Abstract
Propionate is produced in the human large intestine by microbial fermentation and may help maintain human health. We have examined the distribution of three different pathways used by bacteria for propionate formation using genomic and metagenomic analysis of the human gut microbiota and by designing degenerate primer sets for the detection of diagnostic genes for these pathways. Degenerate primers for the acrylate pathway (detecting the lcdA gene, encoding lactoyl-CoA dehydratase) together with metagenomic mining revealed that this pathway is restricted to only a few human colonic species within the Lachnospiraceae and Negativicutes. The operation of this pathway for lactate utilisation in Coprococcus catus (Lachnospiraceae) was confirmed using stable isotope labelling. The propanediol pathway that processes deoxy sugars such as fucose and rhamnose was more abundant within the Lachnospiraceae (based on the pduP gene, which encodes propionaldehyde dehydrogenase), occurring in relatives of Ruminococcus obeum and in Roseburia inulinivorans. The dominant source of propionate from hexose sugars, however, was concluded to be the succinate pathway, as indicated by the widespread distribution of the mmdA gene that encodes methylmalonyl-CoA decarboxylase in the Bacteroidetes and in many Negativicutes. In general, the capacity to produce propionate or butyrate from hexose sugars resided in different species, although two species of Lachnospiraceae (C. catus and R. inulinivorans) are now known to be able to switch from butyrate to propionate production on different substrates. A better understanding of the microbial ecology of short-chain fatty acid formation may allow modulation of propionate formation by the human gut microbiota.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Change history
20 May 2014
This article has been corrected since Advance Online Publication and a corrigendum is also printed in this issue
References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ . (1990). Basic local alignment search tool. J Mol Biol 215: 403–410.
Arora T, Sharma R, Frost G . (2011). Propionate. Anti-obesity and satiety enhancing factor? Appetite 56: 511–515.
Bartosch S, Fite A, Macfarlane GT, McMurdo MET . (2004). Characterization of bacterial communities in feces from healthy elderly volunteers and hospitalized elderly patients by using real-time PCR and effects of antibiotic treatment on the fecal microbiota. Appl Environ Microbiol 70: 3575–3581.
Belenguer A, Duncan SH, Holtrop G, Anderson SE, Lobley GE, Flint HJ . (2007). Impact of pH on lactate formation and utilization by human fecal microbial communities. Appl Environ Microbiol 73: 6526–6533.
Berni Canani R, Di Costanzo M, Leone L . (2012). The epigenetic effects of butyrate: potential therapeutic implications for clinical practice. Clin Epigenetics 4: 4.
Bobik TA, Havemann GD, Busch RJ, Williams DS, Aldrich HC . (1999). The propanediol utilization (pdu) operon of Salmonella enterica serovar Typhimurium LT2 includes genes necessary for formation of polyhedral organelles involved in coenzyme B12-dependent 1,2-propanediol degradation. J Bacteriol 181: 5967–5975.
Bourriaud C, Robins RJ, Martin L, Kozlowski F, Tenailleau E, Cherbut C et al. (2005). Lactate is mainly fermented to butyrate by human intestinal microfloras but inter-individual variation is evident. J Appl Microbiol 99: 201–212.
Bryant MP . (1956). The characteristics of strains of Selenomonas isolated from bovine rumen contents. J Bacteriol 72: 162–167.
Chen J, Cummins CS, Johnson JL, Smibert RM, Smith LD, Wilkins TD . (1977) Anaerobe Laboratory Manual. V.P.I. Anaerobe Laboratory Virginia Polytechnic Institute and State University: Blacksburg, Virginia, USA.
Cole JR, Wang Q, Cardenas E, Fish J, Chai B, Farris RJ et al. (2009). The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Res 37: D141–D145.
Counotte GHM, Prins RA, RHAM Janssen, De Bie MJA . (1981). Role of Megasphaera elsdenii in the fermentation of DL-[2-13C]lactate in the rumen of dairy cattle. Appl Environ Microbiol 42: 649–655.
Dabek M, McCrae SI, Stevens VJ, Duncan SH, Louis P . (2008). Distribution of β-glucosidase and β-glucuronidase activity and of β-glucuronidase gene gus in human colonic bacteria. FEMS Microbiol Ecol 66: 487–495.
Daly K, Shirazi-Beechey SP . (2003). Design and evaluation of group-specific oligonucleotide probes for quantitative analysis of intestinal ecosystems: their application to assessment of equine colonic microflora. FEMS Microbiol Ecol 44: 243–252.
Del Dot T, Osawa R, Stackebrandt E . (1993). Phascolarctobacterium faecium gen. nov, spec. nov., a novel taxon of the Sporomusa group of bacteria. Syst Appl Microbiol 16: 380–384.
Derrien M, Vaughan EE, Plugge CM, de Vos WM . (2004). Akkermansia municiphila gen. nov., sp. nov., a human intestinal mucin-degrading bacterium. Int J Syst Evol Microbiol 54: 1469–1476.
Dimroth P, von Ballmoos C . (2008). ATP synthesis by decarboxylation phosphorylation. Results Probl Cell Differ 45: 153–184.
Duncan SH, Louis P, Thomson JM, Flint HJ . (2009). The role of pH in determining the species composition of the human colonic microbiota. Environ Microbiol 11: 2112–2122.
Ezaki T . (2009a). Genus VII. Coprococcus Holdemann and Moore 1974, 260AL. In: Bergey’s Manual of Systematic Bacteriology, Volume Three the Firmicutes. Springer: Dordrecht, Heidelberg, London, New York, pp 940–941.
Ezaki T . (2009b). Genus I. Ruminococcus Sijpesteijn 1948, 152AL. In: Bergey's Manual of Systematic Bacteriology, Volume Three the Firmicutes. Springer, pp 1016–1018.
Flint HJ, Scott KP, Duncan SH, Louis P, Forano E . (2012a). Microbial degradation of complex carbohydrates in the gut. Gut Microbes 3: 289–306.
Flint HJ, Scott KP, Louis P, Duncan SH . (2012b). The role of the gut microbiota in nutrition and health. Nat Rev Gastroenterol Hepatol 9: 577–589.
Gilmour M, Flint HJ, Mitchell WJ . (1994). Multiple lactate dehydrogenase activities of the rumen bacterium Selenomonas ruminantium. Microbiology 140: 2077–2084.
Gottschalk G . (1979) Bacterial Metabolism. Springer Verlag: New York, Heidelberg, Berlin.
Hamer HM, Jonkers D, Venema K, Vanhoutvin S, Troost FJ, Brummer R- . (2008). Review article: the role of butyrate on colonic function. Aliment Pharmacol Ther 27: 104–119.
Hetzel M, Brock M, Selmer T, Pierik AJ, Golding BT, Buckel W . (2003). Acryloyl-CoA reductase from Clostridium propionicum: an enzyme complex of propionyl-CoA dehydrogenase and electron-transferring flavoprotein. Eur J Biochem 270: 902–910.
Hino T, Kuroda S . (1993). Presence of lactate dehydrogenase and lactate racemase in Megasphaera elsdenii grown on glucose or lactate. Appl Environ Microbiol 59: 255–259.
Hosseini E, Grootaert C, Verstraete W, Van de Wiele T . (2011). Propionate as a health-promoting microbial metabolite in the human gut. Nutr Rev 69: 245–258.
Huder JB, Dimroth P . (1993). Sequence of the sodium ion pump methylmalonyl-CoA decarboxylase from Veillonella parvula. J Biol Chem 268: 24564–24571.
Janssen PH . (1992). Growth yield increase and ATP formation linked to succinate decarboxylation in Veillonella parvula. Arch Microbiol 157: 442–445.
Kandasamy V, Vaidyanathan H, Djurdjevic I, Jayamani E, Ramachandran KB, Buckel W et al. (2013). Engineering Escherichia coli with acrylate pathway genes for propionic acid synthesis and its impact on mixed-acid fermentation. Appl Microbiol Biotechnol 97: 1191–1200.
Kilner J, Waby JS, Chowdry J, Khan AQ, Noirel J, Wright PC et al. (2012). A proteomic analysis of differential cellular responses to the short-chain fatty acids butyrate, valerate and propionate in colon epithelial cancer cells. Mol BioSyst 8: 1146–1156.
Kumar Bhat R, Berger S . (2007). New and easy strategy for cloning, expression, purification, and characterization of the 5S subunit of transcarboxylase from Propionibacterium f. shermanii. Prep Biochem Biotechnol 37: 13–26.
Liu C, Finegold SM, Song Y, Lawson PA . (2008). Reclassification of Clostridium coccoides, Ruminococcus hansenii, Ruminococcus hydrogenotrophicus, Ruminococcus luti, Ruminococcus productus and Ruminococcus schinkii as Blautia coccoides gen. nov., comb. nov., Blautia hansenii comb. nov., Blautia hydrogenotrophica comb. nov., Blautia luti comb. nov., Blautia producta comb. nov., Blautia schinkii comb. nov. and description of Blautia wexlerae. Int J Syst Evol Microbiol 58: 1896–1902.
Lombard J, Moreira D . (2011). Early evolution of the biotin-dependent carboxylase family. BMC Evol Biol 11: 232.
Lopez-Siles M, Khan TM, Duncan SH, Harmsen HJM, Garcia-Gil LJ, Flint HJ . (2012). Cultured representatives of two major phylogroups of human colonic Faecalibacterium prausnitzii can utilize pectin, uronic acids, and host-derived substrates for growth. Appl Environ Microbiol 78: 420–428.
Louis P, Duncan SH, McCrae SI, Millar J, Jackson MS, Flint HJ . (2004). Restricted distribution of the butyrate kinase pathway among butyrate-producing bacteria from the human colon. J Bacteriol 186: 2099–2106.
Louis P, Flint HJ . (2009). Diversity, metabolism and microbial ecology of butyrate-producing bacteria from the human large intestine. FEMS Microbiol Lett 294: 1–8.
Louis P, Young P, Holtrop G, Flint HJ . (2010). Diversity of human colonic butyrate-producing bacteria revealed by analysis of the butyryl-CoA:acetate CoA-transferase gene. Environ Microbiol 12: 304–314.
Lyons E, Freeling M . (2008). How to usefully compare homologous plant genes and chromosomes as DNA sequences. Plant J 53: 661–673.
Macfarlane GT, Gibson GR . (1997). Carbohydrate fermentation, energy transduction and gas metabolism in the human large intestine. In: Gastrointestinal Microbiology Vol. I. Chapman and Hall: London, pp 269–318.
Macy JM, Probst I . (1979). The biology of gastrointestinal bacteroides. Annu Rev Microbiol 33: 561–594.
Marchandin H, Teyssier C, Campos J, Jean-Pierre H, Roger F, Gay B et al. (2010). Negativicoccus succinicivorans gen. nov., sp. nov., isolated from human clinical samples, emended description of the family Veillonellaceae and description of Negativicutes classis nov., Selenomonadales ord. nov. and Acidaminococcaceae fam. nov. in the bacterial phylum Firmicutes. Int J Syst Evol Microbiol 60: 1271–1279.
McIntosh FM, Maison N, Holtrop G, Young P, Stevens VJ, Ince J et al. (2012). Phylogenetic distribution of genes encoding β-glucuronidase activity in human colonic bacteria and the impact of diet on faecal glycosidase activities. Environ Microbiol 14: 1876–1887.
Miyazaki K, Martin JC, Marinsek-Logar R, Flint HJ . (1997). Degradation and utilization of xylans by the rumen anaerobe Prevotella bryantii (formerly P. ruminicola subsp. brevis) B(1)4. Anaerobe 3: 373–381.
Morrison DJ, Mackay WG, Edwards CA, Preston T, Dodson B, Weaver LT . (2006). Butyrate production from oligofructose fermentation by the human faecal flora: what is the contribution of extracellular acetate and lactate? Br J Nutr 96: 570–577.
Qin J, Li R, Raes J, Arumugam M, Burgdorf KS, Manichanh C et al. (2010). A human gut microbial gene catalogue established by metagenomic sequencing. Nature 464: 59–65.
Rainey FA, Hollen BJ, Small A . (2009). Genus I. Clostridium Prazmowski 1880, 23AL. In: Bergey’s Manual of Systematic Bacteriology, Volume Three the Firmicutes. Springer: Dordrecht, Heidelberg, London, New York, pp 738–828.
Richardson AJ, Calder AG, Stewart CS, Smith A . (1989). Simultaneous determination of volatile and non-volatile acidic fermentation products of anaerobes by capillary gas chromatography. Lett Appl Microbiol 9: 5–8.
Satokari RM, Vaughan EE, Akkermans ADL, Saarela M, de Vos WM . (2001). Bifidobacterial diversity in human feces detected by genus-specific PCR and denaturing gradient gel electrophoresis. Appl Environ Microbiol 67: 504–513.
Saxena RK, Anand P, Saran S, Isar J, Agarwal L . (2010). Microbial production and applications of 1,2-propanediol. Indian J Microbiol 50: 2–11.
Scott KP, Martin JC, Campbell G, Mayer C-, Flint HJ . (2006). Whole-genome transcription profiling reveals genes up-regulated by growth on fucose in the human gut bacterium ‘Roseburia inulinivorans’. J Bacteriol 188: 4340–4349.
Seeliger S, Janssen PH, Schink B . (2002). Energetics and kinetics of lactate fermentation to acetate and propionate via methylmalonyl-CoA or acrylyl-CoA. FEMS Microbiol Lett 211: 65–70.
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S . (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731–2739.
Ulven T . (2012). Short-chain free fatty acid receptors FFA2/GPR43 and FFA3/GPR41 as new potential therapeutic targets. Front Endocrinol (Lausanne) 3: 111.
Vinolo MAR, Rodrigues HG, Nachbar RT, Curi R . (2011). Regulation of inflammation by short chain fatty acids. Nutrients 3: 858–876.
Vogt JA, Pencharz PB, Wolever TMS . (2004). L-rhamnose increases serum propionate in humans. Am J Clin Nutr 80: 89–94.
Walker AW, Duncan SH, McWilliam Leitch EC, Child MW, Flint HJ . (2005). pH and peptide supply can radically alter bacterial populations and short-chain fatty acid ratios within microbial communities from the human colon. Appl Environ Microbiol 71: 3692–3700.
Watanabe Y, Nagai F, Morotomi M . (2012). Characterization of Phascolarctobacterium succinatutens sp. nov., an asaccharolytic, succinate-utilizing bacterium isolated from human feces. Appl Environ Microbiol 78: 511–518.
Wood J, Scott KP, Avguštin G, Newbold CJ, Flint HJ . (1998). Estimation of the relative abundance of different Bacteroides and Prevotella ribotypes in gut samples by restriction enzyme profiling of PCR- amplified 16S rRNA gene sequences. Appl Environ Microbiol 64: 3683–3689.
Acknowledgements
The Rowett Institute of Nutrition and Health receives financial support from the Scottish Government Rural and Environmental Sciences and Analytical Services. Alvaro Belenguer received financial support from the Spanish Ministry of Education and Science. Nicole Reichardt is funded by a Scottish Government Strategic Partnership on Food and Drink Science. We thank Wolfgang Buckel, Douglas Morrison, Tom Preston, Graeme Milligan, Lynda Williams and Janice Drew for helpful discussions, Freda Farquharson for help with bacterial cultures and supply of genomic DNA, Graham Calder and Gerald Lobley for help with stable isotope analysis and Tony Travis for help with bioinformatic analysis.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on The ISME Journal website
Supplementary information
Rights and permissions
About this article
Cite this article
Reichardt, N., Duncan, S., Young, P. et al. Phylogenetic distribution of three pathways for propionate production within the human gut microbiota. ISME J 8, 1323–1335 (2014). https://doi.org/10.1038/ismej.2014.14
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/ismej.2014.14
Keywords
This article is cited by
-
Gut Microbiota Modulation and Its Implications on Neuropathic Pain: A Comprehensive Literature Review
Pain and Therapy (2024)
-
Dietary Supplementation of Compound Probiotics Improves Intestinal Health by Modulated Microbiota and Its SCFA Products as Alternatives to In-Feed Antibiotics
Probiotics and Antimicrobial Proteins (2024)
-
Lactiplantibacillus plantarum, lactiplantibacillus pentosus and inulin meal inclusion boost the metagenomic function of broiler chickens
Animal Microbiome (2023)
-
Gut microbiota as a target in the bone health of livestock and poultry: roles of short-chain fatty acids
Animal Diseases (2023)
-
Feeding an unsalable carrot total-mixed ration altered bacterial amino acid degradation in the rumen of lambs
Scientific Reports (2023)


