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

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Abstract
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 H2and CO2production online and to ensure complete coverage of all metabolites produced. Method accuracy was confirmed through the calculation of electron and carbon recoveries. Fermentations withAnaerostipes caccaeDSM 14662T,Roseburia faecisDSM 16840T,Roseburia hominisDSM 16839T, andRoseburia intestinalisDSM 14610Trevealed similar patterns of metabolite production with butyrate, CO2, and H2as the main metabolites.R. faecisDSM 16840TandR. intestinalisDSM 14610Twere able to degrade oligofructose, displaying a nonpreferential breakdown mechanism. Lactate consumption was only observed withA. caccaeDSM 14662T.Roseburia inulinivoransDSM 16841Twas 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 H2production, indicating an energy metabolism distinct from that of otherRoseburiaspecies. Oligofructose degradation was nonpreferential. In a coculture ofR. inulinivoransDSM 16841Twith the highly competitive strainBifidobacterium longumsubsp.longumLMG 11047 on inulin, hardly any production of butyrate and CO2was 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 H2formation.

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