Understanding the molecular architecture of bacterial microcompartments to enable their rational exploitation in synthetic biology

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Jon Marles-Wright, Adam D. Crawshaw, Alexandra Faulds-Pain, Mingzhi Liang, Alison C. Pitts, Michael B. Prentice, Juri Rappsilber, Laura R. Tuck, Juan Zhou

Edinburgh University, United Kingdom

Bacterial microcompartments (BMCs) are proteinaceous metabolic compartments that are found in a wide variety of bacterial species and enable the efficient breakdown and utilisation of various environmental carbon and nitrogen sources. These compartments have a modular organisation, with an external shell constructed from a family of structurally related proteins. The shell proteins recruit and encapsulate the enzymes required for their function, while creating a semi-permeable barrier between the BMC lumen and the cytosol. The diversity and modularity of BMCs presents a platform with great potential for exploitation in synthetic biology. BMCs could be used as containers for heterologous biosynthetic pathways; as containers for the production of toxic proteins; or as scaffolds for the production of metallic nanoparticles. A major barrier to the use of BMCs in synthetic biology is the lack of understanding of the principles underlying their formation and substrate transport across the shell. A combination of 3D-proteomics and biophysical characterisation is being used to map the interactions that specify BMC formation and enzyme recruitment. The specificity and substrate selectivity of the BMC shell proteins and the encapsulated enzymes is being studied using biochemical and biophysical methods. Using microcompartments with different protein composition and function, taken from various source organisms, conserved and substrate specific features will be determined. The ultimate outcome of this research will be a model of BMC organisation and function that can be used as the basis for the rational design of synthetic BMCs.