Preparation method of glucose oxidase nanogel with good thermal stability

A technology of glucose oxidase and thermostability, applied in biochemical equipment and methods, oxidoreductase, immobilized on/in organic carriers, etc., can solve the problems of limited stabilization effect and complicated operation, and achieve expanded use Scope and use environment, high relative enzyme activity, effect of preventing changes in enzyme structure

Active Publication Date: 2017-09-15
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But there are certain shortcomings, such as c...

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  • Preparation method of glucose oxidase nanogel with good thermal stability
  • Preparation method of glucose oxidase nanogel with good thermal stability

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Embodiment 1

[0029] A preparation method of glucose oxidase nanogel with good thermal stability, comprising the steps of:

[0030] 1) Weigh 0.02g of glucose oxidase and dissolve it in 10mL of MES buffer solution to prepare a 2mg / mL glucose oxidase solution, measure 1ml of the above solution, and add 200mg of monomeric N-isopropylacrylamide to it and 2 mg of crosslinking agent N, N'-methylenebisacrylamide dissolved to prepare a mixed solution as the water phase;

[0031] 2) Add 1.235 g of the surfactant sodium bis(2-ethylhexyl) succinate sulfonate into 50 mL of isooctane, and dissolve it as the oil phase;

[0032] 3) under constant stirring condition, step 1) gained water phase is joined in step 2) gained oil phase, continue to stir to obtain transparent and stable microemulsion system, the W of final gained microemulsion 0 The value is 20;

[0033] 4) Add 1.5 mg of sodium bisulfite and 1.5 mg of ammonium persulfate to the microemulsion system obtained in step 3) as an initiator, control ...

Embodiment 2

[0036] A preparation method of glucose oxidase nanogel with good thermal stability, comprising the steps of:

[0037] 1) Weigh 0.001g of glucose oxidase and dissolve in 10mL of MES buffer solution to prepare 0.1mg / mL of glucose oxidase solution, measure 2mL of the above solution, add 50mg of monomeric methacrylic acid N,N '-Dimethylaminoethyl ester and 1 mg of cross-linking agent ethylene glycol dimethacrylate were dissolved to prepare a mixed solution as an aqueous phase;

[0038] 2) Add 3.594 g of the surfactant polyethylene glycol p-isooctyl phenyl ether into 20 mL of n-hexane, and dissolve it as the oil phase;

[0039] 3) under constant stirring condition, step 1) gained water phase is joined in step 2) gained oil phase, continue to stir to obtain transparent and stable microemulsion system, the W of final gained microemulsion 0 The value is 20;

[0040] 4) Add 0.25 mg of ferrous sulfate and 0.5 mg of hydrogen peroxide to the microemulsion system obtained in step 3) as a...

Embodiment 3

[0043] A preparation method of glucose oxidase nanogel with good thermal stability, comprising the steps of:

[0044]1) Weigh 0.01g of glucose oxidase and dissolve it in 10mL of MES buffer solution to prepare 1mg / mL of glucose oxidase solution, measure 5mL of the above solution, add 1g of monomeric methacrylic acid N,N' to it -diethylaminoethyl ester and 30 mg of cross-linking agent N,N'-vinylbisacrylamide were dissolved to prepare a mixed solution as the water phase;

[0045] 2) Add 5.062 g of the surfactant cetyltrimethylammonium bromide into 100 mL of cyclohexane, and dissolve it as the oil phase;

[0046] 3) under constant stirring condition, step 1) gained water phase is joined in step 2) gained oil phase, continue to stir to obtain transparent and stable microemulsion system, the W of final gained microemulsion 0 The value is 20;

[0047] 4) Add 5 mg of ammonium bisulfite and 20 mg of potassium persulfate to the microemulsion system obtained in step 3) as an initiator,...

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Abstract

The invention relates to a preparation method of glucose oxidase nanogel with good thermal stability. The preparation method comprises the following steps: preparing a monometer, glucose oxidase and a cross-linking agent into a mixed solution which serves as a water phase, and adding a surfactant into a non-polar or weak-polar organic solvent insoluble in water, thus obtaining an oil phase after dissolving; under the stirring condition, dropwise adding the water phase into the oil phase, carrying out continuous stirring to obtain a transparent and stable microemulsion system, adding an initiator into the microemulsion system, so that the monomer is polymerized and crosslinked in situ in the aqueous phase droplets of the microemulsion, and enabling glucose oxidase to be wrapped inside, so that the nanogel is formed, wherein the three-dimensional network structure of the gel can realize multi-point and multi-face immobilization for glucose oxidase, even when the mutual acting force between two subunits of glucose oxidase is weakened at high temperature, the separation still can be avoided, and therefore, the irreversible enzyme structure change caused by subunit dissociation can be effectively prevented.

Description

technical field [0001] The invention relates to a preparation method of a nanogel, in particular to a preparation method of a glucose oxidase nanogel, and belongs to the technical fields of functional polymer materials, nanomaterials and enzyme immobilization. Background technique [0002] Glucose oxidase (Glucose Oxidase) is widely distributed in organisms, it can specifically catalyze glucose to generate gluconic acid and hydrogen peroxide, and consume oxygen in the process. Therefore, it is used in food, feed, medicine and other fields to remove glucose, deoxidize, sterilize, and measure glucose concentration. Usually, glucose oxidase needs to be kept in a low temperature environment to ensure its enzymatic activity. However, many applications involving glucose oxidase inevitably need to be stored, transported, and implemented at higher temperatures, so improving the thermal stability of glucose oxidase is a technical problem to be solved urgently, which has important ap...

Claims

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Application Information

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IPC IPC(8): C12N11/08C12N11/04
CPCC12N9/0006C12N11/04C12N11/08C12Y101/03004
Inventor 陶倩柏良久李桂英薛众鑫王文香
Owner LUDONG UNIVERSITY
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