Engineering a Synthetic Bacterial Consortium of Escherichia coli and Pseudomonas putida for Mixed Plastic Monomer Bioprocessing
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Date
2025-07-17
Authors
Advisor
Liu, Yilan
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Publisher
University of Waterloo
Abstract
Plastics are indispensable to modern life, but their widespread use has created an environmental crisis due to inefficient waste management. Mixed plastic waste, comprising diverse polymers, presents significant recycling challenges due to the high costs of sorting and processing, leading to ecosystem accumulation and harmful by-product generation. This study addresses this issue by engineering a synthetic bacterial consortium (SBC) designed to degrade mixed plastic monomers. The consortium pairs Escherichia coli Nissle 1917, which uses ethylene glycol (EG), a monomer derived from polyethylene terephthalate (PET), as a carbon source, with Pseudomonas putida KT2440, which metabolizes hexamethylenediamine (HD), a monomer from nylon-6,6, as a nitrogen source. Adaptive evolution of the SBC revealed a novel metabolic interaction where P. putida developed the ability to degrade both EG and HD, while E. coli played a critical role in degrading glycolate, mitigating its by-product toxicity. The evolved cross-feeding pattern enhanced biomass production, metabolic efficiency, and community stability compared to monocultures. The consortium’s performance was validated through constraint-based modeling, high-performance liquid chromatography (HPLC), and comprehensive growth assays. These findings highlight the potential of cross-feeding SBCs in addressing complex plastic waste, offering a promising avenue for sustainable bioremediation and advancing future polymer degradation strategies.