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A method for synthesizing 11α, 15α-dioh-canrenone by using Gibberella ca3-1

A technology of gibberellum and canrenone, applied in the biological field

Active Publication Date: 2019-01-08
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The method for converting canrenone into 11α, 15α-diOH-canrenone by using Gibberella fungus involved in the present invention has not been reported yet

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  • A method for synthesizing 11α, 15α-dioh-canrenone by using Gibberella ca3-1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1 Gibberella intermedia (Gibberella intermedia) CA3-1 transforms canrenone

[0016] (1) Prepare the seed liquid of Gibberella intermedia CA3-1 bacterial strain

[0017] Pick a loop of Gibberella intermedia CA3-1 strain on the solid PDA medium, inoculate it in a 500mL Erlenmeyer flask containing 100mL seed medium, place it on a shaker at 200r / min at 30°C Cultivate for 20-24 hours to the logarithmic phase, and the seed solution of CA3-1 is obtained;

[0018] (2) Insert the seed culture solution into a 250mL shake flask equipped with 30mL fermentation medium with an inoculum size of 4%, and continue to cultivate for 20 hours under the same conditions to obtain a bacterial cell fermentation solution;

[0019] (3) Put canrenone into the bacterial fermentation broth, the feeding concentration is 4.0g / L, transform under the same conditions for 60 hours, extract the product for HPLC analysis, and calculate the conversion rate of canrenone to be 90.1%, 11α, 15α The ...

Embodiment 2

[0021] (1) Seed culture and fermentation culture are the same as in Example 1.

[0022] (2) Put canrenone into the bacterial fermentation broth, the feeding concentration is 6.0g / L, transform for 60 hours under the same conditions, extract the product for HPLC analysis, and calculate the conversion rate of canrenone to be 76.1%, 11α, 15α The yield of -diOH-canrenone was 41.3%.

Embodiment 3

[0024] (1) Seed culture and fermentation culture are the same as in Example 1.

[0025] (2) Put canrenone into the bacterial fermentation broth, the feeding concentration is 8.0g / L, and transform under the same conditions for 60 hours, extract the product for HPLC analysis, and calculate the conversion rate of canrenone to be 54.3%, 11α, 15α The yield of -diOH-canrenone was 34.7%.

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a method for converting substrate canrenone into 11α, 15α-diOH-canrenone by utilizing Gibberella intermedia CA3-1. The strain is deposited in the General Microbiology Center of the China Microorganism Culture Collection and Management Committee, and the preservation number is CGMCC No.4903. Gibberella intermedia CA3-1 can convert canrenone to 11α, 15α-diOH-canrenone, when the substrate concentration is 4g / L, the conversion rate of canrenone is as high as 90.1%, 11α, 15α-diOH ‑ The yield of canrenone was 58.5%. The synthesis of 11α,15α-diOH-canrenone provides more diversified precursors for the research of steroid drugs and their intermediates, and has certain research value and market significance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for converting a substrate canrenone into 11α, 15α-diOH-canrenone by using a strain of Gibberella intermedia CA3-1. Background technique [0002] Canrenone is a steroid hormone cardiovascular drug, which is mainly used as a non-selective aldosterone receptor antagonist to treat cardiovascular diseases clinically. It can be oxidized to generate 11α-OH-canrenone and 11α, 15α-diOH-canrenone. 11α-OH-canrenone, as an essential key precursor compound for the synthesis of eplerenone, a highly effective drug for treating cardiovascular diseases, is one of the hotspots in the field of steroid drug research. 11α, 15α dihydroxy products are rarely reported. Compared with traditional chemical synthesis methods, microbial transformation technology has many advantages such as less reaction steps, mild conditions, high stereoselectivity, and environmental friendliness. Using...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P33/06C12R1/645
CPCC12P33/06C12N1/145C12R2001/645
Inventor 许正宏李会史劲松孙锦
Owner JIANGNAN UNIV