Development and validation of a microarray for the investigation of the CAZymes encoded by the human gut microbiome

PLoS One. 2013 Dec 31;8(12):e84033. doi: 10.1371/journal.pone.0084033. eCollection 2013.

Abstract

Distal gut bacteria play a pivotal role in the digestion of dietary polysaccharides by producing a large number of carbohydrate-active enzymes (CAZymes) that the host otherwise does not produce. We report here the design of a custom microarray that we used to spot non-redundant DNA probes for more than 6,500 genes encoding glycoside hydrolases and lyases selected from 174 reference genomes from distal gut bacteria. The custom microarray was tested and validated by the hybridization of bacterial DNA extracted from the stool samples of lean, obese and anorexic individuals. Our results suggest that a microarray-based study can detect genes from low-abundance bacteria better than metagenomic-based studies. A striking example was the finding that a gene encoding a GH6-family cellulase was present in all subjects examined, whereas metagenomic studies have consistently failed to detect this gene in both human and animal gut microbiomes. In addition, an examination of eight stool samples allowed the identification of a corresponding CAZome core containing 46 families of glycoside hydrolases and polysaccharide lyases, which suggests the functional stability of the gut microbiota despite large taxonomical variations between individuals.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Retracted Publication

MeSH terms

  • Adult
  • Aged
  • Anorexia / genetics
  • Anorexia / microbiology
  • Bacteria / classification
  • Bacteria / enzymology*
  • Bacteria / genetics
  • Biomarkers / metabolism*
  • Case-Control Studies
  • DNA, Bacterial / genetics
  • Feces / chemistry
  • Feces / microbiology
  • Female
  • Gastrointestinal Tract / metabolism*
  • Gastrointestinal Tract / microbiology
  • Gene Expression Profiling*
  • Glycoside Hydrolases / genetics*
  • Glycoside Hydrolases / isolation & purification
  • Glycoside Hydrolases / metabolism
  • Humans
  • Male
  • Metagenome*
  • Middle Aged
  • Obesity / genetics
  • Obesity / microbiology
  • Oligonucleotide Array Sequence Analysis
  • Polysaccharide-Lyases / genetics*
  • Polysaccharide-Lyases / isolation & purification
  • Polysaccharide-Lyases / metabolism
  • Real-Time Polymerase Chain Reaction
  • Thinness / genetics
  • Thinness / microbiology
  • Young Adult

Substances

  • Biomarkers
  • DNA, Bacterial
  • Glycoside Hydrolases
  • Polysaccharide-Lyases

Grants and funding

This work has been supported by the Mediterranean Infection Foundation (http://www.mediterranee-infection.com). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.