Biosynthesis method for improving yield of epothilone B
A technology for epothilone and biosynthesis, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of low yield and high production cost of epothilone B, and achieves improved growth and shortened production. Period, effect of preventing variation
Active Publication Date: 2013-06-12
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 effectively reduce production cost. Biosynthetic method, the yield of epothilone B in the active product produced by the expansion and fermentation of this method can reach as high as 30-45%, which has a good industrial production prospect
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Embodiment 1
[0031] Embodiment 1: a kind of biosynthetic method of improving the productive rate of epothilone B of the present invention, the synthesis step comprises:
[0032] (1), produce starting bacterial classification, select the strong and vigorous starting myxobacterial bacterial classification as production bacterial classification by 3000rpm centrifugal shake flask bacterial liquid;
[0033] (2), solid slant expansion culture: its slant medium components are calculated by mass%, glucose 0.5%, peptone 0.5%, starch 1%, sodium chloride 0.5%, gibberellin 0.002% and agar powder 2%, The balance is water; sterilize at 120°C for 30 minutes, and after 24 hours, spread the well-grown production starting strains on a solid slant medium, and cultivate them at 30°C±2°C and relative humidity of 55% for 8 days;
[0034] (3) Inoculate the well-growing strains into the shake flask expansion medium sterilized at 120°C for 30 minutes. The components of the shake flask expansion medium are starch 5...
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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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IPC IPC(8): C12P17/18C07D493/04C12R1/01
Inventor 陈庆源何福彪张文凯
Owner HUBEI HONCH PHARMA