CN107937453A - A kind of preparation method of II type halogenation polyketides of dichloro substitution and antibacterial activity application - Google Patents
A kind of preparation method of II type halogenation polyketides of dichloro substitution and antibacterial activity application Download PDFInfo
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Abstract
The present patent application discloses a kind of dichloro substitution halogenation II type polyketides zunyimycin A preparation methods, which derives from 2 bacterial strain (bacterial strain preserving numbers of streptomycete Streptomyces sp.FJS31：CGMCC4.7321) secondary metabolite, by the way that culture, separation are activated, expanded to bacterial strain, the final Structural Identification that carries out determines.It can apply to treat the treatment of the gram-positive bacterium such as infectious disease such as staphylococcus aureus (MRSA) and its drug-fast bacteria, staphylococcus epidermis, bacillus subtilis；Apply to treat the treatment of Escherichia coli (ESBL), proteus, Brucella infectious disease that gramnegative bacterium such as produces extended spectrum β lactamases；Apply to the treatment such as Candida albicans of fungal infection disease.
Description
Technical field
The present invention relates to microbial technology field, and in particular to a kind of II type halogenation polyketides of dichloro substitution
Preparation method and antibacterial activity application, more particularly to a kind of dichloro substitution halogenation II type polyketides zunyimycin A
Preparation method and antibacterial activity application.
Background technology
The natural products of Streptomyces often has good bioactivity and patent medicine potentiality, is the important next of antibiotic
Source.But with the abuse of antibiotic, the appearance of many " superbacterias ", the treatment zone to diseases such as clinically pathogenic bacterial infections is come
There is the phenomenon that can be cured without medicine in huge challenge, some superbacterias.In view of current increasingly serious antibacterial situation, increases
Antibiotic R＆D intensity, is explored and research and development antibiotics become the task of top priority, and solves the problems, such as the effective way of antibiotics resistance
Footpath.
The halogenation natural products of Streptomyces has the characteristics that structure is novel, activity is good.Such as：Live with anti-leukocythemia
The chlorination polyketone neocarzillin A of property；The BE-19412A inhibited to mouse tumor cell and its methylate
Product；The polyketone chinikomycins A inhibited to human body difference tumour cell；To more drug resistance tuberculosis branch bars
Bacterium has the chlorophenazine of bactericidal effect.In addition clinically anti-infectious vancomycin, teicoplanin and 2014
The antimicrobial agent infection antibiotic new drug Dalbavancin of U.S. FDA approval listing, oritavancin and safe ground azoles amine, they all belong to
In halogenation natural products.
Separate one plant of identification this seminar early period from the special habitats Mount Fanjing soil of Guizhou and be named as Streptomyces
Sp. FJS31-2 bacterial strains (bacterial strain preserving number：CGMCC4.7321), and by secondary metabolite study, separation identification obtains newly
Compound zunyimycin A, zunyimycin B, such as the zunyimycin C, Chinese patent of type chlorination polyketone series
A kind of halogenation II type polyketides compound, preparation method and applications, halogenation II types disclosed in CN106167495A
Polyketide compound is compound zunyimycin A,
Zunyimycin A structural formulas
But the defects of due to preparation method, the yield of acquisition is smaller, and yield is also low, simultaneously as this yield is seldom (every
A compound about 7mg or so), so only having done resistant Staphylococcus aureus activity.
The content of the invention
It is higher to obtain the invention is intended to provide a kind of preparation method of the II type halogenation polyketides of dichloro substitution
Yield, while other antibacterial activities are studied.
A kind of preparation method of the II type halogenation polyketides of dichloro substitution, it is characterised in that comprise the following steps：
Step 1: the activation of bacterial strain：The streptomycete Streptomyces sp.FJS31-2 bacterial strains of preservation are taken out from -80 DEG C of refrigerators,
The deposit number of bacterial strain is CGMCC NO.4.7321, and preservation day is on June 2nd, 2016, and depositary institution is China Microbiological bacterium
Kind preservation administration committee common micro-organisms center, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.Activation
When：Take the spore of bacterial strain with aseptic inoculation ring, cross streak inoculation stands training on the tablet of basal medium at 28 DEG C
3d is supported, picking single bacterium colony amplifies culture after being passaged to the third generation；
Step 2: the fermented and cultured of tunning：Improvement GYD solid-substrate fermentation cultures are selected, are added in culture medium
Agar powder, 28 DEG C, quiescent culture 17d, thalline is smashed to pieces together with culture medium, is added ethyl acetate and is extracted twice, combining extraction liquid,
It is concentrated under reduced pressure, obtains tunning；
Step 3: tunning purifies and separates：A, after tunning is dissolved with acetone solvent, used after being mixed with silica white
Chloroform-acetone is according to pure chloroform；30:1；15:1；10:1；5:1；2:1；Pure acetone, successively 7 gradient elutions collect eluant, eluent,
After eluant, eluent rotary evaporation is recovered under reduced pressure, it is unfolded with solvent, merging 365nm fluorescence shows yellow green, 8% sulfuric acid ethanol shows yellow
The point of color, migration value (Rf) 0.5, weighs after merging and obtains F5；
B, F5 is separated using normal phase silica gel column chromatography, after acetone solvent dissolving F5 components, is mixed with silica white and is used oil
Ether-acetone is according to 6:1；4:1；2:13 gradient elutions successively, collect eluting solvent, and eluting solvent rotary evaporation is recovered under reduced pressure
Afterwards, it is unfolded with solvent, merges 365nm fluorescence and show yellow green, 8% sulfuric acid ethanol displaing yellow, point of the migration value (Rf) 0.5,
Weighed after merging and obtain F5-1；
C, after F5-1 acetone solutions, using thin-layer chromatography method, the silica white of yellow-green fluorescence will be shown under 365nm fluorescence
Scrape off, the silica gel grinding scraped off is uniform, load splitter, eluted using acetone, recycle acetone solvent, obtain
Zunyimycin A compounds.
The preparation method of the present invention is compared with the prior art, when initial spore amount is the same, by fermentation based component
Compounding, can effectively improve the reproductive capacity of bacterial strain, accelerate the metabolism of bacterial strain, the 6 times of left sides compared to existing technologies obtained with this
Right zunyimycin A compounds.
Further, the preparation method of the II type halogenation polyketides of dichloro substitution, the improvement GYD solid mediums
Middle glucose 4g/L, malt extract 4g/L, dusty yeast 4g/L, calcium carbonate 2g/L, trace element premix liquid 0.5mL, 10g/L
The humic acid of soaked in absolute ethyl alcohol 24h, adds deionized water and is settled to 1L, 121 DEG C of autoclaving 30min.
Further, the trace element premix liquid for 1~2 part of ZnSO47H2O, 1~2 part of FeSO47H2O,
1~2 part of MnCl24H2O, 1~2 part of CuSO45H2O, 1~2 part of Na2B4O410H2O and (NH4) 6MO74H2O
1~2 part, it is settled to 1L with distilled water and is formulated.
Further, the solvent is petroleum ether：Acetone=2:1st, chloroform：Acetone=5:1 or chloroform：Methanol=10：1.
The application of the II type halogenation polyketides of dichloro substitution, the II type halogenation polyketone class chemical combination of the dichloro substitution
Thing is used for resisting gram-positive bacteria activity.The gram-positive bacteria includes staphylococcus epidermis and bacillus subtilis.
The application of the II type halogenation polyketides of dichloro substitution, the II type halogenation polyketone class chemical combination of the dichloro substitution
Thing is used for anti-Gram-negative bacteria activity.Escherichia coli of the Gram-negative bacteria including generation extended spectrumβ-lactamase,
Proteus and Brucella.
The application of the II type halogenation polyketides of dichloro substitution, the II type halogenation polyketone class chemical combination of the dichloro substitution
Thing is used for antifungal activity.The fungi is Candida albicans.
Brief description of the drawings
Fig. 1 isolates and purifies figure for Zunyimycin A；
Fig. 2 is the anti-staphylococcus epidermis activity figures of Zunyimycin A；
Fig. 3 is Zunyimycin A resistance to deformations bacillus activity figure；
Fig. 4 is the anti-Brucella activity figures of Zunyimycin A；
Fig. 5 is Zunyimycin A anti-Candida albicans activity figures；
Fig. 6 is the anti-bacillus subtilis activity figures of Zunyimycin A；
Fig. 7 is the anti-MRSA activity figures of Zunyimycin A；
Fig. 8 is the anti-ESBL activity figures of Zunyimycin A.
Embodiment
Below by embodiment, the present invention is described in further detail：
The preparation method of the II type halogenation polyketides of dichloro substitution
Step 1: the activation of streptomycete Streptomyces sp.FJS31-2 bacterial strains
The strain that glycerine inclined-plane preserves is taken out from -80 DEG C of refrigerators, 1 ring bacterial strain is taken with aseptic inoculation ring
The spore of Streptomyces sp. FJS31-2, in the basal medium tablet of diameter 11cm, 28 DEG C quiet for cross streak inoculation
Culture 3d is put, picking single bacterium colony, which is passaged to the third generation, can amplify culture.
Step 2: the fermented and cultured of tunning
Streptomycete Streptomyces sp.FJS31-2 bacterial strains select improvement GYD solid-substrate fermentation cultures, in specification
500 mL conical flasks fill 150mL fermentation mediums, and every bottle adds fine jade by agar powder quality and the 1.8% of fermentation medium volume
Cosmetics.The activated spawn amount that every bottle of inoculum concentration is 1 × 1cm2 of area on culture plate, 28 DEG C, quiescent culture to 17d, thalline
Smashed to pieces together with culture medium, add ethyl acetate and be extracted twice, every time in 130rpm oscillation extraction 24h, combining extraction liquid, 40 DEG C
It is concentrated under reduced pressure, obtains tunning.Repeat above operation, amount to fermented and cultured 100L merging tunnings and obtain 38g.
Improve GYD culture medium prescriptions：Glucose 4g/L, malt extract 4g/L, dusty yeast 4g/L, calcium carbonate 2g/L are micro-
The humic acid of secondary element premixed liquid 0.5mL, 10g/L soaked in absolute ethyl alcohol 24h, adds deionized water and is settled to 1L, 121 DEG C of height
Pressure sterilizing 30min.
Trace element premix liquid：ZnSO4·7H2O 2g、FeSO4·7H2O 2g、MnCl2·4H2O 2g、CuSO4·
5H2O 2 g, Na2B4O410H2O 2g and (NH4) 6MO74H2O 2g, are settled to 1L with distilled water and are formulated.
Step 3: tunning purifies and separates
A, with the tunning of the dissolving 32g of acetone solvent, with silica gel in mass ratio about 1:1.5 (i.e. in 32g tunnings
Add 100-200 mesh silica white 48g) it is uniformly mixed, river sand shape sample, sample of the sample as upper prop are obtained after the solvent is volatilized
Product；Weigh 100~200 mesh silica white 1200g and be uniformly mixed and (bubble cannot be produced in the process) loading length with chloroform solvent
For 975mm, internal diameter is in 120mm splitters, allows silica white slowly to sink until adding upper prop sample when no longer sinking, uses
Chloroform-acetone [pure chloroform；30:1；15:1；10:1；5:1；2:1；Pure acetone], 7 gradient elutions successively, each gradient elution
3-4 (about each column volume elution 7.2L-9.6L eluting solvents) column volumes, carry out per 1000mL eluting solvents to be a
Collect, collect 80 parts altogether, after every part of rotated evaporation under reduced pressure recycling, it is 20mL to move on to specification after addition 10mL acetone solutions
In cillin bottle, thin-layer chromatography (TCL) contact plate, uses petroleum ether：Acetone=2:1st, chloroform：Acetone=5:1 or chloroform：Methanol=10：
1 solvent is unfolded, and tool 254nm or 365nm fluorescence is seen whether under Conventional UV visible light analysis instrument, then with 8% sulfuric acid second
The color agent colour developing of alcohol colour developing；Merge 365nm fluorescence and show yellow green, 8% sulfuric acid ethanol displaing yellow, migration value (Rf) is 0.5
Point, weighs after merging and obtains F5 3.124g.
B, separation is further purified in F5 3.124g, is separated using normal phase silica gel column chromatography.The dissolving 3.124g of acetone solvent
Component, with silica gel in mass ratio about 1:1.5 (i.e. in 3.124g tunnings plus 100-200 mesh silica white 4.7g) mixing are equal
It is even, river sand shape sample, sample of the sample as upper prop are obtained after the solvent is volatilized；Weigh 140 g of 100-200 mesh silica white with
Petroleum ether：Acetone=6:1 (cannot produce bubble in the process) loads a length of 975mm, and internal diameter is in 50mm splitters, is allowed
Silica white slowly sinks until upper prop sample is added when no longer sinking, using petroleum ether-acetone [6:1；4:1；2:1], successively 3
A gradient elution, each gradient elution 4-5 (about each column volume elution 1L-1.2L eluting solvents) column volumes, often
100mL eluting solvents are collected for portion, collect 40 parts altogether, after every part of rotated evaporation under reduced pressure recycling, add 10mL third
Specification is moved on to after ketone dissolving as in 20mL cillin bottles, thin-layer chromatography (TCL) contact plate, uses petroleum ether：Acetone=2:1st, chloroform：Third
Ketone=5:1 or chloroform：Methanol=10：1 solvent be unfolded, seen whether under Conventional UV visible light analysis instrument tool 254nm or
365nm fluorescence, then the color agent to be developed the color with 8% sulfuric acid ethanol develop the color；Merge 365nm fluorescence and show yellow green, 8% sulfuric acid ethanol
Displaing yellow, point of the migration value (Rf) 0.5, weighs after merging and obtains F5-3 91mg.
C, F5-1 91mg components, after acetone solution, using thin-layer chromatography (TCL) method, the point (specification of plate：20cm
× 20cm) thin layer chromatography board, the volume containing the sample 45mg/ blocks of plate, prepare chloroform：Acetone：Formic acid=6:1:1 drop 140mL solvents,
Solvent is poured into expansion cylinder, thin layer chromatography board is put into expansion cylinder, treats that about at 0.5cm, plate is taken away from edges of boards edge for solvent
Go out, solvent volatilizes, and is placed on 365nm observed under fluorescent light, and the silica white that yellow-green fluorescence is shown under 365nm fluorescence is scraped off, will be scraped
The silica gel grinding that gets off is uniform, loads splitter, is eluted using acetone, recycles the molten machine of acetone, obtains zunyimycin A compounds
41mg。
Zunyimycin A antibacterial activity applications
(1) test strain：Staphylococcus aureus (MRSA), bacillus subtilis, Brucella, staphylococcus epidermis,
Proteus, the Escherichia coli (ESBL) for producing extended spectrumβ-lactamase, Candida albicans.
(2) culture medium：MH (A) culture medium, weighs MH (A) culture medium 36.5g and is mixed with 1L distilled water, 121 DEG C of high temperature height
Pressure sterilizing 15min.Culture medium is poured into plate, each culture dish about 20mL in super-clean bench.Culture dish is placed on 30 DEG C of baking ovens
Interior culture 24h, has seen whether bacterium growth, sterile to use.
(3) bacterial strain activates：The test strain that glycerine inclined-plane preserves is taken out from -80 DEG C of refrigerators, takes 1 with aseptic inoculation ring
The various test strains of ring, cross streak inoculation is in MH (A) culture medium flat plate of diameter 11cm, 37 DEG C of quiescent culture 1d, picking
Single bacterium colony is passaged to the third generation.
(4) making of compound Zunyimycin solution As：Weigh the DMSO that Zunyimycin A 3mg are dissolved into 1mL
In solution, make the limpid no muddiness of its solution.
(4) using agar diffusion method measurement antibacterial activity：Firstth, with distilled water compound concentration be 10-6 or 10-7 tests bacterium
Suspension；Second, supernatant liquid is hung by the feet in MH (A) culture medium, makes its face coat uniform；3rd, the MH of test bacterium is being scribbled
(A) get that the aperture of diameter 8mm is some on culture medium, be inoculated with the solution of the Zunyimycin A of 100uL and right in aperture respectively
According to；Taking-up measures its antibacterial circle diameter the 4th, culture dish to be put into 37 DEG C of incubation insulating box culture 24h after.
6th, applicating example
Tested using the antibacterial activity of agar diffusion method measure Zunyimycin A, with staphylococcus aureus (MRSA),
Bacillus subtilis, Brucella, surface staphylococcus, proteus, the Escherichia coli of ESBLs-producing bacteria
(ESBL), Candida albicans is test bacterium, and DMSO is negative control.The result shows that：Zunyimycin A are to Staphylococcus aureus
Bacterium (MRSA), bacillus subtilis, Brucella, surface staphylococcus, proteus, ESBLs-producing bacteria it is big
Enterobacteria (ESBL), Candida albicans etc. all have good inhibiting effect, its inhibition zone is both greater than 10mm, the results detailed in Fig. 2
~8 and table 1.
Table 1：Zunyimycin A antibacterial activity testing results
Zunyimycin A minimal inhibitory concentrations (MIC)
(1) test strain：Staphylococcus aureus (MRSA), bacillus subtilis, Brucella, staphylococcus epidermis,
Proteus, the Escherichia coli (ESBL) for producing extended spectrumβ-lactamase, Candida albicans.
(2) culture medium：MH (A) culture medium, weighs MH (A) culture medium 36.5g and is mixed with 1L distilled water, 121 DEG C of high temperature height
Pressure sterilizing 15min.Culture medium is poured into plate, each culture dish about 20mL in super-clean bench.Culture dish is placed on 30 DEG C of baking ovens
Interior culture 24h, has seen whether bacterium growth, sterile to use.
(3) bacterial strain activates：The test strain that glycerine inclined-plane preserves is taken out from -80 DEG C of refrigerators, takes 1 with aseptic inoculation ring
The various test strains of ring, cross streak inoculation is in MH (A) culture medium flat plate of diameter 11cm, 37 DEG C of quiescent culture 1d, picking
Single bacterium colony is passaged to the third generation.
(4) making of compound Zunyimycin solution As：The solution of 512 μ g/mL is configured to, filtering with microporous membrane is standby
With.
(5) prepared by bacteria suspension：After bacteria to be tested is activated three generations, physiological saline is injected into slant medium in super-clean bench
It is interior, gently scrape bacterium colony with oese.Bacterium solution, which is poured into 50ml triangular flasks, gently makes bacterium solution fully mix, and is measured with nephelometer
It is configured to Maxwell concentration bacteria suspension.
(6) MIC is measured：First：MH meat soups are added in 96 orifice plates；Second：Prepared 512 μ is added in first hole
G/mL liquids, fully being blown and beaten with liquid-transfering gun makes liquid be sufficiently mixed with meat soup, then draws 100 μ L in the second hole from the first hole
Fully piping and druming mixes.Method to 11-holes are drawn 100 μ L and are discarded like this.Each medicine repeats two rows of parallel laboratory tests；3rd：
100 μ L of bacterium solution are added in per hole；4th step：Result is observed after 96 orifice plates are put into 37 DEG C of constant incubator culture 20h.
Table 2：Minimal inhibitory concentration result
Above-described is only the embodiment of the present invention, and the general knowledge such as known concrete structure and characteristic is not made herein in scheme
Excessive description, technical field that the present invention belongs to owns before one skilled in the art know the applying date or priority date
Ordinary technical knowledge, can know the prior art all in the field, and with applying normal experiment before the date
The ability of means, one skilled in the art can improve and real under the enlightenment that the application provides with reference to self-ability
This programme is applied, some typical known features or known method should not become one skilled in the art and implement this
The obstacle of application.It should be pointed out that for those skilled in the art, without departing from the structure of the invention, may be used also
To make several modifications and improvements, these should also be considered as protection scope of the present invention, these are implemented all without the present invention is influenced
Effect and practical applicability.The scope of protection required by this application should be based on the content of the claims, in specification
The record such as embodiment can be used for the content for explaining claim.
Claims (10)
1. a kind of preparation method of the II type halogenation polyketides of dichloro substitution, it is characterised in that comprise the following steps：
Step 1: the activation of bacterial strain：The streptomycete Streptomyces sp.FJS31-2 bacterium of preservation are taken out from -80 DEG C of refrigerators
Strain, the deposit number of bacterial strain is CGMCC4.7321, the spore of bacterial strain is taken with aseptic inoculation ring, cross streak inoculation is in basis
On the tablet of culture medium, quiescent culture 3d at 28 DEG C, picking single bacterium colony amplifies culture after being passaged to the third generation；
Step 2: the fermented and cultured of tunning：Improvement GYD solid-substrate fermentation cultures are selected, agar is added in culture medium
Powder, 28 DEG C, quiescent culture 17d, thalline is smashed to pieces together with culture medium, is added ethyl acetate and is extracted twice, combining extraction liquid, and decompression is dense
Contracting, obtains tunning；
Step 3: tunning purifies and separates：A, after tunning is dissolved with acetone solvent, chlorine is used after being mixed with silica white
Imitative-acetone is according to pure chloroform；30:1；15:1；10:1；5:1；2:1；Pure acetone, 7 gradient elutions collection eluant, eluents, are washed successively
After de- agent rotary evaporation is recovered under reduced pressure, be unfolded with solvent, merge 365nm fluorescence show yellow green, 8% sulfuric acid ethanol displaing yellow,
Point of the migration value (Rf) 0.5, weighs after merging and obtains F5；
B, F5 is separated using normal phase silica gel column chromatography, after acetone solvent dissolving F5 components, is mixed with silica white and is used petroleum ether-the third
Ketone is according to 6:1；4:1；2:13 gradient elutions successively, collect eluting solvent, after eluting solvent rotary evaporation is recovered under reduced pressure, use exhibition
Agent expansion is opened, merges 365nm fluorescence and shows yellow green, 8% sulfuric acid ethanol displaing yellow, migration value (Rf) in 0.5 point, claim after merging
Measure F5-1；
C, after F5-1 acetone solutions, using thin-layer chromatography method, the silica white that yellow-green fluorescence is shown under 365nm fluorescence is scraped
Come, the silica gel grinding scraped off is uniform, load splitter, eluted using acetone, recycle acetone solvent, obtain zunyimycin
A compounds.
2. the preparation method of the II type halogenation polyketides of dichloro substitution according to claim 1, it is characterised in that：
Glucose 4g/L in the improvement GYD solid mediums, malt extract 4g/L, dusty yeast 4g/L, calcium carbonate 2g/L, micro member
Plain premixed liquid 0.5mL, the humic acid of 10g/L soaked in absolute ethyl alcohol 24h, adds deionized water and is settled to 1L, 121 DEG C of autoclavings
30min。
3. the preparation method of the II type halogenation polyketides of dichloro substitution according to claim 2, it is characterised in that：
The trace element premix liquid for 1~2 part of ZnSO47H2O, 1~2 part of FeSO47H2O, 1~2 part of MnCl24H2O,
1~2 part of 1~2 part of CuSO45H2O, 1~2 part of Na2B4O410H2O and (NH4) 6MO74H2O, with distilled water constant volume
It is formulated to 1L.
4. the preparation method of the II type halogenation polyketides of dichloro substitution according to claim 3, it is characterised in that：
The solvent is petroleum ether：Acetone=2:1st, chloroform：Acetone=5:1 or chloroform：Methanol=10：1.
5. the application of the II type halogenation polyketides substituted according to any dichloro of Claims 1 to 4, its feature exist
In：The II type halogenations polyketides of the dichloro substitution are used for resisting gram-positive bacteria activity.
6. the application of the II type halogenation polyketides of dichloro substitution according to claim 5, it is characterised in that：It is described
Gram-positive bacteria includes staphylococcus epidermis and bacillus subtilis.
7. the application of the II type halogenation polyketides substituted according to any dichloro of Claims 1 to 4, its feature exist
In：The II type halogenations polyketides of the dichloro substitution are used for anti-Gram-negative bacteria activity.
8. the application of the II type halogenation polyketides of dichloro substitution according to claim 7, it is characterised in that：It is described
Gram-negative bacteria includes proteus, Brucella and the Escherichia coli for producing extended spectrumβ-lactamase.
9. the application of the II type halogenation polyketides substituted according to any dichloro of Claims 1 to 4, its feature exist
In：The II type halogenations polyketides of the dichloro substitution are used for antifungal activity.
10. the application of the II type halogenation polyketides of dichloro substitution according to claim 9, it is characterised in that：Institute
It is Candida albicans to state fungi.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109182412A (en) * | 2018-10-11 | 2019-01-11 | 遵义医学院 | The preparation method and its antibacterial activity application of II type polymeric polyketone derivatives |
| CN109875995A (en) * | 2019-04-11 | 2019-06-14 | 延安大学 | II type polyketide of halogenation is inhibiting the application in Colon Cancer Cells |
| CN109875994A (en) * | 2019-04-11 | 2019-06-14 | 延安大学 | II type polyketide of halogenation is inhibiting the application in hepatoma cell proliferation |
| CN109999023A (en) * | 2019-04-11 | 2019-07-12 | 延安大学 | II type polyketide of halogenation is inhibiting the application in Cells Proliferation of Human Breast Cancer |
| CN114903887A (en) * | 2022-06-30 | 2022-08-16 | 延安大学 | Application of halogenated II type polyketone antibiotics in enhancing immunity |
| CN115054597A (en) * | 2022-06-30 | 2022-09-16 | 延安大学 | Application of halogenated II type polyketone compound in intervention of Alzheimer's disease |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106167495A (en) * | 2016-06-03 | 2016-11-30 | 遵义医学院 | A kind of halogenation II type polyketide compound, preparation method and applications |
| CN106432262A (en) * | 2016-06-03 | 2017-02-22 | 遵义医学院 | Streptomyces-derived halogenated polyketide compound, preparation method and application |
-
2017
- 2017-11-22 CN CN201711171668.2A patent/CN107937453B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106167495A (en) * | 2016-06-03 | 2016-11-30 | 遵义医学院 | A kind of halogenation II type polyketide compound, preparation method and applications |
| CN106432262A (en) * | 2016-06-03 | 2017-02-22 | 遵义医学院 | Streptomyces-derived halogenated polyketide compound, preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| LÜ YUHONG等: "Molecular Genetic Characterization of an Anthrabenzoxocinones Gene Cluster in Streptomyces Sp. FJS31-2 for the Biosynthesis of BE-24566B and Zunyimycin Ale", 《MOLECULES》 * |
| LÜ YUHONG等: "Zunyimycins B and C, New Chloroanthrabenzoxocinones Antibiotics against Methicillin-Resistant Staphylococcus aureus and Enterococci from Streptomyces sp. FJS31-2", 《MOLECULES》 * |
| 王荫荫等: "链霉菌Streptomyces sp.FJS31-2产卤化二型聚酮类化合物的发酵条件优化", 《中国酿造》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109182412A (en) * | 2018-10-11 | 2019-01-11 | 遵义医学院 | The preparation method and its antibacterial activity application of II type polymeric polyketone derivatives |
| CN109875995A (en) * | 2019-04-11 | 2019-06-14 | 延安大学 | II type polyketide of halogenation is inhibiting the application in Colon Cancer Cells |
| CN109875994A (en) * | 2019-04-11 | 2019-06-14 | 延安大学 | II type polyketide of halogenation is inhibiting the application in hepatoma cell proliferation |
| CN109999023A (en) * | 2019-04-11 | 2019-07-12 | 延安大学 | II type polyketide of halogenation is inhibiting the application in Cells Proliferation of Human Breast Cancer |
| CN109999023B (en) * | 2019-04-11 | 2022-02-22 | 延安大学 | Application of halogenated II type polyketone antibiotics in inhibition of breast cancer cell proliferation |
| CN114903887A (en) * | 2022-06-30 | 2022-08-16 | 延安大学 | Application of halogenated II type polyketone antibiotics in enhancing immunity |
| CN115054597A (en) * | 2022-06-30 | 2022-09-16 | 延安大学 | Application of halogenated II type polyketone compound in intervention of Alzheimer's disease |
| CN115054597B (en) * | 2022-06-30 | 2023-08-25 | 延安大学 | Application of halogenated type II polyketide in intervention of Alzheimer's disease |
| CN114903887B (en) * | 2022-06-30 | 2023-09-05 | 延安大学 | Application of halogenated type II polyketone antibiotics in enhancing immunity |
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