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Team:Hong Kong HKU
From 2013.igem.org
Bacterial Microcompartments (BMCs) are closed polyhedral macromolecular complexes with a diameter of 100-150 nm, enclosing enzymes and cofactors for various metabolism reactions. One of the example of such endogenous capsid micro-reactors is the Ethanolamine utilization (Eut) BMC from Salmonella enterica spp.
In Salmonella enterica spp., 5 genes (Eut S, M, N, L, K) encodes thousands of copies of shell proteins to form a heterogenous MCP shell. Such empty, recombinant Eut BMCs has been successfully cloned and expressed in Escherichia coli. In addition, a localization signal has been identified which can be fused into the N-terminus of unnaturally-encapsulated enzymes or proteins to facilitate their localization into the microcompartment
Inspired by these studies, we speculated that this Eut BMC can become a versatile tool if we can:
(a) modify the exterior surface to confer various novel physical, chemical, and biochemical properties to the whole capsule;
(b) localize special enzyme into the BMC for specfic functionalization;
(c) store or recycle useful and harmful molecules into the BMC
As a proof of concept, we displayed His tag on the surface of the Eut BMCs (Fig. 1), and at the same time, we fused the localization signal to polyphosphate kinase 1 from Tannerella forsythia (Fig. 2). After cloning both of the above constructs into Escherichia coli, we hope to demonstrate the one application of such integration by recycling phosphates from the polluted waters aided by the activity of polyphosphate kinase and encapsulating characteristics of Eut BMC.
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