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A method for increasing the content of thermus trehalose synthase

A technology of trehalose synthase and Thermus, which is applied in the field of bioengineering to achieve the effects of reducing production costs, realizing production and high catalytic yield

Inactive Publication Date: 2016-01-20
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant reports or patents on the use of pressure to increase the content of trehalose synthase in Thermus bacterium

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A method for increasing the content of Thermus trehalose synthase, the steps are as follows:

[0028] (1) Pick 2 strains of Thermus annulus from the inclined surface of the test tube and inoculate them into the liquid seed medium, and cultivate them at 70°C and 150r / min for 16 hours to obtain activated liquid seeds, and then inoculate the liquid seeds at a volume ratio of 5 % of the inoculum was inoculated into the liquid medium, and cultivated at 70°C and 200r / min for 24h to obtain the strain cell culture medium;

[0029] (2) Put the strain cell culture solution prepared in step (1) in a pressure reactor, and ferment for 4 hours under a pressure of 1.5MPa, the fermentation condition is 70°C, 200r / min, to obtain a fermentation solution;

[0030] (3) Centrifuge 1000mL of the fermentation broth prepared in step (2) at 5000r / min for 20min, collect the bacterial cells, resuspend the bacterial cells in deionized water, and prepare a suspension with a cell mass concentration ...

Embodiment 2

[0036] A method for increasing the content of Thermus trehalose synthase, the steps are as follows:

[0037] (1) Pick 1 Thermus annulus strain from the slant of the test tube and inoculate it into the liquid seed medium, cultivate it at 65°C and 100r / min for 12h to prepare the activated liquid seed, and then inoculate the liquid seed at a volume ratio of 3 % of the inoculum was inoculated into the liquid medium, and cultivated at 60°C and 150r / min for 20h to obtain the cell culture medium of the strain;

[0038] (2) Put the strain cell culture liquid prepared in step (1) in a pressure reactor, and ferment for 1 hour under the pressure of 2.0MPa, and the fermentation conditions are 60°C and 150r / min to obtain the fermentation liquid;

[0039] (3) Centrifuge 1000mL of the fermentation broth prepared in step (2) at 5000r / min for 20min, collect the bacterial cells, resuspend the bacterial cells in deionized water, and prepare a suspension with a cell mass concentration of 35wt%. ...

Embodiment 3

[0044] The method for improving the content of Thermus trehalose synthase as described in Example 1, the difference is:

[0045] In step (2), the strain cell culture solution is placed in a pressure reactor, and fermented at a pressure of 0.5 MPa for 6 hours to obtain a fermentation solution.

[0046] Under these conditions, 862 mg of trehalose synthase was obtained.

[0047] After testing, the activity of trehalose synthase in the cells of Thermus was 78.1U / g wet cells.

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Abstract

The invention relates to a method for increasing the trehalose synthase content of thurmus. The method comprises the following steps: (1) inoculating a liquid seed of a thurmus strain into a liquid culture medium for culturing to prepare a strain cell culture solution; (2) fermenting the strain cell culture solution in a pressure reactor under the pressure to prepare a fermentation solution; (3) centrifuging the fermentation solution, collecting bacterial cells, breaking the cells to prepare a cell breaking solution, and extracting the cell breaking solution by a two-aqueous-phase extraction system to prepare trehalose synthase. According to the method, the trehalose synthase content of the thurmus cells can be increased, the production cost of the trehalose synthase can be reduced, the trehalose synthase can be continuously produced on a large scale, and the production process of the trehalose synthase is simple and convenient.

Description

technical field [0001] The invention relates to a method for increasing the content of thermus trehalose synthase, which belongs to the technical field of bioengineering. Background technique [0002] Trehalose synthase (Trehalosesynthase, EC5.4.99.16) is an intramolecular glucosidic transferase that can specifically catalyze the conversion of α-1,4 glycosidic bonds of maltose into α-1,1 glycosidic bonds to generate trehalose. Trehalose is a non-reducing disaccharide formed by the condensation of two glucose molecules through hemiacetal hydroxyl groups. It has the special function of protecting the biofilm and biological macromolecules of organisms under adversity, and can endow lower animals and plants The ability to resist nutrient deficiency, high temperature, high osmotic pressure, dryness, toxic substances and other harsh environments, and microorganisms, so it has broad application prospects in the fields of biological products, food, medicine, crop breeding and fine c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/90C12R1/01
CPCC12N9/90C12Y504/99016
Inventor 董永胜马蕾段元良
Owner QILU UNIV OF TECH
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