Complex waste stream valorization through combined enzymatic hydrolysis and catabolic assimilation by Pseudomonas putida.

Biogenic waste-derived feedstocks for production of fuels, chemicals, and materials offer great potential supporting the transition to net-zero and greater circularity. However, such feedstocks are heterogeneous and subject to geographical and seasonal variability. Here, we show that, through careful strain selection and metabolic engineering, Pseudomonas putida can be employed to permit efficient co-utilization of highly heterogeneous substrate compositions derived from hydrolyzed mixed municipal-like waste fractions (food, plastic, organic, paper, cardboard, and textiles) for growth and synthesis of exemplar bioproducts. Design of experiments was employed to explore the combinatorial space of nine waste-derived monomers, displaying robust catabolic efficiency regardless of substrate composition. Prospective Life-Cycle Assessment (LCA) and Life-Cycle Costing (LCC) illustrated the climate change (CC) and economic advantages of biomanufacturing compared with conventional waste treatment options, demonstrating a 41-62% potential reduction in CC impact. This work demonstrates the potential for expanding treatment strategies for mixed waste to include engineered microbes.