New method for improving life and stability of glucose oxidase by modifying surface of glucose oxidase with biocompatible polymer

A glucose oxidase, biocompatible technology, applied in a new field of modifying the surface of glucose oxidase with biocompatible polymers to improve its lifespan and stability

Inactive Publication Date: 2012-10-10
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few reports on the modification of the surf

Method used

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  • New method for improving life and stability of glucose oxidase by modifying surface of glucose oxidase with biocompatible polymer
  • New method for improving life and stability of glucose oxidase by modifying surface of glucose oxidase with biocompatible polymer
  • New method for improving life and stability of glucose oxidase by modifying surface of glucose oxidase with biocompatible polymer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1. Take RAFT reagent 0.527g, 2-mercaptothiazoline 0.238g, dissolve in the methylene chloride of 10ml. Further, 0.497 g of a dehydrating agent dicyclohexylcarbodiimide (DCC) and 24.5 mg of a catalyst 4-dimethylaminopyridine (DMAP) were added, and stirred at room temperature for 7 hours. After the reaction, the solid particles were removed by suction filtration, and then the product was purified through a silica gel column. 7.6 mg of RAFT reagent with mercaptothiazoline terminal groups, α-(1-oxo-2-propenyl)-ω-methoxypoly(oxyethylene) (PEG-A) 2 g, azobisisobutyl Cyanide (AIBN) 1 mg was dissolved in 4 ml of dioxane, wherein the molar ratio of RAFT reagent to PEG-A was 1:100. Nitrogen gas was used to remove oxygen for 40 minutes, and then the reaction system was sealed and reacted in an oil bath at 75° C. for 5 hours. After the reaction, the polymer was purified by precipitation with ether to obtain the expected product. Add 5 mg of glucose oxidase solution in ...

Embodiment 2

[0015] Example 2. Change the molar ratio in 1 to RAFT reagent: PEG-A=1:50, and others are as in Example 1.

Embodiment 3

[0016] Example 3. Change the molar ratio in 1 to RAFT reagent: PEG-A=1:200, and others are as in Example 1.

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PUM

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Abstract

The invention provides a new method for modifying surface of glucose oxidase, and aims at improving heat stability and life of glucose oxidase by modifying glucose oxidase utilizing amino group on the surface. The method comprises the steps: synthesizing alpha-(1-oxo-2-propenyl)-omega-methoxyl-poly(ethylene oxide) (PEG-A) high polymer with RAFT (reversible addition-fragmentation chain transfer) reagent with mercaptothiazoline terminal group through RAFT polymerization method; and carrying out amidation reaction between mercaptothiazoline group at the terminal of polymer chain and amino group on the surface of glucose oxidase to connect the mercaptothiazoline group to the surface of glucose oxidase. The polymer chain of the modified glucose oxidase partially wrap the activity center (flavin adenine dinucletide: FAD) and other components of the enzyme, so that stabilizing and protective action on the enzyme structure are realized. Therefore, the modified glucose oxidase has excellent heat stability and longer service life.

Description

technical field [0001] The method relates to a modification method of the glucose oxidase, and a biocompatible high molecular polymer is connected on the surface of the glucose oxidase through multiple amino groups on the surface of the glucose oxidase to be modified. The modified glucose oxidase has better thermal stability and longer service life, and belongs to the field of biological enzyme research. Background technique [0002] High-purity glucose oxidase is light yellow powder, easily soluble in water, completely insoluble in ether, chloroform, butanol, etc. The molecular weight is about 160KDa, and each molecule of glucose oxidase contains two molecules of flavin adenine dinucleotide (FAD). The solid enzyme preparation is stable for at least two years at 0°C and 8 years at -15°C. The stable temperature range of glucose oxidase is 30°C-60°C, and the optimum pH is 5. Glucose oxidase can oxidize glucose to generate D-gluconolactone in the presence of molecular oxygen...

Claims

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Application Information

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IPC IPC(8): C12N9/96
Inventor 刘敬权罗雄王蕊
Owner QINGDAO UNIV
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