Study On Purification And Characterization Of Superoxide Dismutase From Rumex K-1(Rumex Patientia ×R.tianschanicus Cv.Rumex K-1)

This dissertation discussed the purification and characterization of superoxide dismutase from Rumex. A superoxide dismutase from Rumex was purified by the steps including ammonium sulfate fractionation, Sephadex G-75 gel filtration and DEAE Cellulose chromatography. There was a single band using polyacrylamide gel electrophoresis, which indicated that the enzyme had been purified to homogeneity.The specific activity of the purified enzyme was 8565.3U/mg protein, the activity recovery was 29.2 %. Its activity is strongly inhibited by KCN and H2O2 ,which indicated that SOD was CuZn-SOD.The enzyme exhibited one absorption maximum in the ultraviolet at 264.17nm and another in the visible region at 679.14nm. The characterization of purified enzyme was studied,it had two homogenous subunits,the subunit molecular weight of the purified enzyme was estimated at 16 184 daltons on SDS-polyacrylamide gel electrophoresis. The amino terminal residue was alanine(Ala) by using Dansyl-Cl method. Its PI was about 6.7. The experiment indicated that the purified enzyme was stable to heat below 65℃ and at pH 6.0～10.0,and it can keep activity 9 hours to heat 60℃...