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A method for culturing and screening paclitaxel-producing bacteria from Taxus chinensis

A technology of paclitaxel and strains, applied in the biological field, can solve the problems of limited drug sources, low yield, low paclitaxel content, etc., and achieve the effect of practical and simple operation

Inactive Publication Date: 2019-01-22
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since Taxus has been listed as a national first-class protected plant as an endangered species, it is obvious that the extraction of paclitaxel from the plant tissue of Taxus has been greatly restricted. Meet the needs of clinical medicine
Taxus cell tissue culture requires strict conditions, low yield and expensive chemical synthesis, so neither of these two ways is suitable for obtaining paclitaxel in large quantities
The production of paclitaxel by microbial fermentation is a method that does not need to rely on yew and has great potential for industrial production. However, the paclitaxel produced by the paclitaxel bacteria screened by the current methods has a low content and cannot meet industrial requirements.

Method used

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  • A method for culturing and screening paclitaxel-producing bacteria from Taxus chinensis
  • A method for culturing and screening paclitaxel-producing bacteria from Taxus chinensis
  • A method for culturing and screening paclitaxel-producing bacteria from Taxus chinensis

Examples

Experimental program
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Embodiment

[0027] Cut the fresh yew fruit from Huangshan area into 1cm 2 small pieces. Soak the fruit in 75% alcohol for about 5 minutes, and inoculate it on the following plate medium: Potato 20gL -1 , glucose 20gL -1 , agar 20gL -1 and glucose 40gL -1 , peptone 10gL -1 , yeast extract powder 10gL -1 , agar 20gL -1 , and cultured in a 25°C incubator for 8 days. After the colony grows from the incision, observe the hyphae (colonies) growing from the incision of the material in the petri dish, pick the tip of the mycelium at the incision and transfer it to a new plate medium for culture, and after the colony grows, according to the colony According to the difference in shape, color and growth time, the hyphae on the edge were picked for isolation and culture, the shape of the colony and the neatness of the edge of the colony were observed, and corresponding records were made. After repeated purification by mycelium top purification method, they were transferred to PDA slant medium...

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Abstract

The invention provides a method of culturing and screening taxol-producing fungus from taxus chinensis, wherein the method includes the following steps: 1) culturing tissue of the taxus chinensis, and performing separation and purification to obtain endophytic fungus; 2) culturing the endophytic fungus in the step 1), preliminary screening a fermentation metabolite through thin layer chromatography to screen the strains being capable of producing taxol or analogues thereof; 3) with cytotoxicity test, determining the inhibition rate of the metabolite to CHO cells; and 4) performing further analysis through high performance liquid chromatography to determine a taxol-producing strain. The method of separating the taxol-producing fungus from taxus chinensis is simple in process and low in cost. The taxol-producing fungus extracted from through the method is greatly increased in content of produced taxol when being used for microorganism cultivation. The method provides a wider approach for microorganism engineering.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for cultivating and screening taxol-producing bacteria from yew. Background technique [0002] Paclitaxel was first isolated from the bark of Taxus breviifolia and named paclitaxel. Paclitaxel was later confirmed to have a unique anti-cancer mechanism. Due to its unique anti-cancer mechanism, it has good curative effect on some advanced tumors that ordinary anti-cancer drugs are helpless, and has outstanding curative effect on a variety of clinical malignant tumors, which is completely different from the previous marketed drugs. any anticancer drug. The U.S. Food and Drug Administration (FDA) officially approved paclitaxel for clinical use in December 1992. [0003] Since Taxus has been listed as a national first-class protected plant as an endangered species, it is obvious that the extraction of paclitaxel from the plant tissue of Taxus has been greatly restricted. Meet ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12R1/885
Inventor 汪友明樊美珍马忠友胡丰林张袖丽田超陆瑞利
Owner ANHUI AGRICULTURAL UNIVERSITY
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