Diversity of human colonic butyrate-producing bacteria revealed by analysis of the butyryl-CoA:acetate CoA-transferase gene.
Butyrate-producing bacteria play an important role in the human colon, supplying energy to the gut epithelium and regulating host cell responses. In order to explore the diversity and culturability of this functional group, we designed degenerate primers to amplify butyryl-CoA:acetate CoA-transferase sequences from faecal samples provided by 10 healthy volunteers. Eighty-eight per cent of amplified sequences showed >98% DNA sequence identity to CoA-transferases from cultured butyrate-producing bacteria, and these fell into 12 operational taxonomic units (OTUs). The four most prevalent OTUs corresponded to Eubacterium rectale, Roseburia faecis, Eubacterium hallii and an unnamed cultured species SS2/1. The remaining 12% of sequences, however, belonged to 20 OTUs that are assumed to come from uncultured butyrate-producing strains. Samples taken after ingestion of inulin showed significant (P=0.019) increases in Faecalibacterium prausnitzii. Because several of the dominant butyrate producers differ in their DNA % G+C content, analysis of thermal melt curves obtained for PCR amplicons of the butyryl-CoA:acetate CoA-transferase gene provides a convenient and rapid qualitative assessment of the major butyrate producing groups present in a given sample. This type of analysis therefore provides an excellent source of information on functionally important groups within the colonic microbial community.
Louis
Petra
P
Microbial Ecology Group, Gut Health Division, Rowett Institute of Nutrition and Health, University of Aberdeen, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK. p.louis@abdn.ac.uk
Young
Pauline
P
Holtrop
Grietje
G
Flint
Harry J
HJ
D000085:Acetates / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000378:metabolism; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D002087:Butyrates / Q000378:metabolism*; D015256:Coenzyme A-Transferases / Q000235:genetics* / Q000378:metabolism; D003106:Colon / Q000382:microbiology*; D005051:Eubacterium / Q000235:genetics / Q000378:metabolism; D005673:Fusobacterium / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D006094:Gram-Positive Bacteria / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007444:Inulin / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D000085:Acetates; D000214:Acyl Coenzyme A; D002087:Butyrates; C024343:butyryl-coenzyme A; D007444:Inulin; D015256:Coenzyme A-Transferases; C019803:butyryl-CoA acetoacetate CoA transferase