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Extraction method of fumagillin

A technology of fumagillin and an extraction method, applied in directions such as organic chemistry, can solve the problems of complex structure of fumagillin, difficult to control, complicated steps, etc., and achieves the advantages of environmental protection, low production cost and high resource utilization rate. Effect

Inactive Publication Date: 2016-06-01
BEE RES INST CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Due to the complex structure of fumagillin, the cost of organic synthesis is high, so at present, most of fumagillin is extracted and purified from the fermentation broth of fumagillin
In the previous patents and documents, the method of liquid-liquid extraction was mainly used to extract fumagillin from the fermentation broth. During the extraction process, a large amount of toxic reagents such as dichloromethane or chloroform, tert-butanol, acetone, and ethyl acetate were used. A large number of by-products in the process, such as highly toxic gliotoxin, etc. need to be removed by recrystallization and other methods, the steps are cumbersome and difficult to control

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  • Extraction method of fumagillin
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Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1 Optimization experiments of weakly polar and nonpolar macroporous resins.

[0028] 1) Fermentation and packed column

[0029] Aspergillus fumigatus (purchased from the Institute of Microbiology, Chinese Academy of Sciences) was placed in a 50L fermenter for fermentation. After 42 hours of fermentation, it was ventilated for 12 hours. At this time, the liquid volume was about 30L. Adjust the pH to 3.0 with hydrochloric acid, and centrifuge the fermentation broth at 5000 rpm for 10 minutes to obtain about 28 L of supernatant. Take 2 kg of SP850 resin (activated with absolute ethanol for 24 hours before), slowly place it in a chromatography column (column diameter 10 cm, column height 50 cm), after adding, add 5 L of water to balance.

[0030] 2) Filter the sample liquid into the chromatographic column through a funnel with filter paper, the sample liquid flows through the resin at 2mL / min, and then wash with 5L water and 30% ethanol respectively, and the flow r...

Embodiment 2

[0034] The selection of embodiment 2 resin and fermentation broth volume ratio:

[0035] The SP825 resin was selected for testing to optimize the mass-volume ratio of resin and fermentation broth. The details are as follows: take 35mL of fermentation broth in a 50mL centrifuge tube, respectively weigh 10g, 5g, 4g, 3g, 2g, 1g, 0.5g, 0g of the activated resin into the centrifuge tube, rotate and shake for 2h, and perform static adsorption. Stand still, remove the upper layer of fermentation broth, analyze with 10mL90% ethanol, and measure with liquid chromatography.

[0036] The concentration of fumagillin in the stock solution is 13.5 mg / L. After analysis, it is analyzed by liquid chromatography and converted into a volume of 35 mL. The concentration of fumagillin after different amounts of resin adsorption is calculated. 10 g is 13.3 mg / L, and 5 g is 13.3 mg / L. 13.4mg / L, 4g is 13.3mg / L, 3g is 13.1mg / L, 2g is 13.2mg / L, 1g is 11.2mg / L, 0.5g is 5.3mg / L. From the comparison of t...

Embodiment 3

[0038] Aspergillus fumigatus was inserted into a 50L fermenter for fermentation. After 12 hours of fermentation, 2 kg of SP825 resin (activated with absolute ethanol and water before) was taken, slowly added to the fermenter while stirring, and the pH was adjusted to 3.5 with hydrochloric acid. The resin and fermentation The liquid was formed into a suspension, stirred and ventilated for 12 hours, the volume was about 35 L, and the resin balls were obtained by filtration.

[0039] After washing with 5L of water, the resin ball after cleaning is packed on the chromatographic column, the column diameter is 10cm, and the column height is 100cm, and the chromatographic column is cleaned with the flow rate of 0.5 times / hour bed volume with 5L20% ethanol aqueous solution (in the whole process The medium does not flow through at a constant speed, calculated as the average value) and then eluted with 7.5L of 75% ethanol aqueous solution, collected the eluate, protected from light, and ...

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Abstract

The invention provides an extraction method of fumagillin.The method comprises the steps that 1, the pH value of fumagillin fermentation liquor is adjusted to be 1-6.0 and adsorbed with macroporous resin; 2, the resin adsorbing the fermentation liquor is put into a chromatographic column and cleaned with ethanol water with the mass concentration of 15%-30%; 3, the resin is eluted with absolute ethyl alcohol or ethanol water with the mass concentration of 50%-98%, eluent is concentrated and dried, and a coarse fumagillin extraction product is obtained.According to the extraction method, the processes such as various solvent extraction and back extraction in a traditional technology are omitted, the operational reliability is high, simpleness and convenience are achieved, the production process is safe and free of pollution, the resource utilization rate is high, the production cost is low, environmental friendliness is facilitated, and the extraction method is a green and environment-friendly method for extracting the fumagillin.

Description

technical field [0001] The invention belongs to the field of antibiotic production, in particular to a method for extracting antibiotics from fermented liquid. Background technique [0002] Fumagillin is a metabolite with various activities isolated from the fermentation broth of Aspergillus fumigatus. Its structural formula is: [0003] [0004] This substance has antibiotic activity and angiogenesis-inhibiting effect, thus having a therapeutic effect on tumor growth. In addition, a series of derivatives of fumagillin also have the effect of fighting against malaria parasites and sporozoites. Fumagillin is a very effective drug for the treatment of Nosemaapis in beekeeping and has been used for nearly 60 years in countries such as the United States and Canada. Studies in recent years have shown that fumagillin is also a potential weight-loss drug and an effective drug for treating obesity. As a drug with multiple activities, fumagillin will be used more widely in the...

Claims

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Application Information

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IPC IPC(8): C07D407/04
CPCC07D407/04
Inventor 薛晓锋吴黎明倪辉曹葳蕤杜冰赵柳微王妙张中印
Owner BEE RES INST CHINESE ACAD OF AGRI SCI
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