A novel class of CoA-transferase involved in short-chain fatty acid metabolism in butyrate-producing human colonic bacteria

• Published in: Microbiology (Society for General Microbiology) Vol. 152; no. 1; pp. 179 - 185
• Main Authors: Charrier, Cedric, Duncan, Gary J, Reid, Martin D, Rucklidge, Garry J, Henderson, Donna, Young, Pauline, Russell, Valerie J, Aminov, Rustam I, Flint, Harry J, Louis, Petra
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.01.2006; Society for General Microbiology
• Subjects: Acyl Coenzyme A - metabolism; Amino Acid Sequence; Bacteria - enzymology; Bacteria - genetics; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Biological and medical sciences; Butyrates - metabolism; Coenzyme A-Transferases - genetics; Coenzyme A-Transferases - metabolism; Escherichia coli; Fatty Acids - metabolism; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Metabolism. Enzymes; Microbiology; Molecular Sequence Data; Species Specificity; Substrate Specificity; Human; Polymerase chain reaction; Purification; Enzyme; Transferases; Proteomics; CoA-transferases; Gene overexpression; Lipids; Bacteria; Fatty acids; Butyrate
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/mic.0.28412-0

1 Gut Health Division, Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK 2 Scientific Support Division, Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK Correspondence Petra Louis p.louis{at}rowett.ac.uk Bacterial butyryl-CoA CoA-transferase activity plays a key role in butyrate formation in the human colon, but the enzyme and corresponding gene responsible for this activity have not previously been identified. A novel CoA-transferase gene is described from the colonic bacterium Roseburia sp. A2-183, with similarity to acetyl-CoA hydrolase as well as 4-hydroxybutyrate CoA-transferase sequences. The gene product, overexpressed in an Escherichia coli lysate, showed activity with butyryl-CoA and to a lesser degree propionyl-CoA in the presence of acetate. Butyrate, propionate, isobutyrate and valerate competed with acetate as the co-substrate. Despite the sequence similarity to 4-hydroxybutyrate CoA-transferases, 4-hydroxybutyrate did not compete with acetate as the co-substrate. Thus the CoA-transferase preferentially uses butyryl-CoA as substrate. Similar genes were identified in other butyrate-producing human gut bacteria from clostridial clusters IV and XIVa, while other candidate CoA-transferases for butyrate formation could not be detected in Roseburia sp. A2-183. This suggests strongly that the newly identified group of CoA-transferases described here plays a key role in butyrate formation in the human colon. Abbreviations: MALDI-TOF, matrix-assisted laser desorption ionization time-of-flight mass spectrometry The GenBank/EMBL/DDBJ accession numbers for the sequences reported in this paper are: Roseburia sp. A2-183 CoA-transferase, AY796317 ; Eu. hallii L2-7 CoA-transferase, DQ072258 ; F. prausnitzii A2-165 CoA-transferase, DQ072259 ; A. caccae L1-92 CoA-transferase I, DQ151450 ; A. caccae L1-92 4-hydroxybutyrate dehydrogenase and CoA-transferase II, DQ151451 .