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Method for separating and purifying glutamine transaminase activation proteinase inhibitor

A protease inhibition, glutamine technology, applied in the direction of protease inhibitors, peptide sources, animal/human peptides, etc., can solve the problem of unclear regulation mechanism, and achieve the effect of short cycle, simple separation process and reduced operation time.

Inactive Publication Date: 2011-09-07
JIANGNAN UNIV
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  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

This inhibitor plays a regulatory role in the activation process of TGase, but the specific regulatory mechanism is still unclear

Method used

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  • Method for separating and purifying glutamine transaminase activation proteinase inhibitor
  • Method for separating and purifying glutamine transaminase activation proteinase inhibitor
  • Method for separating and purifying glutamine transaminase activation proteinase inhibitor

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Embodiment Construction

[0018] Using hydrophobic chromatography as the main means to separate and purify cutinase, the specific steps are as follows:

[0019] (1) Fermentation of Streptomyces hygroscopicus to produce fermentation broth containing transglutaminase-activating protease inhibitors Streptomyces hygroscopicus (Streptomyces hygroscopicus) WSH03-13 was used as the starting strain, and the seeds cultivated to the logarithmic growth phase were inserted into the fermentation culture culture medium at 32° C. for 30 h to obtain a fermentation broth containing transglutaminase-activating protease inhibitors.

[0020] (2) Ethanol precipitation

[0021] After centrifuging the fermentation broth to remove bacteria, add ethanol to a final concentration of 70%, and slowly stir to dissolve it completely, and let it stand at 4°C for 10-20min, then refrigerate and centrifuge at 10,000rpm for 15-30min, collect the protein precipitate, and reconstitute the precipitate Dissolve in an appropriate amount of H...

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Abstract

A separation and purification method of the inhibitor gene for the transglutaminase activated protease belongs to the separation and purification technological field of the biological products. The invention uses the hygroscopicus streptomyces as the starting bacteria; the inhibitor gene for the transglutaminase activated protease can be prepared after the submerged fermentation of the liquid; after the deposition of the ethanol and the replacement of the cation for the chromatography and hydrophobic chromatography, the inhibitor gene solution for the transglutaminase activated protease can be made and used in the following protein sequencing and property study. The steps of the method are simple; the purity of the enzyme is high; and the method is a simple and effective separation and purification method of the enzyme.

Description

technical field [0001] The invention discloses a method for separating and purifying a transglutaminase-activating protease inhibitor, which belongs to the technical field of separation and purification of biological products. The invention relates to a method for separating and purifying transglutaminase-activating protease inhibitors using cation exchange chromatography and hydrophobic chromatography as main technical means. Background technique [0002] Transglutaminase (TGase for short) is a transferase that catalyzes the acyl transfer reaction, which can catalyze intramolecular and intermolecular crosslinking of proteins, connections between proteins and amino acids, and glutamine groups in protein molecules. The hydrolysis reaction can directly change the structure and functional properties of the protein itself and the cells and tissues to which the protein is attached, improve the nutritional value of the protein, and produce a new type of protein food that meets peo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/81C07K1/36C07K1/16
Inventor 陈坚吴敬堵国成张东旭
Owner JIANGNAN UNIV
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