CN102936609B - Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein - Google Patents
Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein Download PDFInfo
- Publication number
- CN102936609B CN102936609B CN201210434092.5A CN201210434092A CN102936609B CN 102936609 B CN102936609 B CN 102936609B CN 201210434092 A CN201210434092 A CN 201210434092A CN 102936609 B CN102936609 B CN 102936609B
- Authority
- CN
- China
- Prior art keywords
- acid glycoprotein
- hours
- preparation
- peacilomyce hepiahi
- paecilomyces varioti
- 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.)
- Expired - Fee Related
Links
Landscapes
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein. The method includes culturing first-level seeds of swift moth paecilomyces varioti bacterial strains, conducting inoculated fermentation to obtain swift moth paecilomyces varioti fermentation liquid, conducting work procedures including ethanol precipitation, deionized water redissolution, deproteinization, freezing, drying, diethyl-aminoethanol (DEAE)-52 column chromatography and the like on the fermentation liquid to extract the swift moth paecilomyces varioti extracellular acid glycoprotein. The preparation method has the advantages of being simple in process, mild in extraction condition, low in cost, environment-friendly, apt to industrial production and the like. The obtained swift moth paecilomyces varioti extracellular acid glycoprotein can serve as raw materials of medicines and health food.
Description
Technical field
The present invention relates to bat moth bacterium Paecilomyces varioti bacterial strain liquid fermenting and from its fermented liquid the separated technical field that obtains acid glycoprotein born of the same parents outside, be specifically related to a kind of peacilomyce hepiahi born of the same parents preparation method of acid glycoprotein outward.
Background technology
Cordyceps sinensis is traditional famous and precious strong, tonic herb of China, and it is that for human being's production and the real huge value of life they can produce the multiple physiologically active substance that has important value in biotechnology and medicines and health protection.Their physiologically active is rich and varied, can be antibacterial, antitumor, antiplatelet condenses, radioprotective, improve memory, regulate body immunity, calcium ion antagonism, brain and heart are had to provide protection, calmness and analgesia etc. under normobaric hypoxia.
Due to the strict parasitics of natural cordyceps and special ecotope requirement, its resource is extremely limited, expensive.In recent years, along with Anamorph of Cordyceps Sinensis progress of research, the research of relevant Anamorph of Cordyceps polysaccharide is just carried out.Cordyceps sinensis can carry out fermentative production by bionic method, Cordyceps mycelium to artificial culture, through chemistry, pharmacological research, proof is basically identical with natural cs, through clinical application for many years, by the world of medicine, be known as the substitute that can be used as Cordyceps sinensis, the People's Republic of China's existing clear and definite regulation of version pharmacopeia in 2000.Meanwhile, the also various bioactivators such as the trace element such as rich in proteins, VITAMIN, zinc, copper and cordycepic acid (D mono-N.F,USP MANNITOL), cordycepin (3 '-deoxyadenosine), polyose, alkaloids, cyclic peptide class, SOD enzyme in Anamorph of Cordyceps fermented liquid.Wherein, polyose is as one of the abundantest most important monoid in the contained physiologically active substance of Cordyceps sinensis, having the different physiological roles such as anti-oxidant, antitumor, hypoglycemic, reducing blood-fat, is an important breakthrough mouth of Cordyceps sinensis development research, has a good application prospect.
Peacilomyce hepiahi (Paecilomyces hepiali) is a kind of of Anamorph of Cordyceps, and this bacterial strain is separated in the natural cs sporophore of Yushu Regions, Qinghai Province in 2002, and DSMZ of the Bing You Chinese Academy of Sciences identifies.The popularity distributing due to Chinese caterpillar fungus and the diversity of anamorphic strain, there is very big-difference in different scientific research institutions to the research of anamorph Chinese caterpillar fungus, and about utilize this strain liquid fermentation and from fermented liquid outside separated born of the same parents the research of acid glycoprotein there is not yet report.
Summary of the invention
The object of the invention aims to provide a kind of method of preparing the outer acid glycoprotein of peacilomyce hepiahi born of the same parents, realizes concrete scheme of the present invention as follows:
(1) from be stored in the mother's kind potato slope substratum, take mycelium piece (size is 0.5cm * 0.5cm), be inoculated in (every bottled seed culture medium 100mL of 250mL triangle) in seed culture medium, at 17 ℃, static cultivation is after 12 hours, under 120rpm, shaking culture is cultivated 72 hours, can obtain seed liquor.Seed culture medium (g/L): sucrose 30.00, yeast extract paste 6.00, potassium primary phosphate 1.50, magnesium sulfate 0.50, wort 10.00, murphy juice 10.00, pH 6.5.
(2) seed liquor of getting in step (1) is inoculated into (every bottled fermention medium 200mL of 500mL triangle) in fermention medium, inoculum size is 1% (V/V), at 17 ℃, static cultivation is after 12 hours, under 120rpm, shaking culture was cultivated after 120 hours, tunning centrifugation (centrifugation condition: 4000rmp, 15min) is obtained to peacilomyce hepiahi fermented liquid.Fermentative medium formula (g/L): sucrose 50.86, yeast extract paste 2.00, ammonium sulfate 6.61, potassium primary phosphate 1.36, magnesium sulfate 0.05, murphy juice 15.00, wort 15.00, pH 6.5;
(3) by the peacilomyce hepiahi fermented liquid obtaining in step (2), 50 ℃ are evaporated to 1/2 of original volume, the dehydrated alcohol that adds 3 times of volumes, precipitate 12 hours, with after 80% washing with alcohol 3 times, centrifugation (centrifugation condition: 4000rmp, 15min), obtains crude extracellular polysaccharide.
(4) crude extracellular polysaccharide that step (3) made ventilates and flings to ethanol, after adding a small amount of deionized water and redissolving, with Sevag method (chloroform: propyl carbinol=4: 1), after deproteinated 3 times, aqueous phase solution lyophilize is obtained to Crude polysaccharides freezing dry powder.
(5) after step (4) gained Crude polysaccharides is dissolved, be splined on DEAE-52 cellulose column, and with deionized water, 0.1mol/LNaCl and 0.3mol/L NaCl, carry out wash-out respectively, the 0.3mol/L NaCl elution fraction of collection is carried out to concentrating under reduced pressure, deionized water dialysis 72 hours, concentrated postlyophilization, obtains the outer acid glycoprotein component of born of the same parents.The outer acid glycoprotein of born of the same parents of preparation is pale powder, and wherein polysaccharide content 80.56%, protein content 18.76%.
(6) the outer acid glycoprotein of step (5) gained born of the same parents is detected to its molecular weight distribution situation with molecular-exclusion chromatography, with Infrared spectroscopy characteristic group, form, with uv scan, determine whether it has protein component.
2, according to patent, require a kind of preparation method with the outer acid glycoprotein of biological activity born of the same parents described in 1, it is characterized in that: the fermentative medium formula (g/L) of described step (2): sucrose 50.86, yeast extract paste 2.00, ammonium sulfate 6.61, potassium primary phosphate 1.36, magnesium sulfate 0.05, murphy juice 15.00, wort 15.00, pH 6.5.
The outer acid glycoprotein of peacilomyce hepiahi born of the same parents of preparation is pale powder, and wherein polysaccharide content 80.56%, protein content 18.76%, the easily moisture absorption.The infrared absorption spectrum of this acid glycoprotein shows that it has polysaccharide, carboxyl, sulfate and amino charateristic avsorption band.The uv-absorbing scanning of the outer acid glycoprotein of these born of the same parents shows the absorption peak that it contains albumen.Molecular weight to the outer glycoprotein of these born of the same parents is measured, and shows that its molecular weight distribution is between 20-80kD.
Accompanying drawing explanation
Fig. 1 substratum forms the impact on the outer acid glycoprotein output of peacilomyce hepiahi born of the same parents
The infrared absorption pattern of the outer acid glycoprotein of Fig. 2 peacilomyce hepiahi born of the same parents
The UV scanning collection of illustrative plates of the outer acid glycoprotein sugar of Fig. 3 peacilomyce hepiahi born of the same parents
Embodiment
Below by embodiment, the invention will be further described.
Embodiment 1:
(1) from be stored in the mother's kind potato slope substratum, take mycelium piece (size is 0.5cm * 0.5cm), be inoculated in (every bottled seed culture medium 100mL of 250mL triangle) in seed culture medium, at 17 ℃, static cultivation is after 12 hours, under 120rpm, shaking culture is cultivated 72 hours, can obtain seed liquor.Seed culture medium (g/L): sucrose 30.00, yeast extract paste 6.00, potassium primary phosphate 1.50, magnesium sulfate 0.50, wort 10.00, murphy juice 10.00, pH 6.5.
(2) seed liquor of getting in step (1) is inoculated into (every bottled fermention medium 200mL of 500mL triangle) in fermention medium, inoculum size is 1% (V/V), at 17 ℃, static cultivation is after 12 hours, under 120rpm, shaking culture was cultivated after 120 hours, tunning centrifugation (centrifugation condition: 4000rmp, 15min) is obtained to peacilomyce hepiahi fermented liquid.Fermentative medium formula (g/L): sucrose 50.86, yeast extract paste 2.00, ammonium sulfate 6.61, potassium primary phosphate 1.36, magnesium sulfate 0.05, murphy juice 15.00, wort 15.00, pH 6.5;
(3) by the peacilomyce hepiahi fermented liquid obtaining in step (2), 50 ℃ are evaporated to 1/2 of original volume, the dehydrated alcohol that adds 3 times of volumes, precipitate 12 hours, with after 80% washing with alcohol 3 times, centrifugation (centrifugation condition: 4000rmp, 15min), obtains crude extracellular polysaccharide.
(4) crude extracellular polysaccharide that step (3) made ventilates and flings to ethanol, after adding a small amount of deionized water and redissolving, with Sevag method (chloroform: propyl carbinol=4: 1), after deproteinated 3 times, aqueous phase solution lyophilize is obtained to Crude polysaccharides freezing dry powder.
(5) after step (4) gained Crude polysaccharides is dissolved, be splined on DEAE-52 cellulose column, and with deionized water, 0.1mol/LNaCl and 0.3mol/L NaCl, carry out wash-out respectively, the 0.3mol/L NaCl elution fraction of collection is carried out to concentrating under reduced pressure, deionized water dialysis 72 hours, concentrated postlyophilization, obtains the outer acid glycoprotein component of born of the same parents.The outer acid glycoprotein of born of the same parents of preparation is pale powder, and wherein polysaccharide content 80.56%, protein content 18.76%.
(6) the outer acid glycoprotein of step (5) gained born of the same parents is detected to its molecular weight distribution situation with molecular-exclusion chromatography, with Infrared spectroscopy characteristic group, form, with uv scan, determine whether it has protein component.
Embodiment 2:
The fermentation condition optimization experiment of the outer acid glycoprotein of peacilomyce hepiahi born of the same parents
(1) the fermentation condition optimization list Factor screening experiment of the outer acid glycoprotein of peacilomyce hepiahi born of the same parents
1. the screening of some factors design to the remarkable factor of fermentation condition.
The outer acid glycoprotein fermentation condition factor of peacilomyce hepiahi born of the same parents and code are: glucose (X
1), Semen Maydis powder (X
2), yeast extract paste (X
3), ammonium sulfate (X
4), KH
2pO
4(X
5), MgSO
4(X
6), murphy juice (X
7), wort (X
8) central point (0) be respectively (g/L): 30,10,2,6,1,0.5,10,10, step-length is respectively: 10,5,1,1,0.5,0.2,5,5.The output of the outer acid glycoprotein of peacilomyce hepiahi born of the same parents of 20 groups of tests is in Table 1.
Table 1 fermention medium significance conditional filtering
Through Design-Expert 7.0 softwares, carry out variance analysis, obtain table 2:
The variance analysis of table 2 fractional factorial design
As can be seen from Table 2, sucrose (X
1), yeast extract paste (X
3), ammonium sulfate (X
4) and potassium primary phosphate (X5) be the significance influence factors of the outer acid glycoprotein output of peacilomyce hepiahi born of the same parents.
But cannot judge sucrose (X from table
1), yeast extract paste (X
3) and ammonium sulfate (X
4) and the remarkable sex concentration central point of potassium primary phosphate (X5) to the outer acid glycoprotein yield of peacilomyce hepiahi born of the same parents.In order further to optimize sucrose (X
1), yeast extract paste (X
3) and ammonium sulfate (X
4) and potassium primary phosphate (X
5) optimum concn and interaction, the present invention adopts response surface method further to test sucrose (X
1), yeast extract paste (X
3) and ammonium sulfate (X
4) and potassium primary phosphate (X
5) optimum concn.
2. response surface method is optimized the outer acid glycoprotein fermentation condition result of peacilomyce hepiahi high yield born of the same parents
Sucrose (X
1), yeast extract paste (X
3) and ammonium sulfate (X
4) and potassium primary phosphate (X
5) central point be respectively (g/L): 50,3,6 and 1.0.Response surface test design and the results are shown in Table 3.According to table 3 result, with Design EXpert7.0 software analysis, obtain multiple regression equation and be:
Y＝-37.99+1.84X
1-0.14X
3+1.19X
4+3.53X
5-0.014X
1 2-0.97X
3 2-0.04X
4 2-0.79X
5 2+0.085X
1X
3-0.093X
1X
4-0.018X
1X
5
According to Equation for Calculating, obtaining optimal conditions of fermentation is: the outer acid glycoprotein fermentative medium formula (g/L) of peacilomyce hepiahi high yield born of the same parents: sucrose 50.86, yeast extract paste 2.00, ammonium sulfate 6.61, potassium primary phosphate 1.36, magnesium sulfate 0.05, murphy juice 15.00, wort 15.00, pH 6.5, and the outer acid glycoprotein production peak of born of the same parents can reach 5.33g/L.
The test of table 3 response surface and fermentation condition optimization result
Claims (2)
1. a preparation method for the outer acid glycoprotein of peacilomyce hepiahi (Paecilomyces hepiali) HN1 bacterial strain born of the same parents, is characterized in that comprising following operation steps:
(1) from being stored in the mother of peacilomyce hepiahi (Paecilomyces hepiali) the HN1 bacterial strain of potato slope substratum, taking size planting and be about 0.5cm * 0.5cm mycelium piece, be inoculated in and be equipped with in the triangular flask that the capacity of 100mL seed culture medium is 250mL, consisting of of this seed culture medium: sucrose 30.00g/L, yeast extract paste 6.00g/L, potassium primary phosphate 1.50g/L, magnesium sulfate 0.50g/L, wort 10.00g/L, murphy juice 10.00g/L, pH6.5, then static cultivation after 12 hours at 17 ℃, be placed in again on the shaking table of 120 revs/min shaking culture 72 hours, can obtain seed liquor,
(2) getting seed liquor in step (1) is inoculated in and is equipped with in the triangular flask that the capacity of 200mL fermention medium is 500mL by 1% of fermention medium volume, consisting of of this fermention medium: sucrose 50.86g/L, yeast extract paste 2.00g/L, ammonium sulfate 6.61g/L, potassium primary phosphate 1.36g/L, magnesium sulfate 0.05g/L, murphy juice 15.00g/L, wort 15.00g/L, pH6.5, then static cultivation after 12 hours at 17 ℃, be placed in again on the shaking table of 120 revs/min shaking culture 120 hours, after cultivation finishes by tunning under 4000 revs/min of conditions after centrifugal 15min, collect supernatant liquor and be peacilomyce hepiahi (Paecilomyces hepiali) HN1 bacterial strain fermentation liquor,
(3) by peacilomyce hepiahi (Paecilomyces hepiali) the HN1 bacterial strain fermentation liquor obtaining in step (2), 50 ℃ are evaporated to 1/2 of original volume, the dehydrated alcohol that adds 3 times of volumes, precipitate 12 hours, with after 80% washing with alcohol 3 times, under 4000 revs/min of conditions, centrifugal 15min collecting precipitation can obtain crude extracellular polysaccharide;
(4) crude extracellular polysaccharide that step (3) made ventilates and flings to ethanol, after adding a small amount of deionized water and redissolving, after the mixing solutions deproteinated that is 4: 1 with chloroform and propyl carbinol volume ratio 3 times, by aqueous phase solution lyophilize with acquisition Crude polysaccharides freezing dry powder;
(5) after step (4) gained Crude polysaccharides is dissolved, be splined on DEAE-52 cellulose column, and with deionized water, 0.1mol/LNaCl and 0.3mol/L NaCl, carry out wash-out respectively, the 0.3mol/L NaCl elution fraction of collection is carried out to concentrating under reduced pressure, deionized water dialysis 72 hours, concentrates postlyophilization, can obtain the acid glycoprotein component of pale powder, in this component, polysaccharide content is 80.56%, and protein content is 18.76%;
(6) step (5) gained acid glycoprotein is detected to its molecular weight distribution situation with molecular-exclusion chromatography, with Infrared spectroscopy characteristic group, form, with uv scan, determine whether it has protein component.
2. according to patent, require the preparation method of the outer acid glycoprotein of a kind of peacilomyce hepiahi (Paecilomyces hepiali) HN1 bacterial strain born of the same parents described in 1, it is characterized in that: the fermentative medium formula of described step (2): sucrose 50.86g/L, yeast extract paste 2.00g/L, ammonium sulfate 6.61g/L, potassium primary phosphate 1.36g/L, magnesium sulfate 0.05g/L, murphy juice 15.00g/L, wort 15.00g/L, pH6.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210434092.5A CN102936609B (en) | 2012-11-05 | 2012-11-05 | Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210434092.5A CN102936609B (en) | 2012-11-05 | 2012-11-05 | Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102936609A CN102936609A (en) | 2013-02-20 |
| CN102936609B true CN102936609B (en) | 2014-12-17 |
Family
ID=47695530
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210434092.5A Expired - Fee Related CN102936609B (en) | 2012-11-05 | 2012-11-05 | Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102936609B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104762341A (en) * | 2015-04-02 | 2015-07-08 | 河南科技学院 | Preparation method of low-molecular-weight paecilomyces hepialid exopolysaccharides |
| CN104911109A (en) * | 2015-05-28 | 2015-09-16 | 吉林大学 | High-yield paecilomyces hepiali mutant strain and cultivation method |
| CN105147715B (en) * | 2015-06-15 | 2019-01-01 | 河南科技学院 | A kind of new application of Paecilomyces hepiali chen neutrality exocellular polysaccharide |
| CN105177050A (en) * | 2015-10-26 | 2015-12-23 | 河南科技学院 | Method for preparing paecilomyces hepiali purpurin |
| CN105176845B (en) * | 2015-10-26 | 2019-07-26 | 河南科技学院 | A kind of preparation method of Paecilomyces hepiali chen fermentation liquid instant granular |
| CN107343889A (en) * | 2016-05-06 | 2017-11-14 | 元龙生技国际股份有限公司 | A kind of peacilomyce hepiahi bacterium fermentation culture medium for being used to treat the 1st type or diabetes B |
| CN106008662A (en) * | 2016-05-16 | 2016-10-12 | 南京农业大学 | Method for preparing zinc-rich neutral glycoprotein from bee-collected Chinese wolfberry pollen serving as raw material |
| CN111139277A (en) * | 2020-03-03 | 2020-05-12 | 长春工业大学 | Crude polysaccharide and preparation process thereof, health-care oral liquid and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101096641A (en) * | 2007-06-13 | 2008-01-02 | 杭州中美华东制药有限公司 | Culture medium for producing winter worm summer herb mycelium |
| CN101463325A (en) * | 2008-12-16 | 2009-06-24 | 浙江博士园生物技术有限公司 | Industrialized cultivation method for north aweto |
| CN101974095A (en) * | 2010-10-14 | 2011-02-16 | 南京林业大学 | Method for extracting and separating Chinese narcissus polysaccharides |
| CN102154407A (en) * | 2011-05-24 | 2011-08-17 | 河北科技大学 | Corayceps militaris polysaccharide two-stage fermentation synthesis process |
-
2012
- 2012-11-05 CN CN201210434092.5A patent/CN102936609B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101096641A (en) * | 2007-06-13 | 2008-01-02 | 杭州中美华东制药有限公司 | Culture medium for producing winter worm summer herb mycelium |
| CN101463325A (en) * | 2008-12-16 | 2009-06-24 | 浙江博士园生物技术有限公司 | Industrialized cultivation method for north aweto |
| CN101974095A (en) * | 2010-10-14 | 2011-02-16 | 南京林业大学 | Method for extracting and separating Chinese narcissus polysaccharides |
| CN102154407A (en) * | 2011-05-24 | 2011-08-17 | 河北科技大学 | Corayceps militaris polysaccharide two-stage fermentation synthesis process |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102936609A (en) | 2013-02-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102936609B (en) | Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein | |
| CN101245361B (en) | Method for producing cordycepin, breeding of high production cordyceps militaris link bacterial strain BYB-08 and application | |
| CN103263448B (en) | Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application | |
| CN102047814A (en) | Micro ventilation lime wood antrodia camphorate cultivation method | |
| CN110129207A (en) | A kind of liquid fermentation medium of the anti-oxidant CSM treated body of high yield and its production method of anti-oxidant CSM treated body electuary | |
| CN103130909A (en) | Preparation method of selenium-rich Morchella polysaccharide | |
| CN102674929B (en) | Inonotus obliquus submerged fermentation culture medium and submerged fermentation method thereof | |
| CN101463326B (en) | Production method of Guangdong Cordyceps sinensismycelium | |
| CN103477994A (en) | Bacterial strain used for producing ganoderma lucidum polysaccharides by complete feed liquid fermentation of rice bran and wheat bran | |
| CN111334440B (en) | Rhizopus oryzae CH5 and application thereof in extraction of ganoderma sinensis polysaccharide | |
| CN110507681A (en) | A kind of technique of higher value application flower of Panax ginseng | |
| CN110438180A (en) | The preparation of the extracellular active polysaccharide of Ganoderma lucidum submerged fermentation and its enhancing immunization purposes | |
| CN102898210A (en) | Medium for fermentation of Inonotus obliquus, and fermentation method for producing polysaccharide and application thereof | |
| CN104472221B (en) | A kind of method by natural cordyceps fructification fermented hypha | |
| CN102816701A (en) | Strain used for fermenting rice bran and wheat bran extracts for producing grifolan | |
| CN103421861A (en) | Method of manufacturing cordyceps sinensis (Berk.) sacc polysaccharide by liquid state fermentation of rice bran and bran complete feed | |
| CN109294927A (en) | A method of improving Cordyceps sinensis polysaccharide content in Cordyceps militaris | |
| CN103555786B (en) | A kind of glossy ganoderma mutagenic fungi liquid state fermentation rice bran wheat bran complete feed is produced the method for polysaccharide | |
| CN100567318C (en) | Nucleoside active matter in the artificial culture Cordyceps militaris (L.) Link. and its production and use | |
| CN104396571A (en) | High-cordycepin-content rich-selenium cordyceps sinensis cultivation method | |
| CN102899368A (en) | Method for preparing selenium-rich tremella aurantia crude polysaccharide powder by using deep liquid fermentation process | |
| 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 | |
| CN105586269A (en) | Method for producing grifola frondosa mycelium raw material from grifola frondosa strain obtained through mutagenesis | |
| CN109402194A (en) | A kind of Chinese caterpillar fungus polysaccharide extraction and purification process based on liquid fermentation | |
| CN101407767A (en) | Method for producing Chinese caterpillar fungus by fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141217 Termination date: 20171105 |