A Sensitive and Reproducible Spectrophotometric Assay for Glucose Oxidase Activity and Its Application in Conformational Lock

摘要

Glucose-oxidase-horseradish peroxidaseguaiacol coupled system was applied for glucose oxidase assay with high sensitivity and prefect reproducibility. The optimum pH value was 5.8. The Michaelis-Menten constant (Km) and Vmax were determined to be 30.0±1.0mM and 2.49±0.08μM/s, respectively, for the assay system at 25°C. The relative standard deviation was 2.8% for 31 times measurements of GOD activity. The GOD assay could be applied in conformational lock study on GOD. Results of bio-thermodynamics suggested that the number of contact area of GOD is two, which is consistent with the computation results by using Ligplot and Pymol.