A method for the separation and extraction of epothilone b based on molecular imprinting adsorption technology

A technology of epothilone and molecular imprinting, applied in organic chemistry, fermentation, etc., can solve the problems of large-scale production that is difficult to realize industrialization, low operation efficiency, low product yield, etc., and achieve broad application prospects and economic benefits , easy operation, high purity effect

Active Publication Date: 2016-08-17
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Researchers have put a lot of energy into its synthesis and separation and extraction. However, for the separation and purification of epothilone, the methods currently used have the following defects and deficiencies: the steps are too cumbersome, making the efficiency of the operation lower and The process cost is high, the product yield is low, and the cost is high, especially it is difficult to realize industrialized large-scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) Adsorption of the resin to the fermentation broth: ferment S. cellulosus ATCC25532 in a liquid medium, and add the treated macroporous resin XAD-16 to the liquid medium, ferment for 150 hours and filter to obtain the adsorption resin, and then Rinse with distilled water to remove S. cellulosus attached to the surface of the adsorption resin;

[0028] Wherein, the addition of described macroporous resin is 2% of the volume of liquid culture medium;

[0029] The composition of the liquid medium is 2.5-3.0 g / L of potato starch, 0.7-1.0 g / L of sucrose, 0.2-0.5 g / L of glucose, 1.7-2.0 g / L of bean cake powder, MgSO 4 ·7H 2 O2.3~2.5g / L, CaCl 2 3.0~3.5g / L, EDTA-Fe 3+ 2mL / L, trace elements 0.5-1.0mL / L, pH value 7.2.

[0030] The macroporous resin is processed by the following method: first, soak the macroporous resin with methanol 3 times the volume of the macroporous resin, shake it on a shaker for 12 hours, take out and discard the methanol, and then wash with pure wat...

Embodiment 2

[0038] (1) Adsorption of the resin to the fermentation broth: ferment S. cellulosus ATCC25569 in a liquid medium, and add the treated macroporous resin XAD-16 to the liquid medium, and filter to obtain the adsorption resin after 120 hours of fermentation, and then Rinse with distilled water to remove S. cellulosus attached to the surface of the adsorption resin;

[0039] Wherein, the addition of described macroporous resin is 5% of the volume of liquid culture medium;

[0040] The composition of the liquid medium is 2.5-3.0 g / L of potato starch, 0.7-1.0 g / L of sucrose, 0.2-0.5 g / L of glucose, 1.7-2.0 g / L of bean cake powder, MgSO 4 ·7H 2 O2.3~2.5g / L, CaCl 2 3.0~3.5g / L, EDTA-Fe 3+ 2mL / L, trace elements 0.5-1.0mL / L, pH value 7.2.

[0041] The macroporous resin is processed by the following method: first, soak the macroporous resin with methanol 5 times the volume of the macroporous resin, shake it on a shaker for 16 hours, take out the methanol, and then wash it with pure wa...

Embodiment 3

[0049] (1) Adsorption of the resin to the fermentation broth: Ferment S. cellulosus ATCC25532 in a liquid medium, and add the treated macroporous resin XAD-16 to the liquid medium, ferment for 140 hours and filter to obtain the adsorption resin, and then Rinse with distilled water to remove S. cellulosus attached to the surface of the adsorption resin;

[0050] Wherein, the addition of described macroporous resin is 4% of the volume of liquid culture medium;

[0051] The composition of the liquid medium is 2.5-3.0 g / L of potato starch, 0.7-1.0 g / L of sucrose, 0.2-0.5 g / L of glucose, 1.7-2.0 g / L of bean cake powder, MgSO 4 ·7H 2 O2.3~2.5g / L, CaCl 2 3.0~3.5g / L, EDTA-Fe 3+ 2mL / L, trace elements 0.5-1.0mL / L, pH value 7.2.

[0052] The macroporous resin is processed by the following method: first, soak the macroporous resin with methanol 4 times the volume of the macroporous resin, shake it on a shaker for 14 hours, take out and discard the methanol, and then wash with pure wat...

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Abstract

The invention provides a separation and extraction method for epothilone B based on a molecular imprinting adsorption technology. The separation and extraction method comprises the following steps: adopting macroporous resin to adsorb fermented liquid; then extracting the adsorbed macroporous resin by using ethyl acetate; filtering extracting liquid by using an ultra-filtering membrane to obtain a crude extract; adding an epothilone B molecular imprinting polymer into the crude extract to specifically adsorb the epothilone B; vibrating on a shaking table, and centrifuging and collecting lower-layer sediment; eluting the sediment by using methanol for a plurality of times and collecting eluting liquid; re-crystallizing to obtain the epothilone B. The separation and extraction method has the characteristics of simplicity and convenience in operation, good selectivity, low pollution and the like. The separation and extraction method is simple and feasible in operation steps, few in separation steps, low in cost and high in yield; the loss of the epothilone B in other complicated preparation flows is avoided; more importantly, the separation and extraction method is convenient for industrial large-scale production and has very wide application prospect and economic benefits.

Description

technical field [0001] The invention relates to a method for separating and extracting epothilone B by combining membrane filtration technology and molecular imprinting technology, in particular to a method for separating and extracting epothilone B based on molecular imprinting adsorption technology. Background technique [0002] Epothilone B (Epothilone B) is a class of secondary metabolites of macrolides produced by the microorganism S. cellulosus, and its molecular formula is 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 is a research hotspot of anti-tumor drugs at present. [0003] Studies have found that epothilone B not only has the same mechanism of action as paclitaxel, it can promote GTP (guanosine triphosphate)-dependent tubulin polymerization to form microtubules, and has a stabilizing effect on microtubules. Moreover, it has superi...

Claims

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Application Information

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Patent Type & Authority Patents(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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