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Please use this identifier to cite or link to this item: http://repository.li.mahidol.ac.th/dspace/handle/123456789/42891
Title: Enhancement of solubility and specific activity of a cu/zn superoxide dismutase by co-expression with a copper chaperone in escherichia coli
Authors: Warawan Eiamphungporn
Sakda Yainoy
Virapong Prachayasittikul
Mahidol University
Keywords: Biochemistry, Genetics and Molecular Biology
Issue Date: 1-Sep-2016
Citation: Iranian Journal of Biotechnology. Vol.14, No.4 (2016), 243-249
Abstract: © 2016 Kowsar Medical Publishing Company. All rights reserved. Background: Human Cu/Zn superoxide dismutase (hSOD1) is an antioxidant enzyme with potential as a therapeutic agent. However, heterologous expression of hSOD1 has remained an issue due to Cu2+insufficiency at protein active site, leading to low solubility and enzymatic activity. Objectives:The effect of co-expressed human copper chaperone (hCCS) to enhance the solubility and enzymatic activity of hSOD1 in E. coli was investigated in the presence and absence of Cu2+. Materials and Methods: pETDuet-1-hSOD1 and pETDuet-1-hCCS-hSOD1 were constructed and individually transformed into E. coli strain BL21(DE3). The recombinant hSOD1 was expressed and purified using immobilized metal affinity chromatography. The yield and specific activity of hSOD1 in all conditions were studied. Results: Co-expression with hCCS increased hSOD1 solubility at 37°C, but this effect was not observed at 25°C. Notably, the specific activity of hSOD1 was enhanced by 1.5 fold and greater than 3 fold when co-expressed with hCCS at 25°C with and without Cu2+supplement, respectively. However, the chaperone co-expression did not significantly increase the yield of hSOD1 comparable to the expression of hSOD1 alone. Conclusions: This study is the first report demonstrating a potential use of hCCS for heterologous production of hSOD1 with high enzymatic activity.
URI: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85013371982&origin=inward
http://repository.li.mahidol.ac.th/dspace/handle/123456789/42891
ISSN: 23222921
17283043
Appears in Collections:Scopus 2016-2017

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