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Method for continuously extracting glutamine transaminase from fermentation liquor

A technology of glutamine and fermented liquid, applied in the field of bioengineering, can solve the problems of large loss of enzyme activity, high equipment requirements, complicated operation, etc., and achieve the effect of small enzyme activity loss, small personnel requirements, and simple equipment

Active Publication Date: 2014-08-06
长握生物科技(江苏)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large loss of enzyme activity, the yield is not high, and the requirements for equipment are high, and the operation is complicated
Moreover, the separation process of the above-mentioned method is not continuous, the labor required for the operation is large, and the operation cycle is long, resulting in low enzyme activity of the final product obtained, so the above-mentioned methods are not suitable for large-scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The fermented liquid of the present embodiment obtains by following method:

[0033] The fermentation strain used in this example is Streptomyces hygroscopicus NYU-70 (CCTCC M 2014121). The strains were enriched in the seed medium and then inoculated into the fermentation medium with 1 / 3 of the inoculation amount. The fermenter was set at a rotation speed of 220rpm, a temperature of 30°C, a pH of 7.2, and a fermentation period of 64 hours.

[0034] At the end of the fermentation, the enzyme activity in the transglutaminase fermentation broth was 15.9U / ml, and the fermentation broth was introduced into the ceramic composite membrane equipment at a flow rate of 80L / h to remove bacteria and impurities. The membrane separation accuracy of the ceramic composite membrane equipment was 50nm. The separation liquid separated by the ceramic composite membrane is introduced into the membrane separation equipment at a flow rate of 80L / h, and then concentrated by ultrafiltration. T...

Embodiment 2

[0039] The fermented liquid of the present embodiment is obtained by the same method as in Example 1.

[0040]At the end of the fermentation, the enzyme activity in the transglutaminase fermentation broth was 15.9U / ml, and the fermentation broth was introduced into the ceramic composite membrane equipment at a flow rate of 80L / h to remove bacteria and impurities. The membrane separation accuracy of the ceramic composite membrane equipment was 50nm. The separation liquid separated by the ceramic composite membrane is introduced into the membrane separation equipment at a flow rate of 80L / h, and then concentrated by ultrafiltration. Add 1% soy protein (w / w) to the ultrafiltration concentrate obtained by ultrafiltration, stir with electric stirring at a speed of 120rpm for 5min, then continue stirring at a speed of 80rpm and add 1.5 times (V / V ) in the concentrated liquid volume of solvent precipitation for 4h, temperature 6 ± 2 ℃. The mixed solution after solvent precipitation ...

Embodiment 3

[0042] The fermented liquid of the present embodiment is obtained by the same method as in Example 1.

[0043] At the end of the fermentation, the enzyme activity in the transglutaminase fermentation broth was 15.9U / ml, and the fermentation broth was introduced into the ceramic composite membrane equipment at a flow rate of 80L / h to remove bacteria and impurities. The membrane separation accuracy of the ceramic composite membrane equipment was 50nm. The separation liquid separated by the ceramic composite membrane is introduced into the membrane separation equipment at a flow rate of 80L / h, and then concentrated by ultrafiltration. Add 1% β-cyclodextrin (w / w) to the ultrafiltration concentrate obtained by ultrafiltration, and stir at a speed of 120rpm for 5 minutes with electric stirring, then continue stirring at a speed of 80rpm and add 1.5 times ( V / V) Solvent precipitation in the volume of the concentrate for 4 hours at a temperature of 6±2°C. The mixed solution after sol...

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Abstract

The invention discloses a method for continuously extracting glutamine transaminase from fermentation liquor, and belongs to the field of bioengineering. The method comprises the following steps: introducing sterile compressed air into a fermentation tank after fermentation is ended; leading out the fermentation liquor to ceramic composite membrane equipment from the fermentation tank, feeding the fermentation liquor from which thallus and impurities are removed into membrane separation equipment, and carrying out ultra-filtration concentration; connecting all the equipment by a pipeline with a flow control valve; continuously running after setting parameters in absence of people involvement; adding auxiliary materials to the fermentation liquor after concentrating, and then sequentially carrying out solvent participation and centrifugal separation; and adding auxiliary materials again and drying in vacuum, so as to obtain glutamine transaminase powder. The method has the advantages of being reasonable and continuous in process, simple in equipment, simple and convenient to operate, low in cost, and easy to industrially amplify. By adopting the method, the technical problem that the steps such as salt precipitation, dialysis, ion exchange column separation and the like in traditional extraction are not needed, resulting in easy industrial application is solved.

Description

technical field [0001] The invention relates to a method for continuously extracting transglutaminase from fermented liquid, belonging to the field of bioengineering. Background technique [0002] Transglutaminase (protein-glutamic acid-glutaminase, Transglutaminase, referred to as TG, EC2.3.2.13), also known as transglutaminase, is a kind of catalyzing glutamine residue in protein or polypeptide. A transferase for acyl transfer reactions. Because it can catalyze the covalent cross-linking and polymerization of many proteins and improve the functional properties of various proteins, it has broad application prospects and advantages in the food industry, textile industry and pharmaceutical industry, and is considered to be the most important method for the production of new protein foods. One of the enzymes, it has attracted great attention and interest from researchers at home and abroad. The main functions of transglutaminase in food industry processing include: (1) modif...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12R1/55
CPCC12N9/1044C12Y203/02013
Inventor 虞龙李玉燕李市场黄思思刘媛
Owner 长握生物科技(江苏)有限公司
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