Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Expression, Purification, Characterization and In Vitro Activity of Recombinant Mouse Cu/Zn-Binding Superoxide Dismutase (mSOD1)

Zide Zhang^1,2, Luyuan Huang³, Zhihui Luo¹, Yong Liu⁴, Aiyun Li¹, Hua Sun¹, Qiuhong Wu¹, Renwang Jiang^1,2,5, 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:- Feng Wang Email: novelfunction@163.com Tel:+862038375022

Received: 14 October 2012 Accepted: 17 May 2013 Published: 12 June 2013

Citation: Zhang Z, Huang L, Luo Z, Liu Y, Li A, Sun H, et al. Expression, Purification, Characterization and In Vitro Activity of Recombinant Mouse Cu/Zn-Binding Superoxide Dismutase (mSOD1). Trop J Pharm Res 2013; 12(3):329-334 doi: 10.4314/tjpr.v12i3.9

© 2013 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

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 af@257;nity chromatography. The identity of the recombinant protein was con@257;rmed 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 @257;nal yield of about 60 mg of pure protein per liter of culture medium. After puri@257;cation, the target protein was > 95 %. The identity of the recombinant protein was con@257;rmed. 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