Controllable enzymatic hydrolysis in reverse Janus emulsion microreactors.

HYPOTHESIS: The key feature of living cells is multicompartmentalization for enzymatic reactions. Artificial cell-like multicompartments with micro domains are appealing to mimic the biological counterparts. In addition, establishing a sustainable, efficient, and controllable reaction system for enzymatic hydrolysis is imperative for the production of natural fatty acids from animal and plant-based fats.

EXPERIMENTS: Reverse Janus emulsion microreactors, i.e. (W1  + W2 )/O, is constructed through directly using natural fats as continuous phase and aqueous two-phase solutions (ATPS) as inner phases. Enzyme is confined in the compartmented aqueous droplets dominated by the salt of Na2 SO4 and polyethylene glycol (PEG). Enzyme catalyzed ester hydrolysis employed as a model reaction is performed under the conditions of agitation-free and mild temperature. Regulation of reaction kinetics is investigated by diverse droplet topology, composition of inner ATPS, and on-demand emulsification.

FINDINGS: Excellent enzymatic activity toward hydrolysis of plant and animal oils achieves 88.5 % conversion after 3 h. Compartmented micro domains contribute to condense and organize the enzymes spatially. Timely removal of the products away from reaction sites of oil/water interface "pushed" the reaction forward. Distribution and transfer of enzyme in two aqueous lobes provide extra freedom in the regulation of hydrolysis kinetics, with equilibrium conversion controlled freely from 14.5 % to 88.5 %. Reversible "open" and "shut" of hydrolysis is acheived by on-demand emulsification and spontaneous demulsification. This paper paves the way to advancing progress in compartmentalized emulsion as a sustainable and high-efficiency platform for biocatalytic applications.