25-hydroxyvitamin D3 production strain, and screening method and application thereof

A technology for the production of hydroxyvitamins and bacterial strains, which is applied in the field of bioengineering, can solve the problems of long cultivation period and high production cost, and achieve the effects of rapid growth, shortened production period and reduced production cost

Inactive Publication Date: 2020-04-24
ZHEJIANG SHENGDA BIO PHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the long cultivation cycle of t...

Method used

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  • 25-hydroxyvitamin D3 production strain, and screening method and application thereof
  • 25-hydroxyvitamin D3 production strain, and screening method and application thereof
  • 25-hydroxyvitamin D3 production strain, and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 High yield of 25-hydroxyvitamin D 3 Screening of strains

[0029] Take a soil sample from the deep soil layer near the chemical plant, weigh about 1.0 g, and add it to 30 mL of sterile water to prepare a bacterial suspension. Draw 0.5mL of the bacterial suspension and inoculate it into the enrichment medium, and culture it on a constant temperature shaker at 37°C for 3 days. The culture solution was diluted with the mother solution and spread on the separation medium plate, and cultivated at 37°C for 24h. Pick 254 single colonies for transformation experiments, extract the transformation solution and detect 25-hydroxyvitamin D by high performance liquid chromatography 3 concentration. details as follows:

[0030] (1) Pick a single colony and culture it in the seed medium at 37°C for 12 hours to obtain the seed solution;

[0031] (2) The seed solution was inoculated in 50 mL transformation medium and cultivated for 12 hours;

[0032] (3) Take 1mL VD 3 Th...

Embodiment 2

[0035] Example 2 Different VD 3 Dissolving solvent vs. 25OH-VD 3 Yield impact

[0036] Although zju 4-2 can obtain a higher yield (70.4%), VD 3 The added solubility of β-cyclodextrin is low (0.039g / L dissolved in 200g / Lβ-cyclodextrin), which is not conducive to large-scale industrial production.

[0037] In order to improve VD 3 The dispersibility of 10% by volume, under 200rpm constant speed oscillation, VD 3 Five common solvents, ethanol, methanol, ethyl acetate, propylene glycol and ethylene glycol, were evaluated at a concentration of 0.1 mM. As shown in Table 1, the solubility of five solvents to VD3 is much greater than that of β-cyclodextrin. In addition, if Figure 4 As shown, ethanol dissolves VD 3 25OH-VD obtained by transformation 3 The yield (40%) and propylene glycol dissolved VD 3 25OH-VD obtained by transformation 3 The yield (54%) was higher than ethyl acetate (20%), methanol (17%) and ethylene glycol (35%). Therefore, ethanol and propylene glycol ar...

Embodiment 3

[0040] Example 3 VD 3 Propylene Glycol to Ethanol Ratio in Dissolving Solvent vs. 25OH-VD 3 Yield impact

[0041] Dissolving VD due to ethanol and propylene glycol 3 Conversion and synthesis of 25OH-VD 3 The yield is higher, but the two have different solubility to VD3. Further, dissolving VD3 in a mixed solvent of propylene glycol and ethanol is converted into 25OH-VD 3 . The feasibility is evaluated, and the balance between solubility and yield is achieved by adjusting the ratio of the two. Such as Figure 5 As shown, the ratio of propylene glycol to ethanol can obtain higher 25OH-VD between 10:0~5:5 (v / v) 3 Yield and dry cell weight. Especially when the ratio (v / v) of propylene glycol to ethanol was 9:1, the yield was 50.1%, and VD 3 The solubility in the co-solvent mixture was significantly increased to 24.49g / L, at which point the VD3 solubility as well as the 25OH-VD 3 Yield reached the highest, which is the best solvent ratio.

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Abstract

The invention relates to a 25-hydroxyvitamin D3 production strain, and a screening method and an application thereof, and belongs to the technical field of bioengineering. The preservation number of the 25-hydroxy vitamin D3 production strain is CCTCC M 2019385, and the preservation date of the 25-hydroxy vitamin D3 production strain is May 23, 2019. The strain can be applied to efficient conversion of a substrate vitamin D3 to synthesize 25-hydroxy vitamin D3. The strain screened by the method is high in vitamin D3 conversion efficiency, easy to culture and easy to industrially amplify.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to 25-hydroxyvitamin D 3 Production strains and their screening methods and applications. Background technique [0002] 25 hydroxyvitamin D 3 (hereinafter referred to as 25OH-VD 3 ), maintain and regulate the normal metabolism of calcium and phosphorus in the body, promote the absorption of calcium and phosphorus, maintain blood calcium and phosphorus levels, and maintain the normal growth and development of bones. It is clinically used to treat rickets, rickets and other diseases. In aquaculture, 25OH-VD 3 It can promote the development of bones, increase the speed of broiler weight gain, enhance feed conversion efficiency, reduce the death rate, improve muscle tenderness, improve processing performance and meat production rate. [0003] Currently, 25OH-VD 3 The production process mainly includes chemical synthesis method and light irradiation method. The ch...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/04C12P7/02C12R1/085
CPCC12P7/02C12Q1/045C12N1/205C12R2001/085
Inventor 徐志南朱勇刚周斌汤丹丹刘伟黄磊
Owner ZHEJIANG SHENGDA BIO PHARM
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