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Method for increasing fermentation yield of artemisinic acid

A technology for increasing the yield of artemisinic acid, which is applied in the field of increasing the fermentation yield of artemisinic acid, can solve problems such as high cost, low yield, and unstable strain performance, and achieve the goals of increasing fermentation yield, reducing fermentation cost, and increasing fermentation titer Effect

Active Publication Date: 2018-10-02
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the third method, artemisinin, the key intermediate of artemisinin, has a great influence on the yield and cost of the final product
[0007] Aiming at the shortcomings of low output, high cost, inability to realize industrialized production, and unstable strain performance in the artemisinic acid fermentation production in the prior art, it is necessary to find a method with high output, low cost, harmless to the human body and friendly to the environment. , the strain has high genetic stability, and an artemisinic acid fermentation process that can realize industrial production is necessary

Method used

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  • Method for increasing fermentation yield of artemisinic acid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] (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%, and cultivate for 16 hours to obtain the strain liquid;

[0066] (2) Resistance screening of bacterial classification: the bacterial classification liquid gained in the step (1) is diluted 10 -7 After doubling, draw 100 μL and spread it on the plate solid medium containing 200 μg / L hygromycin B, culture for 40 hours, and obtain a single colony of Saccharomyces cerevisiae engineered bacteria;

[0067] The plate solid medium formula is: glucose 19.5g / L, (NH4) 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.

[0068] (3) Seed culture: Pick a single colony from the plate solid medium containing antibiotics, inoculate it into the seed medium, and cu...

Embodiment 2

[0076] (1) Activation of strains: inoculate the Saccharomyces cerevisiae engineered strains frozen at -80°C into the seed medium according to the inoculation amount of 0.1% (V / V), cultivate for 30 hours, and obtain the strain liquid;

[0077] (2) Resistance screening of bacterial classification: the bacterial classification liquid gained in the step (1) is diluted 10 -5 After doubling, draw 100 μL and spread it on the plate solid medium containing 300 μg / L knorsmycin, cultivate for 30 hours, and obtain a single colony of Saccharomyces cerevisiae engineering bacteria;

[0078] The plate solid medium formula is: glucose 19.5g / L, (NH4) 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 O40ug / L, succinic acid buffer 100ml / L (0.5M, pH 5.0), agar 20g / L, and the rest are water.

[0079] (3) Seed culture: Pick a single colony from the plate solid medium containing antibiotics, inoculate it into the seed mediu...

Embodiment 3

[0087] (1) Preparation of the seed liquid: Inoculate the Saccharomyces cerevisiae engineered strains frozen at -80°C into the seed culture medium according to the inoculum size of 0.3%, and cultivate for 20 hours to obtain the seed liquid;

[0088] (2) Resistance screening of bacterial classification: the bacterial classification liquid gained in the step (1) is diluted 10 -6 After doubling, draw 100 μL and spread it on the plate solid medium containing 250 μg / L geneticin, culture for 35 hours, and obtain a single colony of Saccharomyces cerevisiae engineered bacteria;

[0089] The plate solid medium formula is: glucose 19.5g / L, (NH4) 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.

[0090] (3) Seed culture: Pick a single colony from the plate solid medium containing antibiotics, inoculate it into the seed me...

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Abstract

The invention provides a method for increasing a fermentation yield of artemisinic acid. The method comprises the following steps: using an engineered strain of Saccharomyces cerevisiae capable of producing the artemisinic acid as a production strain; and fermenting in a fermentation culture medium containing a lactose hydrolysate capable of hydrolyzing into glucose and galactose, mevalonic acid and / or citral and other exogenous factors. The method provided by the invention increases the fermentation yield of artemisinic acid, reduces the fermentation cost of artemisinic acid, adopts a simpleand feasible technical solution, and has advantages of environmental protection, high genetic stability of the strain, and easy industrial production and application.

Description

technical field [0001] The invention relates to the technical field of industrial microbial fermentation, in particular to a method for increasing the fermentation yield 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. It was discovered by Chinese pharmacist Tu Youyou in 1971. Its molecular formula is C 15 h 22 o 5 . 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". Artemisinin has been widely used for more than 40 years. According to statistics, the annual sales of artemisinin and its derivatives worldwide are as high as 1.5 billion US dollars. In recent years, artemisinin has also shown attractive prospects in the treatment of other diseases such as ant...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/865
CPCC12P17/181
Inventor 陈伟滕云徐彬徐美冬陈亚军郑玲辉
Owner ZHEJIANG HISUN PHARMA CO LTD
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