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Method for separating and purifying glutamine transaminase activation proteinase inhibitor

A technology of transglutaminase and protease inhibition, applied in the field of separation and purification of biological products, can solve problems such as unclear regulation mechanism, and achieve the effects of short cycle, quick effect and mild action conditions

Inactive Publication Date: 2008-02-13
JIANGNAN UNIV
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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

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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

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 present invention uses Streptomyces hygroscopicus as the starting strain, and the transglutaminase activating protease inhibitory factor prepared by liquid submerged fermentation obtains electrophoretic pure transglutaminase after ethanol precipitation, cation exchange chromatography and hydrophobic chromatography Enzyme-activating protease inhibitor solution for subsequent protein sequencing and characterization studies. The method has simple steps and high enzyme purity, and is a simple and effective enzyme separation and purification method.

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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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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