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A method for continuously extracting transglutaminase from fermentation broth

A technology of glutamine and fermentation broth, applied in the field of bioengineering, can solve problems such as large loss of enzyme activity, high equipment requirements, complicated operation, etc., and achieve the effects of small loss of enzyme activity, small personnel requirements, and simple equipment

Active Publication Date: 2016-09-21
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 fermentation broth of the present embodiment is obtained by the following methods:

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

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

Embodiment 2

[0039] The fermentation broth of this example was 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 15.9 U / ml. The fermentation broth was introduced into the ceramic composite membrane device at a flow rate of 80 L / h to remove bacteria and impurities. The membrane separation accuracy of the ceramic composite membrane device was 50 nm. The separation liquid obtained by the ceramic composite membrane was introduced into the membrane separation equipment at a flow rate of 80L / h, and concentrated by ultrafiltration. The molecular weight cut off of the membrane was 6000, and the temperature of the enzyme liquid was controlled by circulating water to 6±2℃. Add 1% soy protein (w / w) to the ultrafiltration concentrate obtained by ultrafiltration, stir at a speed of 120rpm for 5min with electric stirring, continue stirring at a speed of 80rpm and add 1.5 times (V / V) at a flux of 500mL / mi...

Embodiment 3

[0042] The fermentation broth of this example was 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 15.9 U / ml. The fermentation broth was introduced into the ceramic composite membrane device at a flow rate of 80 L / h to remove bacteria and impurities. The membrane separation accuracy of the ceramic composite membrane device was 50 nm. The separation liquid obtained by the ceramic composite membrane was introduced into the membrane separation equipment at a flow rate of 80L / h, and concentrated by ultrafiltration. The molecular weight cut off of the membrane was 6000, and the temperature of the enzyme liquid was controlled by circulating water to 6±2℃. Add 1% β-cyclodextrin (w / w) to the ultrafiltration concentrate obtained by ultrafiltration, stir with electric stirring for 5 min at a speed of 120 rpm, continue stirring at a speed of 80 rpm and add 1.5 times ( V / V) precipitated...

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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 glutamine transaminase from fermentation broth, and belongs to the field of biological engineering. Background technique [0002] Transglutaminase (protein-glutamate-glutaminyltransferase, Transglutaminase, referred to as TG, EC2.3.2.13), also known as transglutaminase, is a catalyzing protein or polypeptide in glutamine residues acyl transferase for the acyl transfer reaction. Because it can catalyze the covalent cross-linking polymerization of many proteins and improve the functional properties of various proteins, it has a wide range of application prospects and advantages in the food industry, textile industry and pharmaceutical industry, and is considered to be the most important for the production of new protein foods. It is one of the enzyme species, which has attracted great attention and interest of domestic and foreign researchers. The main functions of transglutaminase in food i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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