Transforming Crowded Coacervates into Multi-Compartmental Hybrid Microreactors for Practical Enzymatic Catalysis

Advancing the design and construction of artificial protocells with organized complexity, diverse functionality and practical applicability is urgently demanded in vitro synthetic biology and bioengineering but remains a grand challenge. Here, we present a versatile Pickering emulsion-based encapsulation approach to transform membraneless coacervate compartments into robust multicompartmental hybrid microreactors, which concurrently assimilate the expected attributes of hierarchically compartmentalized structure, molecularly crowded environment, selectively permeable ability and mechanically reinforced stability. Single or multiple biological and non-biological catalytic species can be spatially sequestered in specific domains of the hybrid microreactor, enabling spatiotemporal regulation of individual biocatalysis or divergent cascades with high reaction efficiency. As proof of concept, we not only demonstrate the markedly improved catalytic activity (1.9-9.2 folds enhancement), strengthened thermostability (up to 100 °C) and impressive long-term durability (1600 h) of the obtained microreactors in lipase-driven kinetic resolution of alcohol medicine intermediates, but also showcase their superior capability in processing chemo-enzymatic cascade of ketone hydrogenation-kinetic resolution and multi-enzymatic cascade of oxidation reactions. Macromolecular crowding and confinement effects arising from structural features of the hybrid microreactors are identified as the dominant factors for the promotion of catalytic functions.