Method of inhibiting proteolytic enzyme in enzyme preparation obtained in glutamin transaminase fermentation method

A technology of transglutaminase and enzyme preparation, which is applied in the field of protease in the enzyme preparation of fermentation method for inhibiting transglutaminase, which can solve the problems of scarce sources, high price, and difficult industrial application, so as to avoid MTG Enzyme activity loss, the effect of simple method

Inactive Publication Date: 2007-05-30
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sources of transglutaminase derived from animals and plants are scarce, and the extraction process is complicated, so the price of this enzyme is relatively high, and it is difficult to be used in industrial applications

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] The MTG solid enzyme preparation product 1 obtained by fermentation has an MTG enzyme activity of 80 U / g and a protease enzyme activity of 3000 U / g. Casein was used as a substrate for application experiments. Add 8mmol sodium hexametaphosphate / g enzyme powder (0.0027mmol sodium hexametaphosphate / U protease) to the enzyme solution. The protease in it was completely inhibited, and the MTG enzyme activity was not affected. And electrophoresis experiments confirmed the formation of cross-linked proteins.

Embodiment 2

[0010] The MTG solid enzyme preparation 2 obtained by fermentation has an MTG enzyme activity of 40 U / g and a protease enzyme activity of 80 U / g. Casein is used as the substrate for the applied experiment, 0.8mmol sodium hexametaphosphate / g enzyme powder (0.01mmol sodium hexametaphosphate / U protease) is added to the enzyme solution, all the proteases are inhibited, and the MTG enzyme activity is not affected , Electrophoresis experiments confirmed the formation of cross-linked proteins, and the formation of an invertible gel with greater strength.

Embodiment 3

[0012] In the extraction process of the MTG fermentation broth obtained by fermentation, sodium hexametaphosphate is added before ultrafiltration or drying, and a metal ion chelating agent is added before the ultrafiltration, so that the metal ions in the fermentation broth are chelated. Remove; or add a metal ion chelating agent before drying to chelate the metal ions in the enzyme preparation. The added amount is 0.002-0.01mmol chelating agent / U protease, the activity of protease in the prepared solid enzyme preparation is all inhibited, and the yield of MTG is not affected. Application experiments with casein as a substrate, electrophoresis experiments confirmed the formation of cross-linked proteins, and quickly formed an invertible gel with greater strength.

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Abstract

The invention relates to a method to make protease in enzyme by restrain glutamine aminopherase fermenting method that belongs to enzyme technology field. The invention uses glutamine aminopherase fermenting to make enzyme. And during the manufacturing process, the metal ion inhibitor would be added and the activity of protease would be restrained. The invention is easy to operate and high efficiency.

Description

Technical field [0001] The invention relates to a method for inhibiting a protease in an enzyme preparation prepared by a transglutaminase fermentation method, and belongs to the technical field of enzyme preparations. Background technique [0002] Transglutaminase (Transglutaminase EC 2.3.2.13), the full name is R-glutaminyl-peptide: aminee γ-glutamyltransferase, or TG for short, is a transferase that catalyzes the acyl transfer reaction. It can catalyze the formation of ε-(γ-glutamyl)lysine covalent bonds between the γ-carboxamide group on the glutamine residues of most proteins and the γ-amino group on the lysine residues. Transglutaminase can catalyze the intramolecular or intermolecular cross-linking of most food proteins, the connection between proteins and amino acids, and the hydrolysis of glutamine amide groups in protein molecules, which greatly improves the performance of various proteins. The structural and functional properties are known as the "21st Century Super Ad...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N9/99
Inventor 陈坚刘和崔莉张东旭堵国成
Owner JIANGNAN UNIV
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