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Journal of Bacteriology, April 2004, p. 2099-2106, Vol. 186, No. 7
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.7.2099-2106.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Restricted Distribution of the Butyrate Kinase Pathway among Butyrate-Producing Bacteria from the Human Colon

Petra Louis,* Sylvia H. Duncan, Sheila I. McCrae, Jacqueline Millar, Michelle S. Jackson,{dagger} and Harry J. Flint

Division of Gut Microbiology and Immunology, Rowett Research Institute, Bucksburn, Aberdeen AB21 9SB, United Kingdom

Received 19 August 2003/ Accepted 11 December 2003

The final steps in butyrate synthesis by anaerobic bacteria can occur via butyrate kinase and phosphotransbutyrylase or via butyryl-coenzyme A (CoA):acetate CoA-transferase. Degenerate PCR and enzymatic assays were used to assess the presence of butyrate kinase among 38 anaerobic butyrate-producing bacterial isolates from human feces that represent three different clostridial clusters (IV, XIVa, and XVI). Only four strains were found to possess detectable butyrate kinase activity. These were also the only strains to give PCR products (verifiable by sequencing) with degenerate primer pairs designed within the butyrate kinase gene or between the linked butyrate kinase/phosphotransbutyrylase genes. Further analysis of the butyrate kinase/phosphotransbutyrylase genes of one isolate, L2-50, revealed similar organization to that described previously from different groups of clostridia, along with differences in flanking sequences and phylogenetic relationships. Butyryl-CoA:acetate CoA-transferase activity was detected in all 38 strains examined, suggesting that it, rather than butyrate kinase, provides the dominant route for butyrate formation in the human colonic ecosystem that contains a constantly high concentration of acetate.

* Corresponding author. Mailing address: Division of Gut Microbiology and Immunology, Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, United Kingdom. Phone: 44-01224-712751. Fax: 44-01224-716687. E-mail: p.louis{at}rowett.ac.uk.

{dagger} Present address: CSIRO Health Sciences and Nutrition, Adelaide SA 5000, Australia.

Journal of Bacteriology, April 2004, p. 2099-2106, Vol. 186, No. 7
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.7.2099-2106.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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