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2018 | 65 | 2 | 235-240
Article title

Cloning, purification and enzymatic characterization of recombinant human superoxide dismutase 1 (hSOD1) expressed in Escherichia coli

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EN
Abstracts
EN
Superoxide dismutase 1 (SOD1) is a metalloenzyme that catalyzes the disproportionation of superoxide into molecular oxygen and hydrogen peroxide. In this study, the human SOD1 (hSOD1) gene was cloned, expressed and purified. The hSOD1 gene was amplified from a pool of Bxpc3 cell cDNAs by PCR and cloned into expression vector pET-28a (+). The recombinant soluble hSOD1 was expressed in E. coli BL21 (DE3) at 37°C and purified using nickel column affinity chromatography. Soluble hSOD1 was produced with a yield of 5.9 μg/mL medium. As metal ions can have a certain influence on protein structure and activity, we researched the influences of different concentrations of Cu2+ and Zn2+ on hSOD1 activity at induction and the time of activity detection. The results implied that Cu2+ and Zn2+ do not enhance SOD1 expression and solubility; they can, however, improve the catalytic activity at induction. Meanwhile, Cu2+ and Zn2+ also enhanced the enzyme activity at the time of detection. Furthermore, most other bivalent cations had the potential to replace Zn2+ and Cu2+, and also improved enzyme activity at the time of detection.
Publisher

Year
Volume
65
Issue
2
Pages
235-240
Physical description
Dates
published
2018
received
2017-11-08
revised
2018-02-15
accepted
2018-02-17
(unknown)
2018-05-30
Contributors
author
  • School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, P. R. China
  • Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, P. R. China
  • Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, P. R. China
author
  • Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, P. R. China
author
  • Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, P. R. China
author
  • Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, P. R. China
author
  • Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, P. R. China
author
  • College of Biological and Pharmaceutical Engineering, West Anhui University, Moon Island, Lu'an 237012, P. R. China
author
  • Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, P. R. China
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Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv65p235kz
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