The focus of this project is to evaluate the potential of cold shock promoters for the expression of recombinant proteins in Escherichia coli following a shift to a low temperature culturing condition. The primary objective of this project is to characterize the expression of different model recombinant proteins placed under the control of the cspA cold shock promoter. The strength and inducibility of the cold shock promoter in directing the low temperature synthesis of proteins which are produced in a biologically active conformation under normal growth conditions will be first compared and contrasted to those of the widely used tac promoter. Potential advantages of cold shock promoters are facilitated folding, and a reduced degradation of recombinant proteins. The cspA-controlled production of a recombinant protein susceptible to aggregation, and a recominant protein susceptible to degradation under normal growth conditions will be specifically investigated. The secondary objective of this project is to extend initial shake flask studies to pilot scale fermentations in order to demonstrate the usefulness of cold shock promoters for the large-scale production of useful proteins.
