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

doi:10.1007/s10126-002-0058-1

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

Department of Biology

Research output: Contribution to journal › Article › peer-review

31 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 language	English
Pages (from-to)	167-173
Number of pages	7
Journal	Marine Biotechnology
Volume	5
Issue number	2
DOIs	https://doi.org/10.1007/s10126-002-0058-1
Publication status	Published - 2003 Mar 1

All Science Journal Classification (ASJC) codes

Aquatic Science
Biotechnology

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title = "Characterization of fish Cu/Zn-superoxide dismutase and its protection from oxidative stress",

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.",

author = "Chuian-Fu Ken and Lin, {Chi Tsai} and Shaw, {Jei Fu} and Wu, {Jen Leih}",

year = "2003",

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doi = "10.1007/s10126-002-0058-1",

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T1 - Characterization of fish Cu/Zn-superoxide dismutase and its protection from oxidative stress

AU - Ken, Chuian-Fu

AU - Lin, Chi Tsai

AU - Shaw, Jei Fu

AU - Wu, Jen Leih

PY - 2003/3/1

Y1 - 2003/3/1

N2 - 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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