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Immobilized glutamine transaminase preparation method

A technology of glutamine and transaminase, applied in biochemical equipment and methods, acyltransferase, transferase and other directions, can solve problems such as easy loss of enzyme activity, and achieve the effects of low price, improved stability and reduced production cost

Active Publication Date: 2019-10-25
TAIXING DONGSHENG FOOD TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention overcomes the above problems existing in the prior art. The present invention innovatively proposes to combine the protective agent formula of the enzyme preparation with the immobilization of the enzyme to solve the problem that the enzyme activity is easily lost at room temperature and relatively high temperature, so that the prepared The stability of immobilized transglutaminase is greatly improved compared with the original original enzyme

Method used

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  • Immobilized glutamine transaminase preparation method
  • Immobilized glutamine transaminase preparation method
  • Immobilized glutamine transaminase preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1) Weigh a certain amount of transglutaminase proenzyme, add an appropriate amount of pure water to dissolve, and prepare an enzyme solution with an enzyme activity of 600U / mL, the measured pH=6.26, and the water activity is 0.97, and then sterilized for later use.

[0039] 2) Prepare natural high molecular compounds such as dextran gel into a suspension, heat and melt to sterilize.

[0040] 3) Cool the sterilized carrier suspension to room temperature, mix it with 100 U / mL free glutamyl transaminase in a certain proportion, and place it in a refrigerator at 4°C for 30 minutes.

[0041] 4) Wash twice with deionized water, add 1% glutaraldehyde solution to enhance immobilization and cross-linking for 1 hour, remove glutaraldehyde, and wash twice with deionized water.

[0042] 5) Add the corresponding formula to the immobilization system according to the following table 2 to prepare immobilized transglutaminase.

[0043] Table 2 Immobilized transglutaminase formula

[0...

Embodiment 2

[0047] 1) Weigh a certain amount of transglutaminase proenzyme, add an appropriate amount of pure water to dissolve, and prepare an enzyme solution with an enzyme activity of 600U / mL, the measured pH=6.26, and the water activity is 0.97, and then sterilized for later use.

[0048] 2) Prepare natural high molecular compounds such as agarose gel into a suspension, heat and melt to sterilize.

[0049] 3) Cool the sterilized carrier suspension to room temperature, mix it with 100 U / mL free glutamyl transaminase in a certain proportion, and place it in a refrigerator at 4°C for 30 minutes.

[0050] 4) Wash twice with deionized water, add 1% glutaraldehyde solution to enhance immobilization and cross-linking for 1 hour, remove glutaraldehyde, and wash twice with deionized water.

[0051] 5) Add the corresponding formula to the immobilization system according to Table 3 to prepare immobilized transglutaminase.

[0052] Table 3 Immobilized transglutaminase formula

[0053]

[005...

Embodiment 3

[0056] 1) Weigh a certain amount of transglutaminase proenzyme, add an appropriate amount of pure water to dissolve, and prepare an enzyme solution with an enzyme activity of 600U / mL, the measured pH=6.26, and the water activity is 0.97, and then sterilized for later use.

[0057] 2) Prepare natural high molecular compounds such as agarose gel into a suspension, heat and melt to sterilize.

[0058] 3) Cool the sterilized carrier suspension to room temperature, mix it with 100 U / mL free glutamyl transaminase in a certain proportion, and place it in a refrigerator at 4°C for 30 minutes.

[0059] 4) Wash twice with deionized water, add 1% glutaraldehyde solution to enhance immobilization and cross-linking for 1 hour, remove glutaraldehyde, and wash twice with deionized water.

[0060] 5) Add the corresponding formula to the immobilization system according to Table 4 to prepare immobilized transglutaminase.

[0061] Table 4 immobilized transglutaminase formula

[0062]

[006...

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Abstract

The invention discloses an immobilized glutamine transaminase preparation method. The method includes that natural macromolecular compounds such as sephadex, agarose gel, carrageenan and gelatin are adopted for enzyme immobilization, an immobilized embedded substance is immersed into a liquid mixture of alcohols, carbohydrates, an antioxidant and the like, and accordingly stability of an immobilized enzyme preparation is greatly improved. The immobilized glutamine transaminase preparation method has advantages that preservation time of glutamine transaminase at a low temperature, the normal temperature and a high temperature is greatly prolonged, and stability of glutamine transaminase is remarkably improved.

Description

technical field [0001] The invention relates to the technical field of immobilization of biological enzyme preparations, in particular to a preparation method of immobilized transglutaminase. Background technique [0002] Transglutaminase, a transferase that catalyzes the acyl transfer reaction, can promote protein intramolecular cross-linking, protein intermolecular cross-linking and cross-linking between proteins and amino acids, which can greatly improve protein The functional properties of protein can improve the nutritional value of protein, and it has a wide range of applications in the food processing industry. Transglutaminase was first discovered in the liver of guinea pigs. It was first isolated from Streptomyces maoyuan in 1989. It was produced on a large scale in 1997. It was officially listed as a food additive in my country in 2001. Transglutaminase widely exists in nature, and it can be divided into three categories according to the source: animal-derived tra...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/02C12N9/96C12N9/10
CPCC12N9/1044C12N9/96C12N11/02C12N11/10C12Y203/02013
Inventor 步国建甘李张玉如寇雪蕊和超楠刘艳贺晓洁
Owner TAIXING DONGSHENG FOOD TECH
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