Since the use of solubility enhancer proteins is one of the effective methods to produce active recombinant proteins within Escherichia coli, the development of a novel fusion expression partner that can be applied to various aggregation-prone proteins is of crucial importance. In our previous work, two-dimensional electrophoresis (2-DE) was employed to systematically analyze the E. coli BL21 (DE3) proteome profile in response to heat treatment, and KDPG aldolase (EDA) was identified as a heat-responsive and aggregation-resistant protein. When used as fusion expression partner, EDA significantly increased the solubility of seven aggregation-prone heterologous proteins in the E. coli cytoplasm. The efficacy of EDA as a fusion expression partner was evaluated through the analysis of bioactivity or secondary structure of several target proteins: EDA-fusion expression resulted in the synthesis of bioactive human ferritin light chain and bacterial arginine deiminase and the formation of correct secondary structure of human granulocyte colony stimulation factor.
Bibliographical noteFunding Information:
This study was supported by the 2012 NLRL (National Leading Research Lab.) Project (grant no. 2012R1A2A1A01008085 ) (the main project that supported this work) and the Basic Science Research Program (ERC program, grant no. 2010-0029409 ) of the National Research Foundation of Korea (NRF) . We also appreciate the kind donation of cDNA clone of Mycoplasma arginine deiminase from Prof. Bon Hong Min (College of Medicine, Korea University).
© 2014 Elsevier B.V.
- Escherichia coli BL21
- KDPG aldolase (EDA)
- Solubility enhancer
- Stress-responsive protein
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology