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A kind of 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: 2018-06-26
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

Method used

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  • A kind of extraction method of fumagillin
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Experimental program
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Effect test

Embodiment 1

[0027] Example 1 Optimization experiment of weakly polar and non-polar macroporous resins.

[0028] 1) Fermentation and packing column

[0029] Aspergillus fumigatus (purchased from the Institute of Microbiology, Chinese Academy of Sciences) was fermented in a 50L fermenter, and after 42h of fermentation, aeration was performed for 12h, and the liquid volume was about 30L at this time. The pH was adjusted to 3.0 with hydrochloric acid, and the fermentation broth was centrifuged at 5000 rpm for 10 min to obtain about 28 L of the supernatant. Take SP850 resin (previously activated with absolute ethanol for 24h) 2kg, slowly put it in a chromatography column (column diameter 10cm, column height 50cm), after adding, add 5L water to balance.

[0030] 2) Filter the sample solution into the chromatography column through a funnel with filter paper. The sample solution flows through the resin at 2mL / min, and then washes with 5L of water and 30% ethanol respectively. The flow rate is co...

Embodiment 2

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

[0035] The SP 825 resin was selected for testing to optimize the mass-to-volume ratio of resin and fermentation broth. The details are as follows: respectively take 35mL of fermentation broth into a 50mL centrifuge tube, 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. After standing, the upper fermentation broth was removed, analyzed with 10 mL of 90% ethanol, and measured by liquid chromatography.

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

Embodiment 3

[0038] Aspergillus fumigatus was fermented in a 50L fermenter. After 12h fermentation, 2kg of SP825 resin (previously activated with absolute ethanol and water) was slowly added to the fermenter while stirring, and the pH was adjusted to 3.5 with hydrochloric acid. The liquid forms a suspension, which is stirred and aerated for 12 hours, and the volume is about 35 L, and the resin balls are obtained by filtration.

[0039] After washing with 5L of water, the cleaned resin spheres were loaded onto a chromatography column, the diameter of the column was 10cm, the column height was 100cm, and the column was washed with 5L of 20% aqueous ethanol at a flow rate of 0.5 times the bed volume per hour (during the entire process). flow through at a constant speed in the average) and then elute with 7.5L of 75% ethanol aqueous solution, collect the eluate, protect from light, reduce the pressure to less than 50mL at a temperature of less than 50 degrees, and place it in a glass watch dish...

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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 fermentation broth. 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 inhibits angiogenesis, so it has a therapeutic effect on tumor growth. In addition, a series of derivatives of fumagillin also have the effect of fighting against Plasmodium and Sporozoites. In beekeeping, fumagillin is a very effective drug for the treatment of Western honeybee sporozoites (Nosema apis) and has been used in countries such as the United States and Canada for nearly 60 years. In recent years, studies have shown that fumagillin is also a potential weight loss drug and an effective drug for the treatment of obesity. As a drug with multiple activities, fumagillin will ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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