Wood-derived dietary fibers promote beneficial human gut microbiota

Woody biomass is a sustainable and virtually unlimited source of hemicellulosic polysaccharides. The predominant hemicelluloses in softwood and hardwood are galactoglucomannan (GGM) and arabinoglucuronoxylan (AGX), respectively. Based on the structure similarity with common dietary fibers, GGM and AGX may be postulated to have prebiotic properties, conferring a health benefit on the host through specific modulation of the gut microbiota. In this study, we evaluated the prebiotic potential of acetylated GGM (AcGGM) and highly acetylated AGX (AcAGX) obtained from Norwegian lignocellulosic feedstocks in vitro. In pure culture, both substrates selectively promoted the growth of Bifidobacterium, Lactobacillus, and Bacteroides species in a manner consistent with the presence of genetic loci for the utilization of β-manno-oligosaccharides/β-mannans and xylo-oligosaccharides/xylans. The prebiotic potential of AcGGM and AcAGX was further assessed in a pH-controlled batch culture fermentation system inoculated with healthy adult human feces. Results were compared with those obtained with a commercial fructo-oligosaccharide (FOS) mixture. Similarly to FOS, both substrates significantly increased (P < 0.05) the Bifidobacterium population. Other bacterial groups enumerated were unaffected with the exception of an increase in the growth of members of the Bacteroides-Prevotella group, Faecalibacterium prausnitzii, and clostridial cluster IX (P < 0.05). Compared to the other substrates, AcGGM promoted butyrogenic fermentation whereas AcAGX was more propiogenic. Although further in vivo confirmation is necessary, these results demonstrate that both AcGGM and AcAGX from lignocellulosic feedstocks can be used to direct the promotion of beneficial bacteria, thus exhibiting a promising prebiotic ability to improve or restore gut health

Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls.

The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links.This work was supported by the European Union Seventh Framework Programme (FP7 2007-2013) under the WallTraC project (Grant Agreement number 263916). (This article reflects the authors’ views only and the European Union is not liable for any use that may be made of the information contained herein). The work was also supported by the United Kingdom Biotechnology and Biological Research Council (BBSRC, Grant BB/K017489/1). JX acknowledges support from the Chinese Scholarship Council, TAT from a BBSRC studentship and MGR from the Danish Strategic Research Council and The Danish Council for Independent Research, Technology and Production Sciences as part of the GlycAct project (FI 10-093465). We acknowledge kind gifts of enzymes from Harry Gilbert and oligosaccharides from Sanna Koutaniemi. We thank Theodora Tryfona for mass spectrometry analysis.This is the final version of the article. It first appeared from Springer via http://dx.doi.org/10.1007/s00425-015-2375-

Secondary lactic acid bacteria fermentation with wood-derived xylooligosaccharides as a tool to expedite sour beer production

acceptedVersio

Molecular and biochemical tool for the analysis of glycans in plant cell walls

Molecular and biochemical tool for the analysis of glycans in plant cell walls. WallTraC Symposiu

New insight into fine structure of red wine RGII

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<i>Kluyveromyces marxianus</i> Secretes a Pectinase in Shiraz Grape Must That Impacts Technological Properties and Aroma Profile of Wine

Since Saccharomyces cerevisiae strains display no to weak pectinase activity, the utilization of external pectinase is a common practice in winemaking to enhance the extraction of compounds located in the grape berry skins during maceration. In this study, the activity of the native endopolygalacturonase of a Kluyveromyces marxianus strain, isolated from grape juice, was characterized in Shiraz grape must during alcoholic fermentation with or without prefermentative cold maceration. The wines made with K. marxianus had a higher methanol concentration, more free-run wine, an altered volatile compound profile, and displayed pectinase activity in cell-free wine samples. Moreover, the results strongly suggest that K. marxianus’ pectinase released polygalacturonic acid soluble fragments, unlike fungal pectinases, which mostly release monomers. Overall, this study shows that K. marxianus is an effective pectinase producer in wine with potential benefits for wine properties

Recovery and fine structure variability of RGII sub-domains in wine (Vitis vinifera Merlot)

International audienceBackground and Aims Rhamnogalacturonan II (RGII) is a structurally complex pectic sub-domain composed of more than 12 different sugars and 20 different linkages distributed in five side chains along a homogalacturonan backbone. Although RGII has long been described as highly conserved over plant evolution, recent studies have revealed variations in the structure of the polysaccharide. This study examines the fine structure variability of RGII in wine, focusing on the side chains A and B obtained after sequential mild acid hydrolysis. Specifically, this study aims to differentiate intrinsic structural variations in these RGII side chains from structural variations due to acid hydrolysis. Methods RGII from wine (Vitis vinifera Merlot) was sequentially hydrolysed with trifluoroacetic acid (TFA) and the hydrolysis products were separated by anion-exchange chromatography (AEC). AEC fractions or total hydrolysates were analysed by MALDI-TOF mass spectrometry. Key Results The optimal conditions to recover non-degraded side chain B, side chain A and RGII backbone were 0.1 M TFA at 40 degrees C for 16 h, 0.48 M TFA at 40 degrees C for 16 h (or 0.1 M TFA at 60 degrees C for 8 h) and 0.1 M TFA at 60 degrees C for 16 h, respectively. Side chain B was particularly prone to acid degradation. Side chain A and the RGII GalA backbone were partly degraded by 0.1 M TFA at 80 degrees C for 1-4 h. AEC allowed separation of side chain B, methyl-esterified side chain A and non-methyl-esterified side chain A. The structure of side chain A and the GalA backbone were highly variable. Conclusions Several modifications to the RGII structure of wine were identified. The observed dearabinosylation and deacetylation were primarily the consequence of acidic treatment, while variation in methyl-esterification, methyl-ether linkages and oxidation reflect natural diversity. The physiological significance of this variability, however, remains to be determined

Production of low branched rhamnogalacturonan I oligosaccharides from sugar beet pulp

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Production and fine characterisation of new monoclonal antibodies against rhamnogalacturonan I

National audienceThe functional role of the complex pectic rhamnogalacturonane I (RGI) in the context of cell biology is still unclear. RGI is mainly composed of a repeating disaccharide unit [,2)-a-L-rhamnosep-(1,4)-a-D-galacturonic acidp-(1,]n (RU)n decorated primarily with arabinan and (arabino)-galactan side-chains. Monoclonal antibodies (mAbs) are useful to probe pectin structural domains in situ. Several mAbs to the RGI backbone and to arabinan and (arabino)-galactan side-chains have been developed but up to now, there is no mAb against rhamnogalacturonan stretches bearing arabinose or galactose units. Here, we report on the production and fine characterisation of new mAbs against RGI side chains connected to the rhamnogalacturonan backbone. The study first focused on the generation of adequate antigens. A pool of low-branched RU oligosaccharides was prepared and purified from potato pulp. mAbs were raised against low-branched RU oligosaccharides-ovalbumin conjugates. Promising clones producing mAbs recognizing low-branched RU oligosaccharides but recognizing neither unbranched RU oligosaccharides nor galactan oligosaccharides were recovered. The combination of glycan microarray and sub-fractionation of the low-branched RU oligosaccharides pool by anion exchange chromatography prior to competitive ELISA studies allowed fine mAbs characterisation. The use of these new mAbs in immunocytochemistry is expected to give a better understanding of RGI role in planta