Strain for biosynthesis of 3beta, 7alpha, 15alpha-trihydroxyandrost-5-ene-17-one and application thereof

A technology of hydroxyandrosteroids and microbial strains, applied in the biological field, can solve the problems of low feed concentration, low general conversion rate, high industrial cost, etc.

Inactive Publication Date: 2013-03-13
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Currently, there are reports of Colletotrichum, Fusarium, Acremonium (Acremonium) and Nigrospora (Nigrospora) have 7α and 15α hydroxylation ability, which can convert DHEA into 3β, 7α, 15α -Trihydroxyandrost-5-en-17-one, but the general conversion rate is not high, only 40~70%, and the concentration of substrate feeding is low, generally 1~5g / L, and the industrial cost is high

Method used

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  • Strain for biosynthesis of 3beta, 7alpha, 15alpha-trihydroxyandrost-5-ene-17-one and application thereof
  • Strain for biosynthesis of 3beta, 7alpha, 15alpha-trihydroxyandrost-5-ene-17-one and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Activation and screening of strains.

[0019] Add the glycerol tubes of strains stored at -80°C in the laboratory to 10 mL of seed medium for activation culture. Then transfer to the seed medium, the liquid volume is 250 mL Erlenmeyer flask filled with 30 mL of seed liquid cultured for 2 days. Inoculate 4% of the inoculum into the fermentation medium, fill 30 mL in a 250 mL Erlenmeyer flask, and incubate at 30°C and 220 r / min for 24 hours. Accurately weigh 3g / L of DHEA and put it into the cell culture solution, and transform under the same conditions for 60 hours. The conversion solution was extracted and analyzed by HPLC. The results showed that only Gibberella ( Gibberella intermedia ) CA3-1 has the ability to transform DHEA, and can generate the product 3β, 7α, 15α-trihydroxyandrost-5-en-17-one.

Embodiment 2

[0020] Embodiment 2 Gibberella ( Gibberella intermedia ) CA3-1 converts DHEA.

[0021] (1) Place the PDA slant of the inoculated strain in a constant temperature incubator at 30°C, and cultivate it for 4 days to obtain mature spores. Use an inoculation loop to pick a ring of mycelium and inoculate it into a 500mL shake flask containing 100mL of seed medium , at 28°C, cultivated on a shaker at 220rpm for 1 day to obtain a seed culture solution suitable for inoculation;

[0022] (2) Put the seed culture solution into a 250mL shake flask with 30mL of fermentation medium at an inoculum amount of 4%, and continue to cultivate for 20 hours under the same conditions to obtain the cell fermentation solution;

[0023] (3) Put dehydroepiandrosterone into the fermentation broth of the bacteria, the feeding concentration is 3.0g / L, transform under the same conditions for 60 hours and put in the bottle. The extracted product was analyzed by HPLC, and the conversion rate was 90.08%.

Embodiment 3

[0025] (1) Slope culture, seed culture and fermentation culture are the same as in Example 1.

[0026] (2) Put dehydroepiandrosterone into the fermentation broth of the bacteria, the feeding concentration is 5.0g / L, transform under the same conditions for 60 hours and release the bottle. The extracted product was analyzed by HPLC, and the conversion rate was 84.08%.

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Abstract

Belonging to the field of biotechnologies, the invention relates to a strain of Gibberella intermedia CA3-1 for high-efficiency conversion of dehydroepiandrosterone (DHEA) and its application. The strain is preserved in China General microbiological culture collection center and has a preservation number of CGMCCNo.4903. The Gibberella intermedia CA3-1 can convert DHEA into 3beta, 7alpha, 15alpha-trihydroxyandrost-5-ene-17-one. Under the condition of a substrate feeding amount of 5.0g/L, the substrate conversion rate can reach about 90%, and the product yield can be about 80%, so that the strain provided in the invention is of great significance for steroid drug production by bioconversion.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a strain of Gibberella ( Gibberella intermedia ) CA3-1 and its method for converting dehydroepiandrosterone to 3β,7α,15α-trihydroxyandrost-5-en-17-one. Background technique [0002] Yasmin is a compound short-acting oral contraceptive composed of estrogen and progesterone. The main ingredient drospirenone can effectively combat the obesity problem caused by estrogen in oral contraceptives. 3β,7α,15α-trihydroxyandrost-5-en-17-one is an important pharmaceutical intermediate for the production of drospirenone, and its production methods mainly include traditional chemical synthesis, semi-synthesis, and biotransformation . The biotransformation method generally generates 3β,7α,15α-trihydroxyandrost-5-en-17-one through microbial conversion of hydroxylated dehydroepiandrosterone (DHEA), which reduces the synthesis steps and energy compared with chemical methods. Consumption, and greatly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P33/12C12P33/06C12R1/645
Inventor 李会许正宏付珍珍史劲松张旦旦
Owner JIANGNAN UNIV
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