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Fermentation additive capable of changing generation rate of epothilone compound and improving yield of epothilone A

A technology of epothilones and additives, which is applied in the field of fermentation and preparation of antineoplastic drugs, can solve the problems of changing the product ratio, changing the biological ratio of epothilones, etc., and achieve the effects of increasing production, good water solubility, and reducing costs

Active Publication Date: 2015-12-30
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the use of fermentation additives to control the secondary metabolism of epothilone and change the product ratio
By adding specific substances to maintain the reducibility of metal ions or epothilone synthesis intermediates, thereby changing the biological ratio of epothilones and specifically increasing the yield of an epothilone compound has not been reported in the patent literature so far

Method used

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  • Fermentation additive capable of changing generation rate of epothilone compound and improving yield of epothilone A
  • Fermentation additive capable of changing generation rate of epothilone compound and improving yield of epothilone A
  • Fermentation additive capable of changing generation rate of epothilone compound and improving yield of epothilone A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] (1) Selection of fermentation strains: Sorangium cellulosum So0157-2 among the myxobacteria strains.

[0044] (2) Activation of strains: the above strains were inoculated on a solid plate medium, cultured at 30° C. for 7 days, then inoculated into a liquid medium, and cultured at 30° C. at 200 rpm for 5 days.

[0045] Then pour the obtained bacterial solution into a spinner bottle containing glass beads and disperse for 2 minutes, and set aside.

[0046] in,

[0047] The composition of the solid plate culture medium is: KNO 3 0.5g / L, Na 2 HPO 4 0.25g / L, MgSO 4 ·7H 2 O1g / L, FeCl 3 0.01g / L, trace element solution 1ml / L, agar 15g / L, pH7.2 (after pouring the plate to solidify, paste a 1.5×1.5cm square filter paper piece).

[0048] The composition of described liquid medium is: potato starch 8g / L, soybean peptone 2g / L, glucose 2g / L, yeast powder 2g / L, MgSO 4 1g / L, CaCl 2 1g / L, EDTA-Fe3+1ml / L, trace element solution 1.0ml / L, pH7.2.

[0049] The trace element solutio...

Embodiment 2

[0064] (1) Selection of fermentation strains: Sorangium cellulosum So0157-2 among the myxobacteria strains.

[0065] (2) Activation of strains: the above strains were inoculated on a solid plate medium, cultured at 30° C. for 7 days, then inoculated into a liquid medium, and cultured at 30° C. at 200 rpm for 5 days.

[0066] Then pour the obtained bacterial solution into a spinner bottle containing glass beads and disperse for 2 minutes, and set aside.

[0067] in,

[0068] The composition of the solid plate culture medium is: KNO 3 0.5g / L, Na 2 HPO 4 0.25g / L, MgSO 4 ·7H 2 O1g / L, FeCl 3 0.01g / L, trace element solution 1ml / L, agar 15g / L, pH7.2 (after pouring the plate to solidify, paste a 1.5×1.5cm square filter paper piece).

[0069] The composition of described liquid medium is: potato starch 8g / L, soybean peptone 2g / L, glucose 2g / L, yeast powder 2g / L, MgSO 4 1g / L, CaCl 2 1g / L, EDTA-Fe3+1ml / L, trace element solution 1.0ml / L, pH7.2.

[0070] The trace element solutio...

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Abstract

The invention discloses a fermentation additive capable of changing the generation rate of an epothilone compound and improving the yield of epothilone A. The additive is 2,3,5,6-tetrahydroxy-2-hydrosorbic acid-4-lactone; and the final adding concentration in an epothilone liquid fermentation culture medium is 100+ / -15 mu mol / L. An experiment proves that the additive disclosed by the invention is capable of keeping the reducibility of metal ions and metabolic intermediates and adjusting the secondary metabolism flow direction of sorangium cellulosum, so that the gross of the epothilone A is averagely improved by 62%-105% in comparison with that when the additive is not added; the ratio of the epothilone A to the epothilone B in a fermentation liquid is significantly changed; and (1.83+ / -0.16):1 (the epothilone A to the epothilone B) before addition is changed into (5.67+ / -0.71):1 (the epothilone A to the epothilone B). According to the fermentation additive, regulation and control on the secondary metabolism flow direction and the product type of the epothilone compound in the fermentation process are achieved; the fermentation cost of the epothilone A is reduced; and a way is also provided for production of the epothilone A.

Description

technical field [0001] The invention relates to an additive for fermentative production of epothilones and its application, in particular to an additive capable of changing the production ratio of epothilones and increasing the production of epothilones A and its application, which belongs to the fermentation of antitumor drugs Preparation technology field. Background technique [0002] The microtubule polymerization-promoting compound paclitaxel (Paclitaxel, ) and its analog Docetaxel It is currently the most successful anti-tumor chemotherapy drug in clinical practice. It is used in the treatment of solid cancers such as ovarian cancer, thymus cancer, colon cancer, lung cancer and liver cancer. It is the first choice drug for solid tumor chemotherapy approved by the US FDA. Among the natural compounds with the same or similar biological functions as paclitaxel, only Epothilones (Epothilones) is derived from the fermentation product of the microorganism S. cellulosus, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/01
Inventor 李越中胡玮郑连帅
Owner SHANDONG UNIV
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