An enhanced phage-derived lytic platform for programmable autolysis in E. coli

Engineered bacteria are increasingly being harnessed as programmable micromachines capable of sensing specific environmental cues and triggering targeted responses, such as local payload release through autolysis. However, the expression of toxic autolysis-inducing proteins imposes strong selective pressure on bacteria, often leading to circuit inactivation and lysis escape. Here, we developed a multi-input phage-derived lytic architecture based on the principle of division of labor, where lytic strength, and therefore toxicity, is spread across two inducible modules encoding different cell wall–disrupting proteins, increasing bacterial fitness in growth-permissive conditions and eliciting efficient lysis upon co-expression. Following functional and morphological characterization of lysis through optogenetic induction, we demonstrate the scalability of our platform to respond to inputs of the tumor microenvironment. Our work establishes a novel lytic architecture enabling the on-demand release of diverse payloads, offering a versatile and controllable platform for broad targeted applications in synthetic biology, medicine and environmental biotechnology.