting the method for third stage culture to obtain artificial cicada fungus thalline, spraying dry is carried out to the culture medium filtrate produced in fermenting process simultaneously, the effective constituent of the artificial cicada fungus of final acquisition, for fields such as anti-ageing and skin cares.Described method effectively can obtain cicada fungus microbial activity composition, is used simultaneously, significantly improves production efficiency to the activeconstituents in substratum.
Technical scheme of the present invention is: the cultural method of a kind of artificial cicada fungus, and culturing process is divided into three grades:
Step one: the cultivation of first step seed liquor.Triangular flask is adopted to carry out the cultivation of liquid shaking table.Use dextrose peptone medium, each 500ml triangular flask is loaded 150ml substratum, seals bottleneck with tampon, (121 DEG C) sterilizing 25 minutes under 0.1Mpa.Taking-up is cooled to room temperature, and Bechtop is inoculated.First choose appropriate slant strains in the triangular flask that granulated glass sphere and substratum are housed with inoculating needle, mycelia is smashed.Then bacterium liquid being accessed other is equipped with in the triangular flask of substratum, and inoculum size is every bottle of 10ml ~ 20ml.(27 DEG C) shaking culture on shaking table, oscillation frequency 180 revs/min.Shaking culture can stop cultivating for 6 days, proceeds to the second stage and cultivates.
Step 2: secondary is cultivated.The second stage is cultivated and is used cow's milk substratum, is cooled to 27 DEG C, adds the seed liquor of 0.5% (v/v), shake up after sterilizing.(27 DEG C) shaking culture on shaking table, oscillation frequency 180 beats/min.Shaking culture stops cultivating for 6 days, proceeds to third stage enlarged culturing.
Step 3: the third stage is cultivated and uses containing glucose cow's milk substratum, is cooled to 27 DEG C, adds the secondary culture of 1% (v/v), shake up after medium sterilization.Fermentation culture stops cultivating for 9 days.
Step 4: be separated as follows with mycelium mixture by cicada fungus fermentation culture after third stage cultivation and fermentation terminates: decompress filter, collects ferment filtrate and mycelium filter cake.Ferment filtrate carries out drying by spray-drier and obtains filtrate dry powder, and mycelium filter cake carries out drying by dryer, and then pulverizes 80 orders, obtains mycelium dry powder.
Further, step one dextrose peptone medium composition used is (by 1L): glucose 30g, peptone 5g, yeast powder 3g, KH
2pO
40.3g, K
2hPO
43H
2o0.92g, MgSO
47H
2o0.61g, pH5.5.
Further, the cow's milk medium component that step 2 is used is (by 1L): cow's milk 850ml (total milk solid 12%, fat 3%), glucose 15g, KH2PO40.1g, K2HPO43H2O0.3g, MgSO47H2O0.5g, pH5.5.
Further, described step 3 is the pure cow's milk (total milk solid 12%, fat 3%) containing 15g/L glucose containing glucose cow's milk medium component.
The invention has the beneficial effects as follows, the method for the artificial cicada fungus of preparation of the present invention effectively can obtain cicada fungus microbial activity composition, is used simultaneously, significantly improves production efficiency to the activeconstituents in substratum.In addition, using cow's milk as second and the main medium component of third stage fermentation culture, not only improve the mouthfeel of artificial cicada fungus goods, too increase nutritive value.
Embodiment
Being easier to make goal of the invention of the present invention, technique means, technique effect etc. understand, setting forth the present invention further below in conjunction with specific embodiment
A cultural method for artificial cicada fungus, culturing process is divided into three grades:
Step one: the cultivation of first step seed liquor.Triangular flask is adopted to carry out the cultivation of liquid shaking table.Use dextrose peptone medium, medium component is (by 1L): glucose 30g, peptone 5g, yeast powder 3g, KH
2pO
40.3g, K
2hPO
43H
2o0.92g, MgSO
47H
2o0.61g, pH5.5.Each 500ml triangular flask is loaded 150ml substratum, seals bottleneck with tampon, (121 DEG C) sterilizing 25 minutes under 0.1Mpa.Taking-up is cooled to room temperature, and Bechtop is inoculated.First choose appropriate slant strains in the triangular flask that granulated glass sphere and substratum are housed with inoculating needle, mycelia is smashed.Then bacterium liquid being accessed other is equipped with in the triangular flask of substratum, and inoculum size is every bottle of 10ml ~ 20ml.(27 DEG C) shaking culture on shaking table, oscillation frequency 180 revs/min.Shaking culture can stop cultivating for 6 days, proceeds to the second stage and cultivates.
Step 2: secondary is cultivated.The second stage is cultivated and is used cow's milk substratum, and medium component is (by 1L): cow's milk 850ml (total milk solid 12%, fat 3%), glucose 15g, KH2PO40.1g, K2HPO43H2O0.3g, MgSO47H2O0.5g, pH5.5.Be cooled to 27 DEG C after sterilizing, add the seed liquor of 0.5% (v/v), shake up.(27 DEG C) shaking culture on shaking table, oscillation frequency 180 beats/min.Shaking culture stops cultivating for 6 days, proceeds to third stage enlarged culturing.
Step 3: the third stage is cultivated and uses containing glucose cow's milk substratum, medium component is the pure cow's milk (total milk solid 12%, fat 3%) containing 15g/L glucose.Be cooled to 27 DEG C after medium sterilization, add the secondary culture of 1% (v/v), shake up.Fermentation culture stops cultivating for 9 days.
Step 4: be separated as follows with mycelium mixture by cicada fungus fermentation culture after third stage cultivation and fermentation terminates: decompress filter, collects ferment filtrate and mycelium filter cake.Ferment filtrate carries out drying by spray-drier and obtains filtrate dry powder, and mycelium filter cake carries out drying by dryer, and then pulverizes 80 orders, obtains mycelium dry powder.
Described mycelium dry powder and filtrate dry powder can be used for the anti-oxidant anti-aging composition of preparation cicada fungus etc.
Claims (5)
1. a cultural method for artificial cicada fungus, comprises third stage culture process:
Step one: the cultivation of first step seed liquor.Triangular flask is adopted to carry out the cultivation of liquid shaking table.Use dextrose peptone medium, each 500ml triangular flask is loaded 150ml substratum, seals bottleneck with tampon, (121 DEG C) sterilizing 25 minutes under 0.1Mpa.Taking-up is cooled to room temperature, and Bechtop is inoculated.First choose appropriate slant strains in the triangular flask that granulated glass sphere and substratum are housed with inoculating needle, mycelia is smashed.Then bacterium liquid being accessed other is equipped with in the triangular flask of substratum, and inoculum size is every bottle of 10ml ~ 20ml.(27 DEG C) shaking culture on shaking table, oscillation frequency 180 revs/min.Shaking culture can stop cultivating for 6 days, proceeds to the second stage and cultivates.
Step 2: secondary is cultivated.The second stage is cultivated and is used cow's milk substratum, is cooled to 27 DEG C, adds the seed liquor of 0.5% (v/v), shake up after sterilizing.(27 DEG C) shaking culture on shaking table, oscillation frequency 180 beats/min.Shaking culture stops cultivating for 6 days, proceeds to third stage enlarged culturing.
Step 3: the third stage is cultivated and uses containing glucose cow's milk substratum, is cooled to 27 DEG C, adds the secondary culture of 1% (v/v), shake up after medium sterilization.Fermentation culture stops cultivating for 9 days.
2. the artificial culture method of cicada fungus according to claim 1, also comprises step 4: be separated as follows with mycelium mixture by cicada fungus fermentation culture after third stage cultivation and fermentation terminates: decompress filter, collects ferment filtrate and mycelium filter cake.Ferment filtrate carries out drying by spray-drier and obtains filtrate dry powder, and mycelium filter cake carries out drying by dryer, and then pulverizes 80 orders, obtains mycelium dry powder.
3. the method for claim 1-2, is characterized in that, the dextrose peptone medium composition in described step one is (by 1L): glucose 30g, peptone 5g, yeast powder 3g, KH
2pO
40.3g, K
2hPO
43H
2o0.92g, MgSO
47H
2o0.61g, pH5.5.
4. the method for claim 1-3, is characterized in that, described step 2 cow's milk medium component used is (by 1L): cow's milk 850ml (total milk solid 12%, fat 3%), glucose 15g, KH2PO40.1g, K2HPO43H2O0.3g, MgSO47H2O0.5g, pH5.5.
5. the method for claim 1-4, is characterized in that, what described step 3 was used is the pure cow's milk (total milk solid 12%, fat 3%) containing 15g/L glucose containing glucose cow's milk medium component.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543104A (en) * | 2016-01-05 | 2016-05-04 | 江苏农林职业技术学院 | Optimization method of solid culture medium for artificial acclimation and cultivation of wild Isaria cicadae Miquel |
| WO2017202293A1 (en) * | 2016-05-23 | 2017-11-30 | 浙江泛亚生物医药股份有限公司 | Artificial cultivation method for cordyceps sobolifera |
| CN108782022A (en) * | 2018-07-03 | 2018-11-13 | 淮北师范大学 | A method of it is primary raw material culture cicada fungus conidia powder to process waste residue using pomegranate |
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2015
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Patent Citations (3)
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543104A (en) * | 2016-01-05 | 2016-05-04 | 江苏农林职业技术学院 | Optimization method of solid culture medium for artificial acclimation and cultivation of wild Isaria cicadae Miquel |
| WO2017202293A1 (en) * | 2016-05-23 | 2017-11-30 | 浙江泛亚生物医药股份有限公司 | Artificial cultivation method for cordyceps sobolifera |
| CN108782022A (en) * | 2018-07-03 | 2018-11-13 | 淮北师范大学 | A method of it is primary raw material culture cicada fungus conidia powder to process waste residue using pomegranate |
| CN108782022B (en) * | 2018-07-03 | 2020-10-20 | 淮北师范大学 | Method for culturing cordyceps sobolifera spore powder by using pomegranate processing waste residues as main raw materials |
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