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Although giant unilamellar vesicles (GUVs) have been extensively studied as synthetic cell-like microcompartments, their applicability as cytomimetic models is severely compromised by low levels of membrane permeability, low encapsulation efficiencies, and high physicochemical instability. Here, we develop an integrated cytomimetic model comprising a macromolecularly crowded interior with high sequestration efficiency and enclosed within a phospholipid membrane that is permeable to molecules below a molecular weight cutoff of ca. 4 kDa. The protocells are readily prepared by spontaneous assembly of a phospholipid membrane on the surface of preformed polynucleotide/polysaccharide coacervate microdroplets and are designated as giant coacervate vesicles (GCVs). Partial anchoring of the GCV membrane to the underlying coacervate phase results in increased robustness, lower membrane fluidity, and increased permeability compared with GUV counterparts. As a consequence, enzyme and ribozyme catalysis can be triggered in the molecularly crowded interior of the GCV but not inside the GUVs when small molecule substrates or inducers are present in the external environment. By integrating processes of membrane-mediated compartmentalization and liquid- liquid microphase separation, GCVs could offer substantial advantages as cytomimetic models, synthetic protocells, and artificial biomolecular microreactors.
Bibliographical noteFunding Information:
We thank the National Natural Science Foundation of China (21735002, 21778016, 22074030, and 21874035) for financial support. The work was partly supported by the BBSRC (BB/P017320/1), the ERC Advanced Grant Scheme (EC-2016-ADG 740235), and BrisSynBio, a BBSRC/EPSRC Synthetic Biology Research Centre (BB/L01386X/1).
© 2021 American Chemical Society
Copyright 2021 Elsevier B.V., All rights reserved.
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1/07/17 → 30/06/22
Project: Research, Parent
1/12/17 → 1/08/21