Abstract
Superoxide dismutase (SOD), which plays a very important role in protecting organisms from oxygen toxicity, has therapeutic importance. It was purified from sewage isolated E. coli and characterized. Eukaryotic cells also produce SOD but culturing and maintenance of eukaryotic cells for production of SOD is costly as well as difficult. Using prokaryotic cells i.e. bacteria, production cost can be reduced. A rich bacterial source was identified. Bacterial membrane was ruptured in the presence of lysozyme and glass bead. Following ammonium sulphate precipitation, SOD-containing solution was applied to DEAE-cellulose and then Sephadex G- 75 gel columns. SOD was purified 63.91-fold with a specific activity of 3835U/ mg. The molecular weight was estimated to be 35.713 kDa by SDSPAGE gel. Maximum SOD activity was observed between pH 7.0 to 7.5 at temperature range 37–50oC. This enzyme has fair thermal stability. The enzyme was found to be stable in presence of 1% salt only. The activity found to be gradually reduced approximately 50% at higher concentrations. It was totally inactivated above 9% salt concentration.
Highlights
The superoxide radical is an intermediate reduction product of oxygen produced in a variety of biological reactions
Superoxide dismutase (SOD) are divided into three classes on the basis of their active site metals: copper and zinc (Cu/Zn-SOD), manganese (Mn-SOD)/ iron (Fe-SOD) and in some bacteria one more variety is found named as Ni-SOD [6]
CuZn-SOD was found widely in the cytoplasm and certainly in the mitochondrial intermembrane space of the eukaryotic cells and chloroplasts of plants. It is reported in bacteria like S. aureus, Pseudomonas spp., E. coli etc [7,8,9,10] Mn-SOD are located in prokaryotes and in the mitochondria of eukaryotes [11], while Fe-SOD has been found in bacteria, blue-green algae and protozoa [12,13]
Summary
The superoxide radical is an intermediate reduction product of oxygen produced in a variety of biological reactions. Most organisms produce defense systems, such as metallo-enzymes, to protect themselves from reactive oxygen species. CuZn-SOD was found widely in the cytoplasm and certainly in the mitochondrial intermembrane space of the eukaryotic cells and chloroplasts of plants. It is reported in bacteria like S. aureus, Pseudomonas spp., E. coli etc [7,8,9,10] Mn-SOD are located in prokaryotes and in the mitochondria of eukaryotes [11], while Fe-SOD has been found in bacteria, blue-green algae and protozoa [12,13]. Recent reports indicated that the enzyme was present in higher land plants [2]
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