一种新型双酶体系快速检测葡萄糖的方法

A Rapid Determination of Glucose Method based on Novel Double Enzyme System

建立一种基于过氧化物模拟酶/葡萄糖氧化酶体系快速检测葡萄糖的方法。重复的富含G碱基结构单元的DNA片段能形成堆积的G-四链体结构(G4),分子内平行G4-DNA与氯化血红素(hemin)室温孵育成过氧化物模拟酶,将其与葡萄糖氧化酶偶合成双酶体系,葡萄糖氧化酶可催化样品中葡萄糖生成过氧化氢,过氧化物模拟酶可进一步催化生成的过氧化氢与ABTS底物产生颜色变化,即样品中葡萄糖浓度与最终反应后溶液颜色呈正比。结果表明,5 min后通过肉眼观察溶液从无色变成绿色,可快速对葡萄糖的存在进行定性。10 min后通过比较标准色阶卡颜色的深浅,可半定量样品中葡萄糖含量。该方法具有成本较低,方法简便,稳定性和重复性良好等优点,可满足食品和血清中葡萄糖的定性和半定量分析。

Establish a rapid determination of glucose method based on mimetic peroxidase/glucoseoxidase system.Repeated G-base-rich DNA fragments can form a stacked G-quadruplex structure(G4).Mimetic peroxidases were formed by incubating intramolecular parallel G4 DNA with hemin and glucoseoxidase at room temperature.Mimetic peroxidases/glucoseoxidase system was assembled through incubating mimetic peroxidases and glucoseoxidase.Hydrogen peroxide could be catalyzed,when glucose met with glucoseoxidase.Peroxide mimetic enzyme could further catalyze the hydrogen peroxide oxidize ABTS substrate,then the color of solution could be changed from colourless to green.The concentration of glucose in the sample was proportional to the color of the solution after catalytic reaction.The results showed that after 5 min,glucose could be quickly identified with the color of sample solutions changed from colorless to green.It could be quickly quantified with the depth of the standard color scale after 10 min.The established method was cheap,simple,stable and repetitive.It could be used for qualitative analysis and semi-quantitative analysis of glucose in food and blood serum samples.