High-Throughput Fingerprinting of Polymer Biodegradation across Phylogenetically Diverse Microorganisms

Abstract With the growing accumulation of plastic waste, focus has been directed toward biodegradable polymers; however, understanding their performance across diverse microbial communities requires species-level degradation data, which throughput-limited studies of undefined or limited consortia have only partially provided, preventing the cross-species and cross-polymer comparisons driving mechanistic understanding. This work presents an integrative framework fingerprinting polymer biodegradation across phylogenetically diverse microorganisms. A modified high-throughput clear-zone assay was adapted to physiologically diverse species through minimal media formulations and used to characterize fragmentation across 19 microorganisms and 14 commercial polymers. Paucimonas lemoignei, Priestia megaterium, and Pseudomonas stutzeri showed expansive fragmentation capability. Poly(1,4-butylene adipate), degraded by 11 of 19 species, proved most universally degradable, while poly(butylene adipate-co-terephthalate), poly(ε-caprolactone), and poly(lactic acid) were degraded more selectively. Plate-based screening of protease, esterase, and urease activity supported the aliphatic-ester selectivity pattern. MicroResp quantification, addressing the often-conflated distinction between fragmentation and mineralization, confirmed mineralization in 63.3% (19 of 30) fragmentation-positive pairs. Comparative genomics identified 465 depolymerase homologues, 187 with predicted signal peptides. Distinct strategies─P. lemoignei’s four near-identical PhaZ paralogs versus P. stutzeri’s divergent broad-substrate α/β hydrolases─explain their comparable polyester breadth. Overall, this framework establishes a foundation for systematic accumulation of microbial biodegradation fingerprints.