Elucidation of the biodegradation pathways of bis(2-hydroxyethyl) terephthalate and dimethyl terephthalate under anaerobic conditions revealed by enrichment culture and microbiome analysis

[Display omitted] •BHET- and DMT-degradation successfully took place under anaerobic condition.•BHET- and DMT-degrading enzymes were predicted from members of Spirochaeota.•BHET-derived byproducts were utilized by putative acetogens of the class Negativicutes.•Methanol produced from DMT degradation was utilized by methanol-utilizing methanogens. With the globally rising usage of plastics, including polyethylene terephthalate (PET), the environmental risk that disposal of waste plastics to landfills and discharge of microplastics to the marine environment pose have also increased. For example, observation of animal ingestion of fragmented waste plastics (micro- and nano-plastics) has driven awareness for the need of proper environmental risk assessment. In evaluating the biodegradability of PET-derived byproducts and their precursors, most work has focused on hydrolytic enzymes and aerobic organisms that possess such genes, but only few reports on biodegradation in the absence of oxygen (i.e., anaerobic) are available. Here, to elucidate the fate of PET-derived materials under anaerobic environments, a sludge-derived microbial community was cultured with bis(2-hydroxyethyl) terephthalate (BHET) as a model substrate for byproducts of PET degradation and dimethyl terephthalate (DMT) as a potential environmental pollutant discharged from the PET manufacturing process. Metagenome- and metabolome-informed microbiome analyses identified anaerobic BHET and DMT degradation pathways, uncultured organisms affiliated with Spirochaetota and Negativicutes predominant in the BHET-fed cultures, and Methanomethylovorans and Treponema_G predominant in the DMT-fed cultures. Metagenomic analyses newly identified three BHET-degrading and two DMT-degrading enzymes from the genomes of Spirochaeota. In addition, the Negativicutes in the BHET enrichment cultures possessed genes for acetogenically metabolizing EG and/or ethanol. Overall, this study successfully established anaerobic BHET- and DMT-degrading microbial consortia and newly proposed these degradation mechanisms under anaerobic conditions. This study indicated that the cultivation, microbiome, and metabolome analyses can be powerful tools for elucidating consortia capable of degrading plastics-associated waste compounds and the relevant metabolic mechanisms.