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Process for producing recombinant bacillus subtilis of UPD glycosyltransferase

A Bacillus subtilis, glycosyltransferase technology, applied in the field of biological research, can solve problems such as slow fermentation speed, achieve the effect of fast fermentation speed, high efficiency, and shorten the time of comparison

Inactive Publication Date: 2019-12-20
JIANGSU SHIYUTIAN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the deficiencies of the prior art, the present invention provides a process for producing recombinant Bacillus subtilis for UPD glycosyltransferase, which has the advantages of faster fermentation speed and solves the problem of relatively slow fermentation speed

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment one: a kind of technology that is used to produce the recombinant Bacillus subtilis of UPD glycosyltransferase comprises the following steps:

[0022] (1) Use UPD as the raw material to produce UPD glycosyl, then add an enzyme-producing accelerator, and then fuse with the active enzyme to produce UPD-transferase; the enzyme-producing accelerator includes calcium magnesium phytate (phytin), polysorbate -80 (Tween-80), ethylenediaminetetraacetic acid (EDTA), detergent LS (sodium fatty amide sulfonate), polyvinyl alcohol, etc.

[0023] (2) Using recombinant Bacillus subtilis as the starting strain, optimize the medium and fermentation process to obtain a sample of recombinant Bacillus subtilis; insert the recombinant Bacillus subtilis strains stored in a liquid nitrogen tube at -20°C into the medium , and then use a manual shaker with a rotation speed of 20-40rpm / s and a culture temperature of 10-40°C for 5 hours to obtain a seed liquid sample, which is then div...

Embodiment 2

[0033] Embodiment two: a kind of technology that is used to produce the recombinant Bacillus subtilis of UPD glycosyltransferase comprises the following steps:

[0034] (1) Use UPD as the raw material to produce UPD glycosyl, then add an enzyme-producing accelerator, and then fuse with the active enzyme to produce UPD-transferase; the enzyme-producing accelerator includes calcium magnesium phytate (phytin), polysorbate -80 (Tween-80), ethylenediaminetetraacetic acid (EDTA), detergent LS (sodium fatty amide sulfonate), polyvinyl alcohol, etc.

[0035] (2) Using recombinant Bacillus subtilis as the starting strain, optimize the medium and fermentation process to obtain a sample of recombinant Bacillus subtilis; insert the recombinant Bacillus subtilis strains stored in a liquid nitrogen tube at -20°C into the medium , and then use a manual shaker with a rotation speed of 20-40rpm / s and a culture temperature of 10-40°C for 5 hours to obtain a seed liquid sample, which is then div...

Embodiment 3

[0045] Embodiment three: a kind of technology that is used to produce the recombinant Bacillus subtilis of UPD glycosyltransferase comprises the following steps:

[0046] (1) Use UPD as the raw material to produce UPD glycosyl, then add an enzyme-producing accelerator, and then fuse with the active enzyme to produce UPD-transferase; the enzyme-producing accelerator includes calcium magnesium phytate (phytin), polysorbate -80 (Tween-80), ethylenediaminetetraacetic acid (EDTA), detergent LS (sodium fatty amide sulfonate), polyvinyl alcohol, etc.

[0047] (2) Using recombinant Bacillus subtilis as the starting strain, optimize the medium and fermentation process to obtain a sample of recombinant Bacillus subtilis; insert the recombinant Bacillus subtilis strains stored in a liquid nitrogen tube at -20°C into the medium , and then use a manual shaker with a rotation speed of 20-40rpm / s and a culture temperature of 10-40°C for 5 hours to obtain a seed liquid sample, which is then div...

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Abstract

The invention discloses a process for producing recombinant bacillus subtilis of UPD glycosyltransferase. The process comprises the following steps: producing UPD glycosyl by using UPD as a raw material, adding an enzyme production promoter, and fusing with an active enzyme to produce UPD-transferase; by taking recombinant bacillus subtilis as an original strain, optimizing a culture medium and afermentation process to obtain a recombinant bacillus subtilis sample; inoculating the recombinant bacillus subtilis into a seed culture medium, carrying out manual shake cultivation to a logarithmicphase, and optimizing to obtain a seed culture solution; adding UDP-glycosyltransferase; then testing initial enzyme activity parameters; and then placing fused materials at the temperature of 10-40 DEG C, and recording the production speed. The enzyme activity can be adjusted by adjusting different temperatures, so that the efficiency is higher, the comparison time can be shortened by setting a plurality of seed culture solution samples and UDP-glycosyltransferase samples, and the optimal scheme can be found in the shortest time.

Description

technical field [0001] The invention relates to the technical field of biological research, in particular to a process for producing recombinant Bacillus subtilis for UPD glycosyltransferase. Background technique [0002] Bacillus subtilis is a kind of Bacillus genus, single cell 0.7~0.8×2~3 microns, uniform coloring, no capsule, perinatal flagella, able to move. Gram-positive bacteria, spores 0.6-0.9×1.0-1.5 microns, oval to columnar, located in the center of the bacterium or slightly biased, the bacterium does not expand after spore formation. The surface of the colony is rough and opaque, stained white or yellowish, and often forms wrinkles when growing in liquid medium. Aerobic bacteria. It can use protein, various sugars and starch to decompose tryptophan to form indole. It is widely used in genetic research, and the synthesis pathway and regulation mechanism of purine nucleotides in this bacterium are relatively clear. Widely distributed in soil and decaying organi...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N1/21C12R1/125
CPCC12N1/20C12N9/1051
Inventor 陈宇杰
Owner JIANGSU SHIYUTIAN BIOTECHNOLOGY CO LTD
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