Purification, crystallization and properties of iron-containing superoxide dismutase from Pseudomonas ovalis.

Three electrophoretically distinct superoxide dismutases (EC 1.15.1.1) were observed in the crude extracts from Pseudomonas ovalis. One of these was isolated as an iron-containing superoxide dismutase. It contained 1.4 gatoms of Fe per mol of enzyme, and had a specific activity of 3900 units per mg of protein. A crystallized enzyme contained 1.1 gatoms of Fe per mol of enzyme, and had a specific activity of 3100 units per mg of protein. The results of sedimentation equilibrium and gel filtration indicated a molecular weight of 40,000. S020,W was estimated as 3.18 by sedimentation velocity study. Sodium dodecyl sulfate gel electrophoresis indicated that the enzyme was composed of two subunits, and had a molecular weight of 19,500. Analysis for sulfhydryl groups showed that there were four such groups per mol of enzyme. The spectrum of visible and ultraviolet region, the amino acid composition, the CD spectrum of the enzyme, and the effect of certain compounds on the enzyme, were studied and compared with iron-containing superoxide dismutases isolated from other organisms.