Abstract 1025 The devil is in the (molecular) details: Engineering bacterial microcompartments for sustainable chemical production

One promising route to sustainable bioproduction of fuels and chemicals is the engineering of organisms to efficiently convert abundant and low-cost carbon sources to desirable products at high efficiency and low cost.However, many biochemicals are not yet produced biologically due to roadblocks in the cellular biosynthesis process.These roadblocks can include toxicity of intermediates, redox imbalances, and/or loss of product to off pathway reactions.Nature uses spatial organization strategies, such as sequestration in organelles, to alleviate these issues.Even in bacteria, there are organelle-like structures encapsulated by proteins known as bacterial microcompartments that are hypothesized to serve in these functions.However, much is still unknown about the systems, hindering their application in metabolic engineering efforts.With this presentation, I will present how we used a combination of cell-free metabolic engineering, in vivo studies, protein engineering, and modeling to explore the native regulation, assembly and function of microcompartments from a model organism, Salmonella enterica.In so doing, we identified the types of pathways that may benefit from encapsulation, demonstrated the protein shell serves as a barrier to both sequester cofactors and direct flux toward specific enzyme pathway branches, and established design rules for repurposing these organelles for engineered metabolic pathways (e.g., the importance of using a specific encapsulation peptide).Moreover, we were able to show how this may be applied in the context of an acetogen to convert carbon dioxide and carbon monoxide to value-added products more efficiently.