Preparation method of compound biomimetic mineralization nano biocatalyst

A biocatalyst, biomimetic mineralization technology, applied in organic compound/hydride/coordination complex catalysts, physical/chemical process catalysts, chemical instruments and methods, etc., can solve the difficult recovery of nano-microcapsules and the rapid loss of enzyme activity and other problems, to achieve the effects of increasing solvent stability and use stability, less enzyme damage, and reducing mass transfer resistance

Active Publication Date: 2017-05-17
HEBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the adoption of the step of forming the ZIF-8 mineralized layer by armoring the MOF material on the outside of the microcapsule, and using 2-methylimidazole and zinc acetate dihydrate as the core raw materials, the loss of enzyme activity in the extreme environment of the immobilized enzyme of the MOF material is solved. Faster and difficult recovery of nano-microcapsules are two key technical issues

Method used

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  • Preparation method of compound biomimetic mineralization nano biocatalyst
  • Preparation method of compound biomimetic mineralization nano biocatalyst
  • Preparation method of compound biomimetic mineralization nano biocatalyst

Examples

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Embodiment 1

[0039] According to the molar ratio of 30mg catalase: N-acryloyloxysuccinamide (NASM)=1:5, mix 30mg (0.5μmol) catalase with N-acryloyloxysuccinamide (NASM) (2.5μmol) was dissolved in 20mL of PBS to start the acylation reaction. The reaction vessel was a 100mL three-neck flask, kept in a constant temperature water bath at 4°C, and simultaneously reacted with 520r / min magnetic stirring for 2h. Add the pre-modified PGA to the monomer acrylamide (AAM) and N-(3-aminopropyl)-methacryl ester (APM) sequentially at a molar ratio of 1:2500:500:200, crosslinking agent N, N '-methylene acrylamide (BIS). After stirring evenly, 60 mg of ammonium persulfate (APS) and 50 μL of tetramethylethylenediamine (TEMED) were added as triggers, and the reaction was carried out in a constant temperature water bath at 4°C while magnetically stirring at 520 r / min for 4 h. Transfer 20mL of the obtained brown-yellow solution to a 50mL dialysis bag, place it in a beaker of 1000mL pure water to start dialysi...

Embodiment 2

[0046] Dissolve 30 mg of catalase (0.5 μmol) and N-acryloyloxysuccinamide (NASM) (2.5 μmol) in 20 mL of PBS at a molar ratio of 1:5 to start the acylation reaction. The flask was placed in a constant temperature water bath at 4°C while magnetically stirring at 520r / min for 2h. Add the pre-modified PGA to the monomer acrylamide (AAM) and N-(3-aminopropyl)-methacryl ester (APM) sequentially at a molar ratio of 1:2500:500:200, crosslinking agent N, N '-methylene acrylamide (BIS). After stirring evenly, 60 mg of ammonium persulfate (APS) and 50 μL of tetramethylethylenediamine (TEMED) were added as triggers, and the reaction was carried out in a constant temperature water bath at 4°C while magnetically stirring at 520 r / min for 4 hours. Transfer the resulting 20mL brown-yellow solution to a 50mL dialysis bag, place it in a beaker of 1000mL pure water to start dialysis, the dialysis should be carried out in a water bath at 15°C, give 200r / min stirring to accelerate the dialysis, d...

Embodiment 3

[0050]Dissolve 30 mg of catalase (0.5 μmol) and N-acryloyloxysuccinamide (NASM) (2.5 μmol) in 20 mL of PBS at a molar ratio of 1:5 to start the acylation reaction. The flask was placed in a constant temperature water bath at 4°C while magnetically stirring at 520r / min for 2h. Add the pre-modified PGA to the monomer acrylamide (AAM) and N-(3-aminopropyl)-methacryl ester (APM) sequentially at a molar ratio of 1:2500:500:200, crosslinking agent N, N '-methylene acrylamide (BIS). After stirring evenly, 60 mg of ammonium persulfate (APS) and 50 μL of tetramethylethylenediamine (TEMED) were added as triggers, and the reaction was carried out in a constant temperature water bath at 4°C while magnetically stirring at 520 r / min for 4 hours. Transfer the resulting 20mL brown-yellow solution to a 50mL dialysis bag, place it in a beaker of 1000mL pure water to start dialysis, the dialysis should be carried out in a water bath at 20°C, give 200r / min stirring to accelerate the dialysis, du...

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Abstract

The invention discloses a preparation method of a compound biomimetic mineralization nano biocatalyst. According to the method, two immobilized methods adopting MOF material immobilized enzyme and microcapsule immobilized enzyme are combined through steps of (1) enzyme modification; (2) formation of single-enzyme microcapsules; (3) formation of a ZIF-8 mineralization layer and the like, so that an MOF material is armored outside microcapsules. Respective defects of the two immobilized enzyme methods are overcome, and two key technical problems that enzyme activity loss of the MOF material immobilized enzyme at an extreme environment is quicker and nano microcapsules are difficult to recover are solved.

Description

technical field [0001] The invention belongs to the field of immobilized enzymes, and in particular proposes a method for preparing composite biomimetic mineralization nano biocatalysts by using polyacrylamide and metal organic framework materials as protective layers. Background technique [0002] As a biocatalyst, enzymes are favored by people for their advantages such as mild reaction conditions, wide range of sources, no pollution, high catalytic efficiency, and specific catalytic performance. However, the disadvantages of poor stability, difficulty in recycling and easy inactivation of free enzyme not only limit its wide application but also increase the application cost. In order to solve this problem and overcome the shortcomings of free enzymes that are difficult to separate and recover, the concept of immobilized enzymes came into being. After the enzymes are immobilized, they can be separated by simple centrifugation and other operations, which can be reused many t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J31/00B01J31/22
CPCB01J31/003B01J31/22B01J35/1023
Inventor 姜艳军杜英杰高静
Owner HEBEI UNIV OF TECH
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