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Method for promoting accumulation of artemisinic acid

A technology of artemisinic acid and volume is applied in the field of promoting the accumulation of artemisinic acid, which can solve the problems of complex fermentation process, long fermentation period and high production cost, and achieve the effects of increasing fermentation yield, reducing fermentation cost and increasing yield

Active Publication Date: 2019-09-20
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the disadvantages of complex fermentation process, high production cost, long fermentation period, and addition of organic solvents in the fermentation process in the prior art of artemisinic acid fermentation, the present invention provides a method for promoting the accumulation of artemisinic acid

Method used

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  • Method for promoting accumulation of artemisinic acid
  • Method for promoting accumulation of artemisinic acid
  • Method for promoting accumulation of artemisinic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Activation of strains: inoculate the Saccharomyces cerevisiae engineered strains frozen at -80°C into the seed medium according to the inoculum size of 0.5% (V / V), cultivate for 16 hours, and obtain the strain liquid;

[0042] (2) Screening of bacterial classification: the bacterial classification liquid gained in step (1) is diluted 10 -7 After doubling, draw 100 μL and spread it on the solid medium of the plate, cultivate for 40 hours, and obtain a single colony;

[0043] The plate solid medium formula is: glucose 19.5g / L, (NH 4 ) 2 SO 4 15g / L, KH 2 PO 4 8g / L, MgSO 4 ·7H 2 O 6.2g / L, vitamin solution 12ml / L, metal ion solution 10ml / L, CuSO 4 ·5H 2 O 40ug / L, succinic acid buffer 100ml / L (0.5M, pH 5.0), agar 20g / L, and the rest are water.

[0044] (3) Seed culture: Pick a single colony from the plate solid medium, inoculate it into the seed medium, and cultivate it for 24 hours to obtain the seed solution;

[0045] (4) Fermentation culture: inoculate the se...

Embodiment 2

[0066] (1) Activation of strains: inoculate the Saccharomyces cerevisiae engineered strains frozen at -80°C into the seed medium according to the inoculum size of 0.5% (V / V), cultivate for 16 hours, and obtain the strain liquid;

[0067] (2) Screening of bacterial classification: the bacterial classification liquid gained in step (1) is diluted 10 -7 After doubling, draw 100 μL and spread it on the solid medium of the plate, cultivate for 40 hours, and obtain a single colony;

[0068] The plate solid medium formula is: glucose 19.5g / L, (NH 4 ) 2 SO 4 15g / L, KH 2 PO 4 8g / L, MgSO 4 ·7H 2 O 6.2g / L, vitamin solution 12ml / L, metal ion solution 10ml / L, CuSO 4 ·5H 2 O 40ug / L, succinic acid buffer 100ml / L (0.5M, pH 5.0), agar 20g / L, and the rest are water.

[0069] (3) Seed culture: Pick a single colony from the plate solid medium, inoculate it into the seed medium, and cultivate it for 24 hours to obtain the seed solution;

[0070] (4) Fermentation culture: inoculate the se...

Embodiment 3

[0091] (1) Activation of strains: inoculate the Saccharomyces cerevisiae engineered strains frozen at -80°C into the seed culture medium according to the inoculum size of 0.5% (V / V), cultivate for 16 hours, and obtain the strain liquid;

[0092] (2) Screening of bacterial classification: the bacterial classification liquid gained in step (1) is diluted 10 -7 After doubling, draw 100 μL and spread it on the solid medium of the plate, cultivate for 40 hours, and obtain a single colony;

[0093] The plate solid medium formula is: glucose 19.5g / L, (NH 4 ) 2 SO 4 15g / L, KH 2 PO 4 8g / L, MgSO 4 ·7H 2 O 6.2g / L, vitamin solution 12ml / L, metal ion solution 10ml / L, CuSO 4 ·5H 2 O 40ug / L, succinic acid buffer 100ml / L (0.5M, pH 5.0), agar 20g / L, and the rest are water.

[0094] (3) Seed culture: Pick a single colony from the plate solid medium, inoculate it into the seed medium, and cultivate it for 24 hours to obtain the seed solution;

[0095] (4) Fermentation culture: inoculate ...

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Abstract

The invention provides a method for promoting accumulation of artemisinic acid. Exogenous regulating factors and cheap and easily available plant oil are added into a culture medium, magnesium salt and amino acid are added into the culture medium in the fermentation culture process of saccharomyces cerevisiae engineered strains capable of producing the artemisinic acid, fermentation temperature and fermentation dissolved oxygen are regulated by stages, so that accumulation of the artemisinic acid is greatly promoted based on guarantee of plasmid stability of the saccharomyces cerevisiae engineered strains, the fermentation yield of the artemisinic acid can be at most increased by 50% or more, fermentation period is effectively shortened, and effects are remarkable. The method is simple and easy, the production efficiency of the artemisinic acid is further improved, and industrial production and application of the artemisinic acid are easily facilitated.

Description

technical field [0001] The invention relates to the technical field of industrial microbial fermentation, in particular to a method for promoting the accumulation of artemisinic acid. Background technique [0002] Artemisinin is a sesquiterpene lactone drug with peroxy groups extracted from the stems and leaves of the compound inflorescence plant Artemisia annua, and its molecular formula is C 15 h 22 o 5 , discovered by Chinese pharmacist Tu Youyou in 1971. Artemisinin is the most effective anti-malarial drug after pyrimethamine, chloroquine, and primaquine, especially for cerebral malaria and anti-chloroquine malaria. The only effective malaria treatment in the world". Its anti-malarial mechanism mainly lies in the activation of artemisinin to generate free radicals during the treatment of malaria. The free radicals combine with malarial proteins and act on the membrane structure of Plasmodium, causing the vesicle membrane, nuclear membrane and plasma membrane to be da...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/865
CPCC12P17/181
Inventor 陈伟滕云陈亚军胡栋毛亮亮郑玲辉徐彬应雪肖
Owner ZHEJIANG HISUN PHARMA CO LTD
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