Polyethylene transformation by a psychrotolerant Rhodococcus strain assessed by transcriptomics and 13C-isotope tracing

Polyethylene is increasingly accumulating in nature, including remote places like the Arctic. While abiotic processes fragment polyethylene in situ , biotic transformation by microorganisms is assumed to occur. However, the enzymes and pathways involved remain poorly characterized. In this study, we used an in-house biobank from cold environments to screen for potential bacteria capable of degrading polyethylene by screening the strains in silico using the database PlasticDB and in vivo using a fluorescence-based assay. Using transcriptomic and proteomic analyses to identify genes in promising candidate strains that encode extracellular enzymes potentially capable of degrading PE, we selected a Rhodococcus erythropolis strain and two of its enzymes: a hypothetical protein (Hypr1) and a lipase family protein (Lip2). Expressing the candidate genes heterologously in Escherichia coli resulted in positive results in the fluorescence-based assay for polyethylene transformation. Applying 13 C-labelled polyethylene for assessing and estimating polyethylene transformation and carbon assimilation, we found that R. erythropolis and both untransformed and recombinant E. coli extracellularly transformed the initially added polyethylene after 70 days. In addition, untransformed E. coli and R. erythropolis converted small, but significant amounts of polyethylene-derived carbon to carbon dioxide. The 13 C-label was also traced into the bacterial biomass of R. erythropolis . Overall, our results provide evidence for biotic transformation of untreated polyethylene and suggests a hypothetical protein and a lipase family protein as two novel enzyme candidates associated with PE transformation.