Separating and purifying method of taxol synthesized by biological fermentation of nodulisporium sylviforme entophytic fungi as well as precursors of taxol

A technology of polyarthrospora dendriticum and biological fermentation, applied in the direction of microorganism-based methods, biochemical equipment and methods, peptide preparation methods, etc., can solve the problems of low recovery rate of paclitaxel precursor, low paclitaxel content, low purity, etc. problems, to achieve the effect of high product purity, simple operation process, and small solvent usage

Inactive Publication Date: 2015-05-27
HEILONGJIANG UNIV
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Problems solved by technology

[0013] The purpose of the present invention is to solve the problems of low paclitaxel content in the fermentation broth, low recovery rate and low purity of paclitaxel precursors in the existing method for synthesi

Method used

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  • Separating and purifying method of taxol synthesized by biological fermentation of nodulisporium sylviforme entophytic fungi as well as precursors of taxol
  • Separating and purifying method of taxol synthesized by biological fermentation of nodulisporium sylviforme entophytic fungi as well as precursors of taxol
  • Separating and purifying method of taxol synthesized by biological fermentation of nodulisporium sylviforme entophytic fungi as well as precursors of taxol

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specific Embodiment approach 1

[0037] Specific embodiment one: In this embodiment, the separation and purification method of paclitaxel and its precursors synthesized by the endophytic fungus HDFS4-26 of the dendritic endophyte HDFS4-26 is realized according to the following steps:

[0038] 1. Strain activation and seed culture: Inoculate Polyarthrospora dendriticum HDFS4-26 on PDA slant medium, activate at 28°C for 2-3 days, and then inoculate the activated Polyarthrospora dendriticum HDFS4-26 Cultivate in PDA liquid medium at 28°C, 120r / min shaker for 3 days to obtain seed culture solution;

[0039] 2. Fermentation culture: transfer the seed culture solution into the improved S-7 liquid medium with an inoculation amount of 5% by volume, and ferment and cultivate at 28°C and 120r / min for 12 days, and add 4g on the 4th day of the fermentation culture / L sucrose and 3mL / L vitamin concentrate, add 0.4g / L peptone and 0.2mL / L bean cake hydrolyzate on the 6th day of fermentation culture, add 2g / L sucrose on the ...

specific Embodiment approach 2

[0055] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that in step 2, the vitamin concentrate consists of 15 mg tyrosine, 50 mg phenylalanine, and 10 mg VB per 100 mL. 6 , 10mg VB 1 , 10mg V H , 10mg calcium pantothenate, 10mg linoleic acid and the remainder of distilled water. Others are the same as in the first embodiment.

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Abstract

The invention discloses a separating and purifying method of taxol synthesized by biological fermentation of nodulisporium sylviforme entophytic fungi as well as precursors of the taxol, relates to a separating and purifying method and aims at solving the problems that taxol in fermentation liquor is low in content and the taxol precursors are low in the recovery rate and low in purity in the traditional method for synthesizing by biological fermentation of entophytic fungi. The separating and purifying method disclosed by the invention comprises the following steps of: activating a bacterial strain and culturing seeds; 2, carrying out fermentation culture; 3, separating and recovering fermentation liquor and mycelium, decoloring the fermentation liquor, and obtaining a sample through extracting and concentrating; 4, carrying out primary chromatography; 5, carrying out secondary chromatography; and 6, separating by using a reversed phase chromatography method to finish separation and purification. According to the separating and purifying method, the content of taxol in the fermentation liquor is 556.80mug/L, the taxol purity is 98 percent, the precursor baccatin purity is 97 percent, and the precursor cephalomannine purity is 98 percent; the separating and purifying method has the characteristics of simple operation process, high product purity and low use amount of a solvent; and meanwhile, the production cost of the taxol is indirectly reduced.

Description

technical field [0001] The invention relates to a method for separating and purifying paclitaxel and its precursors through biological fermentation of endophytic fungus HDFS4-26. Background technique [0002] Malignant tumors seriously threaten human health, and cancer is the second leading cause of death in human diseases after cardiovascular and cerebrovascular diseases. [0003] Paclitaxel is a diterpene alkaloid drug extracted from Taxus genus plants with various anti-tumor effects. Because yew (also known as yew) is a rare tree species, its growth rate is slow, and its biological characteristics have high requirements on the ecological environment, and its natural regeneration ability is poor, so the population is extremely limited. It is an endangered plant and its resources are very limited. Therefore, scholars at home and abroad have made various efforts in the research and development of new drug sources of paclitaxel. [0004] The unique anticancer activity of pa...

Claims

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Application Information

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IPC IPC(8): C12P17/02C07K1/22C07K1/20C12R1/645
Inventor 赵凯
Owner HEILONGJIANG UNIV
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