Integrating incompatible tandem photobiocatalysis in artificial cells enables metabolic modulation of natural cells.

Sustaining biological reactions in artificial cells is crucial for their practical integration into living systems, which relies on continuous cofactor supply. Although photocatalysis enables cofactor regeneration in synthetic biological systems, the generated reactive oxygen can deactivate enzymes. Here, we engineer photobiocatalytic artificial cells that modulate hepatocyte metabolism through alleviating alcohol-induced oxidative stress. These artificial cells feature nano-organelles that segregate incompatible modules: one for photocatalytic cofactor regeneration and another for biocatalytic alcohol metabolism. This spatial separation ensures sustainable cofactor provision and protects enzymes from oxidative damage. Co-compartmentalization of alcohol dehydrogenase and aldehyde dehydrogenase within a single nano-organelle enhances cascade reaction efficiency while inhibiting intermediate leakage. When cocultured with hepatocytes, these artificial cells demonstrate excellent biocompatibility and efficiently mitigate oxidative stress from alcohol metabolism. This work advances artificial cells from proof of concept to practical application in living systems. The successful connection of photocatalysis and enzymatic reactions broadens the range of strategies available for chemical synthesis, synthetic biology, and biomedical applications.