Fermentation production process of Epothilone B

A technology for epothilone and fermentation broth, applied in the directions of fermentation, microorganisms, microorganism-based methods, etc., can solve the problem of high production cost of epothilone, difficulty in increasing epothilone fermentation yield, and high cost of separation and purification of target products. The problem is to eliminate the glucose inhibitory effect and improve the yield.

Inactive Publication Date: 2013-11-27
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are 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; the second reason is that the production costs of epothilones are high. It is because there are a lot of epothilone homologues in the product after the fermentation, which leads to very high cost of separation and purification of the target product

Method used

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  • Fermentation production process of Epothilone B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The preparation of molecularly imprinted polymer and the fermentation of epothilone B include the following steps:

[0031] 1) Preparation of molecularly imprinted polymers

[0032] According to the following proportions: mix 1 mmol of epothilone B, 4 mmol of methacrylic acid, 20 mmol of ethylene glycol dimethacrylate, 10 mg of azobisisobutyrocyanide, and 3 ml of acetonitrile into a reaction tube, pass nitrogen and oxygen for 30 minutes, and Under the protection of nitrogen, the reaction tube was sealed, placed 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 a polymerization reaction product;

[0033] Crushing, grinding, and sieving the obtained reaction product to obtain polymer particles with a particle size of 40-60 μm;

[0034] After the obtained polymer particles were washed with water, they were extracted with a Soxhlet extractor in a mixed solution of acetic acid:methanol (volume ratio 1:9) for 48 hours, ...

Embodiment 2

[0049] The preparation of molecularly imprinted polymer and the fermentation of epothilone B include the following steps:

[0050] 1) Preparation of molecularly imprinted polymers

[0051] According to the following proportions: mix 1 mmol of epothilone B, 3 mmol of methacrylic acid, 25 mmol of ethylene glycol dimethacrylate, 10 mg of azobisisobutyronitrile, and 8 ml of methanol into a reaction tube, pass nitrogen and exhaust oxygen for 30 minutes, and Under the protection of nitrogen, the reaction tube was sealed, placed in a 45°C water bath for 24 hours of polymerization, and then polymerized in a 65°C water bath for 12 hours to obtain a polymerization reaction product;

[0052] Crushing, grinding, and sieving the obtained reaction product to obtain polymer particles with a particle size of 40-60 μm;

[0053] After the obtained polymer particles were washed with water, they were extracted with a Soxhlet extractor in a mixed solution of acetic acid:methanol (volume ratio 3:7...

Embodiment 3

[0065] The preparation of molecularly imprinted polymer and the fermentation of epothilone B include the following steps:

[0066] 1) Preparation of molecularly imprinted polymers

[0067] According to the following proportions: mix 1 mmol of epothilone B, 3.5 mmol of methacrylic acid, 30 mmol of ethylene glycol dimethacrylate, 5 mg of azobisisobutylcyanide, and 3 ml of acetonitrile into the reaction tube, pass nitrogen and oxygen for 30 minutes, Under the protection of nitrogen, the reaction tube was sealed, placed 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 a polymerization reaction product;

[0068] Crushing, grinding, and sieving the obtained reaction product to obtain polymer particles with a particle size of 40-60 μm;

[0069] After the obtained polymer particles were washed with water, they were extracted with a Soxhlet extractor in a mixed solution of acetic acid:methanol (volume ratio 3:7) for 48 hours, an...

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Abstract

The invention discloses a fermentation production process of Epothilone B, i.e. a process for increasing the fermentation output of Epothilone B of Epothilone B production bacterial strain sprangium cellulosum. The process comprises the following steps: 1) adding a molecular imprinting polymer adsorbent of the Epothilone B in the fermentation process; 2) selecting soybean meal, corn starch and the like suitable for sprangium cellulosum production; and 3) adding glucose pulses in the fermentation process and the like. So the fermentation production output of the sprangium cellulosum of the epothilone B achieves 50mg / L.

Description

technical field [0001] The invention belongs to the technical field of epothilone B production, and relates to a fermentation production process of epothilone B. Background technique [0002] Epothilone B (Epothilone B) is a polyketide secondary metabolite produced by the myxobacterium Sorangium cellulosum, which has the same microtubule-stabilizing activity as paclitaxel, an anti-tumor chemotherapy drug widely used in clinical practice , and has activity against paclitaxel-resistant tumor cells. In addition, the characteristics of epothilone derived from microbial metabolites make it considered to be a more promising anticancer drug than paclitaxel. [0003] The discovery of epothilones can be traced back to extensive research work on myxobacteria in soil by microbiologists at the German National Center for Biotechnology Research (GBF) in the 1980s. In 1987, when they were looking for antifungal drugs, they discovered that a strain of myxobacteria (Sorangium cellulosum So...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18C08F222/14C08F220/06C08J9/26B01J20/26B01J20/30C12R1/645
Inventor 龚国利陈松李慧
Owner SHAANXI UNIV OF SCI & TECH
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