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Fermentation production method based on epothilone B metabolic pathways

A technology of epothilones and metabolic pathways, applied in the field of fermentation engineering, can solve the problems of instability and low yield of epothilone B, and achieve the effects of reduced fermentation costs, environmental friendliness, and simple operation

Inactive Publication Date: 2013-08-14
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the yield of epothilone B is relatively low and unstable, and is greatly affected by factors such as culture conditions and environment.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The fermentation production method based on the epothilone B metabolic pathway comprises the following steps:

[0029] 1) Inoculate Sotrophysia cellulosus ATCC25531 strain into a 250ml Erlenmeyer flask containing 50ml of M26 medium, and culture it on a shaker at 30°C and 170r / min for 72h to obtain a seed solution for fermentation; , M26 medium components are: potato starch 8.0g / L, soybean peptone 2.0g / L, glucose 2.0g / L, yeast powder 2.0g / L, MgSO 4 ·7H 2 O1.0g / L, CaCl 2 1.0g / L, EDTA-Fe 3+ 1mL / L, trace elements 1mL / L, adjust the pH value to 7.2 with KOH;

[0030] 2) Inoculate 5ml of seed solution into 50ml of resin-containing fermentation broth, and inoculate 1×10 in the initial fermentation broth -5 ~3×10 -5 Potassium ferrocyanide was added in an amount of mol / L, the fermentation vessel was a 250ml Erlenmeyer flask, and cultured on a shaking table at 30°C and 170r / min for 96h; the added resin was macroporous resin XAD16, and the amount added was 20ml / L; among them, ...

Embodiment 2

[0033] The fermentation production method based on the epothilone B metabolic pathway comprises the following steps:

[0034]1) Inoculate a 250ml Erlenmeyer flask containing 50ml of M26 culture medium in a 250ml Erlenmeyer flask containing 50ml of M26 medium, and culture it on a shaker at 25°C and 160r / min for 96 hours to obtain a seed solution for fermentation; where M26 Culture medium composition is with embodiment 1;

[0035] 2) Inoculate 5ml of seed solution into 50ml of resin-containing fermentation broth, and in the initial fermentation broth according to 3×10 -5 Add potassium ferrocyanide and biotin in the amount of mol / L and 8ug / L, the fermentation vessel is a 250ml Erlenmeyer flask, and culture on a shaking table at 25°C and 160r / min for 72h; the added resin is macroporous resin XAD16, add The amount is 20ml / L; Among them, the components of the fermentation medium are: yeast powder 25g / L, corn starch 4g / L, sodium dihydrogen phosphate 2g / L, disodium hydrogen phosphate...

Embodiment 3

[0038] 1) Inoculate a 250ml Erlenmeyer flask containing 50ml of M26 culture medium in a 250ml Erlenmeyer flask containing 50ml of M26 medium, and culture it on a shaker at 30°C and 160r / min for 96 hours to obtain a seed solution for fermentation culture; where M26 Culture medium component is with embodiment 1;

[0039] 2) Inoculate 7.5ml of seed liquid into 50ml of resin-containing fermentation medium, and after 24 hours of cultivation, add sodium propionate in an amount of 0.5mmol / L to the fermentation broth. The fermentation vessel is a 250ml Erlenmeyer flask. Under the condition of shaking for 96 hours, the added resin is macroporous resin XAD16, and the added amount is 30ml / L; among them, the components of the fermentation medium are: yeast powder 35g / L, corn starch 5g / L, sodium dihydrogen phosphate 1g / L L, disodium hydrogen phosphate 1g / L, MgSO 4 ·7H 2 O5g / L, FeSO 4 ·7H 2 O0.1g / L, CaCl 2 2g / L, MnCl 2 0.3g / L and glucose 10g / L, adjust the pH value to 7.5 with KOH;

...

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PUM

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Abstract

The invention discloses a fermentation production method based on epothilone B metabolic pathways and belongs to the technical field of fermentation engineering. The method comprises the following steps of: 1) inoculating Sorangium cellulosum into an M26 culture medium, carrying out shake cultivation to obtain a seed solution; 2) inoculating the seed solution into fermentation liquor containing resin, carrying out shake culturing for 72-120 hours at 25-35 DEG C, adding a precursor and small molecular substances into the fermentation liquor containing resin; and 3) filtering the fermentation liquor after the culture is finished, collecting the resin, washing the resin and oscillating and extracting with methyl alcohol to obtain extract liquor containing the epothilone B. According to the invention, the precursor and small molecular substances related to the biosynthetic pathways obtained by screening are added into the fermentation liquor, and the constructive metabolism pathways of the epothilone B are disturbed, so that the excellent bacterial strain gives full play to the ability of anabolism and the fermentation level of the epothilone B is improved greatly, thus the fermentation cost is reduced and the commercial process of the anti-cancer drug is promoted.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering, relates to microbial fermentation pharmaceutical technology, in particular to a fermentation production method of epothilone B. Background technique [0002] Epothilone is a secondary metabolite produced by S. cellulosus. It is a sixteen-membered macrolide anticancer drug. It has the same effect as the anticancer drug paclitaxel by promoting microtubule polymerization and inhibiting tumor cells. Proliferation, and compared with paclitaxel, epothilone has the advantages of less toxicity, better water solubility, stronger effect on drug-resistant cells, and wider anti-tumor spectrum. At the same time, the chemical structure is simple and easy to modify, causing Therefore, it is a new type of anti-tumor drug that is closely watched by the academic circle, has high hopes in the medical circle, and has a broad prospect. At present, epothilone B has entered clinical research. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/01
Inventor 龚国利王娜刘丽丽
Owner SHAANXI UNIV OF SCI & TECH
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