Enzyme loading of polymersomes requires permeability to enable them to interact with the external environment, typically requiring addition of complex functionality to enable porosity. Herein, we describe a synthetic route towards intrinsically permeable polymersomes loaded with functional proteins using initiator-free visible light-mediated polymerization-induced self-assembly (photo-PISA) under mild, aqueous conditions using a commercial monomer. Compartmentalization and retention of protein functionality was demonstrated using green fluorescent protein as a macro-molecular chromophore. Catalytic enzyme-loaded vesicles using horseradish peroxidase and glucose oxidase were also prepared and the permeability of the membrane towards their small molecule substrates was revealed for the first time. Finally, the interaction of the compartmentalized enzymes between separate vesicles was validated by means of an enzymatic cascade reaction. These findings have a broad scope as the methodology could be applied for the encapsulation of a large range of macromolecules for advancements in the fields of nanotechnology, biomimicry and nanomedicine.