Publication: Cloning of active human manganese superoxide dismutase and its oxidative protection in Escherichia coli
3
Issued Date
2007-10-15
Resource Type
ISSN
18125735
10288880
10288880
Other identifier(s)
2-s2.0-36048976355
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Mahidol University
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SCOPUS
Bibliographic Citation
Pakistan Journal of Biological Sciences. Vol.10, No.20 (2007), 3541-3548
Suggested Citation
Sakda Yainoy, Chartchalerm Isarankura-Na-Ayudhya, Tanawut Tantimongcolwat, Virapong Prachayasittikul Cloning of active human manganese superoxide dismutase and its oxidative protection in Escherichia coli. Pakistan Journal of Biological Sciences. Vol.10, No.20 (2007), 3541-3548. doi:10.3923/pjbs.2007.3541.3548 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/23972
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Title
Cloning of active human manganese superoxide dismutase and its oxidative protection in Escherichia coli
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Abstract
Superoxide radical (O2•-) is a toxic byproduct of oxidative metabolism that extensively damages cellular macromolecules and organelles. Superoxide dismutase (SOD) catalyzes the conversion of superoxide radical to hydrogen peroxide (H2O2) and molecular oxygen (O2) thus providing a biological defense against oxygen toxicity. The structural gene of human manganese superoxide dismutase (hMnSOD) was successfully cloned into the pET46Ek/LIC by using a Ligation Independent Cloning (LIC) technique. The recombinant human MnSOD was expressed in E. coli strain BL21(DE3)pLysS and purified to homogeneity by Ni2+-NTA. Supplementation of Mn2+in the bacterial growth media was proven to be crucial for production of enzymatically active hMnSOD. The recombinant enzyme revealed a specific activity up to 2,857 U mg-1as measured by inhibition of photoreduction of nitroblue tetrazolium (NBT). The molecular weight of each subunit was estimated to be 22 kDa by SDS-PAGE. More interestingly, E. coli expressing hMnSOD provides resistance against oxidative stress induced by the herbicide paraquat up to 1.2 mM. These findings gain insights into the biochemical characterization and significant roles of oxidative-protection of the hMnSOD in biological systems. © 2007 Asian Network for Scientific Information.