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Glutamine transaminase separation and purification method

A technology of glutamine and purification method, which is applied in the field of fermentation, can solve problems such as unsatisfactory texture and physical properties, and achieve the effects of promoting cell proliferation and improving enzyme production efficiency

Inactive Publication Date: 2016-12-21
ANHUI ZHENGWEIQI SEASONING FOOD LIMITED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recently, foods with less salt and less phosphoric acid have been widely promoted, but their texture and physical properties are not satisfactory

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] S1. In a sterile environment, use an inoculation loop to spread the preserved Bacillus subtilis on the NA slant medium, and culture at a constant temperature of 32° C. for 48 hours to obtain activated Bacillus subtilis;

[0029] The NA slant medium is beef extract peptone medium, which consists of: beef extract 3g / L, peptone 5g / L, agar 12.5g / L, glucose 22g / L, and the pH is adjusted to 7.2 with 0.1mol / L NaOH solution ;

[0030] In this example, the deposit number of Bacillus subtilis is CGMCC 1.3358;

[0031] S2. Centrifuge the NA slant medium in S1 at 3000r / min for 10min to discard the supernatant, wash the obtained cells twice with pH7.0 phosphate buffer solution, and dissolve the cells in pH7.0 phosphate buffer solution to prepare Bacteria suspension, in which the concentration of bacteria is 1×10 8 a / mL;

[0032] S3. Apply 5 mL of the bacterial suspension prepared in S2 evenly to the bottom of the petri dish for microwave treatment, wherein the diameter of the pet...

Embodiment 2

[0043] S1. In a sterile environment, use an inoculation loop to spread the preserved Bacillus subtilis on the NA slant medium, and culture at a constant temperature of 32° C. for 48 hours to obtain activated Bacillus subtilis;

[0044] The NA slant medium is a beef extract peptone medium, which consists of: beef extract 3g / L, peptone 10g / L, agar 14g / L, glucose 24g / L, and the pH is adjusted to 7.4 with 0.1mol / L NaOH solution;

[0045] In this example, the deposit number of Bacillus subtilis is CGMCC 1.3358;

[0046]S2. Centrifuge the NA slant medium in S1 at 4000r / min for 10min and discard the supernatant, wash the obtained cells with pH 7.0 phosphate buffer solution for 3 times, and dissolve the cells in pH 7.0 phosphate buffer solution to prepare Bacteria suspension, in which the concentration of bacteria is 1×10 8 a / mL;

[0047] S3. Apply 5 mL of the bacterial suspension prepared in S2 evenly to the bottom of the petri dish for microwave treatment, wherein the diameter of ...

Embodiment 3

[0058] S1. In a sterile environment, use an inoculation loop to spread the preserved Bacillus subtilis on the NA slant medium, and culture at a constant temperature of 32° C. for 48 hours to obtain activated Bacillus subtilis;

[0059] The NA slant medium is beef extract peptone medium, which consists of: beef extract 3g / L, peptone 7g / L, agar 13g / L, glucose 21g / L, and the pH is adjusted to 7.3 with 0.1mol / L NaOH solution;

[0060] In this example, the deposit number of Bacillus subtilis is CGMCC 1.3358;

[0061] S2. Centrifuge the NA slant medium in S1 at 3500r / min for 10min to discard the supernatant, wash the obtained cells twice with pH7.0 phosphate buffer solution, and dissolve the cells in pH7.0 phosphate buffer solution to prepare Bacteria suspension, in which the concentration of bacteria is 1×10 8 a / mL;

[0062] S3. Apply 5 mL of the bacterial suspension prepared in S2 evenly to the bottom of the petri dish for microwave treatment, wherein the diameter of the petri d...

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Abstract

The invention discloses a glutamine transaminase separation and purification method which comprises the following steps: coating preserved Bacillus subtilis in a NA slant culture medium, culturing, centrifugating, discarding the supernate, washing the obtained bacterium with a pH-7.0 phosphoric acid buffer solution, and preparing a bacterium suspension; carrying out microwave treatment on the bacterium suspension, inoculating in a seed culture medium, inoculating the bacterium into a fermentation culture medium according to the inoculum size of 5%, culturing for some time, carrying out ultrasonic treatment, putting in a water bath shaking table, and culturing for 48 hours to obtain a fermentation culture fluid; and centrifugating the fermentation culture fluid, filtering through a microfiltration membrane, collecting the enzyme solution, and carrying out freeze-drying to obtain the glutamine transaminase. The phytase is added into the soybean protein extracting solution to cut off the phytic acid connecteed between the soybean 7S globulin and soybean 11S globulin; the whole separation process is carried out under normal temperature conditions, thereby avoiding the problem of longer globulin separation process time caused by the addition of the reducer; and the separated soybean 11S globulin and soybean 7S globulin have higher yield and purity.

Description

technical field [0001] The invention belongs to the technical field of fermentation, in particular to a method for separating and purifying transglutaminase. Background technique [0002] Transglutaminase, also known as transglutaminase (TGase), is a monomeric protein with an active center and a molecular weight of about 38,000 composed of 331 amino groups, which can catalyze intramolecular and intermolecular covalent crosslinking of protein polypeptides , so as to improve the structure and function of protein, and have significant effects on the properties of protein such as: foaming, emulsifying, emulsifying stability, thermal stability, water retention and gel ability, etc., thereby improving the flavor, taste, texture and quality of food Appearance etc. Traditional meat processing processes often add large amounts of salt and phosphoric acid to improve water holding capacity, consistency and texture. Recently, foods with less salt and less phosphoric acid have been wid...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12R1/125
CPCC12N9/1044C12Y203/02013
Inventor 魏志刚魏巍赵彦杰
Owner ANHUI ZHENGWEIQI SEASONING FOOD LIMITED
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