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Method for preparing 25-hydroxyvitamin D by microbial transformation

A technology for the transformation of hydroxyvitamin and microorganisms, which is applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., and can solve the problems of numerous synthesis steps, complicated separation and purification, and complicated operations, and achieves high yield and short process route. , the effect of mild transformation conditions

Active Publication Date: 2012-09-12
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
The production operation is complicated, and the yield is low, about 10%.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Select 1.5% glucose by weight, 1.5% soybean cake powder, 0.5% corn steep liquor, 0.5% NaCl, 0.2% CaCO3 and 95.8% distilled water and mix evenly, adjust the pH to 7.0, take out 100mL of medium and place it in a 500mL shaker flask Sterilize at 121°C for 30 minutes.

[0034] A spore suspension was prepared. Put all the spores on the plate culture into a Erlenmeyer flask (containing glass beads) containing 20ml of sterile water, and shake at 260rpm for about 15min to make 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.

[0035] 4 mL of the spore suspension was inoculated into the medium, and cultured on a shaker with a rotation speed of 240 rpm and a temperature of 27° C. for 4 days.

[0036] (2) Add prepared 50mg cyclodextrin and 10mg vitamin D to the culture medium 3 , 10mg Triton X-100 and 2mL of absolute ethanol were mixed...

Embodiment 2

[0040] (1) Choose 1.5% glucose by weight, 1.5% soybean cake powder, 0.5% corn steep liquor, 0.7% yeast extract, 0.5% NaCl, 0.2% CaCO 3 Mix well with 95.1% distilled water, take out 100mL medium and place it in a 500mL shake flask, and sterilize at 121°C for 30min.

[0041] A spore suspension was prepared, and a spore suspension with the same concentration was prepared in the manner in Example 1.

[0042] 4 mL of the spore suspension was inoculated into the culture medium, and cultured on a shaker with a rotation speed of 200 rpm and a temperature of 28° C. for 60 h.

[0043] (2) Add 1g of β-cyclodextrin and 340mg of vitamin D to the medium 2 , 100mg Triton X-100, 5mL absolute ethanol and 5mL sterilized water were mixed and cultured for 72 hours to obtain a fermentation broth. The mixing procedure for this mixture is: 340 mg vitamin D 2Dissolve in 5ml of absolute ethanol, add 1g of methylated β-cyclodextrin, 100mg of Triton X-100 and 5ml of sterile water, dissolve and mix we...

Embodiment 3

[0047] (1) The weight percentages are 1.5% glucose, 1.5% soybean cake powder, 0.5% corn steep liquor, 0.5% NaCl, 0.2% CaCO 3 Mix well with 95.8% distilled water, adjust the pH to 7.0, take out 50mL medium and place it in a 250mL shaker flask, and sterilize at 121°C for 30min.

[0048] A spore suspension was prepared, and a spore suspension with the same concentration was prepared in the manner in Example 1.

[0049] 4 mL of the spore suspension was inoculated into the culture medium, and cultured on a shaker with a rotation speed of 200 rpm and a temperature of 27° C. for 3 days.

[0050] (2) Add 10mg of β-cyclodextrin, 5mg of γ-cyclodextrin and 30mg of vitamin D to the medium 2 , 5mg of Triton X-100, 2mL of absolute ethanol, and 1mL of sterile water were mixed and cultured for 3 days to obtain a fermentation broth. The mixing procedure for this mixture is: 30 mg vitamin D 2 , 10mg of β-cyclodextrin, 5mg of γ-cyclodextrin and 5mg of Triton X-100 are dissolved in a mixed sol...

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PUM

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Abstract

The invention discloses a method for preparing 25-hydroxyvitamin D by microbial transformation. The method utilizes Streptomycessp. wsbee2009112 which is preserved in the China general microbiological culture collection center and has a preservation number of CGMCC No.5831 to carry out 25-hydroxy transformation of vitamin D. The method comprises the following steps of inoculating a medium with a prepared spore suspension, carrying out culture on a shaking table at a temperature of 22 to 30 DEG C at a rotation rate of 200 to 280rpm for 2 to 5 days, adding a mixture of one or more cyclodextrins, vitamin D, a surfactant and an organic solvent into the medium, carrying out culture for 1 to 10 days, and filtering to obtain a finished product. The method realizes direct hydroxylation at a 25 site of vitamin D, and has a high yield, simple production processes and a low cost.

Description

technical field [0001] The invention relates to a method for transforming and producing 25-hydroxyvitamin D, specifically a method for transforming and producing 25-hydroxyvitamin D by microorganisms. Background technique [0002] 25-Hydroxyvitamin D 3 And other active vitamins are clinically used for osteoporosis and rickets and osteomalacia caused by various reasons, and can also be used as feed additives in the feed industry. [0003] 25-Hydroxyvitamin D 3 , also known as calcifediol, is vitamin D 3 The active metabolite has stronger physiological activity and does not need to be metabolized by the liver. It Has More Than Regular Vitamin D 3 All functions, 25-hydroxyvitamin D 3 As an active substance bypassing the liver transformation, it can be directly absorbed by humans or animals; when used in poultry animals, it can promote the bone development of poultry animals, promote the maximization of bone density, reduce the mortality rate of chicks by 1% to 3%, and redu...

Claims

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Application Information

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IPC IPC(8): C12P33/06C12R1/465
Inventor 敖绍川姚佳陆曹伟周余涛
Owner 四川菲派生物科技有限公司
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