DESIGN, EXPRESSION AND CHARACTERIZATION OF R10-SOD1 FUSION PROTEIN

Superoxide dismutase (SOD) exhibits unique functions (e.g. antioxidation, radiation protection, and anti-aging), and has been widely applied in pharmaceutical, food, and cosmetic industries. SOD can be classified into three types based on its cellular localization, among which copper/zinc superoxide dismutase (SOD1) is the most widely distributed in tissues. Due to cellular and tissue barriers as well as the low permeability of biological membranes, exogenous SOD proteins cannot cross cell membranes, limiting their practical applications. In this study, a membrane-penetrating fusion protein— R10-SOD1—was constructed and expressed through genetic recombination technology. This protein consists of human-derived copper/zinc superoxide dismutase (SOD1) fused with a cell-penetrating peptide containing 10 arginines (R10). The research encompasses the design and construction of the fusion gene, inducible expression in the Escherichia coli system, purification protocols for the recombinant protein, and functional evaluation of its membrane-penetrating capacity and antioxidant activity. The results demonstrate that the R10-SOD1 fusion protein possesses favorable membrane-penetrating ability, and the in vitro enzymatic activity and antioxidant capacity are superior to those of SOD1 alone. This study not only provides novel research tools for related fields but also establishes theoretical and technical foundations for the subsequent development of bioactive protein-based functional molecules.