Fig.1 Overview of the parts involved in our project

In this section, all the components used in our project have been successfully synthesized and constructed. At the same time, the antioxidant capacity of the enzyme preparations was evaluated through various detection methods. The results showed that both of our dual enzyme preparations exhibited excellent antioxidant activity, and the systematic testing methods can also serve as a reference for future project tests.

To achieve the design objectives of our project, we aimed to evaluate the enzymatic activities and antioxidant capacities of superoxide dismutase (SOD) and catalase (CAT). A comprehensive set of experimental assays was employed to assess their antioxidant properties, including SOD and CAT activity measurements, as well as evaluations of the dual-enzyme system’s capacity to scavenge superoxide anions and ABTS⁺ radicals. To systematically investigate the combined effects of key variables—temperature, pH, and the SOD/CAT ratio—on antioxidant performance, an orthogonal experimental design was implemented. This approach enables efficient and accurate assessment of multiple factors and their interaction effects with a reduced number of experimental runs, thereby minimizing time and resource expenditure. Furthermore, it allows for clear identification of dominant factors and optimal parameter levels through analysis of main and partial interaction effects. The standardized nature of the orthogonal design enhances experimental reproducibility and reliability, making it a robust methodology applicable to multi-factor, multi-level studies of functional genes and enzyme systems.

Fig. 2 Overview of project measurement method contributions