Coumarin metabolism by ex vivo cultures of human stool
Bacteria were cultivated from human feces in a cohort of 9 healthy donors to study coumarin metabolism by the human gut microbiome. 16S rRNA amplicon sequencing was used to evaluate uncultured feces and recovery of fecal bacteria in different media types, stability of consortia over time, and changes to community structure upon coumarin treatment. DNA was extracted from the samples with a QIAamp PowerFecal Pro DNA Kit. The V4-V5 region of the 16S rRNA gene was amplified by PCR with barcoded primers. The library was prepared from the PCR amplified products using a Qiagen QIASeq 1-step amplicon kit. The library was sequenced by Illumina MiSeq platform.