Engineering Transcriptional Regulation and Signal Peptides to Control the Catalytic Activity of Bacterial Microcompartments (Rapid Fire)

Bacterial microcompartments are subcellular bacterial organelles bound by a porous protein membrane. Enteric and other bacteria use these organelles to sequester certain metabolic pathways to enhance growth and pathogenicity. We aim to develop a tunable nanobioreactor system based on these protein structures to spatially organize heterologous biosynthetic pathways in bacterial hosts. To this end, we develop tools to control both the formation of the microcompartments and their enzymatic content.

            By engineering the transcriptional regulation of the microcompartments, we can orchestrate their formation and exercise control over the loading of heterologous proteins to the nanobioreactor. We also engineer the N-terminal signal sequences responsible for enzyme targeting to the compartments, allowing fine control over the loading phenotype of the reactors. Our results have implications both for the controlled encapsulation of heterologous synthetic pathways and for Salmonella and E. coli pathogenesis in vivo, to which the bacterial microcompartments natively contribute.