Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for producing 9α-hydroxyandrostenedione by microbial fermentation

A technology for hydroxyandrostenedione and microbial fermentation, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as pollution and unsuitability for industrial production of 9α-OHAD, and achieve simple extraction and high yield High effect with few by-products

Inactive Publication Date: 2018-07-06
宋浩雷
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] No matter from the environmental pollution of the fermentation process, or from the yield and purity of the product, the currently published methods are not suitable for industrial production of 9α-OH AD

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for producing 9α-hydroxyandrostenedione by microbial fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Strain propagation: ZS3-9α was prepared into a bacterial concentration of 10 7 -10 9 Bacterial suspension of individual / ml, insert in the seed tank of seed culture medium with 5%-10% inoculum size afterwards and cultivate, and seed culture medium comprises: glucose 8g / L, MgSO 4 ·7H 2 O 0.8g / L, (NH 4 ) 2 Fe(SO 4 ) 2 ·6H 2 O 0.02g / L, CaCO 3 5.5g / L and citric acid 1.0g / L, the pH of the seed medium is 7.0;

[0038] Fermentation transformation: OD of seed liquid in the seed tank 600 When it is 15-20, insert it into the transformation medium with 20% inoculum size for cultivation, and the fermentation medium includes: glucose 30g / L, peptone 15g / L, bean cake powder 25g / L, MgSO 4 ·7H 2 O 0.5g / L, (NH 4 ) 2 Fe(SO 4 )2·6H 2 O 0.06g / L, CaCO 3 4.5g / L, 3.0g / L citric acid, pH of the fermentation medium is 7.0; wherein, the substrate phytosterol is added to the prepared basic fermentation medium with a mass fraction of 2.0%. The transformation culture conditions are as ...

Embodiment 2

[0042] In Example 1, the inoculum amount of strains during fermentation transformation was changed to 30%, and the transformation culture conditions were changed to: temperature 32°C, tank pressure 0.05MPa, stirring 150-200rpm, dissolved oxygen 30%, cycle 100h.

[0043] The test result is: the final conversion rate of 9α-OH AD is 93.2%.

Embodiment 3

[0045] Change the inoculum amount of the bacterial classification to 50% during fermentation transformation in Example 1, change the mass fraction of phytosterols to 3.0%, and change the transformation culture conditions to: temperature 32°C, tank pressure 0.05MPa, stirring 150-200rpm, cycle 120h .

[0046] The test result is: the final conversion rate of 9α-OH AD is 95.8%.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention proposes a method for producing 9α-hydroxyandrostenedione by microbial fermentation, using recombinant Mycobacterium smegmatis to ferment and transform phytosterols to produce 9α-hydroxyandrostenedione, comprising the following steps: recombinant expression vector pMV261-ksh Construction; construction of recombinant strains; strain propagation and fermentation transformation. The present invention obtains a genetically engineered bacterial strain overexpressing the 3-sterone-9α-hydroxylase gene (ksh) through genetic engineering means, and the bacterial strain can make the conversion rate of phytosterols as high as 95% or more through 120h fermentation, and has high yield, It has the advantages of less by-products, short fermentation time, simple extraction, less pollution, and less pressure on environmental protection, which has the advantages of industrial production.

Description

technical field [0001] The invention relates to a production method of a steroid drug intermediate, in particular to a method for producing 9α-hydroxyandrostenedione by microbial fermentation. Background technique [0002] 9α-hydroxy androstenedione (9α-hydroxy androstenedione) is abbreviated as 9α-OH AD (the structural formula is shown below), which was first isolated from the fermentation broth of Nocardia as a steroid substance. 9α-OH AD can be used to synthesize corticosteroids, antiandrogen, antiestrogens and contraceptive drugs, and is a very important steroid hormone intermediate. [0003] [0004] At present, there are two main methods for microbial fermentation to produce 9α-OH AD substrates: the first method is to use Nocardia, Rhodococcus, Corynebacterium and Cylindrocarponradicicola on the substrate male Dione (AD) undergoes 9α-hydroxylation to generate 9α-hydroxyandrostenedione (9α-OH AD). For example, the patent "Method for preparing 9α-hydroxy-androstened...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/00C12N15/74C12N1/21C12R1/34
Inventor 宋浩雷张敏然张耀勋周利坤
Owner 宋浩雷