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Orderly, directional and co-immobilization enzyme membrane reactor and preparation method and application thereof

An enzyme membrane reactor and a technology for immobilizing enzymes, which are applied in the field of reactors, can solve the problems of complex preparation methods and low conversion efficiency, and achieve the effects of improving transfer efficiency, mild reaction conditions and improving catalytic reaction efficiency.

Pending Publication Date: 2019-12-06
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] One of the purposes of the present invention is to overcome the problem of low conversion efficiency of existing immobilized enzyme reactors, and provide an ordered and oriented co-immobilized enzyme membrane reactor
[0005] The second object of the present invention is to overcome the complex problem of the existing immobilized enzyme reactor preparation method, and provide the preparation method of the ordered directional co-immobilized enzyme membrane reactor

Method used

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  • Orderly, directional and co-immobilization enzyme membrane reactor and preparation method and application thereof
  • Orderly, directional and co-immobilization enzyme membrane reactor and preparation method and application thereof
  • Orderly, directional and co-immobilization enzyme membrane reactor and preparation method and application thereof

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Embodiment 1

[0033] A preparation method of an ordered directional co-immobilized enzyme membrane reactor:

[0034] The first step is to prepare the immobilized enzyme solution of the combined enzyme

[0035] Step 1: Dissolve 5 mg of horseradish peroxidase (HRP) molecule and 5 mg of glucose oxidase molecule (GOD) in 1 mL of deionized water to obtain horseradish peroxidase solution and glucose oxidase solution;

[0036] In step 1 of the present invention, the glucose oxidase molecule contains prosthetic flavin adenine dinucleotide (FAD).

[0037] Step 2: Mix the horseradish peroxidase (HRP) solution and the glucose oxidase (GOD) solution in step 1 with the mixed solution respectively, react at room temperature for 30 minutes and then stand for 3 hours to obtain two kinds of reaction solutions. The mixed solution Including 2-3mL zinc nitrate solution with a concentration of 0.31 mol / L and 20-21mL 2-methylimidazole solution with a concentration of 1.36 mol / L;

[0038] Step 3: Centrifuge the...

Embodiment 2

[0051] A preparation method of an ordered directional co-immobilized enzyme membrane reactor:

[0052] The first step is to prepare the immobilized enzyme solution of the combined enzyme

[0053] Step 1: Dissolve 4.5mg of tyrosinase (TYR) molecule and 4.5mg of alkaline phosphatase (AKP) molecule in 1.2mL deionized water to obtain tyrosinase solution and alkaline phosphatase solution;

[0054] Step 2: Mix the tyrosinase solution and alkaline phosphatase solution in step 1 with the mixed solution respectively, react at room temperature for 30 minutes and then let it stand for 3 hours to obtain two kinds of reaction solutions. The mixed solution includes 2-3 mL of 0.26 mol / L zinc nitrate solution and 20-21mL 2-methylimidazole solution with a concentration of 1.13 mol / L;

[0055] Step 3: Centrifuge the two kinds of reaction solutions obtained in step 2 at 6000r / min for 10min, and collect two kinds of white powders respectively;

[0056] Step 4: Wash the two white powders with d...

Embodiment 3

[0064] A preparation method of an ordered directional co-immobilized enzyme membrane reactor:

[0065] The first step is to prepare the immobilized enzyme solution of the combined enzyme

[0066] Step 1: Dissolve 5.2mg long-chain alcohol oxidase (LCAO) molecule and 5.2mg ω-transaminase (ω-TA) molecule in 1.5mL deionized water respectively to obtain long-chain alcohol oxidase (LCAO) solution and ω-transaminase (ω-TA) -TA) solution;

[0067] Step 2: Mix the long-chain alcohol oxidase solution and ω-transaminase solution obtained in step 1 with the mixed solution respectively, react at room temperature for 30 minutes and then stand for 3 hours to obtain two kinds of reaction solutions. The mixed solution includes 2-3 mL of 0.26 mol / L zinc nitrate solution and 20-21mL 2-methylimidazole solution with a concentration of 1.13 mol / L;

[0068] Step 3: Centrifuge the two kinds of reaction solutions obtained in step 2 at 6000 rpm for 10 minutes, and collect two kinds of white powders r...

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Abstract

The invention discloses an orderly, directional and co-immobilization enzyme membrane reactor and a preparation method and application thereof. On the basis of a pervaporation membrane assembly, the pervaporation membrane assembly comprises a pervaporation membrane composited with a microfiltration membrane, immobilized enzymes of combination enzymes needed for a reaction are sequentially immobilized to the microfiltration membrane according to the enzyme catalysis cascade reaction sequence, the combination enzymes are any one of combination of horse radish peroxidase and glucose oxidase, combination of tyrosinase and alkaline phosphatase, combination of long-chain alcohol oxidase and omega-transaminase, and combination of ethanol dehydrogenase, cyclohexanone monooxygenase and lipase, thecatalytic reaction efficiency is advantageously improved, the immobilized enzymes are integrated on one microfiltration membrane, and the multi-step catalytic intermediate product transferring efficiency is improved; by arranging the microfiltration membrane, reacting and separating are integrated, and reaction balance advantageously moves in the favorable direction; and reaction conditions are mild, and an enzyme immobilization method is simple.

Description

technical field [0001] The invention belongs to the technical field of reactors, and in particular relates to an ordered and oriented co-immobilized enzyme-membrane reactor as well as a preparation method and application thereof. Background technique [0002] Enzymes act as biocatalysts that increase the rate of a broad and diverse range of chemical reactions that occur in living organisms. It participates in important chemical reaction processes necessary for life, such as DNA replication and transcription, protein synthesis, primary and secondary metabolism, signal transduction and cellular defense mechanisms, and is an integral part of living organisms. Enzymes are widely used in biocatalysis, biosensing, and biomedical devices. However, due to the low thermal stability, poor stability of organic solvents, high cost and poor reusability of enzymes, the wide application of enzymes has been greatly hindered. [0003] Enzyme immobilization technology is one of the effectiv...

Claims

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Application Information

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IPC IPC(8): C12M1/40C12M1/24C12M1/12C12M1/00C12Q1/00C12N11/14C12N11/02
CPCC12M21/18C12M23/08C12N11/14C12N11/02C12Y111/01007C12Y101/03004C12N9/0059C12Y110/03001C12Y301/03001C12Y101/0302C12Y206/01C12Y101/01001C12Y114/13022C12N9/20C12Y301/01003C12N9/0006C12N9/0073C12N9/1096C12N9/16
Inventor 叶鹏祝黛莲
Owner ZHEJIANG SCI-TECH UNIV
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