CN104804071B - A kind of depside peptides and its preparation method and application - Google Patents
A kind of depside peptides and its preparation method and application Download PDFInfo
- Publication number
- CN104804071B CN104804071B CN201410042654.0A CN201410042654A CN104804071B CN 104804071 B CN104804071 B CN 104804071B CN 201410042654 A CN201410042654 A CN 201410042654A CN 104804071 B CN104804071 B CN 104804071B
- Authority
- CN
- China
- Prior art keywords
- peptides
- depside
- preparation
- depside peptides
- formula
- 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
- 0 CCC(/C=*/*)(N)[N+] Chemical compound CCC(/C=*/*)(N)[N+] 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention relates to microbe-derived insecticide, specifically a kind of depside peptides and its preparation method and application.The depside peptides such as formula(One)It is shown.Depside peptides involved in the present invention are by muscardine(Beauveria felina AS‑70)(It is preserved in China General Microbiological culture presevation administrative center, preservation date:On September 29th, 2012, preserving number:CGMCC6643)Fermented culture, extraction are separated and obtained, and its chemical constitution has preferable insecticidal activity through the technical appraisement such as nuclear magnetic resonance and mass spectrum.
Description
Technical field
The present invention relates to microbe-derived insecticide, specifically a kind of depside peptides and preparation method thereof
And application.
Background technology
With a large amount of uses of chemical pesticide, plant pest is also more and more obvious to its resistance to the action of a drug, consequently also brings
Environmental pollution and the problems such as person poultry poisoning.In recent years, " green barrier " seriously governs me caused by chemical pesticide remains
The outlet of state's agricultural product, traditional agriculture industry are also faced with bigger risk and challenge.And with traditional chemical synthetic pesticide phase
Than microbial pesticide has to people and animals and non-target organism safety, and environment compatibility is good, is not likely to produce resistance and is easy to scale
The advantages that metaplasia is produced.So sustainable development of the exploitation of microbial pesticide with application to human health, environmental protection and agricultural
Open up significant.
The content of the invention
It is an object of the invention to provide a kind of depside peptides and its preparation method and application.
To achieve the above object, the technical solution adopted in the present invention is:
A kind of depside peptides, shown in depside peptides such as formula (one),
A kind of preparation method of depside peptides,
1)By muscardine(Beauveria felina AS-70)(It is preserved in China Committee for Culture Collection of Microorganisms
Common micro-organisms center, depositary institution address are Datun Road, Chaoyang District, Beijing City, preservation date:On September 29th, 2012, preservation
Number:CGMCC6643, taxology are named as Beauveria felina)It is inoculated in fungus solids culture medium and is left to ferment, through has
Solvent is extracted, and obtains extract, stand-by;
2)Said extracted thing is subjected to silica gel column chromatography, successively using petroleum ether-ethyl acetate and chlorine imitation-carbinol as eluent
Gradient elution is carried out, collects eluent, eluent detects through thin-layer chromatography;
3)By above-mentioned steps 2)Middle chlorine imitation-carbinol volume ratio 20-10:The elution fraction of 1 gradient, reverse phase silica gel is carried out successively
Column chromatography, gel filtration chromatography and preparative high-performance liquid chromatographic isolate and purify, and collect retention time tRIt is worth for 10.8-14.8min's
Component, produce depside peptides shown in formula (one);
The step 3)Collect retention time tRIt is worth the component for 12.8min, produces depside peptides shown in formula (one)
Compound.
The step 1)By muscardine(Beauveria felina AS-70)It is inoculated in fungus solids culture medium and ferments
Culture 35-40 days, is carried through the one or more in ethyl acetate, acetone, chloroform, methanol, ethanol, water as organic solvent
Take, obtain extract,
The step 1)Fungus solids culture medium is rice medium.
The step 2)Petrochina ether-ethyl acetate gradient is 20:1 to 1:1, chloroform-methanol elution gradient is 40:
1 to 1:1.
The step 3)In reversely silica gel column chromatography eluent be volume ratio 1:1 methanol-water;Gel filtration chromatography eluent
For acetone;Preparative high-performance liquid chromatographic condition is volume ratio 9:11 acetonitrile-water, flow velocity 16mL/min, Detection wavelength are
210nm。
A kind of application of depside peptides, depside peptides shown in the formula (one) can be used for preparing agriculture
Use insecticide.
Advantage for present invention:
1)Depside peptides involved in the present invention have preferable insecticidal activity, to artemia LD50It is worth for 26.6 μM.
2)Depside peptides involved in the present invention can be as the novel pesticide composition with insecticidal action or elder generation
Lead compound.
3)Depside peptides involved in the present invention can carry out scale fermentation using microorganism, have production work
The features such as skill is simple, and the cycle is short, product cost is low.
Embodiment
To illustrate the understanding to feature of present invention, the present invention is done into one with reference to some non-limiting embodiments
Step illustrates.
Embodiment 1:Depside peptides such as formula(One)It is shown(Arabic numerals and Greek alphabet in structural formula are
The mark of carbon atom in chemical constitution).
Embodiment 2:The fermenting and producing of depside peptides and isolate and purify:
1) fermented and cultured
Spawn incubation:According to the conventional culture methods of microorganism, picking is stored in agar-malt extract culture medium on a small quantity
Muscardine(Beauveria felina AS-70)Strain, PDA plate surface is inoculated in, 28 DEG C are cultivated 3 days, as scale fermentation
The strain of culture, it is stand-by.
It is appropriate to cut the strain on above-mentioned PDA plate surface, is seeded to conical flask that is sterilized, filling rice medium
In, it is stored at room temperature culture 35 days.Ethyl acetate sterilizing is added, it is stand-by.
The rice medium is rice 100g/ bottles, peptone 0.6g/ bottles, natural sea-water 100mL/ bottles, 1000mL
Erlenmeyer triangular flasks.
2) compound isolates and purifies
By above-mentioned rice medium ethyl acetate ultrasonic extraction 3 times, combined ethyl acetate extract solution, leaching is evaporated under reduced pressure to
Cream.Carried out silica gel VLC(vacuum liquid chromatography)Rapid column chromatography, it is incremented by according to eluent polarity
Sequentially, with volume ratio 20:1 to 1:1 petroleum ether-ethyl acetate(Flow velocity is 150mL/min), volume ratio 40:1 to 1:1 chloroform-
Methanol(Flow velocity is 150mL/min)Gradient elution is carried out successively.Eluent is collected, and through thin-layer chromatography(TLC)Detection, during detection
Using anisaldehyde-concentrated sulfuric acid as developer, same or like part is merged according to Rf values and colour developing situation.Collection chloroform-
Methanol volume ratio 20-10:The elution fraction of 1 gradient, the component of collection is subjected to reversed-phase silica gel column chromatography, with volume ratio 1:9 to
1:0 methanol-water(Flow velocity is 5mL/min)Gradient elution is carried out successively, and collected volume ratio is 1:1 methanol-water elution component.
The component is through gel filtration chromatography, using acetone as elution solution(Flow velocity is 2mL/min), most separated afterwards through preparative high-performance liquid chromatographic
Purifying(Chromatographic separation condition is volume ratio 9:11 acetonitrile-water, flow velocity 16mL/min, Detection wavelength 210nm), collect and protect
Stay time tRIt is worth the component for 12.8min, produces formula(One)Shown depside peptides.
Formula(One)Shown depside peptides, clear crystal;UV(MeOH)λmax(logε)202(4.45)nm;
ESIMS m/z540[M+H]+;HRESIMS m/z540.3392[M+H]+(calcd for C26H46N5O7 +,540.3392);1H
With13C NMR, are shown in Table 1.
The formula of table 1.(One)Shown depside peptides1H (500MHz) and13C NMR (125MHz) spectral data
NMR tests solvent for use:DMSO-d6。
Embodiment 3:Insecticidal activity is tested
Artemia(brine shrimp)Also referred to as salt solution fairy shrimp, belongs to Arthropoda in classification, Crustachia, Anostraca,
Salt solution fairy shrimp section, genus artemia.Artemia has had a relevant report as a kind of insecticide screening model both at home and abroad, and king is strong etc. again
[agricultural chemicals, 2011,50 (4):261-263] insecticidal activity of 14 kinds of common insecticides is have rated using artemia as model organism, is tied
Fruit shows the compound for having insecticidal activity with artemia screening, and method is easy, and quick to the insecticide of a variety of different mechanism of action
Sense;Hu Zhiyu etc. [circulate a notice of, 2000,19 (4) by ocean:36-41] it is quick to screen in marine actinomycete using artemia as instruction biology
Compound with insecticidal activity;[J.Antibiot, 1989,42 (8) such as Blizzard T.A.:1304-1307] it is fast with artemia
The analog of speed screening insecticide AVM.In a word, artemia as the model organism of insecticidal activity assay have wide material sources,
The advantages such as the amount of simple to operate, required compound is few, can improve screening efficiency, the research and development to new agricultural insecticide have important
Meaning.
1) hatching of artemia cysts
Take artemia cysts 100mg to be placed in 500mL beakers, add artificial seawater 400mL, slowly inflated with inflator pump, room temperature
Hatch 24h, remove chorion and unhatched ovum, artemia continues to cultivate 24h, standby.
2) preparation of sample solution
Testing compound is dissolved with DMSO, is formulated as 4mg/mL solution, and is diluted to 2 according to this, 1 with 0.5mg/mL solution,
It is standby.
3) test method
According to Solis improved methods, 96 porocyte culture plates are taken, add 195 artificial seawaters of the μ L containing 10-15 artemia per hole
Liquid, test cultures plate is made.Blank control group and each concentration samples group respectively set three parallel holes, and blank control group adds 5 μ L artificial
Seawater, sample sets add the sample liquid of concentration needed for 5 μ L.After incubated at room temperature 24 hours, the detection counting artemia under binocular anatomical lens
Dead individuals number.
Artemia killing activity represents that calculation formula is as follows with corrected mortality:
Corrected mortality=(control group survival rate-treatment group survival rate)/control group survival rate × 100%, and calculate half
Number fatal rate LD50Value.
Result of the test is formula(One)The LD of shown depside peptides50It is worth for 26.6 μM, there is preferable insecticidal activity.
Above-mentioned the results show compound involved in the present invention has preferable insecticidal activity, and they can be used for preparing agriculture
Use insecticide.
Claims (8)
- A kind of 1. depside peptides, it is characterised in that:Shown in depside peptides such as formula (one),Formula(One).
- A kind of 2. preparation method of the depside peptides described in claim 1, it is characterised in that:1)By muscardine(Beauveria felina)AS-70 bacterial strains(It is preserved in China General Microbiological culture presevation management The heart, preservation date:On September 29th, 2012, preserving number:CGMCC 6643)It is inoculated in fungus solids culture medium and is left to ferment, passes through Organic solvent extracts, and obtains extract, stand-by;2)Said extracted thing is subjected to silica gel column chromatography, carried out successively using petroleum ether-ethyl acetate and chlorine imitation-carbinol as eluent Gradient elution, collects eluent, and eluent detects through thin-layer chromatography;3)By above-mentioned steps 2)Middle chlorine imitation-carbinol volume ratio 20-10:The elution fraction of 1 gradient, reverse phase silica gel is carried out successively Column chromatography, gel filtration chromatography and preparative high-performance liquid chromatographic isolate and purify, and collect retention time tRIt is worth for 10.8-14.8 min Component, produce depside peptides shown in formula (one).
- 3. the preparation method of the depside peptides as described in claim 2, it is characterised in that:The step 3)Collect and protect Stay time tRIt is worth for 12.8 min component, produces depside peptides shown in formula (one).
- 4. the preparation method of the depside peptides as described in claim 2, it is characterised in that:The step 1)Will be white stiff Bacterium(Beauveria felina)AS-70 inoculations fermented and cultured 35-40 days in fungus solids culture medium, through acetic acid second One or more in ester, acetone, chloroform, methanol, ethanol are extracted as organic solvent, obtain extract.
- 5. the preparation method of the depside peptides as described in claim 2, it is characterised in that:The step 1)Fungi is consolidated Body culture medium is rice medium.
- 6. the preparation method of the depside peptides as described in claim 2, it is characterised in that:The step 2)Petrochina Ether-ethyl acetate gradient is 20:1 to 1:1, chloroform-methanol elution gradient is 40:1 to 1: 1.
- 7. the preparation method of the depside peptides as described in claim 2, it is characterised in that:The step 3)Elution group Divide and carry out reversed-phase silica gel column chromatography, with volume ratio 1:9 – 1:0 methanol-water carries out gradient elution successively, and collected volume is than 1: 1 methanol-water elution component;Gel filtration chromatography eluent is acetone;Preparative high-performance liquid chromatographic condition is volume ratio 9: 11 Acetonitrile-water, flow velocity is 16 mL/min, and Detection wavelength is 210 nm.
- A kind of 8. application of the depside peptides described in claim 1, it is characterised in that:Contracting phenol shown in the formula (one) Sour peptides are used to prepare agricultural insecticide.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410042654.0A CN104804071B (en) | 2014-01-29 | 2014-01-29 | A kind of depside peptides and its preparation method and application |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410042654.0A CN104804071B (en) | 2014-01-29 | 2014-01-29 | A kind of depside peptides and its preparation method and application |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104804071A CN104804071A (en) | 2015-07-29 |
CN104804071B true CN104804071B (en) | 2018-01-09 |
Family
ID=53689297
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410042654.0A Active CN104804071B (en) | 2014-01-29 | 2014-01-29 | A kind of depside peptides and its preparation method and application |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104804071B (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106810601B (en) * | 2017-01-13 | 2020-06-16 | 中国科学院海洋研究所 | Destruxin depsipeptide derivative and preparation method and application thereof |
CN107827950A (en) * | 2017-11-16 | 2018-03-23 | 青岛农业大学 | A kind of Trichomide classes Cyclopeptide derivatives and its preparation method and application |
CN112646729B (en) * | 2020-11-26 | 2022-06-10 | 中山大学 | Sea squirt-derived fungus and application thereof |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103045488A (en) * | 2012-12-04 | 2013-04-17 | 中国科学院海洋研究所 | Beauveria felina AS-70 strain and application thereof |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1014427B (en) * | 1986-01-21 | 1991-10-23 | 云南省玉溪市北山林场 | Muscardine culturing method with solid medium |
-
2014
- 2014-01-29 CN CN201410042654.0A patent/CN104804071B/en active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103045488A (en) * | 2012-12-04 | 2013-04-17 | 中国科学院海洋研究所 | Beauveria felina AS-70 strain and application thereof |
Non-Patent Citations (1)
Title |
---|
Beauvericin from the endophytic fungus, Fusarium redolens, isolated from Dioscorea zingiberensis and its antibacterial activity;Xu L等;《Natural Product Communications》;20100501;第5卷(第5期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN104804071A (en) | 2015-07-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Kusari et al. | An endophytic fungus from Azadirachta indica A. Juss. that produces azadirachtin | |
CN103497980B (en) | The preparation method of polyetherin A | |
CN102187870B (en) | Application of diterpene alkaloid compound serving as secondary metabolite of seaweed endophytic fungus | |
CN102311981B (en) | Method for preparing and purifying prodigiosin | |
CN102659912B (en) | Oxygen-rich disesquiterpenes compound, and preparation method and application thereof | |
CN104804071B (en) | A kind of depside peptides and its preparation method and application | |
CN104277982A (en) | Tricyclic sesquiterpenoid compound as well as preparation method and applications thereof | |
CN106810601B (en) | Destruxin depsipeptide derivative and preparation method and application thereof | |
CN108640832B (en) | Cadinane sesquiterpenoids, and preparation and application thereof | |
CN103045488B (en) | Beauveria felina AS-70 strain and application thereof | |
CN102675293B (en) | Indole diketopiperazines derivatives, preparation method and application thereof | |
CN109400444B (en) | Sesquiterpenoids for inhibiting plant pathogenic fungi and preparation method thereof | |
CN102659547B (en) | Ophiobolin sesterterpene compound and preparation and application thereof | |
CN106565639B (en) | A kind of tetrahydrofurans and its preparation method and application | |
CN104630119B (en) | Algistatic activity material violacein and preparation method thereof | |
CN108033877A (en) | A kind of anti-MRSA naphthalene compounds and preparation method thereof | |
CN102190698B (en) | Alga endophytic fungi diterpenoid alkaloid compound, preparation method thereof and application thereof | |
CN102234669B (en) | Biotransformation and purification method of 4-(2,3,5,6-tetramethylpyrazine-1-group)-4'-demethylepipodophyllotoxin | |
CN102031277B (en) | Aurovertin metabolins and application thereof | |
CN107827950A (en) | A kind of Trichomide classes Cyclopeptide derivatives and its preparation method and application | |
CN101830868B (en) | Oligosporol derivatives and application thereof | |
CN108503534A (en) | The extracting method of P-hydroxybenzoic acid and application | |
CN108017528A (en) | A kind of naphthalene compounds and preparation method and application | |
CN109810906B (en) | Endophytic fungi and application thereof in fermentation preparation of phenolic acid compounds | |
CN102190699B (en) | Marine algous endophytic fungus diterpenoid alkaloids and preparation and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
EXSB | Decision made by sipo to initiate substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |