Production of recombinant methionine-containing elastin-like polypeptides in a fermenter using ECPM1 medium

Elastin-like polypeptides (ELPs) are recombinant protein-like polymers whose macromolecular structure can be precisely controlled through genetic manipulation of their sequence and length. Their lower critical solution temperature (LCST) phase behavior facilitates purification via chromatography-free techniques and can be explored for self-assembly. As a result, ELPs are extensively investigated for diverse biological, biomedical, and biotechnological applications. So far, ELPs have mostly been isolated from bacteria grown in flasks or fermenters containing complex media that only yield limited amounts of biomass. We herein explored the use of the semi-defined ECPM1 medium, known to limit the accumulation of toxic metabolites and rich in glycerol as a low energy carbon source, to produce ELPs of different chain lengths and containing oxidation-sensitive methionine residues. We report the optimized bioproduction using ECPM1 of ELP[M₁V₃-n] with n = 20, 40, 80 in a fermenter in good yields and confirm their intact protein sequence using various chemical characterization techniques.