Identification and Metabolic Analysis of Uncultivable Bacterium by means of Metagenomic Analysis

Abstract

Uncultivable bacteria are nearly impossible to identify and characterize, due to the inability to reliably grow them in a laboratory setting. One approach to identifying uncultivable bacteria is to preform genomic shotgun sequencing. Genomic data for uncultured bacteria is growing in availability rapidly, due to the ever lowing cost of sequencing efforts. Genomic data was sequenced from an anoxic sulfide and sulfur-rich spring in southwestern Oklahoma, and a bacterial genome was found to be a candidate for a new phylum based on a phylogenetic analysis of 120 single markers. While this bacterium is uncultivable in the laboratory, it is possible to predict some of its traits from a genomic analysis.  The genome of this bacterium was assembled by first assembling the shotgun sequencing into a MAG, then binning the contigs into individual genomic assemblies, and the genome completeness was assessed. Transcripts were then produced to aid in the prediction of the bactieriums structural properties, and then metabolic patways were constructed using KEGG. This bacterium is predicted to be a slow growing, gram negative rod, with a type four pilis. This bacterium is also predicted to be an anoxic heterotrophic bacterium that is fermentative. There are also detailed prediction on the metabolic processes that the bacterium carries out, and substrate utilization.