CN101701193A - Method for preparing podophyllotoxin and/or astragaloside and special bacterial strain thereof - Google Patents
Method for preparing podophyllotoxin and/or astragaloside and special bacterial strain thereof Download PDFInfo
- Publication number
- CN101701193A CN101701193A CN200910237790A CN200910237790A CN101701193A CN 101701193 A CN101701193 A CN 101701193A CN 200910237790 A CN200910237790 A CN 200910237790A CN 200910237790 A CN200910237790 A CN 200910237790A CN 101701193 A CN101701193 A CN 101701193A
- Authority
- CN
- China
- Prior art keywords
- podophyllotoxin
- weqe
- penicillium
- astragalin
- ateckii
- 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.)
- Granted
Links
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a method for preparing podophyllotoxin and/or astragaloside and a special bacterial strain thereof. The bacterial strain is Penicillium ateckii WEQE of which the collection number is CGMCC NO.3401. The bacterial strain in the invention can be used for preparing podophyllotoxin and/or astragaloside. The method for preparing podophyllotoxin and/or astragaloside in the invention is not influenced by plant resources, has the advantages of simple operation, low cost, easy operation, short production cycle and the like, provides a new path for preparing podophyllotoxin, avoids limitations of other preparation methods in the prior art, has strong maneuverability, is suitable for large-scale industrialized production, and has important significance on protecting ecological diversity of plants.
Description
Technical field
The present invention relates to a kind of method and special strain therefore thereof for preparing podophyllotoxin and/or astragalin.
Background technology
Podophyllotoxin (podophyllotoxin) mainly is present in Berberidaceae Podophyllum emodi var chinense genus and the kindred plant thereof, be the natural compounds that a class has extensive biologic activity, have quasi-estrin sample, estrogen antagonist sample, anticancer, antiviral, antimycotic, calm, suppress plant-growth and effect such as anti-oxidant.Particularly it has stronger antitumour activity, and its mechanism of action is the activity of unzipping to stop cell fission and to suppress the DNA topoisomerase that suppresses tubulin.The glycosides derivatives etoposide of podophyllotoxin and the toxicity of teniposide are lower, kinds of tumors diseases such as small cell lung cancer, lymphatic cancer are all had better curative effect and become the broad-spectrum anti-cancer drug of clinical application.
At present, podophyllotoxin mainly is to extract to obtain from wild Podophyllum emodi var chinense class plant rhizome, and this class plant only has 4 to belong to 15 kinds in the whole world, China has 3 to belong to 12 kinds, mainly be grown in high altitude localitiess such as the Qinling Mountains, Tibet, its poor growth, and the content of podophillotoxines material is lower.Because the podophillotoxines material has broad application prospects at aspects such as medical science and agricultural controls, thereby its demand increases day by day; To a large amount of collections of wild Podophyllum emodi var chinense class plant resources, its resource is seriously damaged, cause the irreversible loss of species diversity and ecotope, what therefore solve the podophillotoxines material comes source problem significant.
Because the plant growth environment complexity be difficult to simulation, and its growth cycle is long, is difficult to carry out large-scale artificial culture, and tissue culture technique is also inapplicable for Podophyllum emodi var chinense class plant; In addition, podophillotoxines material chemistry complex structure, though chemosynthesis can be carried out, cost is too high, and makes up engineering bacteria also owing to the enzyme quantity that participates in such material of formation too much can't be carried out by the genetic engineering means.So utilize endophyte to have to produce and the character of the same or analogous biologically active substance of host, solve the source problem that comes of podophyllotoxin, may be unique also be effective means.
In addition, owing to also contain multiple flavonoid compound in the Podophyllum emodi var chinense class plant, thereby its endophyte also may produce flavonoid compound.Many studies confirm that, flavonoid compound have multiple biological activity in human body, comprise anti-oxidant, anticancer, anti-cancer, remove free radical, antiviral, anti-inflammatory and promotion vasorelaxation etc.In addition, isoflavones (as genistein and hydroxy-isoflavone) also has the oestrogenic hormon characteristic.The multiple physiological action of flavonoid compound makes it be with a wide range of applications in industries such as medicine, food.
Summary of the invention
An object of the present invention is to provide a strain and can produce the bacterial strain and the cultural method thereof of podophyllotoxin and/or astragalin.
Bacterial strain provided by the present invention is Si Shi mould (Penicillium ateckii) WEQE.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on November 5th, 2009 and (is called for short CGMCC, address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number is CGMCC No.3401, classification called after Penicillium ateckii, strain name is WEQE.
The method of cultivation Si Shi mould (Penicillium ateckii) WEQE CGMCC No.3401 provided by the present invention is with potato glucose culture medium culturing Si Shi mould (Penicillium ateckii) WEQE CGMCCNo.3401.
In the described cultural method, described culture condition can comprise that temperature is that 26-30 ℃, hunting speed are 100-150rpm.Described temperature specifically can be 28 ℃; Described hunting speed specifically can be 120rpm.
Another object of the present invention provides a kind of method for preparing podophyllotoxin and/or astragalin.
The method for preparing podophyllotoxin and/or astragalin provided by the present invention comprises the steps: fermentation culture Si Shi mould (Penicillium ateckii) WEQE CGMCC No.3401, obtains podophyllotoxin and/or astragalin.
The substratum of using in the described fermentation culture can be the potato glucose substratum.
The condition of described fermentation culture comprises that temperature is that 26-30 ℃, hunting speed are 100-150rpm.The time of described fermentation culture can be 5-9 days.Described temperature specifically can be 28 ℃; Described hunting speed specifically can be 120rpm; The described time specifically can be 7 days.
After described fermentation, also comprise the step of extraction among the described preparation method.
Described extraction agent is a chloroform.
The application of Si Shi mould (Penicillium ateckii) WEQE CGMCC No.3401 in preparation podophyllotoxin and/or astragalin also belongs to protection scope of the present invention.
Bacterial strain of the present invention can be used to prepare podophyllotoxin and/or astragalin.The method for preparing podophyllotoxin and/or astragalin of the present invention has advantage simple to operate, with low cost, easy to operate, with short production cycle.The inventive method provides new way for the preparation podophyllotoxin; avoided other various preparation methods' limitation in the prior art; have very strong operability, be suitable for large-scale industrialization production, simultaneously significant to the protective plant ecological diversity again.
Description of drawings
Fig. 1 is the colonial morphology of Si Shi mould.
Fig. 2 is Si Shi mould conidial head and spore shape.Conidium head morphology when A and B are respectively (10 * 40) and (10 * 100), C (10 * 40) is a spore shape
Fig. 3 is the HPLC spectrogram of podophyllotoxin standard substance
Fig. 4 is the HPLC spectrogram of astragalin standard substance
Fig. 5 is the HPLC spectrogram of the chloroform extract of Si Shi mould (Penicillium ateckii) WEQE CGMCC № 3401 fermented liquids
Embodiment
Employed experimental technique is ordinary method if no special instructions among the following embodiment.
Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.
The genetic resources title: Rhizoma et Radix Diphylleiae (Diphylleia sinensis L.) (in Chinese Plants will, is a Diphylleia sinensis; Rhizoma et Radix Diphylleiae is the title in the flora of the Qinling Mountains.)。The acquiring way of genetic resources: I, genetic resources are taken from: plant, II, obtain manner: gather voluntarily.Direct sources: acquisition time: 2007 08 month; Acquisition mode is gathered ground (country, province (city)) Chinese Shaanxi Province Mei County Red River Valley (height above sea level 1560m), picker's title (name): Zhao Changqi, picker's contact method: 010-58805046.Primary source: same direct sources.
The structural formula of podophyllotoxin (podophyllotoxin) is suc as formula shown in the I, and the structural formula of astragalin (Astragalin) is suc as formula shown in the II:
Formula I formula II
Separation and the evaluation of embodiment 1, Si Shi mould (Penicillium ateckii) WEQE CGMCC № 3401
One, separates
To gather from the root and the root stock of the plant Rhizoma et Radix Diphylleiae (Diphyllria sinensisL.) of Mei County, Shaanxi Province Red River Valley (height above sea level 1560m) and clean with tap water respectively, with the Ethanol Treatment 5min of 75% volumn concentration, aseptic water washing is 3-5 time then in Bechtop; Then handle 10min with the clorox of 2.5% quality percentage composition again, aseptic water washing is 3-5 time again.Tweezers and blade with sterilization are peelled off the crust of the root of Rhizoma et Radix Diphylleiae, and the small pieces that are cut into 0.5cm * 0.5cm size are again planted on the PDA solid medium, cultivate 3-7 days for 28 ℃.The root of the Rhizoma et Radix Diphylleiae that same sterilising treatment is crossed is not done peeling and cutting simultaneously, and the root of Rhizoma et Radix Diphylleiae after the week of rolling on the PDA substratum, is cultivated in contrast with condition and to be observed.
Cultivating after several days the incision of finding at the root of Rhizoma et Radix Diphylleiae has mycelia to grow, get the mycelia that incision newly grows, be transferred on the fresh PDA substratum and cultivate, after treating that bacterium colony occurs, form according to bacterium colony, the difference of color and bacterium colony grow the difference of time, the mycelium inoculation at picking substratum edge carries out separation and Culture on new PDA substratum respectively, until filtering out single bacterium colony, single bacterial strain is cultivated, extract product, the product that extracts is carried out high performance liquid chromatography detect, obtain producing the bacterial strain of the podophyllotoxin and the Radix Astragali, this bacterial strain called after WEQE.
Two, identify
Above-mentioned bacterial strains is carried out the evaluation of following Physiology and biochemistry, bacterium form:
(1) colony characteristics:
Colonial morphology as shown in Figure 1.
Single strain WEQE is seeded on the PDA substratum, after cell grows to the logarithmic growth later stage, and the bacterium colony size is stable, observes colonial morphology.The result shows, the bacterial strain WEQE that above-mentioned separation obtains bacterium colony on the PDA substratum is thin, and the growth limitation is produced spore face dark blue green or greyish-green, and is velvet-like, on secrete most colourless so that water droplets of aureus, back side band dark yellow.
(2) mycelia morphological specificity:
The mycelia form as shown in Figure 2.
Conidiophore vertically bears from mycelia, and amacrine has diaphragm, the top life be arranged in the broom shape between the branch, the bottom is branch not; Between branch different in size, scatter, upward give birth to the stigma that majority is arranged in parallel, stigma 8-10 * 2 micron; It is cylindric that the normal work of conidia chain scatters; Conidium is spherical to oval, diameter 2-2.5 micron.
Comprehensive above-mentioned feature, according to data such as " fungi identification handbook ", " Chinese fungi will ", the bacterial strain that above-mentioned separation obtains has the essential characteristic of Si Shi mould, it is accredited as Si Shi mould (Penicillium ateckii), and its classification belongs to deuteromycetes, Moniliales, Moniliaceae, Penicillium.
This bacterial strain Si Shi mould (Penicillium ateckii) WEQE is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on November 5th, 2009 and (is called for short CGMCC, address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number is CGMCC No.3401.
Embodiment 2, utilize Si Shi mould (Penicillium ateckii) WEQE CGMCC № 3401 preparation podophyllotoxin and astragalin
One, cultivates bacterial strain
The potato glucose liquid nutrient medium that 100ml is prepared is loaded in the 250ml triangular flask, sterilization, single colony inoculation of picking Si Shi mould (Penicillium ateckii) WEQE CGMCC № 3401 is in the good potato glucose liquid nutrient medium of above-mentioned sterilization, 28 ℃, the 120r/min shaking table was cultivated 7 days, obtained fermented liquid.
The potato glucose liquid nutrient medium is prepared as follows: take by weighing potato 200g and (remove the peel, be cut into about 2cm
2Fritter), put into water and boil 30min, filter with double gauze then, get its filtrate and add 20g glucose, supply water again to 1000mL, natural pH.
3 repetitions are established in experiment.
Two, the extraction of product
With fermented liquid suction filtration twice, collect mycelium and fermented liquid supernatant with suction filter respectively.Fermented liquid supernatant is used isopyknic chloroform extraction respectively, get extraction liquid at any time and detect down, under UV-detector, do not have fluorescence until extraction liquid in Ultraviolet Detector, combining extraction liquid, underpressure distillation is reclaimed.Obtain the chloroform extract of fermented liquid supernatant, the extract sample carries out the detection of following steps three.
3 repetitions are established in experiment, and the result takes the mean.
Three, the high performance liquid chromatography of product (HPLC) detects
The standard substance podophyllotoxin is available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, and catalog number is 111645;
The standard substance astragalin is separated from plant China unmrellaleaf according to method in the following document and is obtained: Liu Chang, the unmrellaleaf chemical composition analysis research [master's Diplomarbeit] of medicinal plant China, NMR data refer document: Feng Yulin, Li Yunqiu, Xu Lizhen, Yang Shilin, the chemical constitution study of Flower of Hollyhock (II)---flavones ingredient research herbal medicine, 2006,37 (11): 1622-1624[annotates: astragalin and Herba Astragali Melilotoidis (Herba Astragali Sinici) glycosides are with a kind of material].The standard substance astragalin also can be available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.
Before the detection, get the chloroform extraction matter sample of podophyllotoxin standard substance, astragalin standard substance, fermented liquid supernatant, fully dissolve with methyl alcohol respectively, 0.45 μ m filtering with microporous membrane; Adopting external standard method that sample is carried out HPLC analyzes.Each sample is triplicate at least.
Concrete detection method is as follows:
Instrument: Aglient 1100 type high performance liquid chromatographs;
Chromatographic column: Aglient 1100 Eclipse XDB-C
18(5 μ m, 4.6 * 150mm);
Detector: UV-detector; Detect wavelength: 254nm;
Chromatographic condition:
Moving phase: methyl alcohol: water=60: 40 (volume ratio)
Flow velocity: 0.5ml/min
Sample size: 5 μ l
Column temperature: room temperature
Standard substance: podophyllotoxin, astragalin.
Three repetitions are established in experiment, and the result is shown in Fig. 3-5.Wherein, Fig. 3 is the HPLC spectrogram of podophyllotoxin standard substance, and Fig. 4 is the HPLC spectrogram of astragalin standard substance, and Fig. 5 is the HPLC spectrogram of the chloroform extraction matter sample of Si Shi mould (Penicillium ateckii) WEQE CGMCC № 3401 fermented liquid supernatant.The retention time of podophyllotoxin is 8.336 minutes.The retention time of astragalin is 4.965 minutes.
The HPLC spectrogram of the chloroform extraction matter sample of Si Shi mould (Penicillium ateckii) WEQE CGMCC № 3401 fermented liquid supernatant is compared with the HPLC spectrogram of podophyllotoxin standard substance and astragalin standard substance respectively.
The result shows, when retention time is 8.306 minutes, have absorption peak in the HPLC spectrogram of the chloroform extraction matter sample of fermented liquid supernatant, and the podophyllotoxin standard substance had absorption peak in the time of 8.336 minutes, illustrate in the chloroform extraction matter sample of fermented liquid supernatant and contain podophyllotoxin; In the HPLC spectrogram of the chloroform extraction matter sample of fermented liquid supernatant, when retention time is 4.960 minutes, have absorption peak, and the astragalin standard substance had absorption peak in the time of 4.965 minutes, illustrate in the chloroform extract of fermented liquid supernatant and contain astragalin.Above result shows, contains podophyllotoxin and astragalin in the chloroform extract of fermented liquid supernatant.
Four, the thin-layer chromatography of product (TLC) detects
Standard substance are with accepted standard product in above-mentioned " high performance liquid chromatography of product (HPLC) detection ".
At first, get the chloroform extraction matter sample of podophyllotoxin standard substance, astragalin standard substance, Si Shi mould (Penicillium ateckii) WEQE CGMCC № 3401 fermented liquid supernatant, fully dissolve with methyl alcohol respectively, on positive and reversed phase thin layer plate, be total to chromatography then respectively, and under ultraviolet lamp, detect, and further use sulfuric acid-ethanol (15: 85, v/v) colour developing, color after the spot fluorescence color of observation sample and standard substance and the colour developing, and calculate the Rf value respectively.
Concrete testing conditions is as follows:
The positive thin-layer chromatography:
Chloroform layer extract developping agent: chloroform: methyl alcohol=80: 20 (v/v)
Inverse thin layer chromatography:
Chloroform layer extract developping agent: methyl alcohol: water=60: 40 (v/v)
Ultraviolet detection wavelength: 254nm.
The result shows, on the normal-phase chromatography plate, the Rf value of podophyllotoxin standard substance is 0.85, and same in the chloroform extraction matter sample of fermented liquid supernatant to have a Rf value be 0.85 spot, its spot has identical fluorescence color with the podophyllotoxin standard substance under ultraviolet lamp (254nm), and with sulfuric acid-ethanol (15: 85, v/v) colour developing after, also have identical color, illustrate in the chloroform extraction matter sample of fermented liquid supernatant and contain podophyllotoxin; The Rf value of astragalin standard substance is 0.30, and same in the chloroform extraction matter sample of fermented liquid supernatant to have a Rf value be 0.30 spot, its spot has identical fluorescence color with the podophyllotoxin standard substance under ultraviolet lamp (254nm), and with sulfuric acid-ethanol (15: 85, v/v) after the colour developing, also have identical color, illustrate in the chloroform extraction matter sample of fermented liquid supernatant and contain astragalin.
On the reversed phase chromatography plate, the Rf value of podophyllotoxin standard substance is 0.32, and same in the chloroform extraction matter sample of fermented liquid supernatant to have a Rf value be 0.32 spot, its spot has identical fluorescence color with the podophyllotoxin standard substance under ultraviolet lamp (254nm), and with sulfuric acid-ethanol (15: 85, v/v) after the colour developing, also have identical color, illustrate in the chloroform extraction matter sample of fermented liquid supernatant and contain podophyllotoxin; The Rf value of astragalin standard substance is 0.58, and same in the chloroform extraction matter sample of fermented liquid supernatant to have a Rf value be 0.58 spot, its spot has identical fluorescence color with the podophyllotoxin standard substance under ultraviolet lamp (254nm), and with sulfuric acid-ethanol (15: 85, v/v) after the colour developing, also have identical color, illustrate in the chloroform extraction matter sample of fermented liquid supernatant and contain astragalin.
Claims (9)
1. Si Shi mould (Penicillium ateckii) WEQE, its preserving number is CGMCC No.3401.
2. cultivating the method for Si Shi mould (Penicillium ateckii) WEQE CGMCC No.3401, is with potato glucose culture medium culturing Si Shi mould (Penicillium ateckii) WEQE CGMCC No.3401.
3. method according to claim 2 is characterized in that: described culture condition is: 26-30 ℃, 100-150rpm vibration.
4. a method for preparing podophyllotoxin and/or astragalin comprises the steps: fermentation culture Si Shi mould (Penicillium ateckii) WEQE CGMCC No.3401, obtains podophyllotoxin and/or astragalin.
5. method according to claim 4 is characterized in that: the substratum of using in the described fermentation culture is the potato glucose substratum.
6. according to claim 4 or 5 described methods, it is characterized in that: the condition of described fermentation culture is: 26-30 ℃, 100-150rpm shaking culture 5-9 days.
7. according to arbitrary described method among the claim 4-6, it is characterized in that: after described fermentation, also comprise the step of extraction among the described preparation method.
8. method according to claim 7 is characterized in that: described extraction agent is a chloroform.
9. the application of Si Shi mould (Penicillium ateckii) WEQE CGMCC No.3401 in preparation podophyllotoxin and/or astragalin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102377904A CN101701193B (en) | 2009-11-19 | 2009-11-19 | Method for preparing podophyllotoxin and/or astragaloside and special bacterial strain thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102377904A CN101701193B (en) | 2009-11-19 | 2009-11-19 | Method for preparing podophyllotoxin and/or astragaloside and special bacterial strain thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101701193A true CN101701193A (en) | 2010-05-05 |
| CN101701193B CN101701193B (en) | 2011-09-21 |
Family
ID=42156120
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009102377904A Expired - Fee Related CN101701193B (en) | 2009-11-19 | 2009-11-19 | Method for preparing podophyllotoxin and/or astragaloside and special bacterial strain thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101701193B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102168021A (en) * | 2010-12-28 | 2011-08-31 | 北京师范大学 | Diphyllria sinensis L.-derived endophytic fungus Geotrichum candidum and application thereof |
| CN102168022A (en) * | 2010-12-28 | 2011-08-31 | 北京师范大学 | Endophytic fungus Penicillium ateckii from plant Chinese Umbrellaleaf rhizome and application thereof |
-
2009
- 2009-11-19 CN CN2009102377904A patent/CN101701193B/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102168021A (en) * | 2010-12-28 | 2011-08-31 | 北京师范大学 | Diphyllria sinensis L.-derived endophytic fungus Geotrichum candidum and application thereof |
| CN102168022A (en) * | 2010-12-28 | 2011-08-31 | 北京师范大学 | Endophytic fungus Penicillium ateckii from plant Chinese Umbrellaleaf rhizome and application thereof |
| CN102168021B (en) * | 2010-12-28 | 2012-05-23 | 北京师范大学 | Diphyllria sinensis L.-derived endophytic fungus Geotrichum candidum and application thereof |
| CN102168022B (en) * | 2010-12-28 | 2012-05-23 | 北京师范大学 | Endophytic fungus Penicillium ateckii from plant Chinese Umbrellaleaf rhizome and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101701193B (en) | 2011-09-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102168022B (en) | Endophytic fungus Penicillium ateckii from plant Chinese Umbrellaleaf rhizome and application thereof | |
| CN103952362B (en) | One strain is to the oranges and tangerines endogeny rayungus of various plants pathogenic bacteria tool bacteriostatic activity | |
| CN102676392B (en) | Endophytic fungus in salvia miltiorrhiza bunge and application thereof | |
| CN104212725B (en) | Improve rhodioside and the method for butyl alcohol content in Radix Rhodiolae tissue cultured seedling | |
| CN105886405A (en) | Dendrobium officinale Kimura et Migo endophytic fungus and applications thereof | |
| CN104871821A (en) | Phellinus igniarius strain and culturing method for same | |
| CN106085945B (en) | Method for inducing conidia of panax notoginseng orbicularis | |
| CN101575576B (en) | Aspergillus fumigatus and application thereof | |
| CN108102928B (en) | One plant of gingko endogenous fungus and its application | |
| CN101603011B (en) | Method for preparing epipodophyllotoxin diglucoside and special strain used thereby | |
| CN101701193B (en) | Method for preparing podophyllotoxin and/or astragaloside and special bacterial strain thereof | |
| CN102757903B (en) | Endophytic fungi for producing flavones and application thereof | |
| CN103820330B (en) | One strain variable color aspergillus and preparing the application in podophyllotoxin list glucoside and podophyllotoxin | |
| CN103820329B (en) | One strain gibberella and the application in preparation rutin and Quercetin thereof | |
| CN102154123A (en) | Dioscorea nipponica Makino Fusarium sp. and application thereof | |
| CN102168021B (en) | Diphyllria sinensis L.-derived endophytic fungus Geotrichum candidum and application thereof | |
| AU2020102894A4 (en) | A strain of Aspergillus versicolor and its application in the preparation of podophyllotoxin monoglucose and podophyllotoxin | |
| CN110511876B (en) | Epimedium koreanum endophyte, culture method and metabolite thereof | |
| CN103789212B (en) | One plant height produces spore Cordyceps strain | |
| CN102943103B (en) | Penicillium fungus M1 and application thereof to increase of saponins yield in fermentation process of ginseng or Americginseng | |
| CN106282032A (en) | Aspergillus citrimum LB and prepare the application in phillygenol at bioconversion phillyrin | |
| CN102559517B (en) | Endophytic fungi from podophyllum hexandrum plant and application of endophytic fungi | |
| CN103820328B (en) | One strain flavus and the application in preparation rutin and podophyllotoxin list glucoside thereof | |
| CN113773967B (en) | Alternaria tenuissima and application thereof | |
| CN102660466B (en) | Endophytic fungi for improving content of main active ingredients of schisandra chinensis through fermentation method |
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: 20110921 Termination date: 20181119 |