Detection of Ascorbate Oxidase by Regulating the Catalytic Activity of Cerium Oxide Nano-Enzyme Based on Ascorbic Acid
Nanozyme is a kind of simulated enzyme that has both the unique properties of nanomaterials and enzyme like catalytic activity.It owns the high catalytic activity of natural enzymes and the characteristics of economy and stability of simulated enzymes.Nanozymes have been widely applied in the field of bioanalysis.Herein,cerium oxide nanoparticles(CeO2 NPs)with horseradish peroxidase activity were synthesized by simple hydrothermal method.The nano-sized CeO2NPs have both+4 and+3 valence forms,in which+4 valence can react with hydrogen peroxide to oxidize 3,3,5,5'-tetramylbenzidine(TMB),making the solution turning blue.Ascorbic acid(AA)can reduce Ce4+and inhibit its catalytic activity on H2O2,further suppress the oxidation of TMB,resulting in a lighter color of the solution.Ascorbic acid oxidase(AAO),which catalyzes the oxidation of AA,obstructs the reduction of Ce4+and restores the catalytic activity of CeO2NPs.Using this method,the quantitative detection of A AO was achieved in the range of 12 to 400 mU/mL(R2=0.9704),and the detection limit was 3.2 mU/mL.This study provides a basis for the colorimetric method based on CeO2 NPs to detect the activity of AAO and its related applications.
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