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Characterization of Fish Cu/Zn–Superoxide Dismutase and Its Protection from Oxidative Stress

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Abstract

Copper/zinc superoxide dismutase was cloned from the zebrafish (Danio rerio). The full coding region of the zebrafish superoxide dismutase (ZSOD) complementary DNA was ligated with pET-20b(+) and successfully expressed in Escherichia coli strain AD494(DE3)pLysS. The active enzyme was purified by His tagging. The ZSOD yield was 6 mg from 0.2 L of E. coli culture, and the specific activity was 2000 U/mg as assayed using a RANSOD kit. The enzyme stability was characterized by reaction to temperature, pH, and detergent treatment. The results showed enzyme activity was still active after heat treatment at 70°C for 10 minutes, resistant to pH treatment from 2.3 to 12, and resistant to treatment with sodium dodecyl sulfate (SDS) under 4%. In addition, the recombinant ZSOD was used to protect fish from 100 ppm of paraquat-induced oxidative injury by soaking fish larva in 55 µg/ml SOD enzyme. The results were significant.

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Acknowledgements

This work was partially supported by the National Science Council of the Republic of China under Grant NSC 89-2313-B-019-066 to C.-T. L. and supported by the Council of Agriculture, Executive Yuan under Grant 90-BT-2.1-FID-F1 (3) to C.-T. L.

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Authors and Affiliations

  1. Graduate Institute of Life Sciences, National Defense Medical Center, Taiwan

    Chuian-Fu Ken

  2. Institute of Marine Biotechnology, National Taiwan Ocean University, Keelung, Taiwan

    Chuian-Fu Ken, Chi-Tsai Lin, Jei-Fu Shaw & Jen-Leih Wu

  3. Institute of Botany, Academia Sinica, Nankang, Taipei, Taiwan

    Jei-Fu Shaw

  4. Institute of Zoology, Academia Sinica, Nankang, Taipei, Taiwan

    Jen-Leih Wu

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  1. Chuian-Fu Ken
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  2. Chi-Tsai Lin
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Correspondence to Jen-Leih Wu.

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Ken, CF., Lin, CT., Shaw, JF. et al. Characterization of Fish Cu/Zn–Superoxide Dismutase and Its Protection from Oxidative Stress . Mar. Biotechnol. 5, 167–173 (2003). https://doi.org/10.1007/s10126-002-0058-1

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  • DOI: https://doi.org/10.1007/s10126-002-0058-1

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