Elastin-like polypeptides (ELPs) are an attractive, economical alternative to conventional tags for protein purification.  They are comprised of repeating pentapeptides that selectively and reversibly aggregate in response to shifts in osmostic strength and temperature.  These ELPs can be used in conjunction with self-cleaving inteins to form simple, protease-free purification tags.  High cell-density fermentations are important for large scale production of recombinant proteins in general, including those tagged with self-cleaving ELP.  Thus, the development of effective fermentation strategies for these tags is an important goal for the future ofthis technology, where the specific demands of the ELP tag provide unique challenges.  In this paper we report an optimized high cell-density fermentation strategy for the production of ELP-tagged organophosphate hydrolase (OPH).  Specifically, we investigated the protein yields attainable with leaky and induced expression from the T7 promoter at high cell densities in both rich and defined media.  We also looked at the effect of various fermentation parameters on the solubility of this inclusionbody-prone target protein.  These results thus also provide an interesting case study for the production of a highly metabolic-burdensome protein in fermentation cultures.

Large scale production and purification of elastin-like
polypeptide tagged organophophate hydrolase

239th National Meeting (2010)

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Large scale production and purification of elastin-like polypeptide tagged organophophate hydrolase