title: Effect of chemical chaperones in improving the solubility of recombinant proteins in Escherichia coli.
abstract: The recovery of active proteins from inclusion bodies usually involves chaotrope-induced denaturation, followed by refolding of the unfolded protein. The efficiency of renaturation is low, leading to reduced yield of the final product. In this work, we report that recombinant proteins can be overexpressed in the soluble form in the host expression system by incorporating compatible solutes during protein expression. Green fluorescent protein (GFP), which was otherwise expressed as inclusion bodies, could be made to partition off into the soluble fraction when sorbitol and arginine, but not ethylene glycol, were present in the growth medium. Arginine and sorbitol increased the production of soluble protein, while ethylene glycol did not. Production of ATP increased in the presence of sorbitol and arginine, but not ethylene glycol. A control experiment with fructose addition indicated that protein solubilization was not due to a simple ATP increase. We have successfully reproduced these results with the N-terminal domain of HypF (HypF-N), a bacterial protein which forms inclusion bodies in Escherichia coli. Instead of forming inclusion bodies, HypF-N could be expressed as a soluble protein in the presence of sorbitol, arginine, and trehalose in the expression medium.
authors.affiliation: Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, Punjab 160 062, India.
authors.forename: Shivcharan
authors.initials: S
authors.lastname: Prasad
authors.forename: Prashant B
authors.initials: PB
authors.lastname: Khadatare
authors.forename: Ipsita
authors.initials: I
authors.lastname: Roy
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism; D018832:Molecular Chaperones / Q000378:metabolism*; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism*; D015203:Reproducibility of Results; D012995:Solubility
chemical_list: D018832:Molecular Chaperones; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate