Enzymes Immobilized in Coacervates Droplets for Reactions in Organic Solvents

Coacervates have emerged as versatile platforms for encapsulating proteins, including enzymes, to enable enzymatic catalysis in aqueous environments. Despite their potential, the application of coacervates is limited by the solubility of substrates in aqueous media. In this study, we present a novel strategy to disperse and stabilize coacervate droplets within water-immiscible organic solvents, forming a stable emulsion of coacervate microdroplets. These droplets exhibit remarkable resistance to coalescence, maintaining their stability even under phase separation and centrifugal forces. This stability is attributed to the robust interfacial film, which not only preserves droplet integrity but also facilitates their separation and redispersion. Remarkably, this coacervate-based system enables the catalysis of reactions in organic solvents while retaining the enzyme in an aqueous microenvironment, thus overcoming the challenges of protein instability in organic media. The coacervate droplets effectively stabilize the enzyme for extended periods, allowing for sustained catalytic activity over several days. Furthermore, the system's design permits the recycling of coacervate droplets, enabling multiple cycles of enzymatic catalysis in fresh or the same organic solvents. This work opens new avenues for the application of coacervate-based systems in biocatalysis, particularly in non-aqueous environments, with potential implications for industrial processes requiring enzyme-mediated transformations in organic media.