Degradation and Utilization of Xylans by the Rumen AnaerobePrevotella bryantii(formerlyP. ruminicolasubsp.brevis) B14

Miyazaki, Kohji; Martin, Jennifer C.; Marinšek-Logar, Romana; Flint, Harry J.

doi:10.1006/anae.1997.0125

Cited by 166 publications

(158 citation statements)

References 24 publications

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“…All other bacterial strains used were described before (Dabek et al, 2008) or obtained from the German Collection of Microorganisms and Cell Cultures, the American Type Culture Collection, the National Collection of Industrial and Marine Bacteria (as indicated by DSM, ATCC and NCIMB numbers) or from the Rowett Institute strain collection (V. parvula L59). Bacteria were grown anaerobically in M2GSC (Miyazaki et al, 1997) or YCFAGSC medium (Lopez-Siles et al, 2012) apart from Akkermansia muciniphila DSM 22959 (M2GSC þ 0.2% procine mucin, Sigma-Aldrich, Gillingham, UK) and V. parvula L59 (M2GSC medium with maltose replacing starch þ 1% lactate). Growth experiments on specific substrates were performed in triplicate on basal YCFA medium supplemented with substrates as detailed in the results (at 0.5% substrate, unless specified otherwise).…”

Section: Methodsmentioning

confidence: 99%

Phylogenetic distribution of three pathways for propionate production within the human gut microbiota

et al. 2014

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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 lactoylCoA 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.

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Section: Methodsmentioning

confidence: 99%

Phylogenetic distribution of three pathways for propionate production within the human gut microbiota

et al. 2014

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“…Routine anaerobic culturing of R. inulinivorans A2-194 was in M2GSC medium (39). Growth on single carbon sources used basal YCFA medium (YCFA -yeast extract-casein hydrolysate-fatty acids) (3) supplemented with 0.5% wt/vol of the specific substrate (glucose, FOS, Dahlia inulin, fructose or amylopectin starch from Sigma).…”

Section: Methodsmentioning

confidence: 99%

Substrate-driven gene expression in Roseburia inulinivorans : Importance of inducible enzymes in the utilization of inulin and starch

Scott

Martin

Chassard

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

120

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Roseburia inulinivorans is a recently identified motile representative of the Firmicutes that contributes to butyrate formation from a variety of dietary polysaccharide substrates in the human large intestine. Microarray analysis was used here to investigate substratedriven gene-expression changes in R. inulinivorans A2-194. A cluster of fructo-oligosaccharide/inulin utilization genes induced during growth on inulin included one encoding a β-fructofuranosidase protein that was prominent in the proteome of inulin-grown cells. This cluster also included a 6-phosphofructokinase and an ABC transport system, whereas a distinct inulin-induced 1-phosphofructokinase was linked to a fructose-specific phosphoenolpyruvatedependent sugar phosphotransferase system (PTS II transport enzyme). Real-time PCR analysis showed that the β-fructofuranosidase and adjacent ABC transport protein showed greatest induction during growth on inulin, whereas the 1-phosphofructokinase enzyme and linked sugar phosphotransferase transport system were most strongly up-regulated during growth on fructose, indicating that these two clusters play distinct roles in the use of inulin. The R. inulinivorans β-fructofuranosidase was overexpressed in Escherichia coli and shown to hydrolyze fructans ranging from inulin down to sucrose, with greatest activity on fructo-oligosaccharides. Genes induced on starch included the major extracellular α-amylase and two distinct α-glucanotransferases together with a gene encoding a flagellin protein. The latter response may be concerned with improving bacterial access to insoluble starch particles.anaerobic gut bacteria | differential gene expression | fructooligosaccharides | butyrate | prebiotic P lant cell wall polysaccharides, storage polysaccharides such as resistant starch and inulin, and many oligosaccharides of plant origin remain undigested in the upper gastrointestinal tract and become important substrates for the growth of colonic bacteria. Prebiotics are defined as dietary substrates that reach the colon where they selectively stimulate the growth of beneficial gut bacteria (1), with the most widely used prebiotics being the fructans inulin and fructo-oligosaccharides (FOS). Inulin is a linear polymer [degree of polymerization (DP) = 3-60] of β-2,1-linked fructose monomers with a terminal glucose residue, whereas FOS have the same backbone but a maximal chain length of 8 monomeric units. The increasing use of prebiotics to enhance gut health is driving research into their mode of action. Many studies have shown that both inulin and FOS selectively stimulate the growth of potentially beneficial gut bacteria such as bifidobacteria and lactobacilli (reviewed in ref.2). However, only a few studies have considered the potential for stimulation of other bacterial groups that may also be beneficial (3-6).The bifidogenic dose of inulin has been defined as 5-8 g/d (7), yet the effect of such supplementation on other beneficial groups of gut bacteria and the differences relating to different compositions of inulin...

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“…are the most numerically dominant xylanolytic bacteria in the rumen (2,3); therefore, the mechanism by which they degrade and utilize xylan has been an important topic of investigation (4 -9). Prevotella bryantii B 1 4 is frequently isolated from the rumen microbiome and can grow with xylan as the sole carbohydrate source (6). However, to date only six genes with roles in xylan hydrolysis have been cloned and characterized from P. bryantii B 1 4, including two glycoside hydrolase (GH) 2 family 10 endoxylanases (7,8,10), three GH family 3 ␤-xylosidases (4), and one GH family 43 ␤-xylosidase enzyme (7,8).…”

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confidence: 99%

Transcriptomic Analyses of Xylan Degradation by Prevotella bryantii and Insights into Energy Acquisition by Xylanolytic Bacteroidetes

Dodd

Moon

Swaminathan

et al. 2010

Journal of Biological Chemistry

112

101

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Enzymatic depolymerization of lignocellulose by microbes in the bovine rumen and the human colon is critical to gut health and function within the host. Prevotella bryantii B 1 4 is a rumen bacterium that efficiently degrades soluble xylan. To identify the genes harnessed by this bacterium to degrade xylan, the transcriptomes of P. bryantii cultured on either wheat arabinoxylan or a mixture of its monosaccharide components were compared by DNA microarray and RNA sequencing approaches. The most highly induced genes formed a cluster that contained putative outer membrane proteins analogous to the starch utilization system identified in the prominent human gut symbiont Bacteroides thetaiotaomicron. The arrangement of genes in the cluster was highly conserved in other xylanolytic Bacteroidetes, suggesting that the mechanism employed by xylan utilizers in this phylum is conserved. A number of genes encoding proteins with unassigned function were also induced on wheat arabinoxylan. Among these proteins, a hypothetical protein with low similarity to glycoside hydrolases was shown to possess endoxylanase activity and subsequently assigned to glycoside hydrolase family 5. The enzyme was designated PbXyn5A. Two of the most similar proteins to PbXyn5A were hypothetical proteins from human colonic Bacteroides spp., and when expressed each protein exhibited endoxylanase activity. By using site-directed mutagenesis, we identified two amino acid residues that likely serve as the catalytic acid/base and nucleophile as in other GH5 proteins. This study therefore provides insights into capture of energy by xylanolytic Bacteroidetes and the application of their enzymes as a resource in the biofuel industry.

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Degradation and Utilization of Xylans by the Rumen AnaerobePrevotella bryantii(formerlyP. ruminicolasubsp.brevis) B14

Cited by 166 publications

References 24 publications

Phylogenetic distribution of three pathways for propionate production within the human gut microbiota

Phylogenetic distribution of three pathways for propionate production within the human gut microbiota

Substrate-driven gene expression in Roseburia inulinivorans : Importance of inducible enzymes in the utilization of inulin and starch

Transcriptomic Analyses of Xylan Degradation by Prevotella bryantii and Insights into Energy Acquisition by Xylanolytic Bacteroidetes

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