A method for immobilizing enzymes on nanofibers

Abstract

The invention relates to an enzyme immobilization method, which comprises the following steps: preparing a first mixed solution comprising polysulfone (PSF), polyvinyl alcohol (PVA) and an enzyme: dissolving PSF powder in a solvent, then adding PVA powder and stirring to dissolve , and then mixed with the enzyme solution obtained by dissolving the enzyme in water; nanofibers are prepared from the above-mentioned first mixed solution: the first mixed solution is injected into the capillary, and the solution is extruded through a micro-extrusion pump, and formed under the action of high voltage Electrospinning, collecting nanofibers by rotating collector, vacuum drying the collected nanofibers, and storing; performing glutaraldehyde cross-linking: using hydrochloric acid as a catalyst, preparing a second mixed solution containing hydrochloric acid and glutaraldehyde , the nanofibers prepared above are placed in a steam environment above the second mixed solution, and cross-linked at room temperature; the cross-linked nanofibers are washed with distilled water, and then a desiccant is added for storage.

Description

technical field

[0001] The present application belongs to the field of biochemical industry, and specifically relates to a method for immobilizing enzymes on nanofibers. Background technique

[0002] In the field of biochemical engineering, methods that need to immobilize enzymes are often involved, so that the enzymes can be reused and the cost of separation and purification following the reaction can be reduced. The patent documents using nanofibers as immobilized enzyme materials are, for example:

[0003] Chinese patent document CN1948474A (Application No. 200610154566.5) discloses a preparation method of chitosan nanofiber composite film immobilized enzyme, the method is to dissolve chitosan and polyvinyl alcohol in a mixed solvent of glacial acetic acid and water to form a transparent solution , the chitosan nanofiber composite membrane is made by electrospinning, and it is used as the carrier for enzyme immobilization, and the enzyme is immobilized on the nanofiber co...

Examples

Embodiment 1

[0095] (1) Dissolve PSF powder in DMAC (100 mM, pH 6), at 85 °C, dissolve PSF in the solvent at a mass concentration of 75 g / L PSF while stirring, at 90 °C, according to the mass concentration ratio PSF:PVA=2:1 PVA was added, and the PVA was dissolved in the solution with stirring. The solution was cooled at room temperature and then mixed with the CGTase solution to obtain a PSF / PVA mixed solution with a final enzyme concentration of 1 mg / ml.

[0096]The enzyme was purchased from Japan Amano Company, the enzyme activity preservation rate was 99%, the effective substance content was 99%, white crystal, and the solubility in water increased with the increase of temperature. Not hygroscopic, but easy to form various stable hydrates.

[0097] (2) inject the PSF / PVA solution prepared in step (1) into the syringe respectively, use the flattened needle as the capillary for spinning, fix the syringe on the micro-extrusion pump, and control the solution flow rate to 0.2ml / h. The pos...

Embodiment 2

[0100] (1) Dissolve PSF powder in DMAC (100 mM, pH 6), at 85 °C, dissolve PSF in the solvent at a mass concentration of 75 g / L PSF while stirring, at 90 °C, according to the mass concentration ratio PSF:PVA=2:1 PVA was added, and the PVA was dissolved in the solution with stirring. The solution was cooled at room temperature and then mixed with the CGTase solution to obtain a PSF / PVA mixed solution with a final enzyme concentration of 2 mg / ml.

[0101] The enzyme was purchased from Japan Amano Company, the enzyme activity preservation rate was 99%, the effective substance content was 99%, white crystal, and the solubility in water increased with the increase of temperature. Not hygroscopic, but easy to form various stable hydrates.

[0102] (2) inject the PSF / PVA solution prepared in step (1) into the syringe respectively, use the flattened needle as the capillary for spinning, fix the syringe on the micro-extrusion pump, and control the solution flow rate to 0.2ml / h. The po...

Embodiment 3

[0105] (1) Dissolve PSF powder in DMAC (100 mM, pH 6), at 85 °C, dissolve PSF in the solvent at a mass concentration of 75 g / L PSF while stirring, at 90 °C, according to the mass concentration ratio PSF:PVA=2:1 PVA was added, and the PVA was dissolved in the solution with stirring. The solution was cooled at room temperature and then mixed with the CGTase solution to obtain a PSF / PVA mixed solution with a final enzyme concentration of 3 mg / ml.

[0106] The enzyme was purchased from Japan Amano Company, the enzyme activity preservation rate was 99%, the effective substance content was 99%, white crystal, and the solubility in water increased with the increase of temperature. Not hygroscopic, but easy to form various stable hydrates.

[0107] (2) inject the PSF / PVA solution prepared in step (1) into the syringe respectively, use the flattened needle as the capillary for spinning, fix the syringe on the micro-extrusion pump, and control the solution flow rate to 0.2ml / h. The po...

technical field

[0001] The present application belongs to the field of biochemical industry, and specifically relates to a method for immobilizing enzymes on nanofibers. Background technique

[0002] In the field of biochemical engineering, methods that need to immobilize enzymes are often involved, so that the enzymes can be reused and the cost of separation and purification following the reaction can be reduced. The patent documents using nanofibers as immobilized enzyme materials are, for example:

[0003] Chinese patent document CN1948474A (Application No. 200610154566.5) discloses a preparation method of chitosan nanofiber composite film immobilized enzyme, the method is to dissolve chitosan and polyvinyl alcohol in a mixed solvent of glacial acetic acid and water to form a transparent solution , the chitosan nanofiber composite membrane is made by electrospinning, and it is used as the carrier for enzyme immobilization, and the enzyme is immobilized on the nanofiber co...

Examples

Embodiment 1

[0095] (1) Dissolve PSF powder in DMAC (100 mM, pH 6), at 85 °C, dissolve PSF in the solvent at a mass concentration of 75 g / L PSF while stirring, at 90 °C, according to the mass concentration ratio PSF:PVA=2:1 PVA was added, and the PVA was dissolved in the solution with stirring. The solution was cooled at room temperature and then mixed with the CGTase solution to obtain a PSF / PVA mixed solution with a final enzyme concentration of 1 mg / ml.

[0096]The enzyme was purchased from Japan Amano Company, the enzyme activity preservation rate was 99%, the effective substance content was 99%, white crystal, and the solubility in water increased with the increase of temperature. Not hygroscopic, but easy to form various stable hydrates.

[0097] (2) inject the PSF / PVA solution prepared in step (1) into the syringe respectively, use the flattened needle as the capillary for spinning, fix the syringe on the micro-extrusion pump, and control the solution flow rate to 0.2ml / h. The pos...

Embodiment 2

[0100] (1) Dissolve PSF powder in DMAC (100 mM, pH 6), at 85 °C, dissolve PSF in the solvent at a mass concentration of 75 g / L PSF while stirring, at 90 °C, according to the mass concentration ratio PSF:PVA=2:1 PVA was added, and the PVA was dissolved in the solution with stirring. The solution was cooled at room temperature and then mixed with the CGTase solution to obtain a PSF / PVA mixed solution with a final enzyme concentration of 2 mg / ml.

[0101] The enzyme was purchased from Japan Amano Company, the enzyme activity preservation rate was 99%, the effective substance content was 99%, white crystal, and the solubility in water increased with the increase of temperature. Not hygroscopic, but easy to form various stable hydrates.

[0102] (2) inject the PSF / PVA solution prepared in step (1) into the syringe respectively, use the flattened needle as the capillary for spinning, fix the syringe on the micro-extrusion pump, and control the solution flow rate to 0.2ml / h. The po...

Embodiment 3

[0105] (1) Dissolve PSF powder in DMAC (100 mM, pH 6), at 85 °C, dissolve PSF in the solvent at a mass concentration of 75 g / L PSF while stirring, at 90 °C, according to the mass concentration ratio PSF:PVA=2:1 PVA was added, and the PVA was dissolved in the solution with stirring. The solution was cooled at room temperature and then mixed with the CGTase solution to obtain a PSF / PVA mixed solution with a final enzyme concentration of 3 mg / ml.

[0106] The enzyme was purchased from Japan Amano Company, the enzyme activity preservation rate was 99%, the effective substance content was 99%, white crystal, and the solubility in water increased with the increase of temperature. Not hygroscopic, but easy to form various stable hydrates.

[0107] (2) inject the PSF / PVA solution prepared in step (1) into the syringe respectively, use the flattened needle as the capillary for spinning, fix the syringe on the micro-extrusion pump, and control the solution flow rate to 0.2ml / h. The po...

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