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11beta-hydroxy-1,4-diene-3,20-dione steroid prepared through combined fermentation of absidia and arthrobacter

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

Active Publication Date: 2016-07-20
TIANJIN JINYAO GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • 11beta-hydroxy-1,4-diene-3,20-dione steroid prepared through combined fermentation of absidia and arthrobacter
  • 11beta-hydroxy-1,4-diene-3,20-dione steroid prepared through combined fermentation of absidia and arthrobacter
  • 11beta-hydroxy-1,4-diene-3,20-dione steroid prepared through combined fermentation of absidia and arthrobacter

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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 investigated. Incline, seed cultivation method as described in the summary of the invention.

Embodiment 1-1

[0068] In 1L of water, 10g of fermentation substrate RSA, 15g of glucose, 5g of peptone, 30g of corn steep liquor, 2g of ammonium nitrate, 5g of potassium dihydrogen phosphate, 0.4g of dihydrogen phosphate, and 0.4g of foam enemy in 1L of water, the pH is 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 la...

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 invention discloses a 11beta-hydroxy-1,4-diene-3,20-dione steroid which is prepared by taking a 4-ene-3,20-dione steroid as a starting material through the one-step combined fermentation of absidia and arthrobacter.

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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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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