Connecting the worlds of bio-polymerizations and life mimesis, here we report the development of biocatalytic polymerization-induced self-assembly (bioPISA) and its application as a bio-compatible methodology to synthesise synthetic cell-like structures. The scope of conditions for bioPISA was investigated, demonstrating its robustness in different media useful for biological applications. To achieve biomimetic systems, different enzymes were encapsulated in GUV synthesised by bioPISA, acting as microreactors, thus showing that the activity of the encapsulated enzymes was retained. Other proteins were instead used to trigger the formation of cytoskeleton-like structures and for osteoblast-inspired biomineralisation. Moreover, as PISA achieves high encapsulation efficiencies, it is well-suited for the creation of complex cell mimics. Thus, bacterial cell lysate and different plasmids were encapsulated inside the GUVs, effectively reconstituting a functional cytoplasm with a simplified chromosome within an artificial cell membrane. The resulting artificial cells were capable of expressing several proteins, which were, in turn, used to alter the artificial cell's internal structure.