Abstract

Bacteria capable of dechlorinating the toxic environmental contaminant dichloromethane (DCM, CH2Cl2) are of great interest for potential bioremediation applications. A novel, strictly anaerobic, DCM-fermenting bacterium, “DCMF”, was enriched from organochlorine-contaminated groundwater near Botany Bay, Australia. The enrichment culture was maintained in minimal, mineral salt medium amended with dichloromethane as the sole energy source. PacBio whole genome SMRTTM sequencing of DCMF allowed de novo, gap-free assembly despite the presence of cohabiting organisms in the culture. Illumina sequencing reads were utilised to correct minor indels. The single, circularised 6.44 Mb chromosome was annotated with the IMG pipeline and contains 5,773 predicted protein-coding genes. Based on 16S rRNA gene and predicted proteome phylogeny, the organism appears to be a novel member of the Peptococcaceae family. The DCMF genome is large in comparison to known DCM-fermenting bacteria. It includes an abundance of methyltransferases, which may provide clues to the basis of its DCM metabolism, as well as potential to metabolise additional methylated substrates such as quaternary amines. Full annotation has been provided in a custom genome browser and search tool, in addition to multiple sequence alignments and phylogenetic trees for every predicted protein, http://www.slimsuite.unsw.edu.au/research/dcmf/.

Highlights

  • Dichloromethane (DCM, CH2Cl2) is a toxic environmental contaminant

  • We report the whole genome sequencing and assembly of a novel, DCM-fermenting bacterium, referred to as DCMF

  • The absence of methanogenic populations was confirmed via archaeal specific PCR

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Summary

Introduction

Dichloromethane (DCM, CH2Cl2) is a toxic environmental contaminant. Approximately 70% of all DCM worldwide is of anthropogenic origin, due to its extensive use in industry asHow to cite this article Holland SI, Edwards RJ, Ertan H, Wong YK, Russell TL, Deshpande NP, Manefield MJ, Lee M. 2019. Dichloromethane (DCM, CH2Cl2) is a toxic environmental contaminant. It is currently present at 30% of Superfund National Priority List sites within the United States and its territories (US National Library of Medicine, 2019), and global capacity for DCM continues to steadily increase (Marshall & Pottenger, 2016). As well as being harmful to human health (Agency for Toxic Substances and Disease Registry, 2000), DCM has recently been recognised as a potent greenhouse gas (Hossaini et al, 2017)

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