Function and Phylogeny of Bacterial Butyryl Coenzyme A:Acetate Transferases and Their Diversity in the Proximal Colon of Swine.
Studying the host-associated butyrate-producing bacterial community is important, because butyrate is essential for colonic homeostasis and gut health. Previous research has identified the butyryl coenzyme A (CoA):acetate-CoA transferase (EC 2.3.8.3) as a gene of primary importance for butyrate production in intestinal ecosystems; however, this gene family (but) remains poorly defined. We developed tools for the analysis of butyrate-producing bacteria based on 12 putative but genes identified in the genomes of nine butyrate-producing bacteria obtained from the swine intestinal tract. Functional analyses revealed that eight of these genes had strong But enzyme activity. When but paralogues were found within a genome, only one gene per genome encoded strong activity, with the exception of one strain in which no gene encoded strong But activity. Degenerate primers were designed to amplify the functional but genes and were tested by amplifying environmental but sequences from DNA and RNA extracted from swine colonic contents. The results show diverse but sequences from swine-associated butyrate-producing bacteria, most of which clustered near functionally confirmed sequences. Here, we describe tools and a framework that allow the bacterial butyrate-producing community to be profiled in the context of animal health and disease.



Butyrate is a compound produced by the microbiota in the intestinal tracts of animals. This compound is of critical importance for intestinal health, and yet studying its production by diverse intestinal bacteria is technically challenging. Here, we present an additional way to study the butyrate-producing community of bacteria using one degenerate primer set that selectively targets genes experimentally demonstrated to encode butyrate production. This work will enable researchers to more easily study this very important bacterial function that has implications for host health and resistance to disease.
Food Safety and Enteric Pathogens Research Unit, National Animal Disease Center, Agricultural Research Service, Ames, Iowa, USA Interdepartmental Microbiology Graduate Program, Iowa State University, Ames, Iowa, USA.
Julian
J
Trachsel
Infectious Bacterial Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, Ames, Iowa, USA.
Darrell O
DO
Bayles
Food Safety and Enteric Pathogens Research Unit, National Animal Disease Center, Agricultural Research Service, Ames, Iowa, USA.
Torey
T
Looft
Food Safety and Enteric Pathogens Research Unit, National Animal Disease Center, Agricultural Research Service, Ames, Iowa, USA.
Uri Y
UY
Levine
Food Safety and Enteric Pathogens Research Unit, National Animal Disease Center, Agricultural Research Service, Ames, Iowa, USA heather.allen@ars.usda.gov.
Heather K
HK
Allen
D000085:Acetates / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D002087:Butyrates / Q000378:metabolism*; D015256:Coenzyme A-Transferases / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D003106:Colon / Q000382:microbiology*; D017931:DNA Primers; D005243:Feces / Q000382:microbiology; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D064307:Microbiota / Q000235:genetics / Q000502:physiology; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S; D013552:Swine / Q000382:microbiology*
D000085:Acetates; D000214:Acyl Coenzyme A; D002087:Butyrates; D017931:DNA Primers; D012336:RNA, Ribosomal, 16S; C024343:butyryl-coenzyme A; D015256:Coenzyme A-Transferases