Research On Selection And Analytical Applications Of Peroxidase Mimetics

Peroxidase is a pure natural enzyme that can be involved in and catalyze various metabolism in the organism.There is heme in its structure,which is the auxiliary group of it.Peroxidase,as a catalyst with high efficiency in the physiological reactions of the organism,has strong activation to catalyze the oxidation reaction with oxides or peroxides involved.The pathogenesis of many diseases contain peroxide reaction.Under the reaction,cells,tissues and related organs in the organism may be damaged.Horseradish peroxidase is a natural peroxidase that discovered by people very early.It can be extracted directly from plants.However,it does not meet the demands of food safety detection,clinical examination and other aspects due to the difficulty to obtain,high price,variability,catalytic conditions,harsh storage conditions and other reasons.Therefore,it is of great significance to study the catalytic properties of model enzymes,explore the optimal reaction conditions of them and screen out the best mimetic peroxidase.In recent years,various model enzymes such as DhHP-6,metallic nanometer particles,and nano metal peroxides have been researched and developed in succession.These model enzymes present similar catalytic effect with horseradish peroxidase.This essay intends to replace natural peroxidase and explores the application of detecting hydrogen peroxide and glucose by screening different model enzymes.In this experiment,DhHP-6,Fe₃O₄NPs,CeO₂NPs,PtNPs,Co₃O₄NPs,CuONPs have been screened by comparing the spectral characteristics of model enzymes.Then the optimal reaction conditions for the model enzymes were worked out,and DhHP-6was determined as the best model enzymes by comparing the concentration and reaction time of model enzymes,pH,temperature,the concentration of H₂O₂ and other catalytic conditions,as well as the properties of the enzyme kinetics.This thesis takes DhHP-6 as the alternative enzyme to detect H₂O₂ and glucose and compare the consequence with the detection kits.The result shows that under optimal conditions,DhHP-6's detection linear range of H₂O₂ was 0.69-22.0 mmol/L;the repeatability was:CV=2.09%;stability:better than detection kit in 24 hours;compared with detection kits,R?0.99;the linear range of glucose was 0.35-22.0 mmol/L;the repeatability was:CV=2.60%;stability:within 3 hours,concentration CV?5.0%;compared with commercial kits,R?0.99;the average recovery rate reached 96.4%.The result shows that DhHP-6 is well correlated with the detection kits,and it can replace natural peroxidase in the application of detecting H₂O₂ and glucose.