| Title |
Expression and Characterization of an Iron-Containing Superoxide Dismutase from Burkholderia pseudomallei |
| Author |
Min-Hee Cho1, Yong-Woo Shin1,2, Jeong-Hoon Chun1, Kee-Jong Hong1, Byoung-Kuk Na3, Gi-eun Rhie1, Baik-Lin Seong2, and Cheon-Kwon Yoo1* |
| Address |
1Division of High-Risk Pathogen Research, Center for Infectious Diseases, National Institute of Health, Chungbuk 363-951, Republic of Korea, 2Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea, 3Department of Parasitology and the Institute of Health Sciences, College of Medicine, Gyeongsang National University, School of Medicine, Jinju 660-751, Republic of Korea |
| Bibliography |
Journal of Microbiology, 50(6),1029-1033, 2012,
|
| DOI |
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| Key Words |
Burkholderia pseudomallei, Fe-superoxide dismutase, melioidosis |
| Abstract |
A superoxide dismutase (SOD) gene from Burkholderia pseudomallei, the causative agent of melioidosis, was cloned and expressed in Escherichia coli, and its product was functionally and physically characterized. The gene has an open-reading
frame of 579 bp. The deduced amino acid sequence has 192 residues with a calculated molecular mass of ~22 kDa. Sequence comparison with other bacterial SODs showed that the protein contains typical metal-binding motifs and other
Fe-SOD-conserved residues. The sequence has substantial similarity with other bacterial Fe-SOD sequences. The enzymatic activity of the expressed protein was inhibited by hydrogen peroxide but not by sodium azide or potassium cyanide, attributes that indeed are characteristic of typical bacterial Fe-SODs. Western blotting with antiserum against the recombinant Fe-SOD revealed that it is expressed in B. pseudomallei. Transformed E. coli that expressed the Fe-SOD had significantly increased SOD activity and was highly tolerant to paraquat-mediated replication inhibition, compared to transformed cells carrying an empty vector. Our results
provide a basis for further biochemical characterization of the enzyme and elucidation of its role in the pathogenesis of B. pseudomallei. |
