(724f) Encapsulating Proteins Into Bacterial Microcompartments

Bacteria have been favored for the production of biologically derived chemicals due to their rapid growth, robustness to environmental conditions, simple genetics, and capacity to overexpress recombinant proteins.  However, problems such as cellular toxity and undesired side reactions prohibit or greatly reduce yields for many products.  Targeting the synthesis of these products to bacterial microcompartments can solve these problems by sequestering toxic intermediates and excluding non-relevant enzymes from the compartment.  Furthermore, the enhanced kinetics achieved by concentrating substrates and enzymes can improve yields.  Here, we show the encapsulation of GFP to the propanediol (PDU) microcompartment by fusion of an N-terminal signal peptide and evaluate the efficiency of encapsulation by flow cytometry.  We then investigate the role of charged amino acids in the N-terminal signal peptides for efficient encapsulation.  Understanding the encapsulation of heterologous proteins is the first step in targeting multienzyme pathways to bacterial microcompartments to serve as nanobioreactors for the synthesis of bioproducts in bacteria.