Reversible pH-Responsive Coacervate Formation in Lipid Vesicles Activates Dormant Enzymatic Reactions |
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Authors: | Celina Love Jan Steinkühler David T Gonzales Naresh Yandrapalli Tom Robinson Rumiana Dimova Dr T-Y Dora Tang |
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Institution: | 1. Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstraße 108, 01307 Dresden, Germany;2. Max Planck Institute of Colloids and Interfaces, 14424 Potsdam, Germany;3. Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstraße 108, 01307 Dresden, Germany
Cluster of Excellence Physics of Life, TU Dresden, 01602 Dresden, Germany
Center for Systems Biology Dresden, Pfotenhauerstraße 108, 01307 Dresden, Germany |
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Abstract: | In situ, reversible coacervate formation within lipid vesicles represents a key step in the development of responsive synthetic cellular models. Herein, we exploit the pH responsiveness of a polycation above and below its pKa, to drive liquid–liquid phase separation, to form single coacervate droplets within lipid vesicles. The process is completely reversible as coacervate droplets can be disassembled by increasing the pH above the pKa. We further show that pH-triggered coacervation in the presence of low concentrations of enzymes activates dormant enzyme reactions by increasing the local concentration within the coacervate droplets and changing the local environment around the enzyme. In conclusion, this work establishes a tunable, pH responsive, enzymatically active multi-compartment synthetic cell. The system is readily transferred into microfluidics, making it a robust model for addressing general questions in biology, such as the role of phase separation and its effect on enzymatic reactions using a bottom-up synthetic biology approach. |
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Keywords: | Flüssig-flüssig-Phasentrennung Koazervate Mikrofluidik pH-Responsivität Protozellen |
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