| Literature DB >> 12876653 |
Chuian-Fu Ken1, Chi-Tsai Lin, Jei-Fu Shaw, Jen-Leih Wu.
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 degrees 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 micro g/ml SOD enzyme. The results were significant.Entities:
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Year: 2003 PMID： 12876653 DOI： 10.1007/s10126-002-0058-1
Source DB: PubMed Journal: Mar Biotechnol (NY) ISSN： 1436-2228 Impact factor: 3.619