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Method for immobilizing phospholipase A2 by using sodium alginate-chitosan

A technology for immobilizing phospholipase and sodium alginate, applied in the direction of being fixed on/in an organic carrier, can solve the problems of difficult recovery, easy deactivation and denaturation, etc., and achieves good mechanical strength, little loss of enzyme activity, and high reaction efficiency. mild conditions

Inactive Publication Date: 2012-11-28
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the problem of free phospholipase A in the process of enzymatic degumming 2 Easily inactivated, denatured and difficult to recover, but now phospholipase A 2 The immobilization of phospholipase A with sodium alginate-chitosan was proposed to immobilize phospholipase A 2 Methods

Method used

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Effect test

specific Embodiment approach 1

[0007] Specific embodiment one: use sodium alginate-chitosan to immobilize phospholipase A 2 The method realizes through the following steps: one, immobilized phospholipase A 2 Preparation: Weigh a certain amount of sodium alginate and dissolve it in a water bath at 40°C. At the same time, weigh a certain amount of chitosan and dissolve it in a 5% acetic acid solution in a water bath at 40°C. A certain amount of CaCl was added to the acetic acid solution 2 solution, mix thoroughly. Use a pipette gun to accurately pipette 1ml of the 10-fold diluted enzyme solution into a certain concentration of 10mL sodium alginate solution, and stir evenly. After standing still for a period of time, inhale the above mixed solution with a sterilized 10mL syringe, and inject a certain concentration of 10mL chitosan acetic acid solution, CaCl 2 In the mixed solution, stir at a speed of 180r / min to form smooth microcapsule balls, then add a certain concentration of glutaraldehyde, stir rapidly...

specific Embodiment approach 2

[0008] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that in step 4, phospholipase A will be carried out under the condition that sodium alginate with a concentration of 1.5% to 2.5% is dissolved in a 40°C water bath. 2 immobilization. Other steps are the same as in the first embodiment.

specific Embodiment approach 3

[0009] Specific embodiment three: the difference between this embodiment and specific embodiment one is that in step four, the chitosan with a concentration of 1.5% to 2.5% is subjected to phospholipid under the condition that 5% acetic acid solution is insulated and dissolved in a water bath at 40°C. Enzyme A 2 immobilization. Other steps are the same as in the first embodiment.

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Abstract

The invention provides a method for immobilizing phospholipase A2 by using a sodium alginate-chitosan composite carrier as material through a physical embedding method. According to the invention, the influences of different immobilization conditions such as sodium alginate concentration, chitosan concentration, calcium ion concentration, glutaraldehyde concentration, cross-linking time and the like on the immobilization efficiency and catalysis effect of the phospholipase A2 are researched; and a response surface analysis method is used for optimizing the researched factors. Through immobilizing an SEM (Scanning Electron Microscope) image of the phospholipase A2, the advantage of fixing the phospholipase A2 by using sodium alginate-chitosan as a carrier is further explained. According to the embedding characteristic of the composite carrier in the invention, the phospholipase A2 is immobilized, so that the activity and stability of enzyme in a reaction system are improved, the activity and selectivity of the enzyme are adjusted and controlled and then benefit is brought to the recovery of enzyme and the production of products; and the activity recovery rate of the finally obtained immobilized enzyme is 74.8% and important guiding significance is provided to the enzymatic degumming process in oil and fat refining.

Description

Technical field: [0001] The present invention relates to a kind of fixed phospholipase A 2 Methods. Background technique: [0002] Phospholipase A 2 (PLA 2 ) is a class of enzymes that can selectively break the ester bond at the Sn-2 site of the phospholipid glycerol part, which can hydrolyze phospholipids into a series of biologically active media; due to PLA 2 The enzyme source comes from porcine pancreas, which is relatively in short supply and has not been popularized. The German Lurgi company was the first to use enzymatic degumming in industrial production. There are also many related reports in China. The enzyme used for degumming today is mainly Lecitase 10L (PLA 2 ), Lecitase Novo (PLA 1 ) and Lecitase Ultra (PLA 1 ). Among them, the phospholipase Lecitase10L derived from porcine pancreas is no longer used for oil degumming, but is replaced by the more dominant microorganism Rohalase MPL (PLA 2 ) instead. [0003] However, free Rohalase MPL (PLA 2 ) is exp...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/04
Inventor 于殿宇江连洲胡立志李宝昌张佳宇邹小雨刘鑫李万振宋鹏王玥宋云花张春艳
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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