Hydrophilicity and Electrostatic Complementarity Controlled Substrate Selectivity of Enzymes Inside Reverse Micelles

Harnessing enzymes for synthetic processes in non‐native environments while maintaining selectivity across different substrates is challenging. Here, we report amphiphilic random copolymers that self‐assemble into reverse micelles, which can then sequester enzymes to form nanoreactors and maintain catalytic activity in organic solvents. Photo‐responsive functional groups in scaffold design offer spatial and temporal control over crosslinking density and reaction kinetics. Variations in p K a and density of charged functional groups were explored to improve protein‐polymer complexation as well as induce charge‐based selectivity. A library of peptide substrates varying in partition coefficient, charge, and specificity constants has been synthesized to further delineate the driving force for selectivity. The findings here provide insights on structural factors for polymeric assemblies to enhance substrate selectivity without altering enzyme structure under non‐native conditions.