Genome Mining and Pangenome Analysis of the Stutzerimonas Genus: a Novel Source of Plastic-Degrading Enzymes

Abstract Nowadays, finding new sustainable ways to combat plastic pollution is a pressing challenge. Here, we provide a comprehensive genome mining analysis of 284 publicly available Stutzerimonas genomes for potential PET-active enzymes (PETases). While Stutzerimonas is a relatively newly established genus, it emerges as an interesting candidate in the search for novel biocatalysts. Hence, the first pangenome assessment of this genus based on its high-quality publicly available genomes was performed. An increasingly open pangenome was revealed, suggesting the versatility and adaptability of these strains to a variety of ecological niches. Moreover, functional characterisation of a new isolate, Stutzerimonas frequens VG-9, was carried out, confirming that enzymes found via in silico analyses may indeed display activity towards different polyesters. In summary, this study provides insights into the diversity of PETase homologues within still underexplored bacterial hosts, offering new perspectives for enzyme discovery in the Pseudomonadaceae family. Impact Statement Microbial enzymes known as PETases have emerged as promising candidates for the biological degradation of PET. This study investigated the potential of underexplored bacterial genera by genome mining of PETase homologues. Our findings provide new insights into the distribution of PETase-like enzymes in the Pseudomonadaceae family, offering a more comprehensive view of their plastic degradation capacity. These results hold practical implications for the development of optimized enzyme discovery strategies, while also highlighting the vast genetic plasticity of Pseudomonadaceae. We also provided the first report on the Stutzerimonas pangenome and insights into the enzymatic activity towards polyesters of a newly isolated strain. Hence, the role of this genus as a highly adaptable and versatile entity was reinforced, further disclosing it as a potential source of novel biocatalysts. Data Summary The genome of S. frequens VG9 has been deposited in Genbank under the accession number SAMN49487720. The accession numbers of all analyzed genomes are listed in Tables S2 and S3 (available in the online Supplementary Material).