CN106632230B - A kind of marine fungi pawl aspergillus bromo contracting phenol naphthenic acid ether compound and its preparation method and application - Google Patents
A kind of marine fungi pawl aspergillus bromo contracting phenol naphthenic acid ether compound and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of marine fungi pawl aspergillus bromo contracting phenol naphthenic acid ether compounds and its preparation method and application.The marine fungi pawl aspergillus bromo contracting phenol naphthenic acid ether compound chemical structural formula such as following formula () shown in:The compound is left to ferment by the way that pawl aspergillus DLEP2008001 to be inoculated in fungi culture medium, fermentation liquid is extracted through ethyl acetate, mycelium is extracted by organic solvent, crude extract carries out silicagel column gradient chromatography with organic solvent after merging, it is detected through silica gel thin-layer chromatography, the elution fraction containing target substance is continued what silicagel column, gel filtration chromatography and positive preparative liquid chromatography isolated and purified.The compound can inhibit fungi Candida albicans, Gram-negative bacteria pseudomonas aeruginosa and the gram-positive bacteria penicillin of resistance to methoxyl group staphylococcus aureus, and artemia larva is killed, there is the potential use as antifungal antibiotic, antibacterium antibiotic or insecticide.
Description
Technical field
The invention belongs to microbial fermenters extractive technique fields, and in particular, to a kind of marine fungi pawl aspergillus bromo
Depsidone class compound and its preparation method and application.
Background technique
The drug resistance of the current microorganism that finds the cause of disease in clinical treatment is more and more stronger, especially the penicillin of resistance to methoxyl group gold
Staphylococcus aureus (MRSA), pseudomonas aeruginosa (being commonly called as Pseudomonas aeruginosa) and Candida albicans (being commonly called as Candida albicans) are made
At infection it is increasingly severe, many conventional antibiotics substantially lose curative effect, it is therefore necessary to find novel antibiotic come
Cope with this challenge.In addition, agricultural production because excessively use various chemical synthetic pesticides, cause serious food, soil and
Water pollution, and jeopardize human health and the ecological balance by food chain, the agricultural product that pesticide residue also seriously hinders China go out
Mouthful；And biological pesticide has many advantages, such as to be not likely to produce drug resistance, safe to non-target organism, environmental-friendly and be easy to natural degradation,
Have great importance for human health, environmental protection and agricultural sustainable development.It the antibiotic in marine fungi source and kills
Worm agent has many advantages, such as that being easy to large scale fermentation produces, is efficient, being easy to overcome drug resistance, it is easy to accomplish medicine source stable supplying, tool
There are preferable economic, society and environmental benefit.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the defect of existing antibiotic and insecticide and deficiency, one kind is provided
Extract from the bromo depsidone class compound of marine fungi pawl aspergillus, the compound have antimycotic, antibacterial activity and
Insecticidal activity is with a wide range of applications in preparing antifungal agent, antibacterial agent and agricultural insecticide.
The object of the present invention is to provide a kind of marine fungi pawl aspergillus bromo depsidone class compounds.
It is a further object of the present invention to provide the preparation methods of above compound.
Another object of the present invention is to provide the application of above compound.
Above-mentioned purpose of the invention is that used following technical scheme gives realization.
Marine fungi pawl aspergillus that the present invention uses (Aspergillus unguis) CGMCC No.3372 strain, in
On October 28th, 2009 is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), and preservation is compiled
Number be CGMCC No.3372, strain number be DLEP2008001.
A kind of marine fungi pawl aspergillus (Aspergillus unguis) bromo depsidone class chemicals, feature exists
In, chemicals chemical structural formula such as () shown in:
()
The preparation method of above-mentioned marine fungi pawl aspergillus bromo depsidone class chemicals includes the following steps:
S1. it ferments: pawl aspergillus DLEP2008001 being inoculated in after being left to ferment in fungi culture medium, collect mycelia respectively
Body and fermentation liquid；；
S2. it slightly mentions: the fermentation liquid of step S1 being concentrated after ethyl acetate extracts, mycelium is after organic solvent extracts
Concentration obtains crude extract after merging；
S3. it isolates and purifies: the crude extract of step S2 being subjected to silicagel column gradient chromatography, is detected through silica gel thin-layer chromatography, it will
Elution fraction containing target substance continues silicagel column, gel filtration chromatography and positive preparative liquid chromatography and isolates and purifies to obtain
Target compound；
Wherein, the formula of fungi liquid culture medium described in step S1 are as follows: in every liter contain following component: murphy juice 400~
600 mL, NaBr 15~25g, 15~25g of sucrose, 400~600 mL of pure water or tap water.
Preferably, the formula of the fungi liquid culture medium are as follows: contain following component: 500 mL of murphy juice in every liter,
20 g of NaBr, 20 g of sucrose, 500 mL of pure water or tap water.
Specifically, the preparation method of above-mentioned marine fungi pawl aspergillus bromo depsidone class chemicals includes following step
It is rapid:
S1. ferment: by pawl aspergillus (Aspergillus unguis) CGMCC No.3372 is inoculated in fungi liquid culture
Be left to ferment in base, after fermentation 2~4 days, the final concentration of preferred 1mM of 0.5~2 mM(be added) Anuject, after supervention
Ferment 18~22 days, mycelium and fermentation liquid were collected in filtering respectively；
S2. it slightly mentions: the fermentation liquid that step S1 is obtained is extracted and is concentrated through ethyl acetate, the mycelia that step S1 is obtained
Body organic solvent is extracted and is concentrated, and gained concentrate is merged, as crude extract；
Wherein, the organic solvent be chloroform, it is acetone, ethyl acetate, ethyl alcohol, one or more of in methanol；
S3. isolate and purify: the crude extract that a. obtains step S2 through silica gel column chromatography, with petroleum ether-methylene chloride or
Petroleum ether-chloroform is eluted in advance, then carries out gradient elution with chloroform-methanol or methylene chloride-methanol, is collected with eluent
The component of the elution of volume ratio 100:0~100:1 gradient；Component characteristics are that solvent is volume ratio on silica gel thin-layer chromatography plate
15:1 chloroform-methanol expansion under, Rf value be 0.29~0.74 component of mixture；
B. elution fraction step a collected carries out silica gel column chromatography again, and eluent is the petroleum ether-the third of volume ratio 5:1
Ketone；
C. elution fraction step b collected carries out sephadex column chromatography, and eluent is methanol, and flow velocity 0.5~
0.7 mL/min；
D. elution fraction step c collected carries out positive preparative liquid chromatography purifying, and eluent is 3~5:1 of volume ratio
Chloroform-the petroleum ether of (preferably 4:1), in silicagel column filling 20~30g(preferably 25, g) silica gel, 2~4 mL/min(of flow velocity are preferred
3 mL/min) in the case of, collect the component that retention time is 14~28 min；
E. the step d elution fraction collected being continued positive preparative liquid chromatography to purify, eluent is pure chloroform,
10~15g(preferably 12 g) silica gel, the preferred 2mL/min of 1~3 mL/min(of flow velocity are filled in silicagel column), it collects and retains
Time is the main peak of 14~35 min, obtains gleanings.
Preferably, the silica gel of silica gel column chromatography described in step a or b is 200~300 mesh.
In addition, specifically, Rf value and chemical colour reaction of the bromo depsidone class compound on thin-layer chromatography
Feature is as follows: when chromatography solvent is pure chloroform, Rf value of the compound on GF254 thin layer silica gel plate is 0.26,254nm
Lower ultraviolet light irradiation is in single, uniform black~skipper spot, and the colour developing of anisaldehyde sulfuric acid is sepia.
Preferably, the single chromatography that compound retention time under the conditions of high-efficient liquid phase analysis is 4.0~5.0 min
Peak.
It is highly preferred that the efficient liquid phase chromatographic analysis condition is Acquity UPLC BEH C18 column, packing material size
1.7 microns, the column dimension mm of 2.1 mm × 50, mobile phase is to add the acetonitrile solution of the modifying agent formic acid of a ten thousandth volume,
0~2.5 min acetonitrile ratio is 60%, and 2.5~2.6 min acetonitrile proportional linearities rise to 100%, 2.6~3.0 min acetonitrile ratios
It is 100%, 3.0~3.1 min acetonitrile proportional linearities are down to 60%, and 3.1~5.0 min acetonitrile ratios are 60%, 0.3 ml/ of flow velocity
min。
Preferably, the bromo depsidone class compound is colorless needles at normal temperatures and pressures.
It is obtained through antibacterial experiment, compound of the present invention is 6.4 μm of ol/L to the MIC value of Candida albicans, is had
Inhibit the activity of Candida albicans；MIC to pseudomonas aeruginosa is 1.6 μm of ol/L, shows that the compound has and inhibits copper
The activity of green pseudomonad growth；MIC value to the staphylococcus aureus for the penicillin of resistance to methoxyl group is 25.6 μm of ol/L, tool
There is the activity for inhibiting MRSA；Half lethal dose LC50 to artemia is 12.0 μm of ol/L, shows it with insecticidal activity.
So application of the above-mentioned bromo depsidone class chemicals on antimycotic, antibacterium or desinsection, and making
Application in standby antifungal agent, antibacterial agent or insecticide, all should be within protection scope of the present invention.
Preferably, the fungi is Candida albicans.
Preferably, the bacterium is Gram-negative bacteria.
It is highly preferred that the Gram-negative bacteria is pseudomonas aeruginosa.
In addition, it is highly preferred that the bacterium is pseudomonas aeruginosa or the penicillin of resistance to methoxyl group staphylococcus aureus.
Preferably, the worm is agricultural pests.
It is highly preferred that the agricultural pests are the evil such as artemia, nematode, beet armyworm, aphid, Ability of Culex Pipiens Larvae or Tetranychus cinnabarinus
Worm.
Compared with the prior art, the present invention has the following beneficial effects:
Present invention obtains a kind of new marine fungi pawl aspergillus bromo depsidone class compounds, have good anti-
Fungi, antibacterial activity and insecticidal activity, and compounds process for production thereof is simple, is easy to be mass produced, it is antimycotic preparing
It is with a wide range of applications in agent, antibacterial agent or agricultural insecticide.
Detailed description of the invention
Fig. 1 is the nuclear magnetic resonance spectroscopy of bromo depsidone class compound of the present invention.
Fig. 2 is the carbon-13 nmr spectra of bromo depsidone class compound of the present invention.
Fig. 3 is the ESI of bromo depsidone class compound of the present invention-Mass spectrum.
Specific embodiment
The present invention is made with specific embodiment with reference to the accompanying drawings of the specification and further being elaborated, the embodiment
It is served only for explaining the present invention, be not intended to limit the scope of the present invention.Test method as used in the following examples is such as without spy
Different explanation, is conventional method；Used material, reagent etc., unless otherwise specified, for the reagent commercially obtained
And material.
Marine fungi pawl aspergillus of the present invention (Aspergillus unguis) CGMCC No.3372 strain, in 2009
On October 28, in is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), and deposit number is
CGMCC No.3372, strain number are DLEP2008001.
The preparation method of 1 marine fungi bromo depsidone class compound of embodiment
1. fermentation
By pawl aspergillus (Aspergillus unguis) CGMCC No.3372 is inoculated in static hair in fungi liquid culture medium
Ferment, fermentation 2 days after, the Anuject of final concentration of 1mM is added, continue fermentation 20 days, filtering, respectively collect mycelium and
Fermentation liquid.
Wherein, the formula of the fungi liquid culture medium are as follows:
Contain following component: murphy juice 500 mL, NaBr 20 g, 20 g of sucrose, pure water or tap water 500 in every liter
mL。
2. slightly mentioning
The fermentation liquid that step 1 is obtained extracts three times through ethyl acetate, is concentrated under reduced pressure, obtains concentrate.Step 1 is obtained simultaneously
The mycelium obtained is concentrated under reduced pressure with there is methanol extract liquid to extract three times, is obtained concentrate.Gained concentrate is merged, is as slightly mentioned
Object；
3. isolating and purifying
A. crude extract step 2 obtained is eluted in advance with petroleum ether-chloroform through silica gel column chromatography, then uses chlorine
Imitation-carbinol carries out gradient elution, collects with the component of the elution of effluent volume ratio 100:0~100:1 gradient；Component characteristics are
Under the chloroform-methanol expansion that solvent is volume ratio 15:1 on silica gel thin-layer chromatography plate, Rf value is 0.29~0.74
Component of mixture；
B. elution fraction step a collected carries out silica gel column chromatography again, and eluent is the petroleum ether-the third of volume ratio 5:1
Ketone；
C. elution fraction step b collected carries out sephadex column chromatography, and eluent is methanol, and flow velocity 0.5~
0.7 mL/min；
D. elution fraction step c collected carries out positive preparative liquid chromatography purifying, and eluent is volume ratio 4:1's
Chloroform-petroleum ether, when silicagel column fills 25 g silica gel, 3 mL/min of flow velocity, collection retention time is 14~28 min
Component；
E. the step d elution fraction collected being continued positive preparative liquid chromatography to purify, eluent is pure chloroform,
When silicagel column fills 12 g silica gel, 2 mL/min of flow velocity, the main peak that retention time is 14~35 min is collected；It is collected
Target substance also is compliant with following characteristics:
(1) it is in single, uniform black~skipper spot, anisaldehyde that TLC detection, which is collected under the irradiation of 254nm ultraviolet light,
Sulfuric acid colour developing is the substance of sepia；
(2) substance that Rf value is 0.26 under the conditions of the thin-layer chromatography of detection；The detection is with thin layer chromatography board
GF254 silica gel plate, solvent are pure chloroform；
(3) group for the single chromatographic peak that retention time is 4.36 min under the conditions of the ultra high efficiency liquid phase analysis of detection
Point；The detection is Acquity UPLC BEH C18 column with efficient liquid phase chromatographic analysis condition, 1.7 microns of packing material size,
The column dimension mm of 2.1 mm × 50, mobile phase are to add the acetonitrile solution of the modifying agent formic acid of a ten thousandth volume, 0~2.5
Min acetonitrile ratio is 60%, and 2.5~2.6 min acetonitrile proportional linearities rise to 100%, and 2.6~3.0 min acetonitrile ratios are 100%,
It is 60% that 3.0~3.1 min acetonitrile proportional linearities, which are down to 60%, 3.1-5.0 min acetonitrile ratio, 0.3 ml/min of flow velocity.
4, the compound obtained through above-mentioned separation process, is determined as pure compound.
Through Spectrum Analysis, Structural Identification is the results show that it is a kind of bromo depsidone class natural products, and structural formula is such as
(I) shown in:
。
(I)
The compound has following physical and chemical and spectral characteristic:
Colorless needles, nuclear magnetic resonance spectroscopy (1H NMR, CDCl3, 500 MHz) δ6.76 (s), 5.38 (q,
6.8), 2.46 (s), 2.33 (s), 1.95 (s), 1.82 (dd, 6.8,1.0), as shown in Figure 1.
Carbon-13 nmr spectra (13C NMR, CDCl3, 125 MHz) δ162.0 (s), 160.2 (s), 156.5 (s),
148.8 (s), 145.3 (s), 142.9 (s), 142.5 (s), 136.1 (s), 132.1 (s), 128.1 (d),
116.1 (s), 115.3 (d), 114.5 (s), 108.0 (s), 99.6 (s), 21.7 (q), 17.9 (q),
14.5 (q), 10.6 (q), as shown in Figure 2.
ESI-Mass spectrum shows that the isotopic peak cluster mass-to-charge ratio of quasi-molecular ion peak [M-H]-is m/z 481/483/485,
Peak intensity is 1:2:1, be typical two bromos feature and with molecular formula C19H17O5Br2It is consistent, as shown in Figure 3.
The fungicidal activities of 2 bromo depsidone class compound of embodiment are tested
1, Candida albicans (Candida albicans) it is a kind of common opportunistic fungus, human body can be invaded and permitted
Multiple location, cause cutaneous candidiasis, candidiasis of the mucous membranes (thrush, angular stomatitis, vaginitis are most common) and internal organ and in
Pivot nerve candidiasis is the most common nosomycosis.Clinical symptoms are intricate, suddenly delay different.Recently as antibiotic, swash
The large dosage application and the development of transplant operation of element, immunosuppressor, candidiasis disease incidence gradually increases, and can endanger
And life causes serious consequence.The bacterium has become the important for target of antifungal drug research and development.
2, experimental method:
Micro broth dilution method (the National Committee for Clinical to adopt international standards
Laboratory Standards. 1997. Reference methed for broth dilution antifungal
susceptibility testing of yeasts: approved standard M27-A, 7th ed. NCCLS,
Wayne, Pa.) compound is measured to the minimal inhibitory concentration (MIC value) of Candida albicans.
Control group is set simultaneously: amphotericin B.
3, experimental result:
The results show that MIC of the bromo depsidone class compound of the acquisition of above-described embodiment 1 to Candida albicans
Value is 6.4 μm of ol/L, has the significant activity for inhibiting Candida albicans.
Moreover, amphotericin B is 12.8 μm of ol/L, bromo depsidone of the present invention to the MIC value of Candida albicans
Class compound is significantly stronger than positive control amphotericin B to the inhibitory activity of Candida albicans.
The suppression gram prolapse of uterus activity experiment of 3 bromo depsidone class compound of embodiment
1, pseudomonas aeruginosa (Pseudomonas aeruginosa) it is also known as Pseudomonas aeruginosa, it is a kind of conditioned pathogen,
It is also one of the main pathogenic fungi of inside-hospital infection.The patient that suffers from metabolic disease, blood disease and malignant tumour and it is postoperative or
Patient susceptible after certain treatments contaminates this bacterium.The bacterium often causes postoperative wound infection, can also cause bedsore, abscess, it is suppurative in
Otitis etc..Its caused infection focus can lead to hematogenous extension, and bacteremia and septicemia occurs.Infections after burn verdigris color
Pseudomonad can cause death.Moreover, with the extensive clinical use of broad-spectrum antibiotic, hormone and immunosuppressor, verdigris
Pseudomonad quickly produces drug resistance to Multiple Classes of Antibiotics, so that clinical anti-infective therapy seems more and more difficult.Mesh
Before, the medicament research and development for the bacterium is also an emphasis of antibiotic research and development.
2, experimental method:
Micro broth dilution method (the National Committee for Clinical to adopt international standards
Laboratory Standards. 2003. Methods for dilution antimicrobial susceptibility
Tests for bacteria that grow aerobically, 5th ed. Approved standard M7-A6.) it surveys
Minimal inhibitory concentration (MIC value) of the fixed compound to pseudomonas aeruginosa.
3, experimental result:
The bromo depsidone class compound that above-described embodiment 1 obtains is 1.6 μ to the MIC of pseudomonas aeruginosa
Mol/L shows that the compound has the activity for inhibiting P. aeruginosa growth well.
The suppression Gram-positive drug-fast bacteria activity experiment of 4 bromo depsidone class compound of embodiment
1, staphylococcus aureus (Staphylococcus aureus) it is very common germ, sometimes it can be into
Enter in human body and causes to infect.The slight meeting of this infection papula and papule on the skin, it is serious, pneumonia or blood can be caused
Liquid inductance dye.Infection caused by staphylococcus is usually treated with the antibiotic methicillin of penicillins, in most cases
It is highly effective.But some aureus strains form drug resistance, that is, the gold for the penicillin of resistance to methoxyl group to methicillin
Staphylococcus aureus (methicillin-resistantStaphylococcus aureus, MRSA).1961 in Britain
It has found the first MRSA, worldwide spreads at an amazing speed later, be the clinical most common Gram-positive drug resistance
Bacterium, it is estimated that annual about 100,000 people important target of hospitalization and antibiotics research and development because of infection MRSA
Pathogenic microorganism.
2, experimental method:
Micro broth dilution method (the National Committee for Clinical to adopt international standards
Laboratory Standards. 2003. Methods for dilution antimicrobial susceptibility
Tests for bacteria that grow aerobically, 5th ed. Approved standard M7-A6.) it surveys
Compound is determined to the minimal inhibitory concentration (MIC value) of the staphylococcus aureus (MRSA) for the penicillin of resistance to methoxyl group.
3, experimental result:
The MIC value for the bromo depsidone class compound that above-described embodiment 1 obtains is 25.6 μm of ol/L, is had very
The activity of good inhibition MRSA.
The insecticidal activity of 5 bromo depsidone class compound of embodiment is tested
1, artemia (brine shrimp) it is also known as salt water fairy shrimp, it is rich to belong to Arthropoda, Crustachia, Anostraca, salt water
Nian Chong section, genus artemia are that a kind of fish food with higher economic value is biological and a kind of more sensitive for toxin
Important experimental animal, the activity that 2 ~ 3 instar larvaes of laboratory cultures can be used for evaluating insecticide are strong and weak.
2, insecticidal activity experimental method (be detailed in applicant paper Toxicology Mechanisms and Methods,
2012,22 (1), 23 ~ 30).
3, a dual chamber linker artemia hatching and collection: is added by the artemia eggs that freezes of Indoor Natural light stimulus recovery
It is cultivated in the hatchery of formula culture apparatus, the seawater (30 g/L) that natural coarse sea salt is prepared is filled in linker, ovum injected volume is 1
G/L first closes linker channel.After hatching in about 2000 lux illumination 24 hours at 28 DEG C, linker channel is opened, and will
Hatchery shading, and collecting pit keeps 12 ~ 24 h of illumination, carries out photoinduction；It is then shut off linker channel, with opening smoothness
Liquid transfer gun head draws the artemia larva in collecting pit and carries out follow-up test.
4, the lethal method of artemia biology: sample is diluted to series of concentrations with the continuous sesquialter of anhydrous methanol in 96 orifice plates, very
Sky dries and removes organic solvent, and 200 microlitres of artemia larva suspensions (containing 20 ~ 30 polypides) is added in every hole, and blank control is arranged
Group counts larva sum and death toll in every hole after 28 DEG C of 24 h of culture under binocular stereo microscope, calculates the correction in every hole
The death rate,
Corrected mortality=(the control group death rate-processing hole death rate)/control group survival rate × 100%；
Figure is done with corrected mortality-concentration log2 logarithm, in linear preferable 10% ~ 90% death rate section line taking
Trendline calculates half lethal dose LC50.
5, experimental result:
The half lethal dose LC50 of the bromo depsidone class compound of above-mentioned acquisition is 12.0 μm of ol/L, is shown
It is with insecticidal activity.
Moreover, copper sulphate is 102.4 μm of ol/L to the LC50 of artemia, bromo depsidone class compound is to artemia
Insecticidal activity is better than positive control copper sulphate.
In addition, the studies have shown that compound is to pests such as nematode, beet armyworm, aphid, Ability of Culex Pipiens Larvae or Tetranychus cinnabarinus,
There is preferable insecticidal activity.
Claims (4)
1. a kind of preparation method of marine fungi pawl aspergillus bromo depsidone class compound, which is characterized in that the chemical combination
Shown in the chemical structural formula of object such as formula (I):Preparation method includes the following steps:
S1. it ferments: pawl aspergillus DLEP2008001 being inoculated in after being left to ferment in fungi liquid culture medium, collect mycelia respectively
Body and fermentation liquid；
S2. it slightly mentioning: the fermentation liquid of step S1 is concentrated after ethyl acetate extracts, mycelium is concentrated after organic solvent extracts,
Crude extract is obtained after gained concentrate is merged；
S3. it isolates and purifies: the crude extract of step S2 being subjected to silicagel column gradient chromatography, detects, will contain through silica gel thin-layer chromatography
The elution fraction of target substance continues silicagel column, gel filtration chromatography and positive preparative liquid chromatography and isolates and purifies, and obtains mesh
Mark compound；
Wherein, the formula of fungi liquid culture medium described in step S1 are as follows: in every liter contain following component: murphy juice 400~
15~25g of 600mL, NaBr, 15~25g of sucrose, 400~600mL of pure water or tap water；
Fermentation described in step S1 is that the Anuject of final concentration of 0.5~2mM is added after standing for fermentation 2~4 days, is continued
Fermentation 18~22 days；Ethyl acetate extraction times described in step S2 are 2~4 times, and the organic solvent extraction times are 2~4 times；
The organic solvent is chloroform, acetone, ethyl acetate, ethyl alcohol, one or more of in methanol；
Isolate and purify that the specific method is as follows described in step S3:
A. crude extract step S2 obtained carries out prewashing through silica gel column chromatography, with petroleum ether-methylene chloride or petroleum ether-chloroform
It is de-, gradient elution then is carried out with chloroform-methanol or methylene chloride-methanol, is collected with effluent volume ratio 100:0~100:1
The component of the elution of gradient；Component characteristics are the chloroform-methanol that solvent is volume ratio 15:1 on silica gel thin-layer chromatography plate
Under expansion, Rf value be 0.29~0.74 component of mixture；
B. elution fraction step a collected carries out silica gel column chromatography again, and eluent is petroleum ether-acetone of volume ratio 5:1；
C. elution fraction step b collected carries out sephadex column chromatography, and eluent is methanol, 0.5~0.7mL/ of flow velocity
min；
D. elution fraction step c collected carries out positive preparative liquid chromatography purifying, and eluent is the chlorine of 3~5:1 of volume ratio
Imitative-petroleum ether, when silicagel column fills 20~30g silica gel, 2~4mL/min of flow velocity, collect retention time be 14~
The component of 28min；
E. the step d elution fraction collected is continued positive preparative liquid chromatography to purify, eluent is pure chloroform, in silica gel
In the case of column fills 10~15g silica gel, 1~3mL/min of flow velocity, the main peak that retention time is 14~35min is collected, is collected
Object.
2. bromo depsidone class compound described in claim 1 is preparing the application in antifungal agent or antibacterial agent；Institute
Stating bacterium is Gram-negative bacteria.
3. application according to claim 2, which is characterized in that the fungi is Candida albicans.
4. application according to claim 2, which is characterized in that the bacterium is pseudomonas aeruginosa.
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| US5350763A (en) * | 1993-06-11 | 1994-09-27 | Merck & Co., Inc. | Unguinol and analogs are animal growth permittants |
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| US5350763A (en) * | 1993-06-11 | 1994-09-27 | Merck & Co., Inc. | Unguinol and analogs are animal growth permittants |
| CN102634551A (en) * | 2012-03-06 | 2012-08-15 | 大连交通大学 | Algal epiphytic fungus chlorinated depside cyclic ether compound, and preparation and application thereof |
Non-Patent Citations (2)
| Title |
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| 7-Chlorofolipastatin, an inhibitor of sterol O-acyltransferase, produced by marine-derived Aspergillus ungui NKH-007;Ryuji Uchida等;《The Journal of Antibiotics》;20160316;第69卷(第8期);647-651页 |
| Directed biosynthesis through biohalogenation of secondary metabolites of the marine-derived fungus Aspergillus unguis;Sanya Sureram等;《RSC Advances》;20121123;第3卷(第6期);1781-1788页 |
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