Expression, Purification,
Characterization and In Vitro Activity of Recombinant
Mouse
Cu/Zn-Binding Superoxide Dismutase (mSOD1)
Zide Zhang 1,2,
Luyuan Huang 3, Zhihui Luo 1, Yong
Liu 4, Aiyun Li 1, Hua Sun 1,
Qiuhong Wu1, Renwang Jiang1,2,5
and Feng Wang 1,2*
1College of Pharmacy, 2Guangdong
Province Key Laboratory of Pharmacodynamic Constituents
of TCM and New Drugs Research, Jinan University,
Guangzhou 510632, 3South China Institute for
Stem Cell Biology and Regenerative Medicine, Guangzhou
Institute of Biomedicine and Health, Chinese Academy of
Sciences, Guangzhou 510633, 4Guangdong Key
Laboratory of Agro-Environment Integrated Control,
Guangdong Institute of Eco-Environmental and Soil
Sciences, Guangzhou 510650, 5Institute of
Traditional Chinese Medicine and Natural Products, Jinan
University, Guangzhou 510632, China
*For correspondence:
Email:
novelfunction@163.com Tel: +86-20-38375022;
Fax: +86-20-85224766
Received: 14 October 2012
Revised accepted:
17 May 2013
Tropical Journal of
Pharmaceutical Research, June 2013;
12(3):
329-334
http://dx.doi.org/10.4314/tjpr.v12i3.9
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-D-thiogalactopyranoside
(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
