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Method for preparing 11Alpha,17Alpha-hydroxyprogesterone

A technology of hydroxyprogesterone and culture medium, which is applied in the field of preparing 11α, 17α-hydroxyprogesterone and steroid intermediates, can solve the problems of bacterial cell damage, inability to recycle, less application of steroid drugs, etc., and achieves low production cost , the effect of saving production costs and high industrial application prospects

Active Publication Date: 2017-01-11
SHANGHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the addition of organic solvents or surfactants in the fermentation broth has the following disadvantages: ① damage to the bacteria
③One-time input, cannot be recycled
[0009] With the rapid development of nanotechnology, it is now used in materials and preparation, microelectronics and computer technology, medicine and health, aerospace and aviation, environment and energy, biotechnology and agricultural products, but in the field of biotransformation of steroid drugs still less applications

Method used

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  • Method for preparing 11Alpha,17Alpha-hydroxyprogesterone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Aspergillus ochraceus was cultured on a slant to obtain the strains required for fermentation.

[0027] 1. Preparation of Aspergillus ochraceus slant strain and seed solution

[0028] (1) Incline cultivation

[0029] Incline medium: peeled potato 200g / L, glucose 20g / L, agar 20g / L, natural pH, sterilized at 121℃ for 30min.

[0030] The bacterial species preserved in the glycerol tube is aseptically drawn a certain amount of bacterial liquid and spread evenly on the slant medium, and cultured at 28°C for 4-6 days.

[0031] (2) Seed cultivation

[0032] Seed medium: Glucose 10g / L, corn steep liquor 10g / L, yeast extract 10g / L, natural pH 7.0, sterilized at 121°C for 30min.

[0033] With the strain cultivated in step (1), under aseptic conditions, use an inoculation loop to pick 6 ring spores into 100ml seed medium, inoculate 4 bottles in the same way, and cultivate them at 28°C and 200r / min for 24h , to obtain the seed liquid.

[0034] 2. Fermentation

[0035] Ferment...

Embodiment 2

[0041] The Aspergillus ochraceus bacterial strain is made into seed liquid (the method is the same as in Example 1), and then fermented.

[0042] Fermentation medium: glucose 20g / L, corn steep liquor 20g / L, yeast extract 10g / L, pH value 5.0. The fermenter containing the fermentation medium was sterilized by high-pressure steam at 121° C. for 15 minutes.

[0043]Use a 5L fermenter for biotransformation, with a liquid volume of 3.5L (70%). Transfer the seed liquid into the fermentation medium according to the 10% inoculum volume, at a temperature of 28°C. According to the concentration of the bacteria, the stirring effect and the dissolved oxygen level, Adjust the air ratio and stirring speed, the air ratio control range is 0.7-1.2v / v / min, the speed control range is 200-600r / min. When the incubation time is 18 hours, add dimethylformamide (DMF) 5g / L, substrate 17α-hydroxyprogesterone 20g / L, and take samples at intervals of 12 hours for TLC to detect the conversion. The color d...

Embodiment 3

[0048] The Aspergillus ochraceus bacterial strain is made into seed liquid (the method is the same as in Example 1), and then fermented.

[0049] Fermentation medium: glucose 20g / L, corn steep liquor 20g / L, yeast extract 10g / L, pH value 5.0. The fermenter containing the fermentation medium was sterilized by high-pressure steam at 121° C. for 15 minutes.

[0050] Use a 5L fermenter for biotransformation, with a liquid volume of 3.5L (70%). Transfer the seed liquid into the fermentation medium according to the 10% inoculum volume, at a temperature of 28°C. According to the concentration of the bacteria, the stirring effect and the dissolved oxygen level, Adjust the air ratio and stirring speed, the air ratio control range is 0.7-1.2v / v / min, the speed control range is 200-600r / min. When the incubation time was 18 hours, 5 g / L of nanometer ferric oxide powder with an average particle size of 30 nm and 20 g / L of the substrate 17α-hydroxyprogesterone were added. During this period...

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Abstract

The invention discloses a method for preparing 11Alpha,17Alpha-hydroxyprogesterone, comprising: using 17Alpha-hydroxyprogesterone as a substrate, adding magnetic nano ferroferric oxide powder into a culture medium, and preparing 11Alpha,17Alpha-hydroxyprogesterone by Aspergillus ochraceus fermentation process; the method has the advantages that feeding concentration is high, substrate molar conversion rate is high, the feeding concentration is up to 30 g / L, and the highest molar conversion ratio is up to 91.9%; the magnetic nano ferroferric oxide powder is recyclable; by using the magnetic nano ferroferric oxide powder, it is possible to greatly increase hydroxyl conversion rate, the process is simple, the production cost is simple, and the method has great industrial application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a steroid intermediate, in particular to a method for preparing 11α, 17α-hydroxyprogesterone. Background technique [0002] Steroids, also known as steroids, are a class of compounds containing cyclopentane polyhydrophenanthrene nuclei. Its basic structure is shown in the figure below. It consists of three six-membered rings and one five-membered ring, called A, B, C, and D rings, and generally has an angular methyl group on the 10th and 13th positions of the steroid nucleus. (-CH 3 ), there may be a hydroxyl group (-OH) or a keto group (-C=O) at the 3rd and 11th positions, some double bonds exist in the A ring or the 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 the differences in substituents, double bond positions or stereo configurations on the steroidal core. [0003] ...

Claims

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Application Information

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IPC IPC(8): C12P33/10C12R1/66
CPCC12P33/10
Inventor 荣绍丰李迎光管世敏蔡保国李茜茜张硕
Owner SHANGHAI INST OF TECH
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