Computational protocol: Draft Genome Sequence of Sphingobacterium sp. CZ-UAM, Isolated from a Methanotrophic Consortium

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[…] Methane (CH4) is produced by natural and human-related activities; it is one of the main greenhouse gases, with a 25-fold greater global warming potential than CO2 (). CH4-consuming bacteria are known as methanotrophs, and they can grow using CH4 as their only carbon source. Biological treatments are the best option to eliminate CH4 as an atmospheric contaminant when its percentage is below the explosive limit (5% in air) (, ). There are many sources of cheap and diluted CH4 that can be used as feedstock (). Therefore, it is environmentally urgent to study new methanotrophs with higher CH4 uptake rates and to explore their metabolic potential to convert CH4 into added-value products (, , ). Here, we report the genome sequence of the novel potential methanotroph Sphingobacterium sp. CZ-UAM, which was isolated from a methanotrophic consortium from a wastewater treatment plant (). The isolated strain is able to grow on mineral medium with a 5% CH4 atmosphere as its sole carbon source.Genomic DNA from Sphingobacterium sp. CZ-UAM was extracted by an in-house phenol-chloroform protocol, RNase treated, and gel verified for quality. Then, a paired-end (average size, 527 bp) genomic DNA library was generated using commercial kits according to the manufacturers’ instructions (TruSeq Nano, Illumina, San Diego, CA, USA). The DNA library quality was verified using a 2100 BioAnalyzer (Agilent Genomics, Santa Clara, CA, USA). The genome sequencing was performed using an Illumina HiSeq 2500 system through the standard rapid sequencing protocol for cluster generation and sequencing by synthesis, which resulted in a total of 3,891,197 reads for a depth coverage of 300×. Genome assembly was performed using SPAdes version 3.9.0 () and IDBA_UD version 1.1.1 (). Then, SSPACE version 3.0 () and Metassembler version 1.5 () were used for merging and scaffolding the obtained contigs. After genome assembly and scaffolding, 12 contigs were obtained. Phylogenetic analysis of the 16S rRNA was performed by extracting the previously annotated 16S ribosomal gene sequence, as well as the annotated Sphingobacterium sequences in the NCBI database. All the sequences were aligned using Muscle software () and edited using Gblocks () prior to phylogenetic tree construction using Bayesian inference. The analyses indicate that the isolated strain is a member of the Sphingobacterium genus. The analysis of the full 16S rRNA gene sequence with the EZBioCloud database indicates higher identity with S. pakistanence (97.1%) and S. canadense (96.8%), S. ginsenosidimutans (96.46%), and S. detergens (95.39%) (). Of the few available Sphingobacterium sequencing projects, none of them have been reported as having the potential to be a methanotroph. The availability of the Sphingobacterium sp. CZ-UAM genome will contribute to the study of the Sphingobacterium genus and will provide information on the molecular basis of this strain’s ability to consume CH4 as a sole carbon source. […]

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