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Functional genomics in a microbe that degrades and metabolizes PET plastic
Summary
This study applies functional genomics tools to characterize the molecular mechanisms by which a plastic-degrading microbe breaks down and assimilates synthetic polymers. By mapping the genes and metabolic pathways involved, the research provides a foundation for engineering microorganisms with enhanced plastic biodegradation capabilities.
Abstract Piscinibacter sakaiensis (formerly Ideonella sakaiensis ) was the first bacterial species known to both completely degrade and assimilate polyethylene terephthalate (PET). However, the absence of efficient genetic tools has limited direct engineering of this organism, forcing most efforts to rely on heterologous expression of PET-degrading enzymes in model hosts. Here, we establish foundational genetic tools to engineer P. sakaiensis . We identify a functional plasmid origin of replication, multiple new selectable markers, and a transposon system for the strain. We use these tools to construct a genome-wide, barcoded transposon mutant library for pooled high-throughput functional screens. We apply this mutant library to growth on PET and identify metabolic and physiological genes that impact PET biodegradation. We also use this library to reveal mutants with improved DNA uptake for genome engineering. Together, these advances provide a platform for functional genomics in P. sakaiensis and positions this naturally evolved plastic-degrading bacterium as an engineerable chassis for synthetic biology and sustainable materials research.