Spontaneous Structuration in Coacervate-Based Protocells by Polyoxometalate-Mediated Membrane Assembly

Molecularly crowded, polyelectrolyte/ribonucleotide-enriched membrane-free coacervate droplets are transformed into membrane-bounded sub-divided vesicles by using a polyoxometalate-mediated surface-templating procedure. The coacervate to vesicle transition results in reconstruction of the coacervate micro-droplets into novel three-tiered micro-compartments comprising a semi-permeable negatively charged polyoxometalate/polyelectrolyte outer membrane, a sub-membrane coacervate shell, and an internal aqueous lumen. We demonstrate that organic dyes, ssDNA, magnetic nanoparticles and enzymes can be concentrated into the interior of the micro-compartments by sequestration into the coacervate micro-droplets prior to vesicle formation. The vesicle-encapsulated proteins are inaccessible to proteases in the external medium, and can be exploited for the spatial localization and coupling of two-enzyme cascade reactions within single or between multiple populations of hybrid vesicles dispersed in aqueous media.