CN101407767A - Method for producing Chinese caterpillar fungus by fermentation - Google Patents
Method for producing Chinese caterpillar fungus by fermentation Download PDFInfo
- Publication number
- CN101407767A CN101407767A CNA2008102288465A CN200810228846A CN101407767A CN 101407767 A CN101407767 A CN 101407767A CN A2008102288465 A CNA2008102288465 A CN A2008102288465A CN 200810228846 A CN200810228846 A CN 200810228846A CN 101407767 A CN101407767 A CN 101407767A
- Authority
- CN
- China
- Prior art keywords
- seeds
- cultivated
- hours
- cultivation
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of medicine and provides a method for producing aweto by fermentation. The method adopts a MK722 culture medium, places an Erlenmeyer flask containing liquid seeds onto a rotary shaking table, and comprises the liquid seed cultivation that is characterized in that the proportion of the seeds to the culture medium is between 1 to 5 and 1 to 20, the rotation speed is 50 to 300 turns per minute, the cultivation time is 50 to 200 hours, the cultivation temperature is 15 to 30 DEG C, the lighting intensity is 0 to 58.4 Mu mol question mark m<-2> question mark s<-1>, and the lighting time lasts for 1 to 24 hours. After the liquid seeds are cultivated for 50 to 200 hours, the seeds with limpid color and luster, uniform mycelia and the diameter of about 1 to 3 mm in the seed liquid are selected for cultivation, and the selected seeds are inoculated into a bioreactor with the capacity of 0.2 to 5000 L according to the inoculation amount that is provided with the proportion of the liquid seeds to the culture medium between 1 to 1 and 1 to 20, and cultivated under aseptic condition. The cultivation temperature is 15 to 30 DEG C, the cultivation time is 50 to 96 hours, the rotation speed is 50 to 300 turns per minute, the lighting time lasts for 1 to 24 hours, and the lighting intensity is 0 to 58.4 Mu mol question mark m<-2> question mark s<-1>. The cultivated seeds are further freezing dried so as to form the frozen dry powder of fermented aweto. The method has easy operation and low production cost, does not pollute environment, and achieves the industrialization level.
Description
Technical field
The invention belongs to medical technical field, relate to a kind of method by fermentative production Cordyceps sinensis.Be means specifically with the microbial fermentation technology, cultivate and culture is carried out freeze dried method by the aweto in large scale that in the reciprocating type incubator of large-scale constant temperature illumination, carries out.
Background technology
Cordyceps sinensis is that Clavicipitaceae (Clavicepitaceae) fungi Cordyceps fungus (Cordycepssinensis (Berk.) Sacc.) colonizes in stroma and the larva cadaveric complex on Hepialidae insect bat moth (Hepialus armoricanus) larva.Have and invigorate the lung and the kidney, effects such as hemostasis and phlegm.Modern pharmacology shows that Cordyceps sinensis also has anticancer, antiviral, the anti-ageing effect of waiting for a long time.Contain multiple active substance with medicinal health care function in the Cordyceps bacterium, wherein adenosine (adenosine) and cordycepin (cordycepin) are one of main active substances.Cordyceps sinensis just extensively is used as invigorant among the people because of its pharmacologically active widely from ancient times, but strict and parasitic conditions is special because of its growing environment, adds the people for excavating transition, and the natural resource rareness costs an arm and a leg.Mainly concentrate on the determining of asexual shape, artificial culture, liquid submerged fermentation cultivation, chemical ingredients and Pharmacological action study about the research of Cordyceps sinensis in recent years.Through studies show that in a large number the artificial fermentation cultivates the mycelium that obtains, through researchs such as toxicity, pharmacology and plants, proof and natural cs chemical constitution, pharmacological action basically identical, and the content of some compositions is higher than natural cordyceps, and therefore people are devoted to submerged fermentation cultivation Cordyceps mycelium product as an alternative always for a long time.Adenosine and cordycepin are the main active substances of Cordyceps sinensis.Adenosine has the blood circulation of the heart and brain of improvement, prevents irregular pulse, suppresses neurotransmitter and discharges and regulate pharmacological actions such as adenylate cyclase activity, at present as the quality control index of Cordyceps sinensis.Cordycepin promptly 3 '-Desoxyadenosine, have immunomodulatory, antibiotic, antiviral, the anticancer and anti-ageing effect of waiting for a long time.
The research of at present domestic liquid culture about Cordyceps sinensis also is in the investigation of the various physical and chemical factors that carry out at different meta-bolitess basically, some scholars were applied to the liquid culture of Cordyceps sinensis with the research method of bio-transformation in recent years, but about investigating precursor substance the influence for secondary metabolite in the Cordyceps sinensis fermentation culture were also rarely had report.
The Cordyceps mycelium preparation that domestic and international Cordyceps sinensis class healthcare products that gone on the market and medicine are natural extract.Because it has very high pharmaceutical use, liked by the patient.Yet because its complicated component, active constituent content is low, has influenced the clinical efficacy of this medicine and the further expansion in market greatly.Particularly natural cordyceps has been subjected to the focused protection of the world and the Chinese government as endangered plant species, forbids to pluck.Each country all drops into the substitute products of lot of manpower and material resources research Cordyceps sinensis for this reason.The substitute products of generally acknowledging in the world are artificial culture Cordyceps sinensis or cordyceps militaris mycelium at present, and the high bacterial classification of screening active constituent content carries out artificial culture, and the mycelium that obtains carries out some conventional processing and makes medicinal extract, capsule or oral liquid.This still belongs to the traditional industry processing mode of Chinese medicine, and it is about 3 months that the production cycle is about.
Summary of the invention
The purpose of this invention is to provide a kind of method by fermentative production Cordyceps sinensis.
The present invention is achieved through the following technical solutions:
The present invention includes liquid seeds cultivation, large scale fermentation cultivation, freeze-drying.
The present invention is according to Cordyceps mycelium fermentation culture production technology, on the MK722 substratum, cultivate, the growth of bacterium ball and the output of effective constituent have been controlled well, the ratio 1 of seed and substratum: 5-20 during liquid seeds is cultivated, rotating speed: 50-300 rev/min, incubation time: 50-200 hour, culture temperature: 15-30 ℃, intensity of illumination: 0-58.4 μ molm
-2S
-1, light application time: 1-24 hour.
Liquid seeds and substratum ratio 1 during large scale fermentation is cultivated: the 1-20 inoculum size is connected in the 0.2-5000L bio-reactor culture temperature: 15-30 ℃, incubation time: 50-96 hour, rotating speed: 50-300 rev/min, light application time: 1-24 hour, intensity of illumination: 0-58.4 μ molm
-2S
-1
Freeze-drying process is sugared acid anhydride class 0.1-5mg/ml of adding and sulfur-containing amino acid 0.0001-12g/100g in fermented liquid ,-20--70 ℃ following freeze-drying, and then form the Cordyceps fermented product lyophilized powder.
Do not see that from domestic and international document of having delivered and patent utilization MK722 substratum and above-mentioned culture condition carry out the report that the Chinese caterpillar fungus technology is cultivated in submerged fermentation.
The culture process of Cordyceps sinensis is as follows:
One, liquid seeds is cultivated
1.MK722 substratum is formed:
Potato powder 1-4.75%, Semen Maydis powder 0.1-1.5%, peptone 0.1-0.5%, KH
2PO
40.1-0.5%, MgSO
40.1-0.5%, vitamins B
10.001-0.005%, vitamins B
20.001-0.005%, (NH
4)
2SO
40.01-0.10%, purine class 0.05-1%, amides 0.01-1%, all the other are sterile distilled water, the pH value is 5.0-7.0.
2. culture condition:
The Erlenmeyer flask that liquid seeds is housed is put on the rotary shaking table ratio 1 of seed and nutrient solution: 5-20, rotating speed: 50-300 rev/min, incubation time: 50-200 hour.Culture temperature: 15-30 ℃, intensity of illumination: 0-58.4 μ molm
-2S
-1, light application time: 1-24 hour.
Two, fermentation culture
Liquid seeds was cultivated after 50-200 hour, to cultivate when choosing limpid in sight, the mycelium uniformity, the about 1-3mm size of seed diameter in the seed liquor, in liquid seeds and substratum ratio 1: the 1-20 inoculum size, be connected in the 0.2-5000L bio-reactor, under aseptic condition, cultivate.
1. fermention medium is formed:
The same seed liquor substratum is formed.
2. culture condition:
Culture temperature: 15-30 ℃, incubation time: 50-96 hour, rotating speed: 50-300 rev/min, light application time: 1-24 hour, intensity of illumination: 0-58.4 μ molm
-2S
-1
3. check: mycelium uniformity, cordycepin content 0.05-2.6mg/g, adenosine content 0.001-2.0mg/g.
Three, freeze-drying
In fermented liquid, add sugared acid anhydride class 0.1-5mg/ml and sulfur-containing amino acid 0.0001-12g/100g ,-20--70 ℃ following freeze-drying, form Cordyceps sinensis fermentation lyophilized powder.
Four, detect
1. the selection of chromatographic condition:
Chromatographic condition: pump: SHIMADZU LC-10Avp; Detector: SHIMADZUSPD-10Avp; Chromatographic column: DIAMONSIL C-18 alkyl silica gel post (250 * 4.6); 5 μ l quantitatively encircle; Moving phase: acetonitrile: water: methyl alcohol (2: 85: 2); Measure wavelength: 260nm; Flow velocity: 1.0ml/min; Column temperature: 40 ℃.
2. the mensuration of sample:
Precision claims to conclude a contract or treaty the 0.5g lyophilized powder, puts in the ground Erlenmeyer flask, adds 20ml 90% methyl alcohol, refluxing extraction 30 minutes, suction filtration, residue is with 90% methanol wash 2 times, concentrate on the filtrate water bath, 90% methanol constant volume in the 5ml volumetric flask, 0.45 μ m filtering with microporous membrane.The gained trial-product reads peak area respectively at 0h, 2h, 4h, 7h sample introduction, calculates RSD.By substitution typical curve calculation sample cordycepin and adenosine content.
Produce Cordyceps sinensis with the inventive method fermentation culture technology, unchangeability, cost is low, and the cycle is short, the Chinese caterpillar fungus freeze-dried powder of formation, adenosine and cordycepin output height, active good, clinical effectiveness is obvious, has the market competitiveness.
Embodiment:
Embodiment 1: the preparation of Cordyceps sinensis
One, liquid seeds is cultivated
1.MK722 substratum is formed:
Potato powder 2%, Semen Maydis powder 0.8%, peptone 0.4%, KH
2PO
40.3%, MgSO
40.4%, vitamins B
10.003%, vitamins B
20.004%, (NH
4)
2SO
40.05%, purine class 0.65%, amides 0.2%, all the other are sterile distilled water, the pH value is 6.5.
2. culture condition:
The Erlenmeyer flask that liquid seeds is housed is put on the rotary shaking table, and the ratio of seed and nutrient solution 1: 20, was cultivated 100 hours by 100 rev/mins.Culture temperature: 20 ℃, intensity of illumination: 40.7 μ molm
-2S
-1Light application time: 4 hours.
Two, fermentation culture
Liquid seeds was cultivated after 100 hours, to cultivate when choosing limpid in sight, the mycelium uniformity, the about 1mm size of the seed diameter in the seed liquor is pressed 15% inoculum size, is connected in the 0.5L bio-reactor, cultivates under aseptic condition.
1. fermention medium is formed:
The same seed liquor substratum is formed.
2. culture condition:
Culture temperature: 20 ℃, cultivated 85 hours, 150 rev/mins of rotating speeds, light application time: 4 hours, intensity of illumination: 45.5 μ molm
-2S
-1
3. check:
The mycelium uniformity, cordycepin content 1.1mg/g, adenosine content 0.04mg/g.
Three, freeze-drying
In fermented liquid, add vehicle sugar acid anhydride class 3mg/ml and sulfur-containing amino acid 2g/100g,, form Cordyceps sinensis fermentation lyophilized powder-45 ℃ of following freeze-drying.
Embodiment 2: different purine are to the influence of adenosine and cordycepin output in the Cordyceps sinensis fermentation
1, xanthine;
| Xanthine | Cordycepin content (mg/g) | Adenosine content (mg/g) |
| (0.5％) | 1.72 | 0.05 |
2, VITAMIN B4
3, Leucon
4, guanine
Embodiment 3: different acid amides are to the influence of adenosine and cordycepin output in the Cordyceps sinensis fermentation
1, l-asparagine
2, glutamine
3, niacinamide
4, o-dime thyl phosphoroamidothioate
Embodiment 4: the comparison of adenosine and cordycepin output in 3 batches of samples of wild cordyceps and Cordyceps fermented product
Claims (5)
1. method by fermentative production Cordyceps sinensis, it is characterized in that: this method comprises that liquid seeds is cultivated, large scale fermentation is cultivated, freeze-drying.
2. method according to claim 1 is characterized in that: the ratio 1 of seed and substratum: 5-20 during liquid seeds is cultivated, and rotating speed 50-300 rev/min, cultivated 50-200 hour, culture temperature: 15-30 ℃, intensity of illumination: 0-58.4 μ molm
-2S
-1, light application time: 1-24 hour.
3. method according to claim 2 is characterized in that: described substratum is the MK722 substratum.
4. method according to claim 1, it is characterized in that: liquid seeds and substratum ratio 1 during described large scale fermentation is cultivated: 1-20 inoculum size, be connected in the 0.2-5000L bio-reactor, culture temperature: 15-30 ℃, cultivated 50-96 hour, rotating speed: 50-300 rev/min, light application time: 1-24 hour, intensity of illumination: 0-58.4 μ molm
-2S
-1
5. method according to claim 1 is characterized in that: freeze-drying process is sugared acid anhydride 0.1-5mg/ml of adding and sulfur-containing amino acid 0.0001-12g/100g in fermented liquid ,-20--70 ℃ following freeze-drying, and then form the Cordyceps fermented product lyophilized powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008102288465A CN101407767A (en) | 2008-11-17 | 2008-11-17 | Method for producing Chinese caterpillar fungus by fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008102288465A CN101407767A (en) | 2008-11-17 | 2008-11-17 | Method for producing Chinese caterpillar fungus by fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101407767A true CN101407767A (en) | 2009-04-15 |
Family
ID=40570981
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008102288465A Pending CN101407767A (en) | 2008-11-17 | 2008-11-17 | Method for producing Chinese caterpillar fungus by fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101407767A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103211212A (en) * | 2013-04-11 | 2013-07-24 | 天津天狮生物发展有限公司 | Cordyceps mycelia and preparation method thereof |
| CN104904496A (en) * | 2015-06-18 | 2015-09-16 | 上海善力健生物科技有限公司 | Preparation method for cordyceps sinensis mycelium |
| CN105154491A (en) * | 2015-09-02 | 2015-12-16 | 天津现代职业技术学院 | Cordyceps sinensis liquid state fermentation culture medium for producing exopolysaccharides |
| CN106282028A (en) * | 2015-05-29 | 2017-01-04 | 东海大学 | Method for culturing cordyceps sinensis |
-
2008
- 2008-11-17 CN CNA2008102288465A patent/CN101407767A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103211212A (en) * | 2013-04-11 | 2013-07-24 | 天津天狮生物发展有限公司 | Cordyceps mycelia and preparation method thereof |
| CN103211212B (en) * | 2013-04-11 | 2016-01-20 | 天津天狮生物发展有限公司 | A kind of Cordyceps mycelium and preparation method |
| CN106282028A (en) * | 2015-05-29 | 2017-01-04 | 东海大学 | Method for culturing cordyceps sinensis |
| CN106282028B (en) * | 2015-05-29 | 2020-02-18 | 东海大学 | Method for culturing cordyceps sinensis |
| CN104904496A (en) * | 2015-06-18 | 2015-09-16 | 上海善力健生物科技有限公司 | Preparation method for cordyceps sinensis mycelium |
| CN105154491A (en) * | 2015-09-02 | 2015-12-16 | 天津现代职业技术学院 | Cordyceps sinensis liquid state fermentation culture medium for producing exopolysaccharides |
| CN105154491B (en) * | 2015-09-02 | 2018-08-24 | 天津现代职业技术学院 | A kind of culture medium for cordyceps sinensis liquid state fermentation production exocellular polysaccharide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106434373A (en) | High-density fermentation medium formula of sparassis crispa and pharmaceutical grade glucan preparation method of high-density fermentation medium formula | |
| US20210079339A1 (en) | Fermentation production process of paecilomyces hepialis cs-4 | |
| CN111996129B (en) | New strain of cicada fungus and its use in anti-tumor and bacteriostasis | |
| CN101492706A (en) | Method for improving cordyceps sinensis bacterium native volume of production with cordyceps militaris link liquid fermentation | |
| WO2017202293A1 (en) | Artificial cultivation method for cordyceps sobolifera | |
| CN102925527A (en) | Method for mixing and fermenting flammulina velutipes and lucid ganoderma | |
| CN1872995A (en) | Method for raising quality of protocorm of Tiepi stem of noble dendrobium cultivated in liquid through inductor of mycorrhizal fungi | |
| CN104087645B (en) | A kind of solid fermentation flower of Panax ginseng and preparation method | |
| CN106086147A (en) | Fungal elicitor is utilized to induce the method extracting betulic acid from Inonqqus obliquus | |
| CN111334440A (en) | Rhizopus oryzae CH5 and application thereof in extraction of ganoderma sinensis polysaccharide | |
| CN107432135A (en) | Promote the method for cynomorium songaricum seed sprouting using fungi | |
| CN101407767A (en) | Method for producing Chinese caterpillar fungus by fermentation | |
| CN102293122B (en) | Cultivating method for cordyceps militaris by using manyprickle acathopanax root | |
| CN100575475C (en) | The method of wheat cultivation Cordyceps militaris | |
| CN102816701A (en) | Strain used for fermenting rice bran and wheat bran extracts for producing grifolan | |
| CN102172171B (en) | Method for culturing germanium-enriched cordyceps mycelium | |
| CN104472221B (en) | A kind of method by natural cordyceps fructification fermented hypha | |
| CN105557312A (en) | Method for cultivating Cordyceps products by taking Tenebrio molitor (L.) larvae as carriers | |
| CN109392600A (en) | A kind of cultural method of selenium-enriched cordceps militaris | |
| CN100567318C (en) | Nucleoside active matter in the artificial culture Cordyceps militaris (L.) Link. and its production and use | |
| CN105296357A (en) | Method for increasing yield of cordyceps sinensis liquid fermentation mycelia by supplementing materials | |
| TWI385248B (en) | A formula of culturing medium for cordyceps spp. | |
| CN105493899A (en) | Method for cultivating cordyceps sinensis products with mealworm pupae as carriers | |
| CN105535035A (en) | Inonotus obliquus fermentation culture composition and preparation method thereof | |
| CN103798293A (en) | Preparation and applications of salvia miltiorrhiza endophytic fungi hypha water-soluble extract and fractions of salvia miltiorrhiza endophytic fungi hypha water-soluble extract |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| ASS | Succession or assignment of patent right |
Owner name: SHANDONG HUIPENG INVESTMENT CO., LTD. Free format text: FORMER OWNER: JIA JINGMING Effective date: 20091204 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20091204 Address after: Pioneer Park for overseas students in Yantai Development Zone, Shandong Applicant after: Shandong Hui Peng Investment Co., Ltd. Address before: No. 103, Wenhua Road, Shenhe District, Liaoning, Shenyang Applicant before: Jia Jingming |
|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20090415 |