Biosynthesis method for improving yield of epothilone B

A technology for epothilone and biosynthesis is applied in the biosynthesis of industrialized production to improve the yield of epothilone B, and in the field of biosynthesis of epothilone, it can solve the problem of high production cost and low yield of epothilone B and other problems to achieve the effect of shortening the production cycle, increasing growth and increasing productivity

Active Publication Date: 2012-07-18
HUBEI HONCH PHARMA
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Problems solved by technology

[0007] The purpose of the present invention is to solve the problem of high production cost and low yield of epothilone B in the prior art, and provide a kind of epothilone with high yield, which can adapt to industrial production and can

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  • Biosynthesis method for improving yield of epothilone B
  • Biosynthesis method for improving yield of epothilone B

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[0031] Example 1: A biosynthetic method for improving the yield of epothilone B according to the present invention, the synthesis steps include:

[0032] (1) For the production of starting strains, the vigorously growing starting myxobacter strains are selected as the production strains through 3000rpm centrifugal shake flask bacterial liquid;

[0033] (2). Solid slope expansion: The components of the slope culture medium are calculated by mass%, glucose 0.5%, peptone 0.5%, starch 1%, sodium chloride 0.5%, gibberellin 0.002% and agar powder 2%, The remainder is water; sterilize at 120°C for 30 minutes. After 24 hours, spread well-grown production starting strains on a solid slant medium, and cultivate for 8 days at 30°C±2°C and relative humidity 55%;

[0034] (3) Inoculate the well-grown strains in the shake flask expansion base sterilized at 120°C for 30 minutes. The components of the shake flask expansion base are starch 5g / L, yeast extract 2g / L, and glucose 1g / L, MgSO 4 1g / L, C...

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Abstract

The invention relates to a biosynthesis method for improving yield of epothilone B. The biosynthesis method comprises the following steps of: selecting robust strains for producing the epothilone B by using a centrifugal process, inoculating the strains into a sterilized seed medium, performing shaking culture at 30DEG C on a shaker, and fermenting; adding a precursor inductor phenylalanine into a fermentation medium; harvesting a culture solution after culturing at 30DEG C for 10 to 11 days; and collecting, adsorbing and saturating by using macroporous resin, eluting by using ethyl acetate, separating and purifying, performing vacuum concentration, extracting, crystalizing at low temperature, and performing vacuum drying. The purity of the epothilone B can be effectively improved by a liquid preparation column, and is over 99.5 percent; and methanol is adopted for extraction and separation, so that the epothilone B is crystallized in the methanol, and the finished product reaches thepharmaceutical grade. By the method, the yield of the epothilone B is greatly improved and is 30-45 percent, and the method has a good industrial production prospect.

Description

technical field [0001] The invention relates to a biosynthesis method of epothilones, in particular to a biosynthesis method for increasing the yield of epothilone B in industrialized production, and belongs to the field of fermentation and cultivation of antibiotic microorganisms. Background technique [0002] Epothilone B is a new member of macrolide antibiotics, and epothilone is a class of antibiotics with unique structure and unique biological activity. Its molecular structure is as follows: [0003] [0004] Epothilone A [0005] Epothilone B [0006] Among them, epothilone B is a macrolide compound produced by myxobacteria (Pyracystis cellulosus). Epothilone B not only has anti-tumor effects, but also has good antagonistic and therapeutic effects on skin diseases caused by fungal infections. Epothilone B has low toxicity and few side effects, and has good application prospects. However, the current method for preparing epothilone B by fermentation has high produ...

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Application Information

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IPC IPC(8): C12P17/18C07D493/04C12R1/01
Inventor 陈庆源何福彪张文凯
Owner HUBEI HONCH PHARMA
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