Preparation method of chitin nano-fiber composite film immobilized enzyme

Abstract

This invention relates to using chitosan nanometer fabric compound membrane for the immobilized enzyme. It soluts chitosan and polyvinyl alcohol in mixed solvents of acetic acid and water to form a transparent solution, through static electricity spinning to form chitosan nanometer fabric compound membrane, and take it as immobilized enzyme carriers, through chemical bonding in ways to fix enzyme on composite membrane. The invention uses nanometer composite membrane of high specific surface area and high porosity, and chitosan which is the reactive amino biocompatibility and can be set with high-volume, high specific activity, high catalytic efficiency of chitosan nanometer fabric compound membrane immobilized enzyme.

Description

technical field

[0001] The invention relates to a technique for immobilizing enzymes with a chitosan nanofiber composite membrane. Background technique

[0002] Enzyme is a special kind of protein produced by living cells with high-efficiency catalytic function, high specificity and high controllability. It can specifically promote a certain chemical reaction without participating in the reaction itself. The reaction product has the characteristics of less pollution, low energy consumption, and easy control of the reaction, so it has been widely used. However, it is prone to denaturation of the enzyme protein under conditions such as acid, alkali, heat and organic solution, thereby reducing or losing the enzyme activity, and the enzyme reaction is mostly carried out in the solution, which is not easy to recover after the reaction is completed, and the separation and purification of the reaction product is difficult. And it is difficult to achieve continuous and automatic pr...

Examples

example 1

[0021] Immerse 1 gram of chitosan nanofiber composite film in 2% glutaraldehyde aqueous solution, seal it and vibrate for 3 hours at a constant temperature of 25°C, with a shaking speed of 40 rpm; take the nanofiber composite film and use a pH value of 7.0 The phosphate buffer solution was fully washed to remove the residual glutaraldehyde on the membrane; the nanofiber composite membrane activated by glutaraldehyde was immersed in the lipase phosphate buffer solution with a concentration of 2 mg / ml, and sealed at 4 Shake at a constant temperature of ℃ for 2 hours, and the shaking speed is 40 rpm; take out the membrane from the solution, wash it repeatedly with deionized water, and wash it 3 times in total, and then you can get the lipase-immobilized chitosan nanofiber composite; The nanofiber composite membrane with immobilized enzymes was immersed in a phosphate buffer solution with a pH of 7.0, and stored at 4°C for use.

example 2

[0023] Immerse 1 gram of chitosan nanofiber composite membrane in 10% glutaraldehyde aqueous solution, shake it under the constant temperature condition of 25 ℃ for 5 hours after sealing, and the shaking speed is 60 rpm; take the nanofiber composite membrane and use pH 7.0 The phosphate buffer solution was fully washed to remove the residual glutaraldehyde on the membrane; the nanofiber composite membrane activated by glutaraldehyde was immersed in the lipase phosphate buffer solution with a concentration of 6 mg / ml, and sealed at 4 Shake at a constant temperature of ℃ for 4 hours, and the shaking speed is 60 rpm; take out the membrane from the solution, wash it repeatedly with deionized water, and wash it 4 times in total, and then you can get the lipase-immobilized chitosan nanofiber composite; The nanofiber composite membrane with immobilized enzymes was immersed in a phosphate buffer solution with a pH of 7.0, and stored at 4°C for use.

example 3

[0025] Immerse 1 gram of chitosan nanofiber composite film in 5% glutaraldehyde aqueous solution, seal it and vibrate for 8 hours at a constant temperature of 25°C at a speed of 80 rpm; take the nanofiber composite film and use a pH value of 7.0 The phosphate buffer solution was fully washed to remove the residual glutaraldehyde on the membrane; the nanofiber composite membrane activated by glutaraldehyde was immersed in a catalase phosphate buffer solution with a concentration of 8 mg / ml, and after sealing Shake at a constant temperature of 4°C for 6 hours at a shaking speed of 80 rpm; take out the membrane from the solution and wash it repeatedly with deionized water for a total of 5 times to obtain catalase-immobilized chitosan Nanofiber composite: The nanofiber composite membrane with immobilized enzyme was immersed in a phosphate buffer solution with a pH of 7.0, and stored at 4°C for use.

technical field

[0001] The invention relates to a technique for immobilizing enzymes with a chitosan nanofiber composite membrane. Background technique

[0002] Enzyme is a special kind of protein produced by living cells with high-efficiency catalytic function, high specificity and high controllability. It can specifically promote a certain chemical reaction without participating in the reaction itself. The reaction product has the characteristics of less pollution, low energy consumption, and easy control of the reaction, so it has been widely used. However, it is prone to denaturation of the enzyme protein under conditions such as acid, alkali, heat and organic solution, thereby reducing or losing the enzyme activity, and the enzyme reaction is mostly carried out in the solution, which is not easy to recover after the reaction is completed, and the separation and purification of the reaction product is difficult. And it is difficult to achieve continuous and automatic pr...

Examples

example 1

[0021] Immerse 1 gram of chitosan nanofiber composite film in 2% glutaraldehyde aqueous solution, seal it and vibrate for 3 hours at a constant temperature of 25°C, with a shaking speed of 40 rpm; take the nanofiber composite film and use a pH value of 7.0 The phosphate buffer solution was fully washed to remove the residual glutaraldehyde on the membrane; the nanofiber composite membrane activated by glutaraldehyde was immersed in the lipase phosphate buffer solution with a concentration of 2 mg / ml, and sealed at 4 Shake at a constant temperature of ℃ for 2 hours, and the shaking speed is 40 rpm; take out the membrane from the solution, wash it repeatedly with deionized water, and wash it 3 times in total, and then you can get the lipase-immobilized chitosan nanofiber composite; The nanofiber composite membrane with immobilized enzymes was immersed in a phosphate buffer solution with a pH of 7.0, and stored at 4°C for use.

example 2

[0023] Immerse 1 gram of chitosan nanofiber composite membrane in 10% glutaraldehyde aqueous solution, shake it under the constant temperature condition of 25 ℃ for 5 hours after sealing, and the shaking speed is 60 rpm; take the nanofiber composite membrane and use pH 7.0 The phosphate buffer solution was fully washed to remove the residual glutaraldehyde on the membrane; the nanofiber composite membrane activated by glutaraldehyde was immersed in the lipase phosphate buffer solution with a concentration of 6 mg / ml, and sealed at 4 Shake at a constant temperature of ℃ for 4 hours, and the shaking speed is 60 rpm; take out the membrane from the solution, wash it repeatedly with deionized water, and wash it 4 times in total, and then you can get the lipase-immobilized chitosan nanofiber composite; The nanofiber composite membrane with immobilized enzymes was immersed in a phosphate buffer solution with a pH of 7.0, and stored at 4°C for use.

example 3

[0025] Immerse 1 gram of chitosan nanofiber composite film in 5% glutaraldehyde aqueous solution, seal it and vibrate for 8 hours at a constant temperature of 25°C at a speed of 80 rpm; take the nanofiber composite film and use a pH value of 7.0 The phosphate buffer solution was fully washed to remove the residual glutaraldehyde on the membrane; the nanofiber composite membrane activated by glutaraldehyde was immersed in a catalase phosphate buffer solution with a concentration of 8 mg / ml, and after sealing Shake at a constant temperature of 4°C for 6 hours at a shaking speed of 80 rpm; take out the membrane from the solution and wash it repeatedly with deionized water for a total of 5 times to obtain catalase-immobilized chitosan Nanofiber composite: The nanofiber composite membrane with immobilized enzyme was immersed in a phosphate buffer solution with a pH of 7.0, and stored at 4°C for use.