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A method for producing epothilone b

A technology for epothilone and bacteria production, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems such as the inability to meet the needs of large-scale production, and achieves the advantages of subsequent extraction and purification, easy promotion, and industrialization. low cost effect

Active Publication Date: 2020-05-05
LUNAN BETTER PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Prior art fermentation methods still cannot meet the needs of large-scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1. solid plate culture

[0031] The composition of the solid medium is 0.1% of glucose, 0.2% of peptone, 0.1% of magnesium sulfate, 0.2% of calcium chloride, 0.2% of dipotassium hydrogen phosphate and 0.3% of yeast extract. Adjust the pH to 7.2, sterilize at 121°C for 30 minutes, spread on a solid plate medium, and incubate upside down at 30°C for 6-7 days.

Embodiment 2

[0032] Example 2. Seed culture of S. cellulosus

[0033] ① Shake flask expansion

[0034] Inoculate the well-growing plates in shake flasks. The components of the shake flask medium are glucose 0.1%, soluble starch 2%, magnesium sulfate 0.1%, calcium chloride 0.2%, dipotassium hydrogen phosphate 0.2%, yeast extract 0.3 %. Adjust the pH to 7.2, sterilize at 121° C. for 30 minutes, and culture at 30° C. with shaking at 200 rpm for 4 days.

[0035] ②Seed cultivation

[0036] Inoculate a well-growing shake flask into a seed tank with the same culture medium as the shake flask medium, steam sterilize at 121°C for 30 minutes, and inoculate after sterilization. The culture temperature is 30°C, the rotation speed is 200rpm, and the ventilation rate is 0.3vvm The tank pressure is 0.04kg / cm2, and 1M sodium hydroxide is added to keep the pH at 7.2. When the dissolved oxygen drops slowly, transplant the seeds. The amount of bacterial cells was determined to be 8%.

Embodiment 3

[0037] Example 3. Seed culture of epothilone B

[0038] Inoculate a well-growing shake flask into a seed tank with the same culture medium as the shake flask medium, steam sterilize at 121°C for 30 minutes, and inoculate after sterilization. The culture temperature is 30°C, the rotation speed is 200rpm, and the ventilation rate is 0.3vvm The tank pressure is 0.04kg / cm2, and 20% ammonia water is added to keep the pH at 7.2. When the dissolved oxygen drops slowly, transplant the seeds. The amount of bacterial cells was determined to be 13%.

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PUM

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Abstract

The invention concretely discloses a fermentation method for producing epothilone B, and belongs to the field of biological fermentation. According to the method, during culturing of a seeding tank, ammonia hydroxide is added in a flowing mode to adjust the pH value, so that a lot of seed thalli are obtained; during fermentation, ammonia hydroxide or propanoic acid is added in a flowing mode to regulate the pH value, in this way, the yield of the epothilone B is increased, the regulation procedures are simplified, production cost is lowered, industrialized production is easy, and application prospects are good.

Description

technical field [0001] The invention belongs to the field of biological fermentation, and in particular relates to a method for producing epothilone B. Background technique [0002] Epothilones (epothilones) was discovered in 1993 in the secondary metabolites of the myxobacteria Sorangium cellulosum, and its structure is a macrolide compound. The anti-tumor mechanism is similar to that of paclitaxel, which has the effect of inhibiting microtubule depolymerization. In contrast, epothilone is more water-soluble and has a strong effect on both multidrug-resistant tumor cells and paclitaxel-resistant tumor cells. It has anticancer activity and does not have serious side effects such as granulocytopenia, peripheral neuropathy, alopecia, and allergy in the clinical application of paclitaxel. Therefore, the development of epothilones has clinical value and social significance. In 2007, ixabepilone, a derivative of epothilones B, was approved for marketing in the United States, wh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18C12N1/20C12R1/01
CPCC12N1/20C12P17/181
Inventor 张贵民杨家森沈书庆
Owner LUNAN BETTER PHARMA
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