Through the proteome analysis of Escherichia coli BL21(DE3), we previously identified the stress-responsive protein, arsenate reductase (ArsC), that showed a high cytoplasmic solubility and a folding capacity even in the presence of stress-inducing reagents. In this study, we used ArsC as an N-terminal fusion partner to synthesize nine aggregation-prone proteins as water-soluble forms. As a result, solubility of the aggregation-prone proteins increased dramatically by the fusion of ArsC, due presumably to its tendency to facilitate the folding of target proteins. Also, we evaluated and confirmed the efficacy of ArsC-fusion expression in making the fusion-expressed target proteins have their own native function or structure. That is, the self-assembly function of human ferritin light chain, l-arginine-degrading function of arginine deiminase, and the correct secondary structure of human granulocyte colony stimulating factor were clearly observed through transmission electron microscope analysis, colorimetric enzyme activity assay, and circular dichroism, respectively. It is strongly suggested that ArsC can be in general an efficient fusion expression partner for the production of soluble and active heterologous proteins in E. coli.
Bibliographical noteFunding Information:
We thank Professor Hang Chul Shin at Soongsil University for kindly providing the gene clones of hG-CSF. This study was supported by the National Research Laboratory Project of the Ministry of Education, Science and Technology (grant no. ROA-2007-000-20084-0 ). This work was also supported by the Second Brain Korea 21 Project . Additional supports from the Microbial Genomics and Applications Center at KRIBB (grant no. 2010-K000599 ), the Public Welfare & Safety Research Program (grant no. 2010-0020778 ), the Basic Science Research Program (ERC program, grant no. 2010-0029409 ), and the National Research Foundation of Korea (NRF) (grant no. 2010-0027771 ) are also appreciated. This work was also supported by the R&D Program of MKE/KEIT (grant no. 10031969 ).
- Escherichia coli BL21(DE3)
- Folding enhancer
- Fusion partner
- Stress-resistant protein
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology