CN108676731A - Raw curved neck trichoderma strain a kind ofly - Google Patents
Raw curved neck trichoderma strain a kind ofly Download PDFInfo
- Publication number
- CN108676731A CN108676731A CN201810547950.4A CN201810547950A CN108676731A CN 108676731 A CN108676731 A CN 108676731A CN 201810547950 A CN201810547950 A CN 201810547950A CN 108676731 A CN108676731 A CN 108676731A
- Authority
- CN
- China
- Prior art keywords
- cyclosporin
- curved neck
- trichoderma strain
- culture
- yield
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/64—Cyclic peptides containing only normal peptide links
- C07K7/645—Cyclosporins; Related peptides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
Abstract
The present invention relates to a kind of raw curved neck trichoderma strain, belongs to microbial secondary metabolite application field.Curved neck trichoderma strain (Tolypocladium geodes) C41 3 is given birth on ground, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC NO15667.Bacterial strain of the present invention is that Dalian ethnic university fungal studies laboratory is detached from Conifer Forest at North Slope of Changbai Mountain forest soil, can be grown within the scope of 5 DEG C 30 DEG C, and sporulation quantity is maximum at 25 DEG C, can be grown in 9 ranges of pH 4 to pH.Optimum growth temperature is 25 DEG C, and the pH value of the most suitable growth is 6, which has the characteristics that high yield cyclosporin.
Description
Technical field
The invention belongs to microbial secondary metabolite application fields, give birth to curved neck trichoderma strain more specifically a kind ofly.
Background technology
The cyclic peptide that immunosuppressor cyclosporin A (CyclosporineA, CsA) is made of 11 amino acid.Its molecule
Amount is 1202, wherein containing there are one form the amino acid containing ethylene double bond by 9 carbon.By NMR (nuclear magnetic resonance measuring) and MS (matter
Amount spectrometry) measurement result infer its molecular formula be C62H111N11O12。
Cyclosporin A does not have direct inhibiting effect, this point resistance to transplanting B cell and macrophage proliferation and function
Formation by property is meaningful.Existing research shows that when small molecule immune inhibitor is used in combination, the identical medicine of action site
Object is used in combination has antagonism to curative effect, and toxicity increases, and the different drug of mechanism of action is used in combination, then can drop
Hypotoxicity and raising curative effect.
Anti- rejection when the immunosuppressive action of cyclosporin A makes it be mainly used for organ transplant and the anti-place of graft
The treatment of main disease (GVHD).With the continuous understanding to cyclosporin A mechanism of action, and the pathogenesis of clinical certain diseases
It is continuous clear, cyclosporin A has been not only applicable to anti-rejection at present.In recent years, the medicine is related in treatment immune system
Disease in terms of, the evaluation especially also affirmed in terms of autoimmune disease.
Invention content
To make up the defect that cyclosporin A in the prior art underproduces, the present invention provides raw curved neck trichoderma strain a kind ofly,
Curved neck trichoderma strain (Tolypocladium geodes) C41-3 is given birth on ground, is preserved in China Committee for Culture Collection of Microorganisms
Common micro-organisms center, deposit number are CGMCC NO15667, and the deposit date is on April 24th, 2018, the ground of depositary institution
Location is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
The present invention gives birth to the curved mould C41-3 of neck with providing bacterial strain, northern from Changbai Mountain by Dalian ethnic university fungal studies laboratory
It detaches in the forest soil of slope, can be grown within the scope of 5 DEG C -30 DEG C, sporulation quantity is maximum at 25 DEG C, equal energy in 9 ranges of pH 4 to pH
Growth.Optimum growth temperature is 25 DEG C, and the pH value of the most suitable growth is 6.
The method that cyclosporin is prepared using bacterial strain of the present invention is as follows：
(1) the raw curved neck mould C41-3 in activation ground：Curved neck mould C41-3 is given birth on ground to be seeded in the PDA culture medium of sterilizing,
It is placed in 25 DEG C of room temperature incubator, is cultivated 7 days；
The PDA culture medium formula is：Potato 100g, glucose 10g, agar 10g, water 500mL；
(2) seed liquor is prepared：Curved neck mould C41-3 is given birth on ground after activation to be placed in primary liquid culture medium, 28 DEG C,
120r/min cultures 5d obtains seed liquor；
The primary liquid culture medium prescription is：Glucose 50g, peptone 10g, KNO32g, MgSO42g, CaCl2
0.1g, (NH4)3PO4·3H2O 2g, water 1000mL, liquid amount 2/5；Liquid amount of the present invention is that culture fiduciary point contains training
Support the volume ratio of based containers；
(3) zymotic fluid is prepared：Seed liquor is inoculated in by the inoculum concentration of 8vt% in Cha Shi fluid nutrient mediums, 24 DEG C,
Shaking table culture 5d obtains zymotic fluid under the conditions of 120r/min；
The formula of the Cha Shi fluid nutrient mediums is sucrose 30g, NaNO33g, K2HPO41g, MgSO40.5g, KCl
0.5g, FeSO40.01g, water 1000mL, pH 5.0；The liquid amount of the Cha Shi fluid nutrient mediums is 2/5；
(4) preparation of cyclosporin：
Isometric ethyl acetate is added in the zymotic fluid prepared to step (3), zymotic fluid and mycelium are all impregnated,
Under the conditions of 24 DEG C, 120r/min shaking table culture for 24 hours after, ultrasonic vibration 30min, stand after its layering after take out upper organic phase
To sterile conical flask, stand, filter, be concentrated to give cyclosporin, when this step operation, can by zymotic fluid and mycelium by several times plus
Enter ethyl acetate, then the organic phase that separating for several times is obtained merges.Cyclosporin of the present invention is cyclosporin A.
Beneficial effects of the present invention are as follows：Curved neck trichoderma strain (Tolypocladium geodes) C41-3 is given birth on ground, this is true
Bacterium is to like cold fungi, and the secondary metabolite cyclosporin A generated is widely applied to field of medicaments and field of biological pesticide, this
To solving cyclosporin A, supply falls short of demand contributes for invention, probe into obtain a kind of high yield cyclosporin A high producing microbial and
Optimal medium and growth regulating, and cyclosporin A is extracted from zymotic fluid, with ethyl acetate by fermentate (fermentation when extraction
Liquid and mycelium) be completely soaked after, under the conditions of 24 DEG C, 120r/min shaking table culture for 24 hours after, ultrasonic vibration 30min is carried significantly
The high recovery rate of cyclosporin.
Description of the drawings
Fig. 1 is the standard curve of the present invention；
Fig. 2 is influence of the different culture media to cyclosporin yield；
Fig. 3 is influences of the initial pH of culture medium to cyclosporin yield；
Fig. 4 is influence of the inoculum concentration to cyclosporin yield；
Fig. 5 is influence of the incubation time to cyclosporin yield；
Fig. 6 is influence of the liquid amount to cyclosporin yield；
Fig. 7 is influence of the cultivation temperature to cyclosporin yield.
Specific implementation mode
The present invention is described further With reference to embodiment, if without specified otherwise, it is former used in the present invention
Material and equipment are ordinary skill in the art means.
Embodiment 1
Curved neck trichoderma strain (Tolypocladium geodes) C41-3 is given birth on ground, is preserved in Chinese microorganism strain preservation pipe
Reason committee common micro-organisms center, deposit number are CGMCC NO15667.
(1) the raw curved neck mould C41-3 in activation ground：The mould C41-3 mother strains of curved neck are given birth on ground, choose female bacterium with the toothpick of sterilizing
Mycelia is seeded in the PDA culture medium of sterilizing, is inoculated into respectively on the culture dish of 5 sterilizings, is placed in 25 DEG C of room temperature culture
Case is cultivated 7 days；
The PDA culture medium formula is：Potato 100g, glucose 10g, agar 10g, water 500mL；
(2) seed liquor is prepared：Curved neck mould C41-3 is given birth in super-clean bench in ground after activation, with the sterilizing of 6cm and uses alcohol
The card punch of lights flame calcination makes a call to three bacteria cakes, is put into the sterilized primary liquid culture mediums of 200mL with the tweezers of sterilizing,
Seed liquor is obtained in 28 DEG C, 120r/min cultures 5d；
The primary liquid culture medium prescription is：Glucose 50g, peptone 10g, KNO32g, MgSO42g, CaCl2
0.1g, (NH4)3PO4·3H2O 2g, water 1000mL, liquid amount 2/5；
(3) prepared by zymotic fluid：Seed liquor is inoculated in by the inoculum concentration of 8vt% in Cha Shi fluid nutrient mediums, 24 DEG C,
Shaking table culture 5d obtains zymotic fluid under the conditions of 120r/min；
The formula of the Cha Shi fluid nutrient mediums is sucrose 30g, NaNO33g, K2HPO41g, MgSO40.5g, KCl
0.5g, FeSO40.01g, water 1000mL, pH 5.0；The liquid amount of the Cha Shi fluid nutrient mediums is 2/5；
(4) preparation of cyclosporin：
A. mycelium is shredded with knife；
B. isometric ethyl acetate is added into zymotic fluid, ensures that zymotic fluid and mycelium are completely soaked；
C. it after being completely soaked zymotic fluid with ethyl acetate, is sealed with aluminium foil (or sealed membrane), in 24 DEG C, 120r/min
Under the conditions of shaking table culture for 24 hours after, in (28 DEG C) of room temperature ultrasonic wave assisted extraction 30min, stand and take out upper layer after its layering
To sterile conical flask, this extraction step is repeated 6 times organic phase, collects 6 organic phases；
D. it stands, waits for zymotic fluid and the layering of organic phase ethyl acetate, with triangular funnel, it is organic that Medium speed filter paper filters upper layer
Excessive anhydrous magnesium sulfate drying is added in filtrate for phase, and not lumpd with anhydrous slufuric acid magnesium dust is advisable, and collects organic phase (3-5 times)；
E. by the organic phase of collection through reduced pressure (bath temperature≤40 DEG C)；
F. dry up, weigh then Cord blood (2-8 DEG C) up to cyclosporin.
Influence of 1 different culture media of comparative example to cyclosporin yield
8 kinds of different culture media formulas：
Original culture medium (C8)：Glucose 50g, peptone 10g, KNO32g, MgSO42g, CaCl20.1g, (NH4)3PO4·3H2O 2g, water 1000mL；
Potato dextrose medium (PD)：Potato 200g, glucose 20g, water 1000m L；
Potato protein peptone dextrose culture-medium (PPD)：Potato 200g, glucose 20g, peptone 10g, water
1000mL；
Potato dextrose yeast powder culture (PDY)：Potato 200g, glucose 20g, yeast powder 10g, water 1000mL；
Sabouraud medium (SDY)：Peptone 10g, glucose 40g, yeast powder 10g, water 1000m L；
Sa Shi malt sugar culture-medium (SMY)：Peptone 10g, maltose 40g, yeast powder 10g, water 1000mL；
Broth bouillon (LB)：Peptone 10gNaCl 5g, yeast extract 3g, sucrose 20g, water 1000mL；
Czapek's medium (CZ)：Sucrose 30g, NaNO33g, K2HPO41g, MgSO40.5g, KCl 0.5g, FeSO4
0.01g, water 1000m L.
The cultural method of this comparative example is the same as embodiment 1.
The assay method of cyclosporin is specific as follows：
Cyclosporin standard items are diluted to a concentration of 5,10,20,40,80 μ g/mL successively with the methanol of chromatographic grade, are passed through
Up to standard serial solution after 0.22 μm of membrane filtration.
Draw standard curve：Cyclosporin content, mobile phase methanol are surveyed using high performance liquid chromatography：Pure water=85：
15, flow velocity 0.5mL/min, 20 μ L of sample size, 60 DEG C of column temperature, Detection wavelength 210nm.With the concentration x of cyclosporin standard items
For abscissa, corresponding peak area y is ordinate, draws standard curve as shown in Figure 1, obtained linear fit equation is： y
=16.03x+352.39, R=0.9874 shows that cyclosporin linear relationship in the range of 5-80ug is good.Thus linear to return
Return the content of equation calculation sample cyclosporin.
The measurement of zymotic fluid cyclosporin content：8 kinds of zymotic fluids pipette 100 μ L respectively after centrifugal filtration, by
BR5.5-MeOH buffer solutions are diluted to 1.0mL, after crossing 0.22 μm of filter membrane, are measured using above-mentioned chromatographic condition, according to above-mentioned
The calculation formula that gained standard curve provides is calculated separately the zymotic fluid cyclosporin obtained using 8 kinds of different culture medias and contained
Amount, the results are shown in Figure 2.
As shown in Fig. 2, in 8 kinds of liquid fermentation mediums, when using Czapek's medium as fermentation medium, cyclosporin yield
Highest is 2066.86ug/mL；Secondly it is C8 culture mediums, cyclosporin content is 1766.95ug/mL；PD culture mediums yield is most
Low, content is only 99.84ug/mL.The result shows that relative to other 7 kinds of culture mediums, the Czapek's medium that the present invention uses is imitated
Fruit is best.
Influences of the 2 difference pH of comparative example to cyclosporin yield
The preparation of this comparative example Spawn incubation method and cyclosporin, assay method with comparative example 1, differ only in
The pH of Czapek's medium is different：
The formula of the Cha Shi fluid nutrient mediums is sucrose 30g, NaNO33g, K2HPO41g, MgSO40.5g, KCl
0.5g, FeSO40.01g, water 1000mL, pH 4,5,6,7 or 8；The liquid amount of the Cha Shi fluid nutrient mediums is 2/5；
The yield of cyclosporin is as shown in figure 3, as seen from Figure 3 when the initial pH of culture medium is 4-5, zymotic fluid middle ring
For spore rhzomorph content between 2027.6-2083.7ug/mL, content is low；When pH is 6.0, cyclosporin content reaches
4369.53ug/mL；When pH value is more than 6.0, content is begun to decline.Illustrate generation of the environmental benefits in cyclosporin of weak acid,
And cross the generation that alkali condition is unfavorable for cyclosporin；Effect is best when initial pH is 5.0.
Influence of 3 inoculum concentration of comparative example to cyclosporin yield
The preparation of this comparative example Spawn incubation method and cyclosporin, assay method with comparative example 1, differ only in
Inoculum concentration is different, and inoculum concentration 2vt%, 4vt%, 6vt%, 8vt%, 10vt%, the results are shown in Figure 4, as can be seen from Figure 4 when
When inoculum concentration is 2vt%-4vt%, zymotic fluid cyclosporin content is between 2987.42-3099.91ug/mL；Work as inoculum concentration
Content is 3781.54ug/mL when being 6%, and cyclosporin content is 2954.27- when inoculum concentration is 8vt%-10vt%
2743.02ug/mL increases with inoculum concentration, therefore cyclosporin yield reduction trend occurs, and inoculum concentration of the present invention is imitated for 8vt%
Fruit is best.
Influence of 4 incubation time of comparative example to cyclosporin yield
The preparation of this comparative example Spawn incubation method and cyclosporin, assay method with comparative example 1, differ only in
Incubation time is different.
The results are shown in Figure 5, when fermented incubation time is 3d, predominantly vegetative growth phase of thalline, and zymotic fluid middle ring
Spore rhzomorph content gradually rises, and from 944.903ug/mL, when incubation time is 4d, content reaches maximum value 2904.43ug/mL,
Later, thalline starts to move towards decline, and secondary metabolite gradually decomposes, and zymotic fluid cyclosporin content starts to reduce, to the 7th
It when content increase a little.Therefore, most suitable incubation time is 5d.
Influence of 5 liquid amount of comparative example to cyclosporin yield
The preparation of this comparative example Spawn incubation method and cyclosporin, assay method with comparative example 1, differ only in
The liquid amount of Czapek's medium is different, and conical flask volume is 500mL.Liquid amount be 50mL, 100mL, 150mL, 200mL,
250mL, the results are shown in Figure 6, and under the same conditions, when 500mL conical flask liquid amounts are 50mL, cyclosporin yield is
4587.04ug/mL, zymotic fluid are in light brown, and mycelium is in spherical, is relatively large in diameter；When liquid amount is 100mL, mycelium
In spherical, cyclosporin yield reaches up to 4706.05ug/mL；Zymotic fluid takes on a red color；Hereafter with the gradually increasing of liquid amount
Add, mycelium starts autolysis occur；When liquid amount is 250mL, cyclosporin content is low, only 2255.37ug/mL.
Illustrate that the generation of thalline own growth and cyclosporin has higher requirement to ventilatory capacity.Therefore, optimum liquid amount is
2/5。
Influence of 6 cultivation temperature of comparative example to cyclosporin yield
The preparation of this comparative example Spawn incubation method and cyclosporin, assay method with comparative example 1, differ only in
Cultivation temperature is different.Cultivation temperature is 20 DEG C, 24 DEG C, 28 DEG C, 32 DEG C, 36 DEG C, when temperature is 20 DEG C, cyclosporin yield
Up to 4277.85ug/mL, mycelium is in spherical；Then, the content of cyclosporin with temperature a degree of reduction of increase.
When temperature increases to 32 DEG C, cyclosporin yield starts to reduce, cyclosporin content be 2210.8ug/mL, more than 32 DEG C it
Afterwards, mycelial growth is suppressed, and ground is given birth to the mould metabolite cyclosporin of curved neck and reduced, and zymotic fluid cyclosporin yield
It is extremely low.Therefore, the optimum temperature of culture is 24 DEG C.
79 groups of Experimental Comparisons of comparative example
According to the following table 1 design experiment, other condition of culture and cyclosporin preparation, assay method are the same as comparative example 1, experiment knot
Fruit such as the following table 1.Incubation time provided by the invention 5 days, inoculum concentration 8vt%, the initial pH of culture medium are 5 as can be seen from Table 1, training
It supports the ground that temperature is 24 DEG C of cultures and gives birth to curved neck mould C41-3, cyclosporin content highest.
Table 1
Above-described embodiment is only intended to the citing to the present invention and explanation, and is not intended to limit the invention to described
In scope of embodiments.In addition it will be appreciated by persons skilled in the art that the present invention is not limited to the above embodiment, according to this hair
Bright introduction can also make more kinds of variants and modifications, these variants and modifications all fall within present invention model claimed
In enclosing.
Claims (4)
1. raw curved neck trichoderma strain a kind ofly, which is characterized in that for raw curved neck mould (Tolypocladium geodes) C41- in ground
3, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC NO.15667.
2. raw curved neck trichoderma strain as described in claim 1, which is characterized in that growth temperature is 25 DEG C.
3. raw curved neck trichoderma strain as described in claim 1, which is characterized in that growth pH is 6.
4. a kind of cyclosporin prepared by according to claim 1ly raw curved neck trichoderma strain.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810547950.4A CN108676731B (en) | 2018-05-31 | 2018-05-31 | Germination tropicalis strain |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810547950.4A CN108676731B (en) | 2018-05-31 | 2018-05-31 | Germination tropicalis strain |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108676731A true CN108676731A (en) | 2018-10-19 |
| CN108676731B CN108676731B (en) | 2021-02-23 |
Family
ID=63809352
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810547950.4A Active CN108676731B (en) | 2018-05-31 | 2018-05-31 | Germination tropicalis strain |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108676731B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017151684A1 (en) * | 2016-03-01 | 2017-09-08 | Sustainable Bioproducts Holdings, Llc | Filamentous fungal biomats, methods of their production and methods of their use |
-
2018
- 2018-05-31 CN CN201810547950.4A patent/CN108676731B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017151684A1 (en) * | 2016-03-01 | 2017-09-08 | Sustainable Bioproducts Holdings, Llc | Filamentous fungal biomats, methods of their production and methods of their use |
Non-Patent Citations (2)
| Title |
|---|
| KEBEDE B等: "Establishing the Secondary Metabolite Profile of the Marine Fungus: Tolypocladium geodes sp MF458 and Subsequent Optimisation of Bioactive Secondary Metabolite Production", 《MARINE DRUGS》 * |
| 姜娃等: "地生弯颈霉的培养与生长条件", 《菌物研究》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108676731B (en) | 2021-02-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107245457B (en) | The extracting method and application of the exocellular polysaccharide and the exocellular polysaccharide of a kind of dendrobium candidum endogenetic fungal bacterial strain and its generation | |
| LU501210B1 (en) | Streptomyces antibioticus, preparation of metabolite thereof, and use thereof in antibacterial aspect | |
| CN102242069B (en) | Paecilomycescicadae (Miq.)Samson and application thereof | |
| CN102676392B (en) | Endophytic fungus in salvia miltiorrhiza bunge and application thereof | |
| CN107904177A (en) | A kind of extracting method and application of dendrobium candidum endogenetic fungal bacterial strain and its exocellular polysaccharide of generation and the exocellular polysaccharide | |
| CN110093283B (en) | Beauveria bassiana strain and culture method thereof | |
| CN103484500B (en) | Bacterial CD-126 fermentation solution and application thereof | |
| CN105779299A (en) | Paecilomyces hepialid strain capable of realizing high yield of adenosine and mannite type substances and application | |
| CN108841889B (en) | Method for producing griseofulvin serving as major component of tranexamycin by microbial fermentation | |
| CN108823110B (en) | Strain for producing griseofulvin and application thereof | |
| CN105331548B (en) | A kind of Lepista mucla (Bull.:Fr.) Cooke bacterial strain and its liquid spawn and preparation method | |
| CN108315265B (en) | Aspergillus versicolor Av-2 strain and application thereof | |
| CN104611236B (en) | Cunninghamella echinulata FAR3 and fermentation thereof are for the method for gamma-linolenic acid oil | |
| CN102321545B (en) | Endophytic fungus for producing triptolide | |
| CN103283482B (en) | Cordyceps militaris, cultivation method and separation method | |
| CN110257260A (en) | A kind of Rhizoma Atractylodis Macrocephalae endogenetic fungus and its application | |
| CN108676731A (en) | Raw curved neck trichoderma strain a kind ofly | |
| CN108727473A (en) | A method of preparing cyclosporin using microorganism | |
| CN109055242A (en) | A kind of monascus purpureus bacterial strain and its zymotechnique and application | |
| CN105695335A (en) | New aspergillus flavus and use thereof | |
| CN100392062C (en) | Short dense Penicillium and application | |
| CN101280333B (en) | Method for preparing penicillium antibacterial peptide from grey rose penicillium | |
| CN101845396B (en) | Radiation-resistant fungus and crude red pigment preparation prepared therefrom | |
| CN106754430A (en) | Dendritic bacterial strains of cladosporium Cc 106 of high yield monascus purpureus and application thereof | |
| CN106479900A (en) | 25 bacterial strain of high yield monascus purpureus penicillium oxalicum Po and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |