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A group of high-yielding baccatin Ⅲ combination strains and the method for producing baccatin Ⅲ

A technology of bacterial strains and bacterial liquids, applied in the direction of fungi, fermentation, etc., can solve the problem of paclitaxel not being separated, and achieve the effects of reducing production costs, reducing environmental damage, and weakening dependence

Inactive Publication Date: 2020-12-04
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, 10-DAB, the precursor of paclitaxel conversion, is mainly obtained from the bark or branches of yew. Therefore, the production of paclitaxel cannot be separated from yew resources.

Method used

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  • A group of high-yielding baccatin Ⅲ combination strains and the method for producing baccatin Ⅲ
  • A group of high-yielding baccatin Ⅲ combination strains and the method for producing baccatin Ⅲ
  • A group of high-yielding baccatin Ⅲ combination strains and the method for producing baccatin Ⅲ

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 no.79 combined with Z58 to produce baccatin III

[0039] The following are the specific steps for the mixed culture of no.79 and Z58 strains to extract baccatin III:

[0040] (1) Preparation of medium:

[0041] 1) PDA medium: 200g potato, 20g sucrose, 20g agar, 1000mL water, natural pH.

[0042] 2) Fermentation medium: potato 200g, bran 10g, yeast extract 1g / L, sucrose 20g, sodium benzoate 250mg / L, magnesium sulfate 0.4g / L, potassium dihydrogen phosphate 2g / L, water 1000mL, natural pH. Peel and cut the potatoes into strips, boil in water for 30 minutes, filter through 4 layers of gauze; boil the bran in water for 30 minutes, filter through 4 layers of gauze.

[0043] (2) Bacteria pre-cultivation: Pick the endophytic fungus no.79 producing baccatin Ⅲ and the endophytic fungus Z58 producing paclitaxel and inoculate them in 60 mL PDA medium respectively, and culture them at 25°C and 100 rpm / min for 1 day to form seeds fermentation broth.

[0044] (3) Bacteria...

Embodiment 2

[0049] Embodiment 2 no.79 and Gliocladium sp. combine to produce Baccatin III

[0050] The embodiment is consistent with Example 1, the baccatin III-producing strain used is no.79, and the paclitaxel-producing strain is Gliocladium sp. Fermentation medium used: potato 200g, bran 5g, yeast extract 1g / L, sucrose 20g, magnesium sulfate 2g / L, sodium benzoate 250mg / L, zinc sulfate 0.05g / L, ammonium molybdate 0.15g / L, water 1000mL, natural pH. Fermentation conditions were unchanged, mixed fermentation and separate fermentation were carried out. Fermentation product carries out high performance liquid chromatography detection, and the content of mixed fermentation baccatin III is 9.36mg / L (see figure 2 ), the yield of baccatin Ⅲ after mixed fermentation was 42.7 times higher than that of single culture no.79.

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Abstract

The invention discloses a group of combined strains for highly producing baccatin III and a method for producing baccatin III, belonging to the technical field of biology. The combined strains for highly producing baccatin III are combined strains of baccatin III production strains and paclitaxel production strains, wherein the baccatin III production strains are Taxus chinensis var mairei endophytic fungi no.79, and paclitaxel production strains are Taxus media endophytic fungi Z58 or Taxus india endophytic Gliocladium SP. By sequentially carrying out independent culture and mixed culture on the baccatin III production strains and the paclitaxel production strains, the yield of baccatin III in an fermented object is substantially increased; by virtue of metabolic pathway complementation and a feedback inhibition theory, the yield of baccatin III is greatly increased and is far higher than that of an independent culture manner. According to the method, the industrial fermentation culture and production of baccatin III by virtue of endophytic fungi are realized, relatively cheap raw materials are provided for the production of paclitaxel by virtue of a semi-synthesis method, and the production cost is lowered; the dependence on the Taxus chinensis resource is reduced, and the damage to the environment is alleviated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and discloses a group of high-yield baccatin III combination strains and a method for producing baccatin III. Background technique [0002] Cancer is one of the major diseases that seriously affect human health. In 1979, the National Cancer Institute confirmed that paclitaxel has a unique anti-cancer mechanism. Paclitaxel has a good curative effect on ovarian cancer, breast cancer, lung cancer, esophageal cancer, melanoma, and head and neck cancer. However, natural paclitaxel is mainly derived from Taxus (Taxus) plants, and the content is very low. It takes 3-4 large yew trees with 100 years to prepare the medicine required by a patient, and only 3-4 large yew trees from the bark of about 2000 large yew trees. 1kg paclitaxel can be extracted. The yew (Taxus) plant is a rare and endangered species in my country, and it is a national first-class protected plant. [0003] There are many research repo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P17/02
CPCC12N1/14C12P17/02
Inventor 李勇超周修任杨靖简在友孟丽
Owner HENAN INST OF SCI & TECH
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