Method for preparing 1,25-dihydroxyl vitamin D3 by microbial transformation

A technology for the transformation of dihydroxyvitamins and microorganisms, which can be applied to microorganism-based methods, biochemical equipment and methods, microorganisms, etc., and can solve the problems of complex operation, low yield, complicated separation and purification, etc., and achieve mild conversion conditions and process routes. Short, conversion-enhancing effects

Active Publication Date: 2018-05-18
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

Benefits of technology

This technology allows us to use tiny bacteria or yeast cells instead of expensive chemical reagents like calcium oxide (CaO) to produce certain substances called lanthanoid compounds from alpha -oxygenated vitamins. These processes can be carried on by different types of biological agents with specific functions but they have limitations due to their high production costs compared to other methods used nowadays.

Problems solved by technology

The technical problem addressed in this patents relating to improving the efficiency of administering certain medications like calcidiators (calciercalc) and diphosphanides (DRP), which help prevent diseases associated with these therapies. However, current sources lack precise dosage control due to their poor solubility and bioavailability. Current syringe systems cannot accurately deliver high amounts of effective medicine without causing side effects. Therefore, new ways must be explored to develop better formulations containing both calcium salt and DRP.

Method used

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  • Method for preparing 1,25-dihydroxyl vitamin D3 by microbial transformation
  • Method for preparing 1,25-dihydroxyl vitamin D3 by microbial transformation

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 100 mL of culture medium, place it in a 500 mL shake flask, and sterilize at 121°C for 30 minutes.

[0036] Prepare a spore suspension. All the spores on the plate culture were put into an Erlenmeyer flask (containing glass beads) containing 20 ml of sterile water, and the spore suspension was made by shaking the bed at 260 rpm for about 15 minutes. At the same time, the spore liquid was counted by a plate. After three counts, the amount of spores per milliliter was basically stable at 10 8 Pcs / ml.

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

[0038] (2) Add prepared 50mg dextrin, 4ml water, 40mg glycerol, 50mg 1α-hydroxyvitamin D to the medium 3 , 100mg Span 60 and 6mL ...

Embodiment 2

[0043] (1) The 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 are mixed evenly, 100 mL of culture medium is taken out and placed in a 500 mL shake flask, and sterilized at 121°C for 30 min.

[0044] A spore suspension was prepared, and the same concentration of the spore suspension was prepared using the method in Example 1.

[0045] 5 mL of the spore suspension was inoculated into the medium, and cultured on a shaker at a rotation speed of 240 rpm and a temperature of 28°C for 3 days.

[0046] (2) Add prepared 200mg dextrin, 8ml water, 100mg glycerol, 100mg 1α-hydroxyvitamin D to the medium 3 , 200mg Span 80, 8mL diol fatty acid ester mixed to make a mixture, continue to cultivate 4d, to obtain fermentation broth. The mixing step of this mixture is to first dissolve 200mg dextrin in 8ml hot water, then 100mg glycerin, 100mg1α-hydroxyvitamin D 3 Dissolve 200mg Tween...

Embodiment 3

[0051] (1) The 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 shake flask, and sterilize it at 121°C for 30min.

[0052] A spore suspension was prepared, and the same concentration of the spore suspension was prepared using the method in Example 1.

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

[0054] (2) Add prepared 50mg dextrin, 8ml water, 20mg glycerol, 100mg 1α-hydroxyvitamin D to the medium 3 , 150mg stearic acid, 50mg pectin, 8mL absolute ethanol mixed to make a mixture, continue to cultivate 4d, to obtain fermentation broth. The mixing step of this mixture is to first dissolve 50mg dextrin and 50mg pectin in 8ml hot water, then 20mg glycerin, 100mg 1α-hydroxyvitamin D 3 Dissolve 150mg stear...

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PUM

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Abstract

The invention relates to a preparation method of 1,25-dihydroxyl vitamin D3, in particular to a method for preparing 1,25-dihydroxyl vitamin D3 by microbial transformation. According to the method, 1alpha-hydroxyvitamin D3 is subjected to 25-site hydroxyl transformation through nocardia sp.fpsw2013097 collected in a unit specified by the State Intellectual Property Office with the collection number CGMCC No.14165. A transformation method comprises the following specific steps: inoculating a prepared spore suspension into a culture medium, and culturing on a shaking table at the rotating speedof 200 to 250rpm and at the temperature of 24 to 32 DEG C for 2 to 5 days; adding a mixture prepared by mixing dextrin, water, glycerinum, 1alpha-hydroxyvitamin D3, a surfactant and an organic solventinto the culture medium, and continually culturing for 3 to 7 days; filtering to obtain a finished product. The method has the advantages of realization of direct hydroxylation at the 25-bite of the1alpha-hydroxyvitamin D3, high yield, simple production steps, low cost and the like.

Description

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Claims

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Application Information

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Owner 四川菲派生物科技有限公司
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