Purification and Characterization of Manganese Superoxide Dismutase from Staphylococcus sciuri

  • Song, Chi-Hyun (Department of Biotechnology, Taegu University) ;
  • Park, Eun-Kyung (Oriental Brewery Co., LTD) ;
  • Suh, Hyung-Joo (Department of Food and Nutrition, College of Health Science, Korea University) ;
  • Lee, Yong-Se (Department of Natural Resources, College of Natural Resources, Taegu University) ;
  • Choi, Jang-Won (Department of Natural Resources, College of Natural Resources, Taegu University) ;
  • Ra, Kyung-Soo (Department of Food and Nutrition, Taegu Technical College)
  • Published : 1999.06.01

Abstract

The intracellular superoxide dismutase (SOD) from Staphylococcus sciuri was isolated to homogeneity by continuous steps, including ammonium sulfate fractionation, DEAE-ion-exchange chromatography, gel filtration, and phenyl hydrophobic gel chromatography. Pure SOD had a specific activity of 4,625 U/mg and was purified 158-fold with a yield of 31 % from a cell free extract. The molecular weight of the purified SOD was determined to be approximately 35.5 kDa by gel filtration and the enzyme was also shown to be composed of dimeric subunits on denaturing SDS-PAGE. The enzyme activity remained stable at pH 5 to 11 and also to heat treatment of up to $50^{\circ}C$ at pH 7.8, with 80% relative activity. The enzyme was insensitive to cyanide, hydrogen peroxide, and azide, indicating that it is a manganese-containing SOD. The EPR spectrum showed the enzyme containing manganese as a cofactor.

Keywords

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