Enzyme-GO-MOFs nano composite catalyst and preparation method thereof

Abstract

The invention discloses an enzyme-GO-MOFs nano composite catalyst and a preparation method thereof. The preparation method comprises the following steps: (1) adding a modifier into an enzyme liquid so as to obtain a modified enzyme solution, wherein the modifier is an N-vinyl amide polymer or pluronic F-127; the N-vinyl amide polymer is one or a mixture of more of polyvinylpyrrolidone, poly N-vinyl caprolactam and poly N-ethylene formamide; (2) mixing GO and a metallic salt solution, performing ultrasonic treatment, further adding an organic ligand solution into the mixed solution so as to obtain a GO-MOFs composite material synthesized stock solution, and performing ultrasonic treatment for another time; (3) adding a modified enzyme solution into the GO-MOFs composite material synthesized stock solution, and performing ultrasonic treatment so as to obtain a reaction liquid; (4) centrifuging the reaction liquid, collecting precipitate, washing the precipitate, and performing vacuum drying, thereby obtaining the enzyme-GO-MOFs nano composite catalyst. The preparation method disclosed by the invention is simple and feasible, and the nano composite material catalyst is gentle in synthesis condition and high in protein immobilization rate.

Description

technical field

[0001] The invention belongs to the technical field of enzyme catalysts, and in particular relates to an enzyme-GO-MOFs nanocomposite catalyst and a preparation method thereof. Background technique

[0002] Enzyme immobilization technology is a core technology in modern biotechnology, which solves many problems in the process of enzyme application and opens up new prospects for the application of enzymes. The immobilized enzyme has higher catalytic activity and stability; and the enzyme and the product are easy to separate, and the product is easy to purify. The performance of the immobilized enzyme mainly depends on the immobilization method and the carrier used, both of which can directly affect the catalytic activity of the immobilized enzyme. However, common immobilization methods and carriers have certain limitations, such as adsorption, cross-linking, embedding, and covalent bonding. The adsorption method is simple to prepare, and the conditions are m...

Examples

Embodiment 1

[0038] Example 1: Prepare GO-MOFs nanocomposite coated lipase according to the following steps:

[0039] (1) Take 20 mg of lipase, make (20 mg-mL) enzyme solution with 1 mL deionized water, then add 20 mg polyvinylpyrrolidone (PVP) (20 mg-mL) to obtain PVP modified enzyme solution.

[0040] (2) Take zinc nitrate hexahydrate and 2-methylimidazole to make a solution (20mM, 15mL) with methanol respectively, take 1mg of GO and add it to the methanol solution of zinc nitrate hexahydrate, after ultrasonic treatment for 6h, add 2-methylimidazole The methanol solution of imidazole was degassed in an ultrasonic bath for 5 min to obtain the synthetic stock solution of GO-MOFs composites.

[0041] (3) Mix the modified enzyme solution obtained in the above steps with the GO-MOFs composite synthetic stock solution, and then treat it in an ultrasonic bath for 1 min to obtain a uniformly mixed reaction solution.

[0042] (4) Stand for reaction at 50°C for 5h, then collect by centrifugation ...

Embodiment 2

[0048] Example 2: Prepare GO-MOFs nanocomposite coated cytochrome c according to the following steps:

[0049] (1) Take 50 mg of cytochrome c, mix it with 2 mL of deionized water to make (25 mg-mL) enzyme solution, and then add 20 mg of poly-N-vinylformamide (10 mg-mL) to obtain a modified enzyme solution.

[0050] (2) Take zinc nitrate hexahydrate and methanol to make (20mM, 30mL) solution, imidazole and methanol to make (150mM, 25mL) solution, take 48.6mg of GO and add zinc nitrate hexahydrate methanol solution, ultrasonic bath treatment After 8 h, a methanol solution of imidazole was added and degassed in an ultrasonic bath for 10 min to obtain a synthetic stock solution of GO-MOFs composites.

[0051] (3) Mix the modified enzyme solution obtained in the above steps with the GO-MOFs composite synthesis stock solution, and then treat it in an ultrasonic bath for 5 minutes to obtain a uniformly mixed reaction solution.

[0052] (4) Stand for reaction at 25°C for 12 hours, then...

Embodiment 3

[0057] Example 3: Prepare GO-MOFs nanocomposite coated with horseradish peroxidase according to the following steps:

[0058] (1) 90 mg of horseradish peroxidase was mixed with 3 mL of deionized water to make (30 mg-mL) enzyme solution, and then 30 mg of Pluronic F-127 (10 mg-mL) was added to obtain a modified enzyme solution.

[0059] (2) Prepare cobalt chloride hexahydrate with N,N-dimethylformamide (25mM, 54mL), and 2-carboxybenzimidazole with N,N-dimethylformamide (100mM, 18mL) For the solution, take GO90mg and add it to the N,N-dimethylformamide solution of cobalt chloride hexahydrate. After ultrasonic bath treatment for 10h, add the N,N-dimethylformamide solution of 2-carboxybenzimidazole The bath was degassed for 20 min to obtain the synthetic stock solution of GO-MOFs composites.

[0060] (3) Mix the modified enzyme solution obtained in the above steps with the GO-MOFs nanocomposite synthesis stock solution, and then treat it in an ultrasonic bath for 8 minutes to obt...

technical field

[0001] The invention belongs to the technical field of enzyme catalysts, and in particular relates to an enzyme-GO-MOFs nanocomposite catalyst and a preparation method thereof. Background technique

[0002] Enzyme immobilization technology is a core technology in modern biotechnology, which solves many problems in the process of enzyme application and opens up new prospects for the application of enzymes. The immobilized enzyme has higher catalytic activity and stability; and the enzyme and the product are easy to separate, and the product is easy to purify. The performance of the immobilized enzyme mainly depends on the immobilization method and the carrier used, both of which can directly affect the catalytic activity of the immobilized enzyme. However, common immobilization methods and carriers have certain limitations, such as adsorption, cross-linking, embedding, and covalent bonding. The adsorption method is simple to prepare, and the conditions are m...

Examples

Embodiment 1

[0038] Example 1: Prepare GO-MOFs nanocomposite coated lipase according to the following steps:

[0039] (1) Take 20 mg of lipase, make (20 mg-mL) enzyme solution with 1 mL deionized water, then add 20 mg polyvinylpyrrolidone (PVP) (20 mg-mL) to obtain PVP modified enzyme solution.

[0040] (2) Take zinc nitrate hexahydrate and 2-methylimidazole to make a solution (20mM, 15mL) with methanol respectively, take 1mg of GO and add it to the methanol solution of zinc nitrate hexahydrate, after ultrasonic treatment for 6h, add 2-methylimidazole The methanol solution of imidazole was degassed in an ultrasonic bath for 5 min to obtain the synthetic stock solution of GO-MOFs composites.

[0041] (3) Mix the modified enzyme solution obtained in the above steps with the GO-MOFs composite synthetic stock solution, and then treat it in an ultrasonic bath for 1 min to obtain a uniformly mixed reaction solution.

[0042] (4) Stand for reaction at 50°C for 5h, then collect by centrifugation ...

Embodiment 2

[0048] Example 2: Prepare GO-MOFs nanocomposite coated cytochrome c according to the following steps:

[0049] (1) Take 50 mg of cytochrome c, mix it with 2 mL of deionized water to make (25 mg-mL) enzyme solution, and then add 20 mg of poly-N-vinylformamide (10 mg-mL) to obtain a modified enzyme solution.

[0050] (2) Take zinc nitrate hexahydrate and methanol to make (20mM, 30mL) solution, imidazole and methanol to make (150mM, 25mL) solution, take 48.6mg of GO and add zinc nitrate hexahydrate methanol solution, ultrasonic bath treatment After 8 h, a methanol solution of imidazole was added and degassed in an ultrasonic bath for 10 min to obtain a synthetic stock solution of GO-MOFs composites.

[0051] (3) Mix the modified enzyme solution obtained in the above steps with the GO-MOFs composite synthesis stock solution, and then treat it in an ultrasonic bath for 5 minutes to obtain a uniformly mixed reaction solution.

[0052] (4) Stand for reaction at 25°C for 12 hours, then...

Embodiment 3

[0057] Example 3: Prepare GO-MOFs nanocomposite coated with horseradish peroxidase according to the following steps:

[0058] (1) 90 mg of horseradish peroxidase was mixed with 3 mL of deionized water to make (30 mg-mL) enzyme solution, and then 30 mg of Pluronic F-127 (10 mg-mL) was added to obtain a modified enzyme solution.

[0059] (2) Prepare cobalt chloride hexahydrate with N,N-dimethylformamide (25mM, 54mL), and 2-carboxybenzimidazole with N,N-dimethylformamide (100mM, 18mL) For the solution, take GO90mg and add it to the N,N-dimethylformamide solution of cobalt chloride hexahydrate. After ultrasonic bath treatment for 10h, add the N,N-dimethylformamide solution of 2-carboxybenzimidazole The bath was degassed for 20 min to obtain the synthetic stock solution of GO-MOFs composites.

[0060] (3) Mix the modified enzyme solution obtained in the above steps with the GO-MOFs nanocomposite synthesis stock solution, and then treat it in an ultrasonic bath for 8 minutes to obt...

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