CN103173390B - Chromobacterium violaceum strain and application thereof - Google Patents
Chromobacterium violaceum strain and application thereof Download PDFInfo
- Publication number
- CN103173390B CN103173390B CN201310087484.3A CN201310087484A CN103173390B CN 103173390 B CN103173390 B CN 103173390B CN 201310087484 A CN201310087484 A CN 201310087484A CN 103173390 B CN103173390 B CN 103173390B
- Authority
- CN
- China
- Prior art keywords
- chromobacterium
- cgmcc
- bacterial strain
- strain
- compound
- 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.)
- Active
Links
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a new chromobacterium violaceum strain and application thereof. The new chromobacterium violaceum strain is called as Chromobacterium sp. HS-13-94, and the preservation No. is CGMCC (china general microbiological culture collection center) No.6247. The Chromobacterium sp. HS-13-94 disclosed by the invention can generate disulfide-bond depsipeptide type compound as an active substance, the compound has great inhibition effect in tumor cells such as lung cancer epithelial cell A549 and HepG (hepatoma carcinoma cell) of human body and has the characteristic of wide antitumor spectrum, and excellent business development prospect is shown.
Description
Technical field
The present invention relates to microbial engineering field, relate in particular to a strain chromobacterium and contain the application in the depsipeptide compounds of cystine linkage in preparation.
Background technology
Malignant tumour is one of serious disease threatening human health.For a long time, oncotherapy is the key subjects of medical circle research always, and the new drug of searching treatment malignant tumour is pharmaceuticals researcher's top priority especially.In antitumor drug, the medicine with optionally anticancer growth of research and development non-cytotoxicity is the research emphasis of current antitumor drug.In recent years, the dependent metalloprotease of a Zn-like ions---histone deacetylase (histone deacetylases, HDACs) becomes the focus of antitumor drug research.HDACs has participated in the adjusting of many functional protein activity in cell, as: histone, tumor-inhibiting factor p53, heat shock protein 90 (HSP90), alpha-tubulin etc.These protein and apoptosis, tumour occur and shift closely relatedly, and in a lot of tumour cells, the expression level of HDACs is all very high, in the propagation of cancer cells, in shifting, play an important role.The HDACs of take has become an available strategy of exploitation wide spectrum non-cytotoxicity antitumor drug as the micromolecular inhibitor of shot design (HDACi).
In US4977138, disclosing chromobacterium (Chromobacterium sp.) can ferment and produce a kind of depsipeptide anti-tumor compounds romidepsin (romidepsin).In its structure, distinctive disulfide linkage is the active crucial group of performance.Due to stable hydrophobic structure, romidepsin easily sees through the cytolemma of tumour, and in born of the same parents, its disulfide linkage is reduced into the effect of sulfydryl performance histone deacetylase (HDACs) inhibitor by gsh.Within 2004, U.S. FDA approval romidepsin is used for the treatment of skin T-cell lymphoma (CTCL).The present invention is from a kind of new chromobacterium bacterial strain HS-13-94 (Chromobacterium sp.HS-13-94, on June 21st, 2012, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, be called for short CGMCC, deposit number is CGMCC No.6247) fermented liquid in isolate a kind of depsipeptide anti-tumor compounds, Spectrum Analysis proof is basic identical with romidepsin.
Summary of the invention
One of object of the present invention is to provide a kind of new chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94), has the ability of producing containing the depsipeptide compounds of cystine linkage, and its deposit number is CGMCC NO.6247.
The present invention also aims to provide a kind of chromobacterium HS-13-94 (Chromobacterium sp.) (CGMCC NO.6247) being prepared as follows shown in formula I containing the application in the depsipeptide compounds of cystine linkage,
Formula I.
The present invention also provides the method for fermentative production and separation and purification formula I compound.Fermented liquid is prepared this compound by steps such as extraction, concentrated, silicagel column wash-out, crystallizations.
The separated formula I compound obtaining from chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94) (CGMCC NO.6247) bacterial strain fermentation liquor, through MS,
1h-NMR and
13the Spectrum Analysis such as C-NMR, determine it is that a kind of molecular weight is 540.98 depsipeptide compounds.This material is carried out to anti tumor activity in vitro detection, find that it has anti-tumor activity.
The present invention also comprises the secondary metabolite that chromobacterium bacterial strain produces, and it has very strong anti-tumor activity in vitro, and kinds of tumor cells is had to good inhibition, as the IC to Human Lung Cancer epithelial cell A549
50for 3.8nM, the IC to liver cancer cell HepG
50for 5.5nM.
The biological characteristics of this bacterial strain is colonial morphology milk yellow, and circle is smooth, has an even surface, moistening, easily picking; Particularly gelatine liquefication negative (-) in biochemical reactions, Starch Hydrolysis positive (+), milk is solidified as the positive (+), and iced milk turns to the positive (+).
This chromobacterium bacterial strain (called after chromobacterium HS-13-94, Chromobacterium sp.HS-13-94, CGMCC NO.6247) on June 21st, 2012, being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (is called for short: CGMCC), address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, preserving number is CGMCC No.6247, and register on the books, prove survival.
The feature of chromobacterium HS-13-94 provided by the invention (Chromobacterium sp.HS-13-94) (CGMCC NO.6247) on morphology, Physiology and biochemistry and molecular level, produce the morphology of bacterium with known romidepsin, the feature on Physiology and biochemistry compares, can assert that chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94) (CGMCC NO.6247) belongs to chromobacterium and belongs to, but with at document as US Patent No. 4977138; European patent EP 0352646; Microbiotic magazine The Journal of Antibiotics, 1994,47 (3): in 301-310, disclosed romidepsin produces bacterium difference, especially gelatine liquefication negative (-) in chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94) (CGMCC NO.6247) physiological and biochemical property, Starch Hydrolysis positive (+), milk is solidified as the positive (+), and iced milk turns to the positive (+).And general chromobacterium be characterized as gelatine liquefication positive (+), Starch Hydrolysis negative (-), milk is solidified as feminine gender (-), iced milk turns to feminine gender (-), proves that thus chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94) is a new bacterial strain.
Accompanying drawing explanation
Fig. 1 is the HPLC collection of illustrative plates of the antitumor formula I compound of the embodiment of the present invention 6 gained;
Fig. 2 is the ESI/MS collection of illustrative plates of the antitumor formula I compound of the embodiment of the present invention 6 gained;
Fig. 3 is the antitumor formula I compound of the embodiment of the present invention 6 gained
1h-NMR collection of illustrative plates;
Fig. 4 is the antitumor formula I compound of the embodiment of the present invention 6 gained
13c-NMR collection of illustrative plates.
Embodiment
Embodiment 1: bacterium source
Chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94, CGMCC No.6247) is the separated strain marine bacteria obtaining from Taizhou of Zhejiang Floor of The East China Sea mud sample.
In the region of delimiting in the East Sea, Taizhou, carry out ALTERNATE SAMPLING; get at random 5 sampling points; each point is got bottom silt sample 10g; put into Erlenmeyer flask; mix rear sample thief 10g, join in an Erlenmeyer flask that 90ml seawater is housed and (in bottle, have a magnetic stirring apparatus), vortex stirs 30min; it is fully mixed and make suspension liquid, be 10
-1bacterium liquid.By dilution spread flat band method, above-mentioned suspension liquid and sterilized water are diluted to 10 in 1: 9 by volume
-2, 10
-3, 10
-4, 10
-5concentration, gets different extension rate bacterium liquid 0.1ml, coats in beef-protein medium flat board, with aseptic painting rod, in media surface, is coated with gently, and under room temperature, standing 30min is placed on 30 ℃ of constant incubators.After bacterium colony grows, observed and recorded colony colour, transparency, bacterium colony surface, edge configuration, inspection is characterized in that no consistent.Finally choose 1000 strain primary dcreening operation bacterial classifications and be inoculated in beef-protein medium bevel, and the checking of fermenting.Thalline one ring with transfering loop picking slant culture, is inoculated into respectively (every bottle contains 30mL fermention medium) in 250ml Erlenmeyer flask, and under 30 ℃ of conditions, shaking culture is 2 days, and the standby survey of fermented liquid is lived.
The lung cancer epithelial cell of take has the marine microorganism of anti-tumor activity as target sieving.The above-mentioned fermented liquid of 5 μ l is added in 200 μ l lung cancer epithelial cell nutrient solutions (different time is got a blank) respectively, then put into 5%CO
2in incubator, cultivate 48h, by microplate reader, detect each hole photoabsorption (OD
490nm), thereby judgement fermented liquid is active to the inhibition of tumor cell line.The final the highest bacterial strain of 1 strain activity that obtains, is numbered HS-13-94.The method for preserving of obtained strains comprises: adopt beef extract-peptone slant medium preservation strain in-4 ℃ of refrigerators, or adopt glycerine pipe preserving process to carry out long-term preservation in-80 ℃ of Ultralow Temperature Freezers.
Embodiment 2: feature is learned in the morphology of bacterial strain (chromobacterium HS-13-94, Chromobacterium sp.HS-13-94, CGMCC No.6247) and cultivation
With reference to the related content in the books such as < < common bacteria system identification handbook > >, < < molecular cloning experiment guide > > and < < Chinese Pharmacopoeia > > (2010 editions XIH), test.The morphology of bacterial strain and cultivation are learned feature experiment and are adopted ISP1, ISP2, ISP3, ISP4, ISP5, Gause I, calcium malate, nutrient agar medium, YMS and 10 kinds of substratum of Cha Shi substratum, cultivate after 7～10 days, observe mycelial color and pigment situation for 28 ℃.The results are shown in Table 1.
The cultural characteristic of table 1 bacterial strain 13-94 on different culture media
Embodiment 3: the physiological and biochemical property of bacterial strain (chromobacterium HS-13-94, Chromobacterium sp.HS-13-94, CGMCC NO.6247)
The utilization test of carbon source: adopt ISP9 as basic medium, the final concentration of various carbon sources is 1.0%, and it the results are shown in Table 2.
Inorganic nitrogen-sourced utilization test: adopt ISP9 as basic medium, the concentration of saltpetre and ammonium sulfate is 1.0%, and it the results are shown in Table 2.
Degrading experiment and NaCl tolerance experiment: all adopt basic medium GYEA (pH6.0).Degradation experiment the results are shown in Table 3; NaCl tolerance experimental results is: NaCl tolerance is poor, and 1% NaCl exists and just can not grow.
Oxydase and catalase test, pH test and humid test: all adopt nutrient agar.Its result is: bacterial strain all can be grown between 7 ℃ to 45 ℃, and optimum growth temperature is 28 ℃; PH5.5-7.5 all can grow, and optimum scope is pH6.5-7.0; Oxydase and catalase test the results are shown in Table 4.
M.R, V-P and urease test: adopt < < common bacteria system identification handbook > > method.It the results are shown in Table 4.
Physiological and biochemical test: except temperature experiment, be 28 ℃ and cultivate 5～7 days.It the results are shown in Table 4.
The Carbon and nitrogen sources of table 2 bacterial strain HS-13-94 utilize situation
The Degrading experiment result of table 3 bacterial strain HS-13-94
The physiological and biochemical property that table 4 bacterial strain HS-13-94 is main
* note: 0: without growth; 1: grow very weak; 2: can grow; 3: well-grown; 4: grow best; +: the positive;-: feminine gender.
Embodiment 4:16SrDNA sequential analysis and strain identification
Collect the new fresh thalli of bacterial strain to be measured, adopt the improved Pitcher method of solution bacterium enzyme process (Letters in Applied Microbiology, 1989,8:151-156) extract total DNA profiling, adopt universal primer (27F and 1495R) to carry out 16S rRNA gene amplification, PCR product after purifying, directly carries out sequencing after testing, and order-checking is undertaken by Shanghai Sheng Gong biotechnology company limited.The 16S rDNA sequence of surveying is carried out homologous sequence BLAST comparison to the relevant sequence of planting, belonging in GenBank database after check and correction, to determine the classification position of this bacterial strain.
In bacterial strain HS-13-94 (CGMCC NO.6247) 16S rDNA sequence and GenBank, correlated series carries out BLAST comparison, the results are shown in Table 5 (only listing the bacterial strain that homology is higher in table).
The homology of table 5 bacterial strain 13-94 and relevant bacterial strain
Bacterial strain HS-13-94 (CGMCC NO.6247) finds that by the order-checking of 16S rDNA region itself and chromobacterium (Chromobacterium sp.) have very high homology, the highest reaches 99%, bacterial strain HS-13-94 (CGMCC NO.6247) is carried out to appearance features test simultaneously, find that this bacterial strain and chromobacterium (Chromobacterium sp.) classification correlation parameter is very approaching, therefore bacterial strain HS-13-94 (CGMCC NO.6247) is accredited as to chromobacterium (Chromobacterium sp.) bacterial strain.
In sum, HS-13-94 (CGMCC NO.6247) belongs to chromobacterium and belongs to, but be different from known romidepsin on morphological feature, produce bacterium Chromobacterium violaceum968, therefore chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94) (CGMCC NO.6247) is the brand-new bacterial classification of a strain.
Embodiment 5: antineoplastic compound is prepared in fermentation
(1) preparation of inclined-plane thalline and cultivation
Inclined-plane adopts beef-protein medium (g/L): extractum carnis 5, peptone 10, sodium-chlor 5, agar 20, distilled water 1000ml, pH7.0-7.2, sterilizing pressure 1.05kg/cm
2, 20min, is cooled to 50-60 ℃ of pendulum inclined-plane, and access one ring thalline is cultivated 2 days through 30 ℃.
(2) preparation of seed liquor and cultivation
Seed culture based formulas (g/L): sucrose 20, peptone 10, yeast powder 5, NaCl3, water 1000ml, pH7.0, through 121 ℃ of sterilizings 20 minutes.Thalline inoculum size is 10
5-10
6c.f.u./mL, 30 ℃ of culture temperature, 250rpm, shaking table shaking culture 24 hours.
(3) preparation of fermention medium and cultivation
Fermentative medium formula (g/L): sucrose 40, Zulkovsky starch 30, NH
4cl3, K
2hPO
43, pH7.0, through 121 ℃ of sterilizings 20 minutes.Seed liquor inoculum size is 5% (volume percent).If shake flask fermentation, at 30 ℃, 250rpm shaking table shaking culture 3 days, makes the fermented liquid of antineoplastic compound.
Embodiment 6: the separation and purification of antineoplastic compound and Structural Identification
(1) fermented liquid extraction
Fermented liquid 30L adds 30L ethyl acetate, stirs, and the standing 30min of room temperature, collects supernatant liquor organic phase and separately deposit.In water, add 25L ethyl acetate again, stir, the standing 30min of room temperature, collects supernatant liquor and is incorporated in the organic phase of last time extraction.
(2) concentrated organic phase
Organic phase after merging, under 40 ℃ of water-baths ,-0.1Mpa vacuum concentration is to 40ml left and right.
(3) mix sample
After concentrated, organic phase joins in 30g column chromatography silica gel (100 order-200 order), in grinding alms bowl, mixes thoroughly, rolls as without granulated powders after standing about 4h volatilizes naturally.
(4) silicagel column wash-out
In glass chromatography column, add about 120g column chromatography silica gel (100 order-200 order), then will mix sample silica gel and add in chromatography column.With ethyl acetate/acetone (v/v), carry out gradient elution, the gradient adopting is respectively 9: 1, and 8: 2,7: 3,6: 4,5: 5, and each gradient difference wash-out 2BV (with the solvent elution of 2 times of column volumes).With TLC, (developping agent is ethyl acetate: acetone=5: 1) colour developing is followed the tracks of, and when using ethyl acetate: acetone=8: during 2 wash-out, gained elutriant TLC colour developing has obvious principal point, starts to collect this component, is concentrated into Powderedly, and about 8mg weighs.
(5) crystallization
Powdered samples 2ml acetone solution, condensing crystal.Crystals weighed is 6mg, purity 96%.HPLC analyzes collection of illustrative plates as shown in Figure 1.HPLC condition is: chromatographic column: Eclipse XDB-C18 9.4mm * 250mm, moving phase: methyl alcohol: water=7: 3.
(6) Structural Identification of antineoplastic compound
The sterling of antineoplastic compound is 540.98 through its molecular weight of MS Analysis deterrmination.By
1h-NMR and
13c-NMR analyzes, and determines that it is a kind of depsipeptide anti-tumor compounds, and structure is as follows:
Formula I.
Embodiment 7: the pharmacology activity research of antineoplastic compound
Adopt CCK-8 method (the .Anal Commun such as document 1:Tominage H, 36, the 47-50 pages (1999); The .The Journal of Antibiotics such as document 2:Jidong Wang, 62, the 483-487 pages (2009)) cytotoxicity of detection compound.The cell harvesting of taking the logarithm vegetative period in centrifuge tube, the centrifugal 5min of 1000r/min, counting, re-suspended cell concentration to 50000/ml, 100 μ l/ holes add 96 hole microwell plates.Negative control is the cell solution of 50000 every milliliter in 100 μ l/ holes, and blank is 100 μ l cell culture fluids (RPMI1640+10% foetal calf serum).Then be placed in 37 ℃, 5%CO
2in incubator, overnight incubation.With DMSO, respectively each testing compound is diluted to 10mg/ml, then uses cell culture fluid doubling dilution, the testing compound sample liquid of the concentration of successively decreasing successively adds in 96 porocyte culture plates, every hole 25 μ l; Negative control and blank all add the cell culture fluid of 25 μ l.Be placed in again 37 ℃, 5%CO
2in incubator, cultivate 48h.Equal 8 the multiple holes of each sample.In each hole, add the romidepsin reagent of 10 μ l to continue to cultivate after 3.5-4h, under 490nm wavelength, measure absorbancy (OD).Be calculated as follows the inhibiting rate of compound to growth of tumour cell:
Growth of tumour cell inhibiting rate %=(1-OD experiment/OD contrast) * 100%
Different concns with same sample is mapped and can be obtained dose response curve growth of tumour cell inhibiting rate, therefrom obtains the half casualty-producing concentrations IC of sample
50.Result shows, the secondary metabolite that chromobacterium HS-13-94 (Chromobacterium sp.HS-13-94) CGMCC NO.6247 produces has very strong anti-tumor activity, the IC to Human Lung gland cancer epithelial cell A549 in vitro
50for 3.8nM, the IC to liver cancer cell HepG
50for 5.5nM.
Claims (2)
1. chromobacterium (Chromobacterium sp.) HS-13-94, its deposit number is CGMCC NO.6247, has the ability of producing containing the depsipeptide compounds of cystine linkage.
2. chromobacterium HS-13-94(CGMCC NO.6247 according to claim 1) be prepared as follows shown in formula I containing the application in the depsipeptide compounds of cystine linkage,
Formula I.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310087484.3A CN103173390B (en) | 2013-03-16 | 2013-03-16 | Chromobacterium violaceum strain and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310087484.3A CN103173390B (en) | 2013-03-16 | 2013-03-16 | Chromobacterium violaceum strain and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103173390A CN103173390A (en) | 2013-06-26 |
| CN103173390B true CN103173390B (en) | 2014-09-10 |
Family
ID=48633584
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310087484.3A Active CN103173390B (en) | 2013-03-16 | 2013-03-16 | Chromobacterium violaceum strain and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103173390B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104894195B (en) * | 2014-03-07 | 2018-11-13 | 上海医药工业研究院 | A kind of fermentation medium improving FK228 yield |
| CN106146616A (en) * | 2015-04-03 | 2016-11-23 | 浙江海正药业股份有限公司 | Cyclic peptide compound and preparation thereof and purposes |
| CN105801667B (en) * | 2016-03-22 | 2019-04-09 | 浙江海正药业股份有限公司 | A method of preparing unformed romidepsin |
| CN107189974B (en) * | 2017-07-31 | 2022-09-30 | 哈尔滨工业大学 | Low-temperature denitrification bacterium for poor nutrition and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1040054A (en) * | 1988-07-26 | 1990-02-28 | 藤泽药品工业株式会社 | New compound of biologically active and preparation method thereof |
| CN101801994A (en) * | 2006-12-29 | 2010-08-11 | 格洛斯特制药公司 | Preparation Romidepsin |
-
2013
- 2013-03-16 CN CN201310087484.3A patent/CN103173390B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1040054A (en) * | 1988-07-26 | 1990-02-28 | 藤泽药品工业株式会社 | New compound of biologically active and preparation method thereof |
| CN101801994A (en) * | 2006-12-29 | 2010-08-11 | 格洛斯特制药公司 | Preparation Romidepsin |
Non-Patent Citations (5)
| Title |
|---|
| Aacute * |
| M. DURÁ |
| M. DUR& * |
| N ，et al.Chromobacterium violaceum and Its Important Metabolites – review.《Folia Microbiol》.2010,第55卷(第6期),535–547. |
| N,et al.Chromobacterium violaceum and Its Important Metabolites – review.《Folia Microbiol》.2010,第55卷(第6期),535–547. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103173390A (en) | 2013-06-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jing et al. | Newly isolated Streptomyces sp. JBS5-6 as a potential biocontrol agent to control banana Fusarium wilt: genome sequencing and secondary metabolite cluster profiles | |
| Riahi et al. | Genus Pseudonocardia: what we know about its biological properties, abilities and current application in biotechnology | |
| CN103173390B (en) | Chromobacterium violaceum strain and application thereof | |
| CN102311981B (en) | Method for preparing and purifying prodigiosin | |
| US20240093142A1 (en) | Strain for degrading deoxynivalenol and use thereof | |
| CN103451137B (en) | A kind of method of new Halomonas and production tetrahydropyrimidine thereof | |
| CN105130963B (en) | A kind of separation and Extraction, Structural Identification and its application of new quorum sensing inhibitor | |
| CN109735467A (en) | A kind of streptomycete mutagenic strain of high yield tacrolimus and its application | |
| CN103667097B (en) | Bacillus sp.N11-8 and active component thereof having antitumor action | |
| CN102168034B (en) | Marine streptomyces and method for preparing tirandamycin A and B by utilizing strain | |
| CN101857841B (en) | Marine fungi aspergillus unguis strain, active extract thereof and preparation method and use of active extract thereof and active components thereof | |
| CN111646993B (en) | Marine anti-glioma active substance iso-wall carboline base C, preparation and application thereof | |
| CN101455687B (en) | Active extract of facultative anaerobic red marine bacteria and production method and use thereof | |
| Kim et al. | Identification of antifungal niphimycin from Streptomyces sp. KP 6107 by screening based on adenylate kinase assay | |
| CN109280034A (en) | A kind of Benzoxazepin class compound and the preparation method and application thereof with bacteriostatic activity | |
| CN100424168C (en) | Active extract of a strictly anaerobic marine bacterium, and its preparing method and use | |
| CN101285048B (en) | Rhodobacter sphaeroides mutant strain and uses thereof | |
| CN111747955B (en) | Marine anti-glioma active substance isobaric carboline alkali A, preparation and application thereof | |
| CN106047751A (en) | Nocardiopsis, separation method of active metabolites of Nocardiopsis and application of Nocardiopsis | |
| CN101496820B (en) | Active extract of facultative anaerobic sea Pseudomonas stuszeri as well as production method and use thereof | |
| CN101302211A (en) | Large ring lactone compound having double thiazole ring | |
| CN110144303A (en) | A kind of piperazinedione compounds, bacterial strain and preparation and application | |
| CN101455688B (en) | Active extract of facultative anaerobic red marine pseudomonas and production method and use thereof | |
| CN114525224B (en) | Bacillus amyloliquefaciens capable of highly producing antibacterial active substance macrolactinA and application thereof | |
| Meenakshi et al. | Actinomycetes: Isolation, Cultivation and its Active Biomolecules. |
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 |