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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, can solve the problems of high price, scarce source, complicated extraction process, etc. The effect of avoiding the loss of MTG enzyme activity

Inactive Publication Date: 2006-01-25
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 80U / g and a protease activity of 3000U / g. Casein was used as the substrate to carry out the applied experiment, and 8mmol sodium hexametaphosphate / g enzyme powder (0.0027mmol sodium hexametaphosphate / U protease) was added 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 fermented MTG solid enzyme preparation 2 has an MTG enzyme activity of 40 U / g and a protease activity of 80 U / g. Casein was used as the substrate for the applied experiment, and 0.8mmol sodium hexametaphosphate / g enzyme powder (0.01mmol sodium hexametaphosphate / U protease) was added to the enzyme solution, the protease in it was completely inhibited, and the MTG enzyme activity was not affected , electrophoresis experiments confirmed the formation of cross-linked proteins, and the formation of an invertible stronger gel.

Embodiment 3

[0012] For the MTG fermentation broth obtained by fermentation, during the extraction process, sodium hexametaphosphate is added before ultrafiltration or drying, and a metal ion chelating agent is added before ultrafiltration, so that the metal ions in the fermentation broth are chelated. Remove; or add a metal ion chelating agent before drying, so that the metal ions in the enzyme preparation are chelated. The addition 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 at the same time. Casein was used as a substrate for applied experiments, and electrophoresis experiments confirmed the formation of cross-linked proteins, and soon formed an invertible and strong gel.

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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 protease in an enzyme preparation prepared by fermentation of transglutaminase, belonging to the technical field of enzyme preparation. Background technique [0002] Transglutaminase EC 2.3.2.13, full name R-glutaminyl-peptide: aminee γ-glutamyltransferase, TG for short, is a transferase that catalyzes acyl transfer reaction. It can catalyze the formation of ε-(γ-glutamyl)lysine covalent bond between the γ-carboxamide group on the glutamine residue and the γ-amino group on the lysine residue in most proteins. Transglutaminase can catalyze the intramolecular or intermolecular crosslinking 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 stability of various proteins. Structural and functional properties, known as "21st century super adhesive". Therefore, it is widely used in the preparati...

Claims

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Application Information

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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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