TY - JOUR
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.
AB - 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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U2 - 10.1007/s10126-002-0058-1
DO - 10.1007/s10126-002-0058-1
M3 - Article
C2 - 12876653
AN - SCOPUS:0037767194
VL - 5
SP - 167
EP - 173
JO - Marine Biotechnology
JF - Marine Biotechnology
SN - 1436-2228
IS - 2
ER -