Characterization of fish Cu/Zn-superoxide dismutase and its protection from oxidative stress

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

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29 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)167-173
Number of pages7
JournalMarine Biotechnology
Volume5
Issue number2
DOIs
Publication statusPublished - 2003 Mar 1

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All Science Journal Classification (ASJC) codes

  • Aquatic Science
  • Biotechnology

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