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Preparation method for polyacrylonitrile immobilized enzyme

A technology of polyacrylonitrile and immobilized enzymes, which is applied to the fields of immobilized on/in organic carriers, biochemical fiber treatment, non-woven fabrics, etc. Recycling and other issues, to achieve the effect of improving the utilization rate of enzymes, high porosity, and high mechanical strength

Inactive Publication Date: 2013-08-21
TONGJI UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

[0005] Aiming at the problems that some carriers used for immobilizing enzymes tend to reduce the catalytic stability of enzymes, the process of immobilizing enzymes is cumbersome, the mechanical strength of immobilized materials is low, and it is difficult to recycle. , the object of the present invention is to provide a kind of preparation method of a kind of polyacrylonitrile nanofiber membrane immobilized enzyme with high specific surface area, high porosity, simple operation of immobilized enzyme process and high mechanical strength of immobilized material

Method used

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  • Preparation method for polyacrylonitrile immobilized enzyme
  • Preparation method for polyacrylonitrile immobilized enzyme

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Experimental program
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Effect test

Embodiment 1

[0020] The preparation method of the polyacrylonitrile nanofiber membrane is as follows: dissolving polyacrylonitrile in N,N-dimethylformamide solvent to form a solution with a weight percentage of 5% to 10%. Then, inject the prepared solution into the electrospinning device, and perform electrospinning under the conditions of a voltage of 12-20 kV, a flow rate of the spinneret solution of 0.8-1.5 ml / hour, and a receiving distance of 10-20 cm. A fiber membrane with a fiber diameter of 100-600 nanometers is obtained. The obtained fiber membrane is placed in a vacuum drying oven and dried under the condition of 30° C. to 50° C. for 5 hours to 12 hours.

[0021] Immerse 1 gram of polyacrylonitrile nanofiber membrane in pure ethanol solution, and pass through hydrogen chloride gas for 10 minutes. The nanofibrous membrane was taken out and fully rinsed with a phosphate buffer solution with a pH of 7.0 to remove residual ethanol on the membrane. Then, immerse the activated nanofib...

Embodiment 2

[0024] Immerse 1 gram of polyacrylonitrile nanofiber membrane in pure ethanol solution, and pass through hydrogen chloride gas for 20 minutes. The nanofibrous membrane was taken out and fully rinsed with a phosphate buffer solution with a pH of 7.0 to remove residual ethanol on the membrane. Then, immerse the activated nanofibrous membrane in a 1.0 mg / ml laccase phosphate buffer solution, seal it and vibrate for 3 hours at 30° C. with a rotation speed of 80 rpm. The membrane was taken out from the solution and washed repeatedly with deionized water for 5 times to obtain the polyacrylonitrile nanofiber membrane immobilized laccase. The nanofibrous membrane immobilized with laccase was immersed in a phosphate buffer solution with a pH of 5.0, and stored at 4°C for use.

Embodiment 3

[0026] Immerse 1 gram of polyacrylonitrile nanofiber membrane in pure ethanol solution, and pass through hydrogen chloride gas for 15 minutes. The nanofibrous membrane was taken out and fully rinsed with a phosphate buffer solution with a pH of 7.0 to remove residual ethanol on the membrane. Then, immerse the activated nanofibrous membrane in a horseradish peroxidase phosphate buffer solution with a concentration of 1.0 mg / ml, seal it and vibrate for 1.5 hours at 20° C. at a speed of 60 rpm. The membrane is taken out from the solution, washed repeatedly with deionized water for 3 times, and the immobilized horseradish peroxidase and polyacrylonitrile nanofiber membrane can be obtained. The nanofibrous membrane immobilized with horseradish peroxidase was immersed in a phosphate buffer solution with a pH of 5.0, and stored at 4°C for use.

[0027] From figure 2 From the results of the scanning electron microscope, it can be seen that the polyacrylonitrile nanofiber membrane ...

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Abstract

The invention relates to a preparation method for polyacrylonitrile nanofiber membrane immobilized enzyme. The preparation method comprises the following steps of dissolving a polyacrylonitrile polymer into N,N-dimethylformamide and preparing a polyacrylonitrile nanofiber membrane through electrostatic spinning; then soaking the polyacrylonitrile nanofiber membrane into an absolute alcohol solution, aerating hydrogen chloride gas for 10-30 minutes, and fixing enzyme into the nanofiber membrane through chemical bonding. According to the preparation method, high voidage and high mechanical strength of the polyacrylonitrile nanofiber membrane are utilized for preparation of the nanofiber membrane immobilized enzyme which has high enzyme content, is easy to recycle from a reaction system and can be used repeatedly. The immobilized zymophore can be used for increasing the use ratio of enzyme and lowering the production cost, and has the advantage of simple preparation processes, and the like.

Description

technical field [0001] The invention relates to a preparation method for immobilizing enzymes using polyacrylonitrile nanofiber membranes. Background technique [0002] Compared with free enzymes, immobilized enzymes have high storage stability, easy separation and recovery, repeated use, continuous and controllable operation, and simple process while maintaining their high-efficiency, specific and mild enzyme-catalyzed reaction characteristics. features. When the enzyme exists in a free state, some defects such as poor stability and short service life have not been improved. Immobilizing the enzyme on the separation membrane can effectively avoid the above-mentioned shortcomings. In addition, when the enzyme exists in a free state, the enzyme's spatial structure changes or the active site is shielded due to unfavorable adsorption on the surface of the enzyme membrane, which reduces the activity of the enzyme. [0003] There are many materials and methods for immobilizin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08D04H1/43D04H1/728D06M11/11D06M16/00D06M101/28
Inventor 徐冉李风亭迟成龙
Owner TONGJI UNIV
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