Purification and Characterization of Superoxide Dismutase in Sphingomonas sp. KS 301
2007
Kang, H.J. (Dankook University, Cheonan, Republic of Korea) | Jeong, H.J. (Dankook University, Cheonan, Republic of Korea) | Choi, J.H. (Dankook University, Cheonan, Republic of Korea) | Son, S.Y. (Dankook University, Cheonan, Republic of Korea), E-mail: [email protected]
Sphingomonas sp. KS 301, which was isolated from oil contaminated soil, was shown to have five different SODs (SODⅠ, Ⅱ, Ⅲ, Ⅳ, Ⅴ) which can be separated by DEAE-Sepharose chromatography, and SOD Ⅲ was finally purified in this study by ammonium sulfate precipitation, DEAE-Sepharose chromatography, Superose 12 gel filtration and Uno-Q1 ion exchange chromatography. The molecular weight of SOD Ⅲ was 23 kDa as determined by SDS-PAGE and the apparent molecular weight of the native enzyme was estimated to be approximately 71 kDa by Superose-12 gel filtration chromatography. These data suggest that the purified SOD consists of at least two subunits. The specific activity of the SOD Ⅲ was higher than Mn type or Fe type SOD of Escherichia coli by 5 fold. To determine the type of SOD Ⅲ, inhibitory effects of NaN₃, H₂O₂, and KCN were examined. 10 mM NaN₃ was able to inhibit 56% of the SOD Ⅲ activity, which indicates that this SOD is Mn type.
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