ProteInProgress: a cellular assembly line for protein manufacturing

Solutions

Several solutions were explored in this project to potentially improve the industrial production of recombinant proteins.

Self-inducible promoters were considered to avoid the usage of inducible systems especially in large-scale industrial bioprocesses, in which protein production has to be triggered by expensive inducer molecules.

Standard and user friendly integrative vectors for E. coli and S. cerevisiae were designed to stably integrate the expression systems of interest in the microbial host genome and to eliminate the need of expensive selection techniques, such as antibiotics or auxotrophic media.

Finally, an "in-cell" protein purification system was implemented using BioBrick parts: PolyHydroxyAlkanoate (PHA) granules were used as a substrate for PHA-binding peptides (Phasins) fused to the target protein, while a pH-based self-cleaving peptide (Intein) was used instead of a protease cleavage site. This solution can thus replace the usage of expensive affinity resins/columns and proteases.

Self-inducible promoters