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Method for preparing transglutaminase from antarctic krill

A technology of transglutaminase and Antarctic krill, which is applied in the field of transferase, can solve the problems of not finding and preparing transglutaminase, and achieve the effects of fast preparation speed, simple operation and reasonable preparation process

Active Publication Date: 2016-02-24
RONGCHENG GUANGRUN SEAFOOD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on the preparation of transglutaminase from Antarctic krill

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A kind of method utilizing Antarctic krill to prepare transglutaminase, it goes through the following process steps:

[0023] (1) Raw material processing: select Antarctic krill as raw material, wash it with water, chop it, add buffer, send it into a homogenizer, control the speed of 6000r / min, and homogenize for 20 minutes; the homogenized Antarctic krill Send it into a centrifuge, control the speed at 8000r / min, and centrifuge for 10min; remove impurities and collect the supernatant; wherein, the buffer is prepared by dissolving 8mMNaCl and 15mM Tris-HCl in 1000ml deionized water, and the ratio of buffer to Antarctic krill It is 1︰2.5(m / v);

[0024] (2) Filtration: the clear liquid obtained in step (1) is filtered with a cellulose acetate membrane, the upper layer of turbid liquid is removed, and the filtrate is collected;

[0025] (3) Dialysis: the filtrate obtained in step (2) is dialyzed using sodium acetate dialyzate, wherein the sodium acetate dialyzate is disso...

Embodiment 2

[0032] A kind of method utilizing Antarctic krill to prepare transglutaminase, it goes through the following process steps:

[0033] (1) Raw material processing: select Antarctic krill as raw material, wash it with water, chop it, add buffer, send it to a homogenizer, control the speed of 5000r / min, and homogenize for 10 minutes; the homogenized Antarctic krill Send it into a centrifuge, control the speed at 7000r / min, and centrifuge for 12min; remove impurities and collect the supernatant; wherein, the buffer solution is prepared by dissolving 8mMNaCl and 15mM Tris-HCl in 1000ml deionized water, and the ratio of buffer solution to Antarctic krill is 1:3(m / v);

[0034] (2) Filtration: the clear liquid obtained in step (1) is filtered with a cellulose acetate membrane, the upper layer of turbid liquid is removed, and the filtrate is collected;

[0035] (3) Dialysis: the filtrate obtained in step (2) is dialyzed using sodium acetate dialyzate, wherein the sodium acetate dialyza...

Embodiment 3

[0042] A kind of method utilizing Antarctic krill to prepare transglutaminase, it goes through the following process steps:

[0043] (1) Raw material processing: choose Antarctic krill as raw material, wash it with water, chop it, add buffer, send it into a homogenizer, control the speed of 4000r / min, and homogenize for 30 minutes; the homogenized Antarctic krill Send it into a centrifuge, control the speed at 6000r / min, and centrifuge for 15min; remove impurities and collect the supernatant; wherein, the buffer is prepared by dissolving 8mMNaCl and 15mM Tris-HCl in 1000ml deionized water, and the ratio of buffer to Antarctic krill is 1:2(m / v);

[0044](2) Filtration: the clear liquid obtained in step (1) is filtered with a cellulose acetate membrane, the upper layer of turbid liquid is removed, and the filtrate is collected;

[0045] (3) Dialysis: the filtrate obtained in step (2) is dialyzed using sodium acetate dialyzate, wherein the sodium acetate dialyzate is dissolved i...

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PUM

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Abstract

The invention discloses a method for preparing transglutaminase from antarctic krill. According to the method, the cleaned antarctic krill are chopped, a buffer solution is added, the mixture is subjected to homogenate and centrifugation, a clear liquid is taken and filtered by a cellulose acetate membrane, a turbidity solution on the upper layer is removed, a filtrate is dialyzed through a sodium acetate dialysis fluid and subjected to ultrafiltration through ultrafiltration equipment, a filtered fluid is subjected to chromatographic purification through ion exchange resin, the purified fluid is subjected to ultrafiltration, concentration, and freeze drying, and purified transglutaminase is obtained. According to the method, a process is reasonable, the operation is simple, the preparation speed is high, and the purity and the enzyme activity recovery rate of extracted transglutaminase are high.

Description

technical field [0001] The invention relates to transglutaminase, in particular to a method for preparing transglutaminase by using Antarctic krill. Background technique [0002] Transglutaminase is a type of enzyme that widely exists in animal tissues and human body fluids. It can catalyze the covalent cross-linking and polymerization between protein molecules, polypeptides and various primary amines. It is related to blood coagulation, It is related to various biological phenomena such as wound healing, epidermal keratinization, and erythrocyte membrane sclerosis. In recent years, transglutaminase has aroused the interest of many technicians, and it has great application potential in food industry, enzyme immobilization and textile industry. [0003] At present, the extraction methods of transglutaminase mainly include extraction from animal and plant tissues and production by microbial fermentation. Among them, microbial fermentation is the most widely produced, but the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10
CPCC12N9/1044C12Y203/02013
Inventor 杨劲峰吴文惠张依婷
Owner RONGCHENG GUANGRUN SEAFOOD CO LTD
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