Method for preparing deoxycholic acid from phytosterol

A technology of deoxycholic acid and phytosterols, which is applied in the field of preparation of deoxycholic acid compounds in a combination of biological fermentation and chemical synthesis, can solve the problems of unsuitability for industrial production, long reaction steps, complicated process, etc. Less, mild reaction conditions, high reaction yield

Active Publication Date: 2021-06-01
湖南醇健制药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, there are mainly two methods for the preparation of deoxycholic acid, one is to extract and separate from organisms, but the deoxycholic acid obtained from animal sources may contain pathogens; the other is to obtain deoxycholic acid through a synthetic method. The synthesis method of the method is complicated, pollutes the environment, and the cost is relatively high
In rec

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  • Method for preparing deoxycholic acid from phytosterol
  • Method for preparing deoxycholic acid from phytosterol
  • Method for preparing deoxycholic acid from phytosterol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The preparation of embodiment 1 compound 1

[0068] 1. Original strain transformation

[0069] 1 strain

[0070] Mycobacterium sp. NRRL B-3805

[0071] 2 conversion process

[0072] 2.1 Seed culture

[0073] First-class seeds: yeast extract powder 10g / L, glucose 15g / L, sodium nitrate 5.4g / L, diammonium hydrogen phosphate 0.06%, pH=7.5, 100ml medium into 500ml shaker flask, sterilized at 121°C for 30 minute. Inoculate from the slant after cooling, culture at 200rpm, 30°C for 48 hours.

[0074] Secondary seeds: yeast extract powder 10g / L, glucose 15g / L, sodium nitrate 5.4g / L, diammonium hydrogen phosphate 0.06%, pH=7.5, put 500ml medium into 2000ml shaker flask, sterilize at 121℃ for 30 minute. After cooling, inoculate from the primary shaker flask with an inoculum size of 10%, culture at 200 rpm, and 30°C for 48 hours.

[0075] 2.2 Conversion

[0076] 10 liter fermenter, 7 liters of sample, medium: stigmasterol 20g / L, soybean oil 160g / L, corn steep liquor 60g / L,...

Embodiment 2

[0099] The preparation of embodiment 2 compound 2

[0100] 1. Seed cultivation

[0101] (1) Incline cultivation

[0102] Adopt Aspergillus ochraceus ATCC 18500 as the production strain, inoculate the preserved bacterial strain by streaking on the slant, and cultivate it for 6-7 days at 30°C. The slant medium is potato medium: potato pieces 200g / L (boiled for 30 minutes , four layers of gauze filter to get the filtrate), glucose 20g / L, agar 20g / L, pH6.5.

[0103] (2) Seed cultivation

[0104] Preparation of spore suspension: Take a fresh slant that has been cultured for 6-7 days, wash the spores of the slant with 0.05% Tween-80 sterile water to make a spore suspension, and count the spore concentration under the microscope to be 2 to 3×10 7 a / ml;

[0105] Shake flask seed culture: inoculum size 10%, culture at 30°C, 180rpm for 36-48h.

[0106] The composition of the seed medium is as follows: corn steep liquor 10g / L, glucose 30g / L; pH value 7.2±0.2.

[0107] Sterilize by ...

Embodiment 3

[0117] The preparation of embodiment 3 compound 2 '

[0118] 1 strain

[0119] The name of the strain: Mycobacterium smegmatis NK-XHX-118 is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2013544.

[0120] 2 conversion process

[0121] 2.1 Seed culture

[0122] First-class seeds: yeast extract powder 10g / L, glucose 15g / L, sodium nitrate 5.4g / L, diammonium hydrogen phosphate 0.06%, pH=7.5, 100ml medium into 500ml shaker flask, sterilized at 121°C for 30 minute. Inoculate from the slant after cooling, culture at 200rpm, 30°C for 48 hours.

[0123] Secondary seeds: yeast extract powder 10g / L, glucose 15g / L, sodium nitrate 5.4g / L, diammonium hydrogen phosphate 0.06%, pH=7.5, put 500ml medium into 2000ml shaker flask, sterilize at 121℃ for 30 minute. After cooling, inoculate from the primary shaker flask with an inoculum size of 10%, culture at 200 rpm, and 30°C for 48 hours.

[0124] 2.2 Conversion

[0125] 10 liter fer...

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Abstract

The invention provides application of a mutagenic strain of which the preservation number is CCTCC (China Center For Type Culture Collection) NO: M 2020987 in preparation of deoxycholic acid. The invention provides a novel method for preparing deoxycholic acid by taking phytosterol as a raw material and combining biological fermentation and chemical synthesis, animal source extraction is not used in the method, industrial wastewater and waste residues can be used as raw materials, and the method is green and environment-friendly; the carbonyl side chain of the cholic acid compound is constructed on the side chain of the phytosterol in one step by virtue of a mycobacterium sp.NRRL B-3805 mutagenic strain biological fermentation method, so that the operation is simple, the yield is high, isomer impurities are few, and the use amount of an organic solvent is small; according to the method for preparing the deoxycholic acid compound, reaction reagents are simple and easy to obtain, reaction conditions are mild, the reaction yield is high, and the method is suitable for large-scale industrial production.

Description

technical field [0001] The invention relates to the technical field of chemical pharmacy, in particular to a method for preparing deoxycholic acid from phytosterols, in particular to a method for preparing deoxycholic acid compounds by using phytosterols as raw materials by combining biological fermentation and chemical synthesis. Background technique [0002] Deoxycholic acid (DCA), also known as deoxycholic acid, has a chemical name of 3α, 12α-dihydroxy-5β-cholanic acid, which is a free bile acid obtained by losing an oxygen atom in cholic acid. Bile mainly exists in the form of taurine and glycine. Deoxycholic acid and its salts have surface activity, are safe and effective emulsifiers in cosmetics and pharmaceuticals, have antifungal and anti-inflammatory effects, and can be used to treat tooth root diseases. It can be used in the treatment of hypersecretion of sebaceous glands in skin surgery. It can be used in products such as face powder to remove excess sebum and s...

Claims

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Application Information

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IPC IPC(8): C12P33/08C12P33/06C12P33/00C12P39/00C07J9/00C12N15/01C12R1/66C12R1/34C12R1/32
CPCC12P33/08C12P33/06C12P33/00C12P39/00C12N15/01C07J9/005
Inventor 蒋红平唐杰李斌郭志军
Owner 湖南醇健制药科技有限公司
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