TY - JOUR
T1 - Bioreactor droplets from liposome-stabilized all-aqueous emulsions
AU - Dewey, Daniel C.
AU - Strulson, Christopher A.
AU - Cacace, David N.
AU - Bevilacqua, Philip C.
AU - Keating, Christine D.
N1 - Funding Information:
This work was supported by the U.S. Department of Energy, Office of Basic Energy Sciences, Division of Materials Sciences and Engineering under Award # DESC0008633 (development and characterization of liposome-stabilized emulsions) and by the NASA Exobiology program, grant number NNX13AI01G (RNA compartmentalization and cleavage reactions). We thank the M. Rolls laboratory for use of their microscope upright Olympus FV1000 confocal microscope. We thank J. Bingaman of the Bevilacqua Lab for assistance in PAGE data analysis.
PY - 2014/8/20
Y1 - 2014/8/20
N2 - Artificial bioreactors are desirable for in vitro biochemical studies and as protocells. A key challenge is maintaining a favourable internal environment while allowing substrate entry and product departure. We show that semipermeable, size-controlled bioreactors with aqueous, macromolecularly crowded interiors can be assembled by liposome stabilization of an all-aqueous emulsion. Dextran-rich aqueous droplets are dispersed in a continuous polyethylene glycol (PEG)-rich aqueous phase, with coalescence inhibited by adsorbed ∼130-nm diameter liposomes. Fluorescence recovery after photobleaching and dynamic light scattering data indicate that the liposomes, which are PEGylated and negatively charged, remain intact at the interface for extended time. Inter-droplet repulsion provides electrostatic stabilization of the emulsion, with droplet coalescence prevented even for submonolayer interfacial coatings. RNA and DNA can enter and exit aqueous droplets by diffusion, with final concentrations dictated by partitioning. The capacity to serve as microscale bioreactors is established by demonstrating a ribozyme cleavage reaction within the liposome-coated droplets.
AB - Artificial bioreactors are desirable for in vitro biochemical studies and as protocells. A key challenge is maintaining a favourable internal environment while allowing substrate entry and product departure. We show that semipermeable, size-controlled bioreactors with aqueous, macromolecularly crowded interiors can be assembled by liposome stabilization of an all-aqueous emulsion. Dextran-rich aqueous droplets are dispersed in a continuous polyethylene glycol (PEG)-rich aqueous phase, with coalescence inhibited by adsorbed ∼130-nm diameter liposomes. Fluorescence recovery after photobleaching and dynamic light scattering data indicate that the liposomes, which are PEGylated and negatively charged, remain intact at the interface for extended time. Inter-droplet repulsion provides electrostatic stabilization of the emulsion, with droplet coalescence prevented even for submonolayer interfacial coatings. RNA and DNA can enter and exit aqueous droplets by diffusion, with final concentrations dictated by partitioning. The capacity to serve as microscale bioreactors is established by demonstrating a ribozyme cleavage reaction within the liposome-coated droplets.
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U2 - 10.1038/ncomms5670
DO - 10.1038/ncomms5670
M3 - Article
C2 - 25140538
AN - SCOPUS:84907354377
SN - 2041-1723
VL - 5
JO - Nature communications
JF - Nature communications
M1 - 4670
ER -