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Method for increasing yield of paclitaxel in endophytic fungus fermentation product

An endophytic fungus and paclitaxel technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of yew felling, depletion of natural resources, biodiversity and ecological environment damage, etc., to increase the content of yew , high yield, short cycle effect

Active Publication Date: 2016-03-16
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the extremely low content of paclitaxel in plants, about 2,500 adult yew trees are needed to produce 1kg of paclitaxel as raw materials, which also caused a large number of yew trees to be felled, resulting in the depletion of its natural resources, resulting in biodiversity and ecology. Great damage to the environment

Method used

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  • Method for increasing yield of paclitaxel in endophytic fungus fermentation product

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1. Material preparation

[0042] PDA plate medium: potato 200g, glucose 20g and agar 15g, dilute to 1000ml with water, natural pH, sterilize at 121°C for 20min.

[0043] Inoculate the paclitaxel-producing endophytic fungus strains preserved on the slant on the PDA plate medium for activation culture at a culture temperature of 22° C. for 10 days to obtain the activated paclitaxel-producing endophytic fungal strains.

[0044] The composition of liquid MRS medium is: peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, glucose 20.0g, sodium acetate 5.0g, diammonium hydrogen citrate 2.0g, Tween-801.0ml, dihydrogen phosphate Potassium 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g and distilled water 1.0 liter.

[0045] The fermentation basal medium is composed of the following components in mass percentage: glucose 1%, K 2 HPO 4 0.1%, MgSO 4 0.05% and the balance water.

[0046] 1 LPBS buffer: 0.27g potassium dihydrogen phosphate, 1.42g disodium hydrogen phos...

Embodiment 2

[0055] 1. Material preparation

[0056] PDA plate medium: potato 200g, glucose 20g and agar 20g, dilute to 1000ml with water, natural pH, sterilize at 121°C for 20min.

[0057] Inoculate the paclitaxel-producing endophytic fungus strains preserved on the slant on the PDA plate medium for activation culture at a culture temperature of 20° C. for 25 days to obtain the activated paclitaxel-producing endophytic fungal strains.

[0058] The fermentation basal medium is composed of the following components in mass percentage: glucose 3%, K 2 HPO 4 0.2%, MgSO 4 0.1% and the balance water.

[0059] Liquid MRS medium and PBS buffer are the same as in Example 1.

[0060] 2. Preparation of endophytic fungal fermentation products

[0061] (1) Take 1cm 2 The size-activated paclitaxel-producing endophytic fungi were inoculated into 1L liquid MRS medium and cultured at 20°C for 6 hours, and the culture solution was centrifuged at 6000rpm for 10 minutes to collect the first bacteria; th...

Embodiment 3

[0068] 1. Material preparation

[0069] PDA plate medium, liquid MRS medium and PBS damping fluid are the same as embodiment 1.

[0070] Inoculate the paclitaxel-producing endophyte strains preserved on the slant on the PDA plate medium for activation culture at a culture temperature of 30° C. for 3 days to obtain the activated paclitaxel-producing endophyte strains.

[0071] The fermentation basal medium is composed of the following components in mass percentage: glucose 2%, K 2 HPO 4 0.15%, MgSO 4 0.08% and the balance water.

[0072] 2. Preparation of endophytic fungal fermentation products

[0073] (1) Take 4cm 2 The size-activated paclitaxel-producing endophytic fungi were inoculated into 1L liquid MRS medium and cultured at 30°C for 4 hours, and the culture solution was centrifuged at 6000rpm for 10 minutes to collect the first bacteria; the first bacteria were suspended in the mass percentage of bile salts Incubate in 0.5% liquid MRS medium containing bile salts a...

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Abstract

The invention discloses a method for increasing the yield of paclitaxel in an endophytic fungus fermentation product. The method comprises the steps that an endophytic fungus seed solution for producing paclitaxel is inoculated into an endophytic fungus solid state fermentation medium containing taxus media and valeriana jatamansi for culture, the mixture is then put into an alternating magnetic field environment for further culture, and a endophytic fungus solid state fermentation product is obtained; then the endophytic fungus seed solution for producing paclitaxel is inoculated into a liquid fermentation medium containing the endophytic fungus solid state fermentation product for culture, and the final endophytic fungus fermentation product is obtained. According to the method, raw materials are natural, no pollution is caused to the environment, a cycle is short, the yield is high, the method is suitable for industrial production, and compared with a common endophytic fungus fermentation product, the yield of paclitaxel in the endophytic fungus fermentation product is obviously increased.

Description

technical field [0001] The invention relates to the field of biological fermentation, in particular to a method for increasing the yield of paclitaxel in fermentation products of endophytic fungi. Background technique [0002] Paclitaxel (trade name Taxol) is a diterpene derivative, a natural product with a unique anticancer mechanism, and its molecular formula is C 47 h 51 NO 14 , originally isolated from the yew plant. It has been clinically proven to have a good anti-tumor effect, especially for ovarian cancer, uterine cancer and breast cancer, which have a high incidence of cancer. Paclitaxel is one of the most popular anticancer drugs in the international market in recent years. With the increase of the incidence of cancer in the global population, the demand for paclitaxel has increased significantly. Paclitaxel currently required for clinical and scientific research is mainly extracted directly from Taxus chinensis. Due to the extremely low content of paclitaxel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/02C12R1/77
CPCC12P17/02
Inventor 程俊文贺亮魏海龙胡传久方茹李海波
Owner ZHEJIANG FORESTRY ACAD
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