Recently, we demonstrated that superoxide dismutase 3 (SOD3) is a strong biomedicine candidate. SOD3 accomplished anti-oxidant function without cell penetration and inhibited inflammatory responses via non-enzymatic functions. SOD3 has the heparin binding domain associating cell surface. Interestingly, we have found that Zn2+ promotes transduction effects of recombinant human SOD3 (rhSOD3) by increasing uptake via heparin binding domain (HBD). We showed an uptake of rhSOD3 from media to cell lysate via HBD, resulting in an accumulation of rhSOD3 into the nucleus, which is promoted by the presence of Zn2+. As a result, we showed the increased inhibitory effects of rhSOD3 on NF-kB and STAT3 signal under the presence of Zn2+ which is elevated association of rhSOD3 into the cells. These results suggested that the optimized procedure can help to enhance the inflammatory efficacy of rhSOD3, as a novel biomedicine.