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Method for preparing homoharringtonine and special strain

A technology for homoharringtonine and Alternaria thaliana, which is applied in the field of preparing homoharringtonine, can solve the problems of insufficient drug source, slow growth, difficulty in reproduction and the like, and achieves that the fermentation process is easy to master , the effect of low production cost, simple separation and purification steps

Inactive Publication Date: 2011-08-24
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to slow growth, difficult reproduction and over-harvesting, the resources are almost exhausted
Insufficient drug sources have become the bottleneck limiting the clinical application of homoharringtonine

Method used

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  • Method for preparing homoharringtonine and special strain
  • Method for preparing homoharringtonine and special strain
  • Method for preparing homoharringtonine and special strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, strain isolation and identification

[0044] 1. Isolation of strains

[0045] Endophytic fungi were isolated from the bark of Torreya hainanensis in Danzhou Tropical Botanical Garden, Hainan Province. After fermentation experiments, HPLC, LC-MS, and MS-MS analysis, it was proved that homoharringtonine could be produced.

[0046]The separation method is as follows: wash off the dust on the surface of the bark of Torreya hainanensis with sterile water, soak it with 75% alcohol for 15 minutes; soak it with 0.1% mercuric chloride solution for 10 minutes, and rinse it with sterile water to remove the surface mercuric chloride solution; Cut the bark into small pieces of 5×5mm with sterile scissors; inoculate the above-mentioned surface-sterilized bark on potato (PDA) medium, and cultivate at a constant temperature of 25°C; Picking method, pick colonies of different shapes, transfer to PDA slant, purify and cultivate at 25-28°C, and store for later use;

[0047...

Embodiment 2

[0060] Embodiment 2, the method for preparing homoharringtonine

[0061] 1. Slope activation: inoculate the strain in PDA medium, activate it, and culture it at 28°C for 48-72h.

[0062] PDA medium: 200g of potatoes, add 800mL of water, boil for 30 minutes, filter through 4 layers, 8 layers, and 16 layers of gauze successively, 15g of glucose, 18g of agar powder, dilute to 1000mL, natural pH, sterilize at 115°C for 20min.

[0063] 2. Seed culture: Inoculate the strain activated twice on the slope into a conical flask filled with 100 mL of PDA liquid medium, and culture on a shaker at 28° C. and 160 r / min for 48 hours to obtain a seed culture solution.

[0064] 3. Fermentation culture:

[0065] (1) Transfer the seed culture solution into the fermentation medium (200mL medium in a 500mL Erlenmeyer flask) according to the inoculum size of 2%, cultivate at 25°C at 200r / min (radius of rotation 250mm), and cultivate for 4 days;

[0066] The fermentation medium is prepared as follo...

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Abstract

The invention discloses a method for preparing homoharringtonine and a special strain. The strain is Alternaria tenuissima CH1307, and the preservation number of the strain is CGMCC No.4443. The experiment proves that the strain has a high homoharringtonine production capacity. The method for preparing the homoharringtonine has the advantages of high yield and low cost and is easy to operate. Therefore, the method solves the problem of resource shortage of the homoharringtonine, guarantees the efficient production of the homoharringtonine, and also prevents trees from being chopped down and protects natural resources.

Description

technical field [0001] The invention relates to a method for preparing homoharringtonine and a special bacterial strain. Background technique [0002] Homoharringtonine is a tetracycline alkaloid drug with various anti-tumor effects, which is extracted from the plant of the genus Triphacetaxelaceae. It is a cell cycle-specific drug and has the strongest killing effect on cells in the G1 and G2 phases , but little effect on S phase cells. At present, clinical application has been approved in China, but it has not been approved for marketing by FDA. The roots, stems, leaves and fruits of Torreya hainanensis contain homoharringtonine, especially the high content of homoharringtonine in the bark, which is the main raw material for the production of such anticancer drugs. However, due to its slow growth, difficult reproduction and over-harvesting, the resources are almost exhausted. Insufficient drug sources have become a bottleneck limiting the clinical application of homohar...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P17/18C12R1/645
Inventor 李从发刘四新蔡坤袁牧
Owner HAINAN UNIVERSITY