Role of Intestinal Microbiota in Ulcerative Colitis

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Methodology part Introduction The aim of this study was to prepare fractions of arabino‐oligosaccharides with different chain length and feruloyl substitutions. The different oligosaccharides were assessed for prebiotic properties using human fecal in vitro fermentations. Flow diagram The author, Louise K. Vigsnæs, performed the small scale in vitro fermentations, and conducted the quantification of the bacterial taxa after fermentation. Louise K. Vigsnæs took part in the evaluation of the results and in preparation of manuscript regarding the in vitro fermentation study. 110 Paper 4
Received: March 11, 2011 Revised: May 12, 2011 Accepted: May 16, 2011 Published: May 16, 2011 ARTICLE pubs.acs.org/JAFC Feruloylated and Nonferuloylated Arabino-oligosaccharides from Sugar Beet Pectin Selectively Stimulate the Growth of Bifidobacterium spp. in Human Fecal in Vitro Fermentations Jesper Holck, † Andrea Lorentzen, § Louise K. Vigsnæs, # Tine R. Licht, # Jørn D. Mikkelsen, † and Anne S. Meyer* ,† † Center for Bioprocess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark § Department of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, Denmark # National Food Institute, Division of Microbiology and Risk Assessment, Technical University of Denmark, 2860 Søborg, Denmark ABSTRACT: The side chains of the rhamnogalacturonan I fraction in sugar beet pectin are particularly rich in arabinan moieties, which may be substituted with feruloyl groups. In this work the arabinan-rich fraction resulting from sugar beet pulp based pectin production was separated by Amberlite XAD hydrophobic interaction and membrane separation into four fractions based on feruloyl substitution and arabino-oligosaccharide chain length: short-chain (DP 2 10) and long-chain (DP 7 14) feruloylated and nonferuloylated arabino-oligosaccharides, respectively. HPAEC, SEC, and MALDI-TOF/TOF analyses of the fractions confirmed the presence of singly and doubly substituted feruloylated arabino-oligosaccharides in the feruloyl-substituted fractions. In vitro microbial fermentation by human fecal samples (n = 6 healthy human volunteers) showed a selective stimulation of bifidobacteria by both the feruloylated and the nonferuloylated long-chain arabino-oligosaccharides to the same extent as the prebiotic fructooligosaccharides control. None of the fractions stimulated the growth of the potential pathogen Clostridium difficile in monocultures. This work provides a first report on the separation of potentially bioactive feruloylated arabino-oligosaccharides from sugar beet pulp and an initial indication of the potentially larger bifidogenic effect of relatively long-chain arabino-oligosaccharides as opposed to short-chain arabino-oligosaccharides. KEYWORDS: prebiotics, arabino-oligosaccharides, feruloyl substitution, hydrophilic interaction chromatography, Clostridium difficile ’ INTRODUCTION Sugar beet pulp is a large side stream from the industrial production of sugar. Currently, the main utilization of sugar beet pulp is for cattle feed, but pectin can be extracted for a limited number of applications. In sugar beet pectin the side chains of rhamnogalacturonan I (RGI) are especially rich in arabinan composed of R-(1,5)-linked backbones with a high degree of R-(1,2) and/or R-(1,3) arabinofuranosyl substitutions 1 along with some β-(1,4)-linked galactan. The side chains of sugar beet RGI can be feruloyl substituted either on O-2 in the main backbone of R-(1,5)-linked arabinan, on O-5 in the terminal arabinose, 2 or on O-6 in the main backbone of galactan. 3 The content of ferulic acid can be up to 8.3 mg/g pectin. 4 The feruloyl substitutions can either be present as monomers or form dimers with other side chains through oxidative coupling reactions. 5 Diferulic cross-linking plays an important role in plant texture. 6 Sugar beet pectin-derived arabino-oligosaccharides with DP 2 6 and DP < 8 have been reported to selectively stimulate bifidobacteria over clostridia, lactobacilli, and bacteroides in in vitro fecal fermentations. 7,8 Moreover, in single-culture studies it has been shown that linear arabino-oligosaccharides may be utilized by Bifidobacterium adolescentis, Bifidobacterium longum, and Bacteroides vulgatus, but not by other bifidobacteria or the tested lactobacilli. 9 The biological effect of oligosaccharide feruloyl substitution on gut microbiota has so far mainly been investigated for arabino-xylo-oligosaccharides. Feruloylated arabino-xylo-oligosaccharides were shown to stimulate the growth of Bifidobacterium bifidum in a single-culture study, 10 xylo-feruloyl-arabinose from grass cell walls has been reported to be metabolized by rat gut microbiota, 11 and recently ferulic acid was found to be released from durum wheat oligosaccharides during digestion by human intestinal microbiota. 12 Moderate feruloyl substitution was shown not to impede the degradation of maize cell wall by human intestinal microbiota. 13 Probiotic bacteria such as bifidobacteria and lactobacilli have been shown to be able to specifically express feruloyl esterase activity in the presence of feruloylated substrates. Bifidobacterium sp., including B. longum, thus express feruloyl esterase activity in the presence of wheat bran, rye bran, barley spent grain, and larchwood arabinogalactan, 14 and Lactobacillus acidophilus expresses feruloyl esterase activity in the presence of destarched wheat bran. 15 These studies on arabinoxylan-derived oligosaccharides are not directly comparable to feruloyl-substituted arabino-oligosaccharides, because feruloyl substitution in r 2011 American Chemical Society 6511 dx.doi.org/10.1021/jf200996h | J. Agric. Food Chem. 2011, 59, 6511–6519
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Role of Intestinal Microbiota in Ul
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Role of Intestinal Microbiota in Ul
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Preface Preface This thesis present
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Summary Summary The microbiota of t
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Dansk sammendrag Dansk sammendrag M
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Introduction and objectives Introdu
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List of Manuscripts Not included in
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List of contents List of Centents P
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List of Centents Methodology append
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1. The intestinal environment Theor
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Theoretical part 5 1. The intestina
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2. The colonic environment Theoreti
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Theoretical part 9 2. The colonic e
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Table 1: The presence of glycoside
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Theoretical part Figure 3: The colo
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3. Inflammatory Bowel disease Theor
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Theoretical part 17 3. Inflammatory
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Theoretical part 19 4. Modulation o
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Table 4: Clinical trials on the pre
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Theoretical part 5. Production of p
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Methodology part
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Methodology part 6. Methodology, co
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Introduction Methodology part 42 Pa
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Abstract Background Detailed knowle
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depending the level of disease acti
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in 1 x TAE at 60 °C for 16 h at 36
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Statistics PCA were generated by SA
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The PCA of the Gram‐positive bact
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layer of UC patients and found that
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Acknowledgements The authors thank
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Table 2 ‐ 16S rRNA gene and 16S
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Page 87 and 88: Methodology part Paper 2 Fecal lact
Page 89 and 90: Fecal lactobacilli and bifidobacter
Page 91 and 92: Introduction The mucus layer lining
Page 93 and 94: efore enrolment and there was no si
Page 95 and 96: (Bio‐Rad Labs, Hercules, Californ
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Page 99 and 100: difference from the luminal microbi
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Page 103 and 104: Table 1 ‐ 16S rRNA gene of phylum
Page 105 and 106: Table 2 ‐ Preference of bacterial
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Page 177 and 178: 7. Discussion and perspectives Disc
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Discussion and conclusion 161 7. Di
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References References Abreu,M.T., V
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References Derrien,M., Vaughan,E.E.
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References Haarman,M. and Knol,J. (
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References Lantz,P.G., Matsson,M.,
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References Matsuki,T., Watanabe,K.,
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References Pullan,R.D., Thomas,G.A.
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References Tannock,G.W. (2010). Ana
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National Food Institute Technical U
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Role of Intestinal Microbiota in Ulcerative Colitis

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