CN104804071A - Depsipeptide compound, and preparation method and application thereof - Google Patents
Depsipeptide compound, and preparation method and application thereof Download PDFInfo
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- CN104804071A CN104804071A CN201410042654.0A CN201410042654A CN104804071A CN 104804071 A CN104804071 A CN 104804071A CN 201410042654 A CN201410042654 A CN 201410042654A CN 104804071 A CN104804071 A CN 104804071A
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Abstract
The invention relates to a microbe-derived pesticide, and concretely relates to a depsipeptide compound, and a preparation method and an application thereof. The depsipeptide compound is represented by formula (1). The depsipeptide compound is obtained by through fermenting culture of Beauveria felina AS-70 (preserved in China General Microbiological Culture Collection Center on Sep. 29, 2012 with the preservation number of CGMCC NO.6643), and through extraction and separation. A result of identification of the chemical structure of the compound through nuclear magnetic resonance and mass spectrum shows that the compound has good insecticidal activity.
Description
Technical field
The present invention relates to microbe-derived sterilant, specifically a kind of depsipeptide compounds and its preparation method and application.
Background technology
Along with a large amount of uses of chemical pesticide, plant pest is also more and more obvious to its resistance, also brings the problem such as environmental pollution and person poultry poisoning thereupon.In recent years, " green barrier " that cause because chemical pesticide remains seriously governs the outlet of agricultural products in China, and traditional agriculture industry is also faced with larger risk and challenge.And compared with traditional chemical synthetic pesticide, microbial pesticide has people and animals and non-target organism safety, environment compatibility is good, not easily produces resistance and is easy to the advantages such as large-scale production.So the Sustainable development of the development & application of microbial pesticide to human health, environment protection and agricultural is significant.
Summary of the invention
The object of this invention is to provide a kind of depsipeptide compounds and its preparation method and application.
For achieving the above object, the technical solution adopted in the present invention is:
A kind of depsipeptide compounds, depsipeptide compounds such as formula shown in (one),
A preparation method for depsipeptide compounds,
1) muscardine (Beauveria felina AS-70) (is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution address is Datun Road, Chaoyang District, Beijing City, preservation date: on September 29th, 2012, preserving number: CGMCC6643, taxonomy called after Beauveria felina) be inoculated in standing for fermentation in fungus solids substratum, through organic solvent extraction, obtain extract, stand-by;
2) said extracted thing is carried out silica gel column chromatography, successively with Shi You Mi – ethyl acetate and Lv Fang – methyl alcohol for elutriant carries out gradient elution, collect elutriant, elutriant detects through thin-layer chromatography;
3) by above-mentioned steps 2) in the elution fraction of Lv Fang – methyl alcohol volume ratio 20 – 10:1 gradient, carry out reversed-phase silica gel column chromatography, gel filtration chromatography and preparative high-performance liquid chromatographic separation and purification successively, collection retention time t
rvalue is the component of 10.8 – 14.8min, obtains depsipeptide compounds shown in formula ();
Described step 3) collects retention time t
rvalue is the component of 12.8min, obtains depsipeptide compounds shown in formula ().
Muscardine (Beauveria felina AS-70) is inoculated in fermentation culture 35-40 days in fungus solids substratum by described step 1), one or more in ethyl acetate, acetone, chloroform, methyl alcohol, ethanol, water are extracted as organic solvent, obtain extract
Described step 1) fungus solids substratum is rice medium.
Described step 2) PetroChina Company Limited. Mi – eluent ethyl acetate gradient be 20:1 to 1:1, Lv Fang – methanol elution gradient is 40:1 to 1:1.
In described step 3), reverse silica gel column chromatography elutriant is the Jia Chun – water of volume ratio 1:1; Gel filtration chromatography elutriant is acetone; Preparative high-performance liquid chromatographic condition is the Yi Jing – water of volume ratio 9:11, and flow velocity is 16mL/min, and determined wavelength is 210nm.
An application for depsipeptide compounds, shown in described formula (), depsipeptide compounds can be used for preparing agricultural insecticide.
The advantage that the present invention has:
1) depsipeptide compounds involved in the present invention has better insecticidal activity, to halogen worm LD
50value is 26.6 μMs.
2) depsipeptide compounds involved in the present invention can as novel pesticide composition or the lead compound with insecticidal action.
3) depsipeptide compounds involved in the present invention can utilize microorganism to carry out scale fermentation, has production technique simple, the features such as the cycle is short, product cost is low.
Embodiment
For illustrating the understanding to feature of the present invention, the embodiment below in conjunction with some indefinitenesses is further elaborated the present invention.
Embodiment 1: depsipeptide compounds is such as formula (Arabic numerals in structural formula and epsilon are the marks of the carbon atom in chemical structure) (one) Suo Shi.
Embodiment 2: the fermentative production of depsipeptide compounds and separation and purification:
1) fermentation culture
Spawn culture: according to the conventional culture methods of microorganism, picking is stored in muscardine (the Beauveria felina AS-70) bacterial classification of agar-malt extract substratum on a small quantity, is inoculated in PDA planar surface, cultivates 3 days for 28 DEG C, as the bacterial classification that scale fermentation is cultivated, stand-by.
The bacterial classification cutting above-mentioned PDA planar surface is appropriate, be seeded to sterilized, fill in the Erlenmeyer flask of rice medium, room temperature quiescent culture 35 days.Add ethyl acetate sterilizing, stand-by.
Described rice medium is rice 100g/ bottle, peptone 0.6g/ bottle, natural sea-water 100mL/ bottle, 1000mL Erlenmeyer triangular flask.
2) separation and purification of compound
By above-mentioned rice medium ethyl acetate supersound extraction 3 times, combined ethyl acetate extracting solution, underpressure distillation obtains medicinal extract.Carried out silica gel VLC(vacuum liquid chromatography) rapid column chromatography, according to elutriant polarity incremental order, gradient elution is carried out successively with Shi You Mi – ethyl acetate (flow velocity is for 150mL/min) of volume ratio 20:1 to 1:1, volume ratio 40:1 to 1:1 Lv Fang – methyl alcohol (flow velocity is 150mL/min).Collect elutriant, and detect through thin-layer chromatography (TLC), using aubepine-vitriol oil as developer during detection, merge identical or similar portions according to Rf value and colour developing situation.Collect the elution fraction of Lv Fang – methyl alcohol volume ratio 20 – 10:1 gradient, the component of collection is carried out reversed-phase silica gel column chromatography, carry out gradient elution successively with the Jia Chun – water (flow velocity is for 5mL/min) of volume ratio 1:9 to 1:0, collected volume is than the Jia Chun – water elution component for 1:1.This component is through gel filtration chromatography, take acetone as elute soln (flow velocity is 2mL/min), finally by preparative high-performance liquid chromatographic separation and purification, (chromatographic separation condition is the Yi Jing – water of volume ratio 9:11, and flow velocity is 16mL/min, determined wavelength is 210nm), collect retention time t
rvalue is the component of 12.8min, obtains depsipeptide compounds shown in formula ().
Depsipeptide compounds shown in formula (), clear crystal; UV (MeOH) λ
max(log ε) 202 (4.45) nm; ESIMS m/z540 [M+H]
+; HRESIMS m/z540.3392 [M+H]
+(calcd for C
26h
46n
5o
7 +, 540.3392);
1h with
13c NMR, in table 1.
Depsipeptide compounds shown in table 1. formula ()
1h (500MHz) and
13c NMR (125MHz) spectral data
NMR tests solvent for use: DMSO-d
6.
Embodiment 3: insecticidal activity is tested
Halogen worm (brine shrimp) also claims salt solution fairy shrimp, classification belongs to Arthropoda, Crustachia, Anostraca, salt solution fairy shrimp section, genus artemia.Halogen worm is as a kind of insecticide screening model, there is relevant report both at home and abroad, king waits [agricultural chemicals by force again, 2011,50 (4): 261 – 263] insecticidal activity of 14 kinds of common sterilants is have rated using halogen worm as model organism, result shows the compound with the screening of halogen worm with insecticidal activity, and method is easy, and responsive to the sterilant of multiple different mechanism of action; Hu Zhiyu etc. [ocean is circulated a notice of, 2000,19 (4): 36 – 41], using halogen worm as indicator organism, have the compound of insecticidal activity in rapid screening marine actinomycete; Blizzard T.A. etc. [J.Antibiot, 1989,42 (8): 1304 – 1307] are with the analogue of halogen worm rapid screening sterilant Avrmectin.In a word, the advantages such as halogen worm has wide material sources as the model organism of insecticidal activity assay, amount that is simple to operate, required compound is lacked, can improve screening efficiency, significant to the research and development of new agricultural insecticide.
1) hatching of artemia cysts
Get artemia cysts 100mg and be placed in 500mL beaker, add artificial seawater 400mL, slowly inflate with aerator pump, incubation at room temperature 24h, removing chorion and unhatched ovum, halogen worm continues to cultivate 24h, for subsequent use.
2) preparation of sample solution
Testing compound dissolves with DMSO, is formulated as 4mg/mL solution, and is diluted to 2 according to this, 1 with 0.5mg/mL solution, for subsequent use.
3) test method
According to Solis improved method, get 96 porocyte culture plates, every hole adds the artificial seawater liquid of 195 μ L containing 10-15 halogen worm, makes test cultures plate.Blank group and each concentration samples group respectively establish three parallel holes, and blank group adds 5 μ L artificial seawaters, and sample sets adds the sample liquid of 5 μ L desired concns.Incubated at room temperature, after 24 hours, detects counting halogen worm dead individuals number under binocular anatomical lens.
Halogen worm killing activity corrected mortality represents, calculation formula is as follows:
Corrected mortality=(control group survival rate-treatment group survival rate)/control group survival rate × 100%, and calculate median lethal rate LD
50value.
Test-results is the LD of depsipeptide compounds shown in formula ()
50value is 26.6 μMs, has better insecticidal activity.
Above-mentioned the results show compound involved in the present invention has better insecticidal activity, and they can be used for preparing agricultural insecticide.
Claims (8)
1. a depsipeptide compounds, is characterized in that: depsipeptide compounds such as formula shown in (one),
2. a preparation method for depsipeptide compounds according to claim 1, is characterized in that:
1) muscardine (Beauveria felina AS-70) (is preserved in China General Microbiological culture presevation administrative center, preservation date: on September 29th, 2012, preserving number: CGMCC6643) be inoculated in standing for fermentation in fungus solids substratum, through organic solvent extraction, obtain extract, stand-by;
2) said extracted thing is carried out silica gel column chromatography, successively with Shi You Mi – ethyl acetate and Lv Fang – methyl alcohol for elutriant carries out gradient elution, collect elutriant, elutriant detects through thin-layer chromatography;
3) by above-mentioned steps 2) in the elution fraction of Lv Fang – methyl alcohol volume ratio 20 – 10:1 gradient, carry out reversed-phase silica gel column chromatography, gel filtration chromatography and preparative high-performance liquid chromatographic separation and purification successively, collection retention time t
rvalue is the component of 10.8 – 14.8min, obtains depsipeptide compounds shown in formula ().
3. by the preparation method of depsipeptide compounds according to claim 2, it is characterized in that: described step 3) collects retention time t
rvalue is the component of 12.8min, obtains depsipeptide compounds shown in formula ().
4. by the preparation method of depsipeptide compounds according to claim 2, it is characterized in that: muscardine (Beauveria felina AS-70) is inoculated in fermentation culture 35-40 days in fungus solids substratum by described step 1), one or more in ethyl acetate, acetone, chloroform, methyl alcohol, ethanol, water are extracted as organic solvent, obtain extract.
5., by the preparation method of depsipeptide compounds according to claim 2, it is characterized in that: described step 1) fungus solids substratum is rice medium.
6., by the preparation method of depsipeptide compounds according to claim 2, it is characterized in that: described step 2) PetroChina Company Limited. Mi – eluent ethyl acetate gradient be 20:1 to 1:1, Lv Fang – methanol elution gradient is 40:1 to 1:1.
7., by the preparation method of depsipeptide compounds according to claim 2, it is characterized in that: in described step 3), reverse silica gel column chromatography elutriant is the Jia Chun – water of volume ratio 1:1; Gel filtration chromatography elutriant is acetone; Preparative high-performance liquid chromatographic condition is the Yi Jing – water of volume ratio 9:11, and flow velocity is 16mL/min, and determined wavelength is 210nm.
8. an application for depsipeptide compounds according to claim 1, is characterized in that: shown in described formula (), depsipeptide compounds can be used for preparing agricultural insecticide.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106810601A (en) * | 2017-01-13 | 2017-06-09 | 中国科学院海洋研究所 | A kind of Destruxin classes depsipeptide derivative and its preparation method and application |
| 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 |
| CN112646729A (en) * | 2020-11-26 | 2021-04-13 | 中山大学 | Sea squirt-derived fungus and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86100658A (en) * | 1986-01-21 | 1987-10-07 | 云南省玉溪市北山林场 | The solid culture method of white muscardine fungi |
| 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 |
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2014
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86100658A (en) * | 1986-01-21 | 1987-10-07 | 云南省玉溪市北山林场 | The solid culture method of white muscardine fungi |
| 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 |
|---|
| XU L等: "Beauvericin from the endophytic fungus, Fusarium redolens, isolated from Dioscorea zingiberensis and its antibacterial activity", 《NATURAL PRODUCT COMMUNICATIONS》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106810601A (en) * | 2017-01-13 | 2017-06-09 | 中国科学院海洋研究所 | A kind of Destruxin classes depsipeptide derivative and its preparation method and application |
| 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 |
| CN112646729A (en) * | 2020-11-26 | 2021-04-13 | 中山大学 | Sea squirt-derived fungus and application thereof |
| CN112646729B (en) * | 2020-11-26 | 2022-06-10 | 中山大学 | Sea squirt-derived fungus and application thereof |
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