Glycomic profiling of the gut microbiota by Glycan-seq

There has been immense interest in studying the relationship between the gut microbiota and human health. Bacterial glycans modulate the cross talk between the gut microbiota and its host. However, little is known about these glycans because of the lack of appropriate technology to study them.Methods: We previously developed a sequencing-based Glycan profiling method called Glycan-seq, which is based on the use of 39 DNA-barcoded lectins. In this study, we applied this technology to analyze the glycome of the intact gut microbiota of mice. Fecal microbiota was incubated with 39 DNA-barcoded lectins exposed to UV, and the number of released DNA barcodes were counted by next-generation sequencing to obtain a signal for each lectin bound to the microbiota. In parallel, the bacterial composition of the gut microbiota was analyzed by 16S rRNA gene sequencing. Finally, we performed a lectin pull-down experiment followed by 16S rRNA gene sequencing to identify lectin-reactive bacteria.