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Applied and Environmental Microbiology, September 2009, p. 5884-5892, Vol. 75, No. 18
0099-2240/09/$08.00+0 doi:10.1128/AEM.00876-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
In Vitro Kinetics of Prebiotic Inulin-Type Fructan Fermentation by Butyrate-Producing Colon Bacteria: Implementation of Online Gas Chromatography for Quantitative Analysis of Carbon Dioxide and Hydrogen Gas Production
,
Gwen Falony,1
An Verschaeren,1
Feije De Bruycker,1
Vicky De Preter,2
Kristin Verbeke,2
Frédéric Leroy,1 and
Luc De Vuyst1*
Research Group of Industrial Microbiology and Food Biotechnology, Faculty of Sciences and Bioengineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium,1 Department of Gastrointestinal Research and Leuven Food Science and Nutrition Research Centre, University Hospital Gasthuisberg, Katholieke Universiteit Leuven, Herestraat 49, B-3000 Leuven, Belgium2
Received 17 April 2009/ Accepted 17 July 2009
Kinetic analyses of bacterial growth, carbohydrate consumption, and metabolite production of five butyrate-producing clostridial cluster XIVa colon bacteria grown on acetate plus fructose, oligofructose, inulin, or lactate were performed. A gas chromatography method was set up to assess H2 and CO2 production online and to ensure complete coverage of all metabolites produced. Method accuracy was confirmed through the calculation of electron and carbon recoveries. Fermentations with Anaerostipes caccae DSM 14662T, Roseburia faecis DSM 16840T, Roseburia hominis DSM 16839T, and Roseburia intestinalis DSM 14610T revealed similar patterns of metabolite production with butyrate, CO2, and H2 as the main metabolites. R. faecis DSM 16840T and R. intestinalis DSM 14610T were able to degrade oligofructose, displaying a nonpreferential breakdown mechanism. Lactate consumption was only observed with A. caccae DSM 14662T. Roseburia inulinivorans DSM 16841T was the only strain included in the present study that was able to grow on fructose, oligofructose, and inulin. The metabolites produced were lactate, butyrate, and CO2, without H2 production, indicating an energy metabolism distinct from that of other Roseburia species. Oligofructose degradation was nonpreferential. In a coculture of R. inulinivorans DSM 16841T with the highly competitive strain Bifidobacterium longum subsp. longum LMG 11047 on inulin, hardly any production of butyrate and CO2 was detected, indicating a lack of competitiveness of the butyrate producer. Complete recovery of metabolites during fermentations of clostridial cluster XIVa butyrate-producing colon bacteria allowed stoichiometric balancing of the metabolic pathway for butyrate production, including H2 formation.
* Corresponding author. Mailing address: Research Group of Industrial Microbiology and Food Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium. Phone: 32 2 6293245. Fax: 32 2 6292720. E-mail: ldvuyst{at}vub.ac.be
Published ahead of print on 24 July 2009.
Supplemental material for this article may be found at http://aem.asm.org/.
Applied and Environmental Microbiology, September 2009, p. 5884-5892, Vol. 75, No. 18
0099-2240/09/$08.00+0 doi:10.1128/AEM.00876-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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