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Method for flexibly immobilizing papain by using modified polyacrylonitrile resin

A technology of polyacrylonitrile resin and papain, which is applied in the direction of being immobilized on/in an organic carrier, can solve the problems of low recovery rate of immobilized enzyme activity, and achieve the effects of easy separation, good stability and easy preparation

Inactive Publication Date: 2014-09-10
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The technical problem to be solved in the present invention is to provide a method for preparing immobilized papain using a flexible carrier, and the method of the present invention can solve the problem of low recovery rate of immobilized enzyme activity in the covalent bonding method

Method used

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  • Method for flexibly immobilizing papain by using modified polyacrylonitrile resin
  • Method for flexibly immobilizing papain by using modified polyacrylonitrile resin
  • Method for flexibly immobilizing papain by using modified polyacrylonitrile resin

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Embodiment 1

[0062] Embodiment 1, a kind of method for the flexible immobilization papain of modified polyacrylonitrile resin, carries out following steps successively:

[0063] 1), prepare flexible carrier, carry out the following steps in sequence:

[0064] ①, polyacrylonitrile resin modification:

[0065] Accurately weigh 1.00g of polyacrylonitrile resin microspheres (PAN) into a 500mL three-neck bottle, then add 300mL of reaction solvent (ethylene glycol --- ED), soak and swell at room temperature (20-30°C) for 24 hours, Add 11.89g of tetraethylenepentamine (TEPA), and a little catalyst---NaOH (0.12g), and turn on nitrogen, stir at room temperature with a stirring speed of 200rpm for 2h, then rapidly rise to 80°C, magnetic stirring ( 200rpm) for 10 hours, and filtered to obtain modified polyacrylonitrile resin microspheres.

[0066] ②. Soak the modified polyacrylonitrile resin microspheres with 50ml of reaction solvent (ethylene glycol—ED) for 3 hours, then wash with reaction solvent...

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Abstract

The invention discloses a method for flexibly immobilizing papain by using modified polyacrylonitrile resin, which comprises the following steps: 1) preparation of a flexible carrier: modifying polyacrylonitrile resin microspheres by use of tetraethylenepentamine to obtain a modified polyacrylonitrile flexible carrier; 2) activation of the carrier: soaking the modified polyacrylonitrile flexible carrier in a phosphate buffer solution, and performing crosslinking reaction by using glutaraldehyde to obtain a glutaraldehyde modified polyacrylonitrile flexible carrier; 3) preparation of immobilized papain: performing immobilization reaction between the glutaraldehyde modified polyacrylonitrile flexible carrier and papain to obtain immobilized papain. The method disclosed by the invention overcomes the enzyme activity loss caused by rigid collision in the enzyme immobilization process of the carrier synthesized by the prior art, and can maintain the homogeneous catalysis activity of free enzymes to the greatest degree so as to solve the problem of low recovery rate of the immobilized enzyme activity of a covalent binding method.

Description

technical field [0001] The invention relates to a method for preparing an immobilized enzyme, in particular to a method for flexibly immobilizing papain by using a modified polyacrylonitrile resin, and belongs to the technical field of enzyme immobilization. Background technique [0002] As one of the important members of biocatalysts, papain is widely used in the fields of food, daily chemical, and medicine. Although papain has excellent catalytic performance, when free enzyme is directly used as biocatalysis, not only the recovery rate is low, the operation stability is poor, and it cannot be reused, which causes the waste of enzyme, increases the cost, and makes the application subject to certain restrictions. Enzyme immobilization technology can better solve the above problems. The core of immobilized papain lies in the rational selection and design of immobilized carriers with excellent performance and simple and effective immobilization methods. However, in the proc...

Claims

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Application Information

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IPC IPC(8): C12N11/08
Inventor 熊春华贾倩姚彩萍周苏果
Owner ZHEJIANG GONGSHANG UNIVERSITY
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