Engineering Synthetic Operons For Enhanced Acid Resistance In Bacteria

This is a ongoing master's degree project in which combinatory Synthetic Biology approaches are employed to engineer bacteria, aiming the development of strains with improved resistance to acidity. Twenty-seven unique synthetic operons were built by assembling three metagenomic genes originally recovered from the metagenome of an polluted acidic river (https://doi.org/10.1111/1462-2920.12021) and synthetic ribosome binding site sequences from the community collection of prokaryotic constitutive RBS of the iGEM project (http://parts.igem.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Co...), which led to different genic dosages between the constructions that are only dependent on the fine translation regulation of the polycistronic mRNA, since the expressed proteins remain the same. Our combinatory approach resulted in bacterial strains showing a synergistic effect between the expressed proteins, which resulted in higher survival percentages than any of the individual genes. After the initial bench scale characterization of these strains, selected constructs will be evaluated in different industrially relevant hosts, and tested both for the orthogonality of constructed cassettes and their stability under conditions that are more similar to the needed by the industry.