11-alpha hydroxylation reaction preparation method of steride hormone substance important intermediate

A technology for steroid hormones and intermediates, which is applied in the field of bioengineering transformation, can solve the problems of complex and tedious processes and high production costs, and achieve the effects of shortening reaction time, simplifying preparation method and shortening total reaction time.

Active Publication Date: 2014-07-16
GUANGXI WANDE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The process uses natural medium, the production cost is high, and the process involves primary and secondary cultivation, the process is complex and cumbersome, and the highest conversion rate is only 73.2%.

Method used

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  • 11-alpha hydroxylation reaction preparation method of steride hormone substance important intermediate
  • 11-alpha hydroxylation reaction preparation method of steride hormone substance important intermediate
  • 11-alpha hydroxylation reaction preparation method of steride hormone substance important intermediate

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) The first step is to cultivate the seeds of Metarhizium anisopliae. Firstly, the slant medium and seed medium are prepared and sterilized. The mold strain is spread on the slant medium and cultivated at 27 degrees Celsius for 5-7 days. Add sterile water to the slant of the bacteria, and sweep it down with a cotton swab to make a concentration of about 1.2×10 8 spores / mL. Take 0.5ml of spore liquid, inoculate it into 100mL of sterilized seed medium, and cultivate it for about 24 hours at 28 degrees Celsius and 150-200r / min. Among them, the composition of the seed medium (g / L): 0.05 dipotassium hydrogen phosphate, 0.05 magnesium sulfate, 0.005 ferric ammonium citrate, 2 citric acid, 2 ammonium nitrate, 10 calcium carbonate, 20 glycerol, and 5 glucose. Slant medium (g / L): Add 15g of agar on the basis of the seed medium.

[0027] (2) The second step is to carry out the cultivation of the shake flask fermentation medium, transplant 80ml of the cultured seed liquid into...

Embodiment 2

[0032] (1) The first step is to cultivate the seeds of Ochraus japonicus. First, prepare the slant medium and seed medium, and sterilize it. Take the mold strain and spread it on the slant medium, and cultivate it at 25 degrees Celsius for 5-7 days. Add sterile water to the slant of the bacteria, and sweep it down with a cotton swab to make a concentration of about 1.2×10 8 spores / mL. Take 0.5ml of spore liquid, inoculate into 100mL sterilized seed medium, and cultivate for about 24h at 27 degrees, 150-200r / min. Among them, the composition of the seed medium (g / L): 0.05 dipotassium hydrogen phosphate, 0.05 magnesium sulfate, 0.005 ferric ammonium citrate, 2 citric acid, 2 ammonium nitrate, 10 calcium carbonate, 20 glycerol, and 5 glucose. Slant medium (g / L): Add 15g of agar on the basis of the seed medium.

[0033] (2) The second step is to carry out the cultivation of the shake flask fermentation medium, transplant 80ml of the cultured seed liquid into the sterilized 1600ml...

Embodiment 3

[0038] (1) The first step is to cultivate the seeds of Trichoderma and Metarhizium anisopliae. First, configure the slant medium and seed medium, and sterilize it. Take the mold strain and spread it on the slant medium, and cultivate it at 32 degrees Celsius for 5 to 7 days. . Add sterile water to the slant of the bacteria, and sweep it down with a cotton swab to make a concentration of about 1.2×10 8 spores / mL. Take 0.5ml of spore liquid, inoculate into 100mL sterilized seed medium, and cultivate for about 24h at 32 degrees, 150-200r / min. Among them, the composition of the seed medium (g / L): 0.05 dipotassium hydrogen phosphate, 0.05 magnesium sulfate, 0.005 ferric ammonium citrate, 2 citric acid, 2 ammonium nitrate, 10 calcium carbonate, 20 glycerol, and 5 glucose. Slant medium (g / L): Add 15g of agar on the basis of the seed medium.

[0039] (2) The second step is to carry out the cultivation of the shake flask fermentation medium, transplant 80ml of the cultured seed liqu...

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Abstract

The invention relates to a 11-alpha hydroxylation reaction preparation method of a steride hormone substance important intermediate. The reaction preparation method comprises the following steps of: inputting a steride substrate I into a fermentation medium containing mold seeds to ferment, carrying out the hydroxylation reaction under the effect of the mold and a catalyst, and modifying 11 sites and 21 sites of the steride substrate I to obtain the steride hormone substance important intermediate II with 11-alpha-hydroxy and 21 hydroxy. According to the reaction preparation method, a borate catalyst is used, so that the reaction speed is improved, the reaction time is saved, the consumption of air in a fermentation process is reduced, and simultaneously, the conversion rate is improved by about 5%; and the synthetic medium is used for replacing a natural medium, so that the cost is reduced, and the section pressure of subsequent separation is relieved.

Description

technical field [0001] The invention belongs to the technical field of bioengineering transformation, and in particular relates to a method for preparing an important intermediate of steroid hormones through 11-alpha hydroxylation reaction. Background technique [0002] Steroid hormones are the main components of corticosteroids. Corticosteroids play a very important role in regulating the body and have strong pharmacological effects such as anti-infection, anti-allergy, anti-virus and anti-shock. my country is a big exporter of corticosteroid raw materials, among which prednisone acetate accounts for 60% of the total output. Since the 1950s, my country has been using the Rhizopus niger conversion process, and the conversion rate of 2% of the feed is 40% to 50%. , and the production level of 80% to 90% in foreign countries is quite large, so the development of new high-yield strains is expected to improve the utilization rate of raw materials from the overall level, resulting...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/10C12P33/06C12P39/00C12R1/645C12R1/885C12R1/66C12R1/67
Inventor 王远秋朱荣谭桂莉
Owner GUANGXI WANDE PHARMA
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