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Fermentation production method of Epothilone D provided with molecular imprinting polymer

A technology of epothilone and molecular imprinting, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the difficulty of increasing epothilone yield, toxicity and feedback inhibition epothilone fermentation yield Difficult to improve and high production cost of epothilone

Inactive Publication Date: 2014-07-02
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Epothilone is currently expensive to produce for two important reasons
The first is that the toxicity and feedback inhibition of epothilones to the production strains make it difficult to increase the fermentation yield of epothilones. Although this difficulty has been achieved by adding macroporous adsorption resin XAD-16 to the fermentation broth However, the resin’s non-specific adsorption of nutrients in the fermentation broth has brought many new difficulties to the improvement of the yield of epothilone
The second reason for the high production cost of epothilones is that there are a large number of epothilones homologues in the product after the fermentation, which leads to very high cost of separation and purification of the target product (for example: epothilone D)
Molecularly imprinted polymers, as a polymer with specific targeting adsorption capacity, have been successfully applied in drug analysis and separation methods, but molecularly imprinted polymers have not been applied to the fermentation production of epothilone D

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The preparation of molecularly imprinted polymer and the fermentation of epothilone D include the following steps:

[0028] 1) Preparation of molecularly imprinted polymers

[0029] Weigh according to the following ratio: 0.4917g (1mmol) epothilone D, 0.284g (4mmol) acrylamide or 0.228g (4mmol) allylamine, 3.35g (25mmol) N,N-methylenebisacrylamide, 120mg Azobisisobutyronitrile and 3ml deionized water were added into the three-necked flask, and the 2 or A r Under protection, the container was ultrasonically oscillated for 20 minutes to fully dissolve and degas it;

[0030] Nitrogen was introduced into the reactor under stirring for 30 minutes to discharge the oxygen in the reactor, polymerized in a water bath at 50°C for 24 hours, and then polymerized in a water bath at 60°C for 12 hours to obtain the reaction product;

[0031] The obtained reaction product was crushed, ground, and sieved to obtain molecularly imprinted polymer particles containing template molecules ...

Embodiment 2

[0043] The preparation of molecularly imprinted polymer and the fermentation of epothilone D include the following steps:

[0044] 1) Preparation of molecularly imprinted polymers

[0045] Weigh according to the following ratio: 0.4917g (1mmol) epothilone D, 0.344g (4mmol) methacrylic acid, 4.96g (25mmol) ethylene glycol methacrylate and 120mg dibenzoyl peroxide dissolved in 3ml methanol solution, in N 2 or A r Under protection, the container is ultrasonically oscillated for 20-30 minutes to fully dissolve and degas it;

[0046] Nitrogen was introduced into the reactor under stirring for 30 minutes to discharge the oxygen in the reactor, polymerized in a water bath at 50°C for 24 hours, and then polymerized in a water bath at 60°C for 12 hours to obtain the reaction product;

[0047] The resulting reaction product was crushed, ground, and sieved to obtain molecularly imprinted polymer particles containing template molecules with a particle size of 40-60 μm, and then washed ...

Embodiment 3

[0058] The preparation of molecularly imprinted polymer and the fermentation of epothilone D include the following steps:

[0059] 1) Preparation of molecularly imprinted polymers

[0060] Weigh according to the following proportions: Weigh 0.4917g (1mmol) epothilone D, 0.414g (2mmol) 2-acrylamido-2-methylpropanesulfonic acid, 3.82g (25mmol) 4-imidazole ethyl acrylate and 120mg of azobisisobutyronitrile dissolved in 3ml of acetonitrile solution, in N 2 or A r Under protection, the container is ultrasonically oscillated for 20-30 minutes to fully dissolve and degas it;

[0061] Nitrogen was introduced into the reactor under stirring for 30 minutes to discharge the oxygen in the reactor, polymerized in a water bath at 55°C for 24 hours, and then polymerized in a water bath at 65°C for 12 hours to obtain a reaction product;

[0062] The obtained reaction product is crushed, ground, and sieved to obtain molecularly imprinted polymer particles containing template molecules with ...

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Abstract

The invention discloses a fermentation production method of Epothilone D provided with a molecular imprinting polymer. The special molecular imprinting polymer adsorbing the Epothilone D is added in the fermentation production process of the Epothilone D, so the Epothilone D produced in fermentation is continuously adsorbed by the molecular imprinting polymer before reaching absorption balance in fermentation, and on the other hand, the Epothilone D is adsorbed by the molecular imprinting polymer, which is conductive to the separation of the Epothilone D in a fermentation product. The Epothilone D can be separated from fermentation liquor as long as the molecular imprinting polymer used as sediment is centrifugally separated and collected, then the Epothilone D is extracted from the molecular imprinting polymer, and the molecular imprinting polymer can be reused after being dried.

Description

technical field [0001] The invention belongs to the technical field of epothilone D production, and relates to a method for fermenting and producing epothilone D by adding molecularly imprinted polymers. Background technique [0002] Epothilone D (Epothilone D, KOS-862) is an anti-tumor drug produced by the microorganism Cystis cellulosus, which is currently undergoing phase II clinical trials, and is currently a research hotspot of anti-tumor drugs. Epothilone is currently expensive to produce, for two important reasons. The first is that the toxicity and feedback inhibition of epothilones to the production strains make it difficult to increase the fermentation yield of epothilones. Although this difficulty has been achieved by adding macroporous adsorption resin XAD-16 to the fermentation broth However, the non-specific adsorption of the resin to the nutrients in the fermentation broth has brought many new difficulties for the improvement of the yield of epothilones. The...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/16C08J9/28C08F222/38C08F220/14C08F2/44C12R1/01
Inventor 龚国利陈松李慧王娜
Owner SHAANXI UNIV OF SCI & TECH
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