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A directional co-immobilization method for a multi-enzyme system

A technology of multi-enzyme system and covalent bond is applied in the field of directional co-immobilization of multi-enzyme system, which can solve problems such as the decrease of immobilized enzyme activity and achieve the effect of efficiently realizing biological activity.

Inactive Publication Date: 2018-01-02
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immobilized enzyme structure obtained by the traditional enzyme immobilization method is that the enzyme is connected to the carrier at any position. The main disadvantage of this method is that the activity of the immobilized enzyme often decreases significantly.

Method used

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  • A directional co-immobilization method for a multi-enzyme system
  • A directional co-immobilization method for a multi-enzyme system
  • A directional co-immobilization method for a multi-enzyme system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Experimental steps:

[0027] 1. Graphene oxide (GO)-3,5-diaminobenzoic acid (3,5-DABA)

[0028] Weigh 1.0 g of graphene oxide, put it in a beaker, add 200 ml THF, and ultrasonicate for 2 h; then add 1.0 g of 3,5-diaminobenzoic acid (~6.57×10 -3 mol), continue to sonicate for 30min; transfer the sonicated solution to a three-neck flask, seal the reaction flask, vacuumize, fill with nitrogen, heat to 70°C to condense and reflux, and magnetically stir for 2h; centrifuge the reaction solution, and wash the precipitate with absolute ethanol times; the precipitate was dried in a vacuum oven.

[0029] 2. GO-3,5-DABA-polyaniline (PANI)

[0030] Weigh 0.3009g of GO-3,5-diaminobenzoic acid, add 60ml of NMP to mix, ultrasonically disperse; add 0.1509g of polyaniline, stir evenly; -4 mol) was dissolved in 10ml of 1.0M hydrochloric acid solution, and slowly added dropwise to the reaction flask; after the dropwise addition, the reaction was continued to stir for 2h; the reaction s...

Embodiment 2

[0053] Same implementation as Example 1, adjust each material as follows:

[0054] 1. Carbon nanotube (CNT)-3,5-diaminobenzoic acid (3,5-DABA)

[0055] Weigh 1.0 g of carbon nanotubes, put them in a beaker, add 200 ml THF, and ultrasonicate for 1 h; then add 1.0 g of 3,5-diaminobenzoic acid (~6.57×10 -3 mol), continue to sonicate for 30min; transfer the sonicated solution to a three-neck flask, seal the reaction flask, vacuumize, fill with nitrogen, heat to 70°C to condense and reflux, and magnetically stir for 2h; centrifuge the reaction solution, and wash the precipitate with absolute ethanol times; the precipitate was dried in a vacuum oven.

[0056] 2. CNT-3,5-DABA-polyaniline (PANI)

[0057] Weigh 0.2g of CNT-3,5-diaminobenzoic acid, add 60ml of NMP to mix, and ultrasonically disperse; add 0.15g of polyaniline, stir evenly; -4 mol) was dissolved in 10ml of 1.0M hydrochloric acid solution, and slowly added dropwise to the reaction flask; after the dropwise addition, the...

Embodiment 3

[0068] 1. Graphene oxide (GO)-3,5-diaminobenzoic acid (3,5-DABA)

[0069] Weigh 1.0 g of graphene oxide, put it in a beaker, add 200 ml THF, and ultrasonicate for 1 h; then add 1.0 g of 3,5-diaminobenzoic acid (~6.57×10 -3 mol), continue to sonicate for 30min; transfer the sonicated solution to a three-neck flask, seal the reaction flask, vacuumize, fill with nitrogen, heat to 70°C to condense and reflux, and magnetically stir for 2h; centrifuge the reaction solution, and wash the precipitate with absolute ethanol times; the precipitate was dried in a vacuum oven.

[0070] 2. GO-3,5-DABA-polyaniline (PANI)

[0071] Weigh 0.2g of GO-3,5-diaminobenzoic acid, add 60ml NMP to mix, and ultrasonically disperse; add 0.15g of polyaniline, stir evenly; -4 mol) was dissolved in 10ml of 1.0M hydrochloric acid solution, and slowly added dropwise to the reaction flask; after the dropwise addition, the reaction was continued to stir for 2h; the reaction solution was centrifuged (5000rpm, ...

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Abstract

The invention discloses a directed common immobilization method of a multi-enzyme system. By utilizing a biocompatible effect between a prothetic-group enzyme and a prothetic-group-free enzyme, the multi-enzyme system is directionally co-immobilized on a carrier. The directed common immobilization method of the multi-enzyme system, provided by the invention, has the main advantages that the high-degree directed common immobilization of the multi-enzyme system is realized and the synergistic effect between the enzymes of the multi-enzyme system is facilitated very well, so that the catalysis efficiency is extremely improved; compared with the activity of a common adsorption-method immobilized enzyme, the activity of a directed compatible-effect immobilized enzyme is remarkably improved, and a feasible method for further constructing a high-efficiency and low-cost biological catalysis system is provided.

Description

technical field [0001] The invention relates to a directional co-immobilization method of a multi-enzyme system, which belongs to the field of protein immobilization. Background technique [0002] Many enzymes in cells often act in a continuous reaction chain, and a continuous chain reaction means that the product of the previous enzymatic reaction is the substrate of the latter enzymatic reaction. In a certain metabolic process in a complete cell, the reaction chain system formed by several enzymes is called a multi-enzyme system. Multi-enzyme system is also called multi-enzyme complex, multi-enzyme system, multi-enzyme system, multi-enzyme cluster. In a continuous reaction system, the product of the previous reaction is the substrate of the latter reaction, and the reactions are connected in sequence to form a metabolic pathway or a part of a metabolic pathway. Because this sequence reaction is carried out in a highly ordered multi-enzyme complex, the catalytic efficienc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/18C12N11/14
CPCC12N9/0006C12N9/0065C12N11/14C12N11/18C12Y101/03004C12Y101/03006C12Y111/01006C12Y111/01007
Inventor 叶鹏胡玲玲
Owner ZHEJIANG SCI-TECH UNIV