When the pH was below 7, the permeability of the cross-linked membrane allowed for substrate/product diffusion, but at basic pH the membrane collapsed and prevented any transport of small molecules. Two different activity tests were performed: 1) GOx and Myo were both entrapped inside the polymersome lumen; 2) GOx and Myo were individually incorporated into the polymersomes and mixed together in solution; in both of the cases the final product formation was monitored by UV–vis spectroscopy. The control over the pH allowed the regulation of the enzymatic cascade (no product was observed at pH 8 in both of the reactive systems), as the diffusion through the membrane was not possible. Moreover, the crosslinking enabled stabilization of the enzymes, which remained active also after 10 days.