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Epothilone B separating and extracting method based on membrane filtration technology

An epothilone and extraction method technology, applied in the field of antibiotic separation and extraction, can solve the problems of low yield, few separation steps, complicated steps and the like, and achieve the effects of improving yield, few separation steps and good selectivity

Active Publication Date: 2014-05-07
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a method for separating and extracting epothilone B based on membrane filtration technology, which has mild conditions, easy operation, few separation steps and good selectivity, and overcomes the The steps in the prior art are cumbersome, the yield is not high and other defects, and the prepared epothilone B has a higher purity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Adsorption of the resin to the fermentation broth: Ferment the S. cellulosus strain ATCC25532 in the liquid medium, and add the treated macroporous resin XAD-16 with 2% volume of the liquid medium to the 2000mL liquid medium, The macroporous resin XAD-16 exists in the entire fermentation process. After 150 hours of fermentation, the adsorption resin is obtained by filtration, and then washed with distilled water to remove the bacteria adhered to the surface of the adsorption resin; among them, the macroporous resin XAD-16 is processed by the following method : First, soak macroporous resin XAD-16 with 3 times the volume of macroporous resin XAD-16 in methanol, shake it for 12 hours, remove methanol, and wash with water until there is no smell of methanol; then soak in methanol for 16 hours, remove methanol and wash with water until No smell of methanol; liquid medium components are potato starch 2.5-3.0g / L, sucrose 0.7-1.0g / L, glucose 0.2-0.5g / L, bean cake powder 1.7...

Embodiment 2

[0032](1) Adsorption of the resin to the fermentation broth: ferment the S. cellulosus strain ATCC25569 in liquid medium, and add macroporous resin XAD-16 and macroporous resin XAD with 4% volume of liquid medium to the liquid medium. -16 exists in the entire fermentation process. After 120 hours of fermentation, the adsorption resin is obtained by filtration, and then washed with distilled water to remove the bacteria adhered to the surface of the adsorption resin; among them, the macroporous resin XAD-16 is processed by the following method: first, use Soak macroporous resin XAD-16 in methanol with 5 times the volume of macroporous resin XAD-16, shake it for 14 hours, remove methanol, wash with water until there is no smell of methanol; soak in methanol for 12 hours, remove methanol and wash with water until there is no smell of methanol; The composition of the liquid medium in the step (1) is potato starch 2.5-3.0g / L, sucrose 0.7-1.0g / L, glucose 0.2-0.5g / L, bean cake powder ...

Embodiment 3

[0038] (1) Adsorption of the resin to the fermentation broth: ferment the S. cellulosus strain ATCC25532 in liquid medium, and add macroporous resin XAD-16 with 5% volume of liquid medium to 2000mL liquid medium, macroporous resin XAD-16 exists in the entire fermentation process. After 130 hours of fermentation, the adsorption resin is obtained by filtration, and then washed with distilled water to remove the bacteria adhered to the surface of the adsorption resin; among them, the macroporous resin XAD-16 is processed by the following method: first, Soak the macroporous resin XAD-16 with 4 times the volume of macroporous resin XAD-16 in methanol, shake it on a shaker for 16 hours, remove methanol, and wash with water until there is no methanol smell; after soaking in methanol for 14 hours, remove methanol and wash with water until there is no methanol smell ; The composition of the liquid medium in the step (1) is potato starch 2.5-3.0g / L, sucrose 0.7-1.0g / L, glucose 0.2-0.5g / L...

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Abstract

The invention relates to an epothilone B separating and extracting method based on a membrane filtration technology. The method comprises the following steps: adsorbing a fermentation liquid by using a macroporous resin; then leaching the absorbed macroporous resin by using ethyl acetate and collecting a leaching liquid; filtering the leaching liquid by using an ultrafiltration membrane; washing and filtering the leaching liquid by using the ethyl acetate; combining filtrates; concentrating; performing detection through preparative HPLC (High Performance Liquid Chromatography); and crystallizing the epothilone B component obtained in a separating manner, thereby obtaining the epothilone B. The method is moderate in condition, simple and convenient to operate, few in separating steps and good in selectivity, and can be used for obviously increasing the yield of the epothilone B, thereby providing conditions more favorable for industrial production. The method with combination of membrane filtration and crystallization is adopted, so that a part of macromolecular impurities can be effectively removed; meanwhile, the application of an organic reagent is avoided, so that the industrial production pollution is reduced. Thus, the loss of the epothilone B in other complicated preparation processes is avoided. The epothilone B prepared in a crystallization manner has relatively high purity. Thus, the epothilone B is broad in application prospect and economic benefit.

Description

technical field [0001] The invention belongs to the field of antibiotic separation and extraction, and relates to an epothilone B separation and extraction method based on membrane filtration technology. Background technique [0002] Epothilone B (epothilone B) is a class of macrolide compounds with the molecular formula C 27 h 41 NO 6 S, with a relative molecular weight of 506.25 and a melting point of 93°C-94°C, has a variety of biological activities and can be used as an antibiotic drug. It was developed by G. It was first reported by et al. in 1993. [0003] Soon after, the Merck laboratory discovered epothilone again in the experiment of screening paclitaxel analogues from 7,000 natural extracts. And found that it has the same mechanism of action as paclitaxel, can promote GTP (guanosine triphosphate)-dependent tubulin polymerization to form microtubules, and has a stabilizing effect on microtubules. The fact that epothilone has an inhibitory effect on tumor cells...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D493/04
CPCC07D493/04C12P17/181
Inventor 龚国利赵婷峰
Owner SHAANXI UNIV OF SCI & TECH
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