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Method for preparing dehydroepiandrosterone by microbial fermentation

A technology of dehydroepiandrosterone and microbial fermentation, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high production cost of dehydroepiandrosterone, high environmental pressure, long reaction route and the like , to achieve the effect of being conducive to environmental protection, low cost and short route

Active Publication Date: 2013-10-23
HUNAN NORCHEM PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is the method for researching and utilizing phytosterols to prepare DHEA at present. It has a longer reaction route, and the multi-step reaction makes the production cost of DHEA larger; and due to the multi-step reaction, each step loss is relatively large. Large, low total yield, high pressure on environmental protection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Step 1 Protection reaction

[0045] Ingredients ratio:

[0046] name Dosage name Dosage Phytosterols (purchased from Zhengzhou Linuo Biotechnology Co., Ltd.) 10gram Phosphorus pentoxide 5 grams Methylal (CAS No.: 109-87-5) 150ml 1% sodium carbonate solution 40 ml Diatomaceous earth (CAS No.: 61790-53-2) 10gram

[0047] Operation method:

[0048] Add phytosterol and methylal according to the above ratio, heat up to about 25°C, stir until completely dissolved, add diatomaceous earth, then slowly add phosphorus pentoxide, control the temperature during the addition process to not exceed 30°C, and stir at about 25°C 1~1.5h, TLC detection reaction is complete (TLC detection uses benzene: acetone = 4:1 as developing solvent, 20% sulfuric acid ethanol solution spray plate, high temperature drying for color development, if a little unfinished, add a small amount of pentoxide diphosphorus). Raise the temperature to above ...

Embodiment 2

[0067] Step 1 Protection reaction

[0068] Ingredients ratio:

[0069] name Dosage name Dosage Sitosterol 10gram Dinitrogen pentoxide 5 grams Methylal 110ml 1% sodium bicarbonate solution 50 ml Silica gel 10gram

[0070] Operation method is suitable with embodiment 1, yield: 103%.

[0071] Step 2 Fermentation Transformation

[0072] Slope culture, seed culture method are suitable with embodiment 1, and fermentation strain is: Mycobacterium sp . NRRL B-3805.

[0073] 20g of protective material in 1L of water, the transformation medium formula is:

[0074] name Dosage(g / L) name Dosage(g / L) canola oil 140 glucose 11 yeast 9 potassium nitrate 5.5 Potassium dihydrogen phosphate 5.5 Dipotassium phosphate 5.5

[0075] pH= 7.2; inoculum size: 11%; shake flask transformation: 30°C, 180rpm, transformation time: 144 hours; 50 liter tank transformation: sample volume 70%, tank pressu...

Embodiment 3

[0081] Step 1 Protection reaction

[0082] Ingredients ratio:

[0083] name Dosage name Dosage Ergosterol 10gram Dinitrogen pentoxide 14 grams Methylal 160ml 1% sodium monohydrogen phosphate solution 50 ml Silica gel 15 grams

[0084] Operation method is suitable with embodiment 1, and yield: 105%.

[0085] Step 2 Fermentation Transformation

[0086] Slope culture, seed culture method are suitable with embodiment 1, and fermentation strain is: Mycobacterium sp . NRRL B-3683.

[0087] 1L of water contains 20 protectants, and the transformation medium formula is:

[0088] name Dosage(g / L) name Dosage(g / L) Corn oil 147.2 glucose 10 Yeast extract 10 sodium nitrate 5 Sodium dihydrogen phosphate 5 Disodium phosphate 5

[0089] pH= 6.8; inoculum size: 9%; shake flask transformation: 30°C, 180rpm, transformation time: 140 hours;

[0090] 50 liter tank conversion: sample volume...

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Abstract

The invention relates to a production method of midbodies of sterides drugs, in particular to a method for preparing dehydroepiandrosterone by microbial fermentation. The method includes the following steps of firstly, protecting three hydroxy of sterol to obtain protections; secondly, fermenting the protections by bacterial strain Mycobacteriumsp.NRRLB-3683 or 3805; thirdly, hydrolyzing a target object; and fourthly, purifying. The invention further provides a new path for preparing dehydroepiandrosterone from plant sterol, production cost of dehydroepiandrosterone is greatly reduced and quality yield of the dehydroepiandrosterone is improved.

Description

technical field [0001] The invention relates to a production method of a steroid drug intermediate, in particular to a method for preparing dehydroepiandrosterone by fermenting and transforming sterols with microorganisms. Background technique [0002] Dehydroepiandrosterone (dehyroepiandrosterone, DHEA) is a C 19 Steroid carrier compound, chemical name 3β-hydroxyandrost-5-en-17-one, molecular formula C 19 h 28 o 2 , with the following structure: [0003] . [0004] DHEA was first isolated from urine in 1944, and human dehydroepiandrosterone sulfate (DHEAS), discovered in 1954, is the most abundant corticosteroid hormone in human plasma. Dihydrocholesterol is converted into DHEA in the royal layer cells of the adrenal cortex by the action of an enzyme system mainly composed of cytochrome P450, and DHEA is catalyzed by thiokinase into DHEAS. Unlike other steroid hormones, the concentration of DHEA and DHEAS in plasma decreases significantly with age. A large number o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/00C12R1/32
Inventor 刘喜荣孟浩杨坤
Owner HUNAN NORCHEM PHARMACEUTICAL CO LTD
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