Despite global issues with plastic accumulation, including of the versatile plastic polyurethane (PU) found in a multitude of materials, recycling strategies are not meeting need. Bacteria such as Pseudomonas protegens can degrade plastics like PU and could provide innovative strategies designed by nature to help recycle this material. However, the fundamental genetic basis of polyurethane biodegradation is incomplete. Functional genomics techniques, such as transposon insertion sequencing (TIS) allow for high throughput screening of the functional role of every gene in a bacterial genome for a particular selective condition, like plastic exposure. We have employed this powerful approach, to gain a more complete understanding of the regulatory and metabolic pathways involved in bacterial polyurethane degradation.