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Molecular Foundry

HIGHLIGHTSRapid In Vitro Assembly and Loading of Bacterial Microcompartment Shells
March 4, 2025

Scientific Achievement

Schematic representation of the chaotrope-based approach to in vitro BMC shell assembly. (A). BMC-H hexamers form sheets that disassemble into assembly competent tiles upon the addition of urea. (B). Combining solutions of heterologously expressed and purified BMC-H and BMC-T shell proteins results in the IVA of HT shells. (C). Combining solutions of all three BMC shell proteins results in the IVA of HTP shells.

The development of an in vitro assembly method for bacterial microcompartments (BMC) from their protein building blocks, enabling the encapsulation of biotic and abiotic cargo.

Significance and Impact

This work presents a new, efficient, and fast method for in vitro assembly of BMC shells and broadens the scope for bioengineered shells that can be designed for specific applications, including drug delivery or bioengineered nanoreactors.

Research Details

  • The researchers utilized urea as a chaotropic agent to control self-assembly, enabling large scale construction of BMC shells.
  • The team demonstrated the viability of the BMC shells for encapsulating catalytic cargo by loading biotic (a non-native enzyme) and abiotic (a photosynthesizer) molecules.
Range, K.L., Chiang, T.K., Pramanik, A., Landa, J.F., Snyder, S.N., Zuo, Z., Tiede, D.M., Utschig, L.M., Hegg., E.L., Sutter, M., Kerfeld, C.A., Ralston, C.Y. ACS Nano. 19, 12, 11913 – 11923. (2025) DOI:10.1021/acsnano.4c15538

Research Summary

Bacterial microcompartments (BMCs) are proteinaceous organelles that self-assemble into selectively permeable shells that encapsulate enzymatic cargo. BMCs enhance catalytic pathways by reducing crosstalk among metabolites, preventing harmful intermediates from leaking into the cytosol and increasing reaction efficiency via enzyme colocalization. The intrinsic properties of BMCs make them attractive for biotechnological engineering. However, in vivo expression methods for shell synthesis have significant drawbacks that limit the potential design space for these nanocompartments. Here, the researchers describe the development of an efficient and rapid method for the in vitro assembly of BMC shells from their protein building blocks. Their method enables large-scale construction of BMC shells by utilizing urea as a chaotropic agent to control self-assembly and provides an approach for encapsulation of both biotic and abiotic cargo under a broad range of reaction conditions. The researchers demonstrate an enhanced level of control over the assembly of BMC shells in vitro and expand the design parameter space for engineering BMC systems with specialized and enhanced catalytic properties.

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