Method for preparing conducting polyaniline immobilized enzyme with nanometer structure

A technology of conductive polyaniline and nanostructures, applied in the direction of fixation on/in the organic carrier, can solve the problems of severe, enzyme activity loss, etc., to increase specific surface area, simple method, improve transmission efficiency and mass transfer diffusion speed effect

Inactive Publication Date: 2010-09-22
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, usually the immobilization reaction is relatively violent, and the activity of the enzyme is seriously lost during the immobilization process.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] This embodiment includes the following steps:

[0023] 1. First, adopt the "template-free" method to prepare nanostructured conductive polyaniline, ultrasonically disperse the aniline monomer in the aqueous solution, the ratio of the aniline monomer to the aqueous solution is 5mmol: 20mL, and then add 0.25M (mol / L ) in the oxidizing agent aqueous solution, the oxidizing agent is (NH 2 ) 2 S 2 o 8 , the molar ratio of the oxidizing agent to the aniline monomer in the oxidizing agent aqueous solution is 1:1, let it stand for 12 hours to react, centrifuge the mixed solution, wash the centrifuged solid with distilled water, absolute ethanol and ether in turn, and finally dry it under vacuum at 60°C After 18 hours, dark green conductive polyaniline nanofibers were obtained;

[0024] Two, the prepared conductive polyaniline nanofibers are dispersed in the phosphoric acid buffer solution with a pH range of 7, the ratio of the conductive polyaniline nanofibers and the phosp...

Embodiment 2

[0027] This embodiment includes the following steps:

[0028] 1. First, adopt the "template-free" method to prepare nanostructured conductive polyaniline, ultrasonically disperse the aniline monomer in the aqueous solution, the ratio of the aniline monomer to the aqueous solution is 2mmol: 10mL, and then add 2M (mol / L) at one time In the oxidant aqueous solution, the oxidant is Fe 2 (SO 4 ) 3 , the molar ratio of the oxidizing agent to the aniline monomer in the oxidizing agent aqueous solution is 10:1, let the reaction stand for 20 hours, centrifuge the mixed solution, wash the centrifuged solid with distilled water, absolute ethanol and ether in turn, and finally dry it under vacuum at 50°C In 24 hours, dark green conductive polyaniline nanofibers were obtained;

[0029] Two, the prepared conductive polyaniline nanofibers are dispersed in the phosphoric acid buffer solution with a pH range of 8, the ratio of the conductive polyaniline nanofibers and the phosphoric acid bu...

Embodiment 3

[0031] This embodiment includes the following steps:

[0032] 1. First, the "soft template" method is used to prepare conductive polyaniline with nanostructure, and salicylic acid with surfactant properties is selected as the dopant, and the aniline monomer and the acidic dopant are mixed and dispersed at a molar ratio of 1:1. In the aqueous solution, the ratio of the aniline monomer to the aqueous solution is 2mmol: 20mL, and then, it is added to the oxidant aqueous solution with a concentration of 0.2M (mol / L) at one time, and the oxidant is Fe(NO 3 ) 3 , the molar ratio of the oxidizing agent and aniline monomer in the oxidizing agent aqueous solution is 2:1, let it stand for 16 hours to react, centrifuge the mixed solution, wash the centrifuged solid with distilled water, absolute ethanol and ether in turn, and finally dry it under vacuum at 30°C After 20 hours, dark green conductive polyaniline microspheres were obtained;

[0033] Two, the prepared conductive polyanilin...

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Abstract

The invention provides a method for preparing conducting polyaniline immobilized enzyme with a nanometer structure. The method comprises the following steps of: 1, adding aniline monomers serving as a template, or an acid dopant which contains a surface active agent and serves as a soft template, or polystyrene (PS) serving as a hard template into deionized water in which the aniline monomers are dispersed, and at room temperature, magnetically stirring the mixed solution to obtain uniform emulsion; under an action of electrostatic force, adding aqueous solution of oxidant into the emulsion at one time to obtain the conducting polyaniline with the nanometer structure; and 2, dispersing the obtained conducting polyaniline in buffer solution of phosphoric acid, uniformly stirring the mixed solution, dissolving the free enzyme in the same buffer solution, performing covalent coupling of the mixed solution on a shaking table at the speed of 60 to 120rpm, washing the mixed solution and removing supernatant to obtain the immobilized enzyme having the nanometer structure and using the conducting polyaniline as a carrier. The immobilized enzyme prepared by the method can be applied to the decoloring reaction of dyes, and has the advantages of mild reaction conditions, high degradation efficiency and repeated use.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzyme preparation, and in particular relates to a preparation method of conductive polyaniline immobilized enzyme with nanostructure. Background technique [0002] As a special biocatalyst, enzyme has the advantages of high catalytic efficiency, mild reaction conditions and strong specificity. However, the enzyme in the free state has poor stability, is sensitive to external factors such as temperature, ion concentration, pH value, and heavy metal ions, is easily inactivated, and is difficult to separate after the reaction, cannot be reused, and has high cost of use. These shortcomings greatly limit the use of free enzymes. Applications. Enzyme immobilization technology, which binds free enzymes in a certain area by physical or chemical methods, can be used repeatedly and continuously while maintaining catalytic activity. Immobilized enzyme overcomes the deficiency of free enzyme, improves ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12N11/04
Inventor 文湘华丁杭军
Owner TSINGHUA UNIV
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