Separation And Purification Of Superoxide Dismutase And Nature

Different kinds of SOD have dissimilar physical and chemical character about molecular weight, isoelectric points, temperature sensitivity, chemical regeant sensitivity, etc. On the other hand, different kinds of SOD from various materials have different distribution and content. So there are diverse purification processes from various materials. The followed methods comform to the aboved thoughtfulness.Porcine erythrocyte coarse Cu/Zn-SOD was obtained by several steps, which were about pretreatment of fresh blood, washing erythrocyte, hemolysis, removing hemoglobin by ethanol and chloroform, extraction by dipotassium hydrogen phosphate, precipitation by acetone, heat denaturation, etc. Then, through DEAE-Cellulose 52, Sephadex G-75, it was completely purified.Spirulina coarse Fe-SOD was obtained by several steps, which were about breaking by ultrasonication, heat denaturation, salting-out by ammonium sulfate, dialysis, etc. Then, through DEAE-Sepharose Fast Flow, CM-Sepharose Fast Flow, Sephadex G-100, purified Fe-SOD was gained.The separation and purification process of Citrus leaf Fe-SOD and pork liver Mn-SOD was similar to spirulina Fe-SOD. But, it had some differences. First, the former was breaked by scissors, the latter was breaked by ultrasonication; Besides, the gradient of salting-out by ammonium sulfate were different.Purified SOD were testified by PAGE. Furthermore, the experiment of the content of metallic element and absorption spectrum further proved that the purified SOD from porcine erythrocyte, spiralina, citrus leaf, pork liver were Cu/Zn-SOD, Fe-SOD, Fe-SOD, Mn-SOD. Besides, research found that Fe-SOD and Mn-SOD were stable to high temperature, Cu/Zn-SOD was not as good as Fe-SOD and Mn-SOD; Cu/Zn-SOD was sensitive to H2O2 and KCN, whereas Fe-SOD was sensitive to H2O2 but not to KCN. Mn-SOD in contrast, was not inhibited by H2O2 and KCN.