The discovery of membraneless organelles (MLOs) formed by liquid-liquid phase separation raised many questions about the spatial organization of biomolecular processes in cells, but also offered a new tool to mimic cellular media. Since disordered and charged protein domains are often necessary for phase separation, coacervates can be used as models both to understand MLO regulation and to develop dynamic cellular-like compartments. A versatile way to turn passive coacervate droplets into active and dynamic compartments is by introducing enzymatic reactions that affect parameters relevant for complex coacervation, such as the charge and length of the components. However, these reactions strictly take place in a heterogeneous medium, and the complexity thereof is hardly addressed, making it difficult to achieve true control. In this chapter we help close this gap by describing two coacervate systems in which enzymatic reactions endow coacervate droplets with a dynamic character. We further highlight the technical challenges posed by the two-phase systems and strategies to overcome them.
Keywords: Chemically active systems; Coacervates; Enzymatic control; Membraneless organelles; Phase separation; Synthetic cell.
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