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Published:
Jun 26, 2013DOI:
10.4314/tjpr.v12i3.9Keywords:
Article Details
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Main Article Content
Expression, Purification, Characterization and <i>In Vitro</i> Activity of Recombinant Mouse Cu/Zn-Binding Superoxide Dismutase (mSOD1)
Abstract
Purpose: To express, purify and characterize recombinant mouse Cu/Zn-binding superoxide dismutase (mSOD1), and investigate its activity in vitro.
Methods: The protein, mSOD1, was expressed after induction with isopropyl-beta-Dthiogalactopyranoside (IPTG). The target protein was purified by Ni-NTA affinity chromatography. The identity of the recombinant protein was confirmed by Western-blot and peptide mass fingerprinting
(PMF) analysis. Protein activity in vitro was investigated by SOD activity assay kit and DNA damage protective assay.
Results: mSOD1 protein was expressed with a final yield of about 60 mg of pure protein per liter of culture medium. After purification, the target protein was > 95 %. The identity of the recombinant protein was confirmed. SOD activity assay showed that the highest activity of the mSOD1 was 3789.0 ± 80.5 U/mg. The present work showed that mSOD1 was effective in protecting DNA from oxidative damage.
Conclusion: High purity recombinant mSOD1 was obtained and characterized, and had high activity in vitro. The study indicates that the mSOD1 may serve as a potential therapeutic agent for those diseases
caused by oxidative stress.
Keywords: Cu/Zn-binding Superoxide dismutase, Expression, Purification, Metal ions, DNA damage
