Generation, optimisation, validation and implementation of bacterial whole-cell biosensors for the detection of novel antibiotics (360G-Wellcome-102494_Z_13_Z)

Antibiotics remain our primary means of treating bacterial infection. Unfortunately, these agents are increasingly being rendered ineffective by theemergence of antibiotic resistance, and the problem is compounded by the dearth of new antibiotics being discovered. Given the failure of rational/synthetic approaches to antibacterial discovery, drug developers are beginning to revisit the screening of natural products from microorganisms in an attempt to identify new antibiotics. This project will address some of the major deficiencies associated with current natural product screening methodologies, particularly the failure to detect antibiotics at low concentrations or in complex mixtures. This will be achieved through generation of bacterial whole-cell biosensors that produce a quantifiable output when exposed to inhibitors of a target pathway. The O'Neill laboratory has recently created first generation biosensors for detection of inhibitors of the peptidoglycan biosynthesis pathway. This project will seek to optimize the sensitivity of these biosensors using directed evolution and synthetic biology circuits, and will generate, validate and optimize biosensors for other target pathways in the bacterial cell (e.g. protein synthesis). Once established, these biosensors will be used for high-sensitivity screening of actinomycetes and other natural product producers to identify novel antibacterial drug candidates.