Method for preparing 9 alpha-hydroxy-androstane-1,4-diene-3,17-dione

A technology of androstane and diene, applied in the field of microbial pharmacy, can solve problems such as low conversion rate, difficult separation and purification, and achieve the effects of improving utilization efficiency, improving biological efficiency and reducing production cost

Inactive Publication Date: 2016-01-06
SHANGHAI APPLIED TECHNOLOGIES COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] Aiming at the above-mentioned technical problems in the prior art, the present invention provides a method for preparing 9α-hydroxy-androst-1,4-diene-3,17-dione, the preparation of 9α-hydroxy- The method of androst-1,4-diene-3,17-dione solves the problem of using two-step fermentation in the prior art to prepare 9α-hydroxy-androst-1,4-diene-3,17-dione The technical problems of low conversion rate and difficult separation and purification of the method

Method used

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  • Method for preparing 9 alpha-hydroxy-androstane-1,4-diene-3,17-dione

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Mycobacterium Mycobacterium (CICC21097) and Rhodococcus rhodochrous (CGMCC4.1480) were cultured on slant and seeds to obtain the strains required for fermentation. 1. Preparation of mycobacterium Mycobacterium (CICC21097) slant strain and seed solution

[0031] (1) Incline cultivation

[0032] Slant medium: peptone 5.0g / L, beef extract 3.0g / L, sodium chloride 5.0g / L, glucose 10.0g / L, agar 20.0g / L, cultured at 29°C for 5-7 days.

[0033] (2) Seed cultivation

[0034] Seed medium: glucose 15.0g / L, peptone 5.0g / L, beef extract 3.0g / L, sodium chloride 5.0g / L, magnesium sulfate heptahydrate 2.5g / L, ammonium dihydrogen phosphate 3.5g / L, phosphoric acid Dipotassium hydrogen 1.0g / L, potassium dihydrogen phosphate 1.0g / L, pH 7.0, sterilized by moist heat at 121°C for 30min.

[0035] The bacterial strain cultivated in step (1) was inoculated under sterile conditions with an inoculation loop into a 250mL shake flask containing 50mL seed medium, and cultivated on a rotary shaker...

Embodiment 2

[0048] Mycobacterium Mycobacterium (CICC21097) and Rhodococcus rhodochrous (CGMCC4.1480) strains were prepared into seed liquid (the method is the same as in Example 1), and then fermented.

[0049] Fermentation medium: phytosterol 35.0g / L, ethanol 31.0g / L, polyethylene glycol 9.0g / L, glucose 20.0g / L, ammonium dihydrogen phosphate 3.0g / L, ferric ammonium citrate 0.05g / L, Magnesium sulfate heptahydrate 2.5g / L, dipotassium hydrogen phosphate 1.0g / L, potassium dihydrogen phosphate 1.0g / L, beet molasses 6.0g / L, pH7.0.

[0050] The fermenter containing the fermentation medium was sterilized by high-pressure steam at 121° C. for 15 minutes.

[0051] At the beginning of the culture, a compound solubilizing agent was added to the medium: ethanol 31.0 g / L, polyethylene glycol 9.0 g / L. Carry out inoculation again, the inoculum size of mycobacterium MycobacteriumCICC (21097) is 12.0%, inoculate Rhodococcus rhodochrous after mycobacterium transformation culture 24h, the inoculum size of ...

Embodiment 3

[0054] Mycobacterium Mycobacterium (CICC21097) and Rhodococcus rhodochrous (CGMCC4.1480) strains were used to prepare seed liquid (the method is the same as in Example 1), and then fermented.

[0055] Fermentation medium: phytosterol 35.0g / L, glucose 22.0g / L, ammonium dihydrogen phosphate 3.0g / L, ferric ammonium citrate 0.05g / L, magnesium sulfate heptahydrate 2.0g / L, dipotassium hydrogen phosphate 1.0g / L, potassium dihydrogen phosphate 1.0g / L, beet molasses 6.0g / L, pH7.0.

[0056] The fermenter containing the fermentation medium was sterilized by high-pressure steam at 121° C. for 15 minutes.

[0057] At the beginning of the culture, a compound solubilizing agent was added to the medium: ethanol 30.0g / L, polyethylene glycol 9.0g / L. Carry out inoculation again, the inoculum size of mycobacterium MycobacteriumCICC (21097) is 13.0%, inoculate Rhodococcus rhodochrous after mycobacterium transformation culture 48h, the inoculum size of Rhodococcus rhodochrous (CGMCC4.1480) is 17....

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Abstract

The invention discloses a method for preparing 9 alpha-hydroxy-androstane-1,4-diene-3,17-dione. The method takes phytosterol as a substrate, utilizes two microbial mixed bacteria, namely Mycobacterium (CICC 21097) and Rhodococcus rhodochrous (CGMCC 4.1480) for fermentation to prepare 9 alpha-hydroxy-androstane-1,4-diene-3,17-dione. Besides, a compound dissolution promotor is added into a fermentation culture medium to ensure the highest rate of molar conversion from phytosterol to 9 alpha-OH-ADD reaches 84.3 percent. Meanwhile, purification in a two-step 9 alpha-hydroxy-androstane-1,4-diene-3,17-dione preparation method is omitted, and the consumption of a solvent is reduced.

Description

technical field [0001] The invention belongs to the field of microbial pharmacy, and relates to a 9α-hydroxy-androsta-1,4-diene-3,17-dione, specifically a method for preparing 9α-hydroxy-androsta-1,4-diene-3,17-dione by using mixed bacteria to biotransform phytosterols. The method of ster-1,4-diene-3,17-dione. Background technique [0002] Steroids, also known as steroids, are a class of compounds containing cyclopentane polyhydrophenanthrene nuclei. Its basic structure is shown below. It consists of three six-membered rings and one five-membered ring, called A, B, C, and D rings respectively. Generally, there are angular methyl groups on the 10th and 13th positions of the steroid nucleus ( -CH 3 ), there may be hydroxyl (-OH) or ketone (-C=O) at the 3rd, 9th, and 11th positions, some double bonds exist in the A ring or B ring, and there are side chains of different lengths at the 17th position. A series of compounds with unique physiological functions are formed due to t...

Claims

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Application Information

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IPC IPC(8): C12P33/16C12P33/06C07J1/00C12P39/00
Inventor 荣绍丰管世敏王敬文李茜茜蔡宝国杨树林李迎光田勋
Owner SHANGHAI APPLIED TECHNOLOGIES COLLEGE
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