Method for preparing 16alpha, 17alpha-epoxy-11alpha-hydroxy-pregna-1,4-diene-3,20-dione through combined microbial fermentation

A combined fermentation and microorganism technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems that the yield and content of the target product do not exceed 50%, and are not easy to repeat

Inactive Publication Date: 2016-07-20
TIANJIN JINYAO GRP
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Chinese patent CN201210316179.2 reported a method for preparing 11α-OH-ADD by jointly transforming and fermenting 4-AD using ochrax and Arthrobacter simplex microorganisms. more than 50%
[0008] At present, there is no one-step preparation of 16α, 17α-epoxy-11α-hydroxy-pregna-1,4-diene-3,20-dione using 16α,17α-epoxyprogesterone as a substrate by combined transformation and fermentation of microorganisms. Literature report

Method used

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  • Method for preparing 16alpha, 17alpha-epoxy-11alpha-hydroxy-pregna-1,4-diene-3,20-dione through combined microbial fermentation

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Embodiment 1

[0039] The selection of embodiment 1 fermentation medium

[0040] In the following examples, the influence of the fermentation medium on the conversion rate of the target product was investigated. Incline, seed cultivation method as described in the summary of the invention.

Embodiment 1-1

[0042] In 1L of water, ferment substrate 16α, 17α-epoxy progesterone 30g, fermentation medium components glucose 20g, sucrose 20g, silkworm chrysalis powder 5g, corn steep liquor 20g, diammonium phosphate 5g, ammonium dihydrogen phosphate 2g, soak Enemy 0.2g, pH 6.0. For the secondary culture of Arthrobacter simplex, after 24 hours of primary seed culture, insert 10% of the inoculum into the secondary seed medium, and after 22 hours of secondary culture, insert 15% of the inoculum into the above fermentation medium ; After 24 hours of culturing the primary seeds of Ochraus ochratum, they were inserted into the above-mentioned fermentation medium according to 30% inoculum amount. Use a 5-ton fermenter for conversion, with a liquid volume of 80%, a tank pressure of 0.05MPa, and an air flow of 40m 3 / hr, temperature 28°C, stirring 180rpm, conversion for 72 hours, sampling by TLC analysis, the conversion of the substrate was complete, the sample was extracted with ethyl acetate, ...

Embodiment 1-2

[0045] In 1L of water, ferment substrate 16α, 17α-epoxyprogesterone 30g, fermentation medium components glucose 30g, sucrose 20g, silkworm chrysalis powder 5g, corn steep liquor 20g, diammonium phosphate 5g, ammonium dihydrogen phosphate 2g, soak Enemy 0.2g, pH 6.0. For the secondary culture of Arthrobacter simplex, after 24 hours of primary seed culture, insert 10% of the inoculum into the secondary seed medium, and after 22 hours of secondary culture, insert 15% of the inoculum into the above fermentation medium ; After 24 hours of culturing the primary seeds of Ochraus ochratum, they were inserted into the above-mentioned fermentation medium according to 30% inoculum amount. Use a 5-ton fermenter for conversion, with a liquid volume of 80%, a tank pressure of 0.05MPa, and an air flow of 40m 3 / hr, temperature 28°C, stirring 180rpm, conversion for 72 hours, sampling by TLC analysis, the conversion of the substrate was complete, the sample was extracted with ethyl acetate, a...

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Abstract

The invention discloses a method for preparing 16alpha, 17alpha-epoxy-11alpha-hydroxy-pregna-1,4-diene-3,20-dione by taking 16alpha,17alpha-epoxy progesterone as a starting material through one-step mixed fermentation of aspergillus ochraceus and arthrobacter simplex.

Description

technical field [0001] The present invention relates to a production method of steroidal drugs, specifically, 16α, 17α-epoxy-11α-hydroxyl-pregna-1,4-diene- 3,20-diketone method. Background technique [0002] Steroid drugs are an important class of drugs, with anti-inflammatory, anti-immune, contraceptive, anti-cancer, endocrine regulation and other functions, widely used in rheumatism, cardiovascular disease, collagen disease, lymphatic leukemia, human organ transplantation, Treatment of bacterial encephalitis, skin diseases, endocrine disorders, senile diseases, hormone-dependent tumors and other diseases. [0003] In the production of steroid hormone drugs, multiple groups of the steroid nucleus need to be modified by chemical methods and microorganisms, among which C 11 α-hydroxylation and C 1,2 The introduction of double bonds by dehydrogenation is an important steroid transformation reaction, which is of great significance for the synthesis and structural modificatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P33/20C12P33/10C12P33/02C12R1/66C12R1/06
Inventor 赵春霞王淑丽
Owner TIANJIN JINYAO GRP
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