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A microbial transformation to manufacture 1,25-dihydroxyvitamin D 3 Methods

A technology for the transformation of dihydroxyvitamins and microorganisms, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., and can solve problems such as complex operations, difficult synthesis of raw materials, and numerous synthesis steps

Active Publication Date: 2021-04-27
四川菲派生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many synthetic steps, complex separation and purification
Synthetic raw materials are not easy to obtain, the operation is complicated in production, and the yield is low, about 10%

Method used

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  • A microbial transformation to manufacture 1,25-dihydroxyvitamin D <sub>3</sub> Methods
  • A microbial transformation to manufacture 1,25-dihydroxyvitamin D <sub>3</sub> Methods

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] (1) The weight percentage is 1.5% glucose, 0.5% peptone, 0.5% yeast powder, 1% corn steep liquor, 0.5% NaCl, 0.1% CaCO 3 Mix well with 95.9% distilled water, adjust the pH to 7.5, take out 100mL medium and place it in a 500mL shaker flask, and sterilize at 121°C for 30min.

[0036]A spore suspension was prepared. All the spores on the plate culture were inserted into a conical flask (containing glass beads) containing 20 ml of sterile water, and the shaker was set at 260 rpm for about 15 minutes to prepare a spore suspension. Simultaneously this spore liquid adopts the plate counting method, after counting three times, the amount of spores per milliliter is basically stable at 10 8 pieces / ml.

[0037] Inoculate 5 mL of the spore suspension into the culture medium, and cultivate for 4 days on a shaker with a rotation speed of 220 rpm and a temperature of 27°C.

[0038] (2) Add prepared 50mg dextrin, 4ml water, 40mg glycerol, 50mg 1α-hydroxyvitamin D to the culture med...

Embodiment 2

[0043] (1) The selected weight percentage is 1.5% glucose, 1% yeast powder, 0.5% soybean powder, 0.5% corn steep liquor, 0.5% NaCl, 0.1% (NH 4 ) 2 SO 4 , 0.1% CaCO3 and 95.8% distilled water were mixed evenly, and 100mL of medium was taken out and placed in a 500mL shaker flask, and sterilized at 121°C for 30min.

[0044] A spore suspension was prepared, and a spore suspension with the same concentration was prepared in the manner in Example 1.

[0045] Inoculate 5 mL of the spore suspension into the culture medium, and cultivate for 3 days on a shaker with a rotation speed of 240 rpm and a temperature of 28°C.

[0046] (2) Add prepared 200mg dextrin, 8ml water, 100mg glycerol, 100mg 1α-hydroxyvitamin D to the culture medium 3 , 200mg Span 80, and 8mL diol fatty acid ester were mixed and cultured for 4 days to obtain a fermentation broth. The mixing procedure of this mixture is to first dissolve 200mg dextrin with 8ml hot water, then dissolve 100mg glycerin, 100mg 1α-hydro...

Embodiment 3

[0051] (1) The selection of weight percentage is 1.5% glucose, 0.5% peptone, 1.5% corn steep liquor, 0.5% NaCl, 0.2% K 2 HPO 4 , 0.1﹪CaCO 3 Mix well with 95.7% distilled water, adjust the pH to 7.5, take out 100mL medium and place it in a 250mL shaker flask, and sterilize at 121°C for 30min.

[0052] A spore suspension was prepared, and a spore suspension with the same concentration was prepared in the manner in Example 1.

[0053] Inoculate 4 mL of the spore suspension into the culture medium, and cultivate for 3 days on a shaker with a rotation speed of 220 rpm and a temperature of 27°C.

[0054] (2) Add prepared 50mg dextrin, 8ml water, 20mg glycerol, 100mg 1α-hydroxyvitamin D to the culture medium 3 , 150mg of stearic acid, 50mg of pectin, and 8mL of dehydrated ethanol were mixed, and continued to cultivate for 4 days to obtain a fermentation broth. The mixing procedure of this mixture is to first dissolve 50mg dextrin and 50mg pectin with 8ml hot water, then dissolve ...

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Abstract

The present invention relates to 1,25-dihydroxyvitamin D 3 The production method, specifically refers to a microbial transformation to produce 1,25-dihydroxyvitamin D 3 Methods. The present invention mainly adopts the actinomycete Nocardia sp.fpsw2013097 preserved by the unit designated by the State Intellectual Property Office to treat 1α-hydroxyvitamin D 3 The 25-hydroxyl conversion was carried out, and the preservation number was CGMCC No.14165. The specific transformation method of the present invention is as follows: inoculate the prepared spore suspension in the culture medium, and cultivate it on a shaker with a rotating speed of 200-250 rpm and a temperature of 24°C-32°C for 2-5 days; Dextrin, Water, Glycerin, 1α-Hydroxyvitamin D 3 , surface active agent and organic solvent, and continue to cultivate for 3 to 7 days; after filtering and other steps, the finished product is obtained. The present invention has the ability to produce 1α-hydroxyvitamin D 3 The 25-position is directly hydroxylated, the yield is high, the production steps are simple, and the cost is low.

Description

technical field [0001] The present invention relates to a method for converting and manufacturing 1,25-dihydroxyvitamin D 3 A method for the production of 1,25-dihydroxyvitamin D by a microorganism 3 Methods. Background technique [0002] 1,25-Dihydroxyvitamin D 3 , also known as calcitriol, calcitriol. 1,25-Dihydroxyvitamin D 3 is 25-hydroxyvitamin D 3 The active metabolite transformed by the kidney is the substance that promotes the absorption and deposition of calcium and phosphorus and finally works, so it is called fully active vitamin D 3 . as vitamin D 3 The active metabolite of 1,25-dihydroxyvitamin D 3 More Than Just the Routine Vitamin D 3 All functions can also bypass the transformation of the liver and kidneys and be directly absorbed by humans or animals to promote bone development in humans and animals and maximize bone density. [0003] 1,25-Dihydroxyvitamin D 3 And other active vitamins are clinically used to treat osteoporosis, chronic renal insuf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/465
CPCC12P7/02C12N1/205C12R2001/465
Inventor 姚佳陆群敖绍川余霞伍金元
Owner 四川菲派生物科技有限公司
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