Preparation of 11β-hydroxy-1,4-diene-3,20-dione steroids by joint fermentation of Absidia and Arthrobacter

A combined fermentation and compound technology, applied in fermentation, microbial-based methods, microorganisms, etc., can solve difficult problems such as large enzyme activity

Active Publication Date: 2021-02-02
TIANJIN JINYAO GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the joint transformation fermentation is technically very difficult, mainly due to the different growth and fermentation conditions of different bacteria, such as: different temperature, medium, and pH value, it is difficult to make the two bacteria exert their maximum enzyme activity under the same fermentation conditions

Method used

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  • Preparation of 11β-hydroxy-1,4-diene-3,20-dione steroids by joint fermentation of Absidia and Arthrobacter
  • Preparation of 11β-hydroxy-1,4-diene-3,20-dione steroids by joint fermentation of Absidia and Arthrobacter
  • Preparation of 11β-hydroxy-1,4-diene-3,20-dione steroids by joint fermentation of Absidia and Arthrobacter

Examples

Experimental program
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Effect test

Embodiment 1

[0065] The selection of embodiment 1 fermentation medium

[0066] In the following examples, the influence of the fermentation medium on the conversion rate of the target product was examined. Incline, seed cultivation method as described in the summary of the invention.

Embodiment 1-1

[0068] In 1L of water, the fermentation substrate RSA 10g, the components of the fermentation medium glucose 15g, peptone 5g, corn steep liquor 30g, ammonium nitrate 2g, potassium dihydrogen phosphate 5g, foam enemy 0.4g, 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 Absidia indigo, they were inserted into the above-mentioned fermentation medium according to 30% inoculum amount. Use a 30L 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, the upper layer was sent to HPLC, the predn...

Embodiment 1-2

[0071] The components of the fermentation medium are 30 g of glucose, 5 g of peptone, 30 g of corn steep liquor, 2 g of ammonium nitrate, 5 g of potassium dihydrogen phosphate, 0.4 g of dihydrogen phosphate, and the pH is 6.0. Other conditions are the same as in Example 1-1.

[0072] After TLC analysis, the conversion of the substrate was complete, the sample was extracted with ethyl acetate, and the upper layer was sent to HPLC, the prednisolone content was 77.1%.

[0073] After the conversion is complete, the fermentation broth is inactivated at 85°C, cooled to room temperature, and filtered with suction. The filter cake was extracted with ethyl acetate and concentrated to obtain white crystals with a crude product yield of 89.2% and a prednisolone content of 78.4%.

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Abstract

The present invention uses 4-ene-3,20-diketone steroid compound as starting material, and utilizes Agsidiaceae and Arthrobacter to jointly ferment one-step preparation of 11βhydroxy-1,4-diene-3,20-diketone steroid compound.

Description

technical field [0001] The invention relates to a production method of steroidal drugs, in particular to one-step preparation of 11β-hydroxyl-1,4-diene-3,20-dione steroidal compounds by joint fermentation of Absidia and Arthrobacter. 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 β hydroxylation and C 1,2 The introduction of double bonds by dehydrogenation is an important steroid transformation reaction, which is of great si...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P39/00C12P33/02C12R1/65C12R1/06
Inventor 赵春霞王淑丽韩昆颖
Owner TIANJIN JINYAO GRP
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