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Aims In this study, we explored the effects that the prebiotic inulin-type fructans, and prebiotic candidates: 2'fucosyllactose and β-glucan from barley, singular and in combination had on microbial load, microbiome profile, and short-chain fatty acid production. This was carried out as a prescreening tool to determine combinations that could be taken forward for use in a human intervention trial. Methods and results Effects of inulin-type fructans, 2'fucosyllactose and β-glucan from barley in singular and combination on microbial load and profile and short-chain fatty acid production (SCFA) was conducted using in vitro batch culture fermentation over 48 h. Changes in microbial load and profile were assessed by fluorescence in situ hybridization flow cytometry (FISH-FLOW) and 16S rRNA sequencing, and changes in SCFA via gas chromatography. All substrates generated changes in microbial load and profile, achieving peak microbial load at 8 h fermentation with the largest changes in profile across all substrates in Bifidobacterium (Q  < 0.05). This coincided with significant increases in acetate observed throughout fermentation (Q  < 0.05). In comparison to sole supplementation combinations of oligofructose, β-glucan and 2'fuscosyllactose induced significant increases in both propionate and butyrate producing bacteria (Roseburia and Faecalibacterium praunitzii), and concentrations of propionate and butyrate, the latter being maintained until the end of fermentation (all Q  < 0.05). Conclusions Combinations of oligofructose, with β-glucan and 2'fucosyllactose induced selective changes in microbial combination and SCFA namely Roseburia, F. praunitzii , propionate and butyrate compared to sole supplementation. [ABSTRACT FROM AUTHOR]

Carbohydrates in forages constitute an important part of the feed ration for all horses. The aim of the present study was to investigate the effect of harvest time on carbohydrate composition and digestion of various grass species. The experiment was divided into three parts 1) characterization of the chemical composition of experimental feeds (6 grass species: meadow fescue [MF], cocksfoot [CF], perennial ryegrass [PR], smooth bromegrass [SB], tall fescue [TF], and timothy [TI], and 3 harvest times: early, medium, and late first cut), 2) measurements of the in vitro digestion of selected experimental feeds (the 6 grass species, and 2 harvest times [early and late]) measured by in vitro gas production, and 3) in vivo digestion of selected experimental feeds (2 grass species: CF and PR, 2 harvest times [early and late]) measured by the mobile bag technique using caecum cannulated horses. An experimental field was established with plots containing each of the grass species in three replicate blocks. Grass samples were cut between 1200 and 1400 h at 4th of June (early first cut), 17th of June (medium first cut), and 1st of July (late first cut) and analyzed for crude protein (CP), neutral detergent fiber with heat stable amylase and free of residual ash (aNDFom) and water-soluble carbohydrates (WSC). The in vitro fermentation was investigated using the ANKOM RF gas production technique, where feeds were incubated for 48 h using horse caecal fluid as an inoculum. Gas production was modeled, and maximum gas production (MGP) was used to evaluate the potential digestibility of the feeds. Based on the chemical analyses and the in vitro experiment, early and late harvested CF and PR were selected for the in vivo experiment, which was conducted as a randomized 4 × 4 Latin square design including four periods, four horses and four feeds. In general, the CP content decreased whereas the aNDFom content increased as the grasses matured. The content of WSC increased in SB and TI, but decreased in CF, and fructans increased in SB, TI, PR, and TF as they matured. The in vitro MGP showed a clearer difference between harvest times than between grass species. Harvest time had larger effect on digestibility than grass species, and a high precaecal disappearance of the WSC fraction was measured by the mobile bag technique. Cocksfoot was identified as a grass species with potentially low digestibility and low WSC content and could potentially be used more for horses. [ABSTRACT FROM AUTHOR]

Fructans such as inulin and levan accumulate in certain taxonomic groups of plants and are a reserve carbohydrate alternative to starch. Onion (Allium cepa L.) is a typical plant species that accumulates fructans, and it synthesizes inulin-type and inulin neoseries-type fructans in the bulb. Although genes for fructan biosynthesis in onion have been identified so far, no genes for fructan degradation had been found. In this study, phylogenetic analysis predicted that we isolated a putative vacuolar invertase gene (AcpVI1), but our functional analyses demonstrated that it encoded a fructan 1-exohydrolase (1-FEH) instead. Assessments of recombinant proteins and purified native protein showed that the protein had 1-FEH activity, hydrolyzing the β-(2,1)-fructosyl linkage in inulin-type fructans. Interestingly, AcpVI1 had an amino acid sequence close to those of vacuolar invertases and fructosyltransferases, unlike all other FEHs previously found in plants. We showed that AcpVI1 was localized in the vacuole, as are onion fructosyltransferases Ac1-SST and Ac6G-FFT. These results indicate that fructan-synthesizing and -degrading enzymes are both localized in the vacuole. In contrast to previously reported FEHs, our data suggest that onion 1-FEH evolved from a vacuolar invertase and not from a cell wall invertase. This demonstrates that classic phylogenetic analysis on its own is insufficient to discriminate between invertases and FEHs, highlighting the importance of functional markers in the nearby active site residues. [ABSTRACT FROM AUTHOR]

Fructan metabolism in bacteria and plants relies on fructosyltransferases and fructanases. Plant fructanases (fructan exohydrolase, FEH) only hydrolyse terminal fructose residues. Levan (β-2,6 linkages) is the most abundant fructan type in bacteria. Dicot fructan accumulators, such as chicory (Cichorium intybus), accumulate inulin (β-2,1 linkages), harbouring several 1-FEH isoforms for their degradation. Here, a novel chicory fructanase with high affinity for levan was characterized, providing evidence that such enzymes widely occur in higher plants. It is adapted to common microbial fructan profiles, but has low affinity towards chicory inulin, in line with a function in trimming of microbial fructans in the extracellular environment. Docking experiments indicate the importance of an N-glycosylation site close to the active site for substrate specificity. Optimal pH and temperature for levan hydrolysis are 5.0 and 43.7 °C, respectively. Docking experiments suggested multiple substrate binding sites and levan-mediated enzyme dimerization, explaining the observed positive cooperativity. Alignments show a single amino acid shift in the position of a conserved DXX(R/K) couple, typical for sucrose binding in cell wall invertases. A possible involvement of plant fructanases in levan trimming is discussed, in line with the emerging 'fructan detour' concepts, suggesting that levan oligosaccharides act as signalling entities during plant-microbial interactions. [ABSTRACT FROM AUTHOR]

Fructans are the main sugar in agave pine used by yeasts during mezcal fermentation processes, from which Candida apicola NRRL Y-50540 and Torulaspora delbrueckii NRRL Y-50541 were isolated. De novo transcriptome analysis was carried out to identify genes involved in the hydrolysis and assimilation of Agave fructans (AF). We identified a transcript annotated as SUC2 , which is related to β-fructofuranosidase activity, and several differential expressed genes involved in the transcriptional regulation of SUC2 such as: MIG1, MTH1, SNF1, SNF5, REG1, SSN6, SIP1, SIP2, SIP5, GPR1, RAS2 , and PKA. Some of these genes were specifically expressed in some of the yeasts according to their fructans assimilation metabolism. Different hexose transporters that could be related to the assimilation of fructose and glucose were found in both the transcriptomes. Our findings provide a better understanding of AF assimilation in these yeasts and provide resources for further metabolic engineering and biotechnology applications. [ABSTRACT FROM AUTHOR]

Aims: Isolating a novel bacterial source of fructan from a saltern and analysis of its genome to better understand the possible roles of fructans in hypersaline environments. Methods and Results: Bacteria were isolated from crude salt samples originating from Çamaltı Saltern in Western Turkey and screened for fructanogenic traits in high‐salt and sucrose‐rich selective medium. Exopolysaccharide accumulated in the presence of sucrose by isolate OK12 was purified and chemically characterized via HPLC, FT‐IR and NMR, which revealed that it was a levan‐type fructan (β‐2,6 linked homopolymer of fructose). The isolate was taxonomically classified as Bacillus licheniformis OK12 through 16S rRNA gene and whole‐genome sequencing methods. Strain OK12 harbours one levansucrase and two different levanase genes, which altogether were predicted to significantly contribute to intracellular glucose and fructose pools. The isolate could withstand 15% NaCl, and thus classified as a halotolerant. Conclusions: Fructanogenic traits in halotolerant B. licheniformis OK12 are significant due to predicted influx of glucose and fructose as a result of levan biosynthesis and levan hydrolysis, respectively. Significance and Impact of the Study: Fructans from the residents of hypersaline habitats are underexplored compounds and are expected to demonstrate physicochemical properties different from their non‐halophilic counterparts. Revealing fructanogenic traits in the genome of a halotolerant bacterium brings up a new perspective in physiological roles of fructans. [ABSTRACT FROM AUTHOR]

We profiled the grain oligosaccharide content of 154 two-row spring barley genotypes and quantified 27 compounds, mainly inulin- and neoseries-type fructans, showing differential abundance. Clustering revealed two profile groups where the 'high' set contained greater amounts of sugar monomers, sucrose, and overall fructans, but lower fructosylraffinose. A genome-wide association study (GWAS) identified a significant association for the variability of two fructan types: neoseries-DP7 and inulin-DP9, which showed increased strength when applying a novel compound ratio-GWAS approach. Gene models within this region included three known fructan biosynthesis genes (fructan:fructan 1-fructosyltransferase , s ucrose:sucrose 1-fructosyltransferase , and s ucrose:fructan 6-fructosyltransferase). Two other genes in this region, 6(G)-fructosyltransferase and vacuolar invertase1 , have not previously been linked to fructan biosynthesis and showed expression patterns distinct from those of the other three genes, including exclusive expression of 6(G)-fructosyltransferase in outer grain tissues at the storage phase. From exome capture data, several single nucleotide polymorphisms related to inulin- and neoseries-type fructan variability were identified in fructan:fructan 1-fructosyltransferase and 6(G)-fructosyltransferase genes. Co-expression analyses uncovered potential regulators of fructan biosynthesis including transcription factors. Our results provide the first scientific evidence for the distinct biosynthesis of neoseries-type fructans during barley grain maturation and reveal novel gene candidates likely to be involved in the differential biosynthesis of various types of fructan in barley. [ABSTRACT FROM AUTHOR]

Aims: Interventions using prebiotic inulin‐type fructans (ITFs) are widely prescribed to modulate the gut microbiota composition and activity to promote health. However, the impacts of ITFs on post‐antibiotic reconstitution of the gut microbiome remain incompletely understood. The aim of the present study was to investigate the effects of ITFs supplementation on intestinal inflammation, the composition of the intestinal microbiota and the colonic transcriptome after antibiotic treatment. Methods and Results: Male BALB/c mice were subjected to an antibiotic cocktail (ABx) treatment for 7 days, and their microbiomes were then reconstituted either spontaneously or with ITFs supplementation (5%) for 14 days. Our data showed that ITFs supplementation delayed the recovery of antibiotic‐induced colitis compared with the spontaneous recovery. Neither ITFs supplementation nor spontaneous recovery could restore the microbial community composition at the genus level back to its initial composition. ITFs supplementation increased the relative abundance of some beneficial bacteria and butyrate levels, but resulted in selective blooms of some opportunistic pathogens and elevated the pathways associated with diseases linked to gut microbiota function. Both ITFs supplementation and spontaneous recovery could restore the colonic transcriptome nearly to the initial profile to a certain extent; however, ITFs supplementation delayed the restoration of the immunoglobulin genes compared to spontaneous recovery. Conclusion: These data showed that post‐antibiotic ITFs consumption did not always lead to beneficial effects but might lead to potential adverse effects in the context of dysbiosis. Significance and Impact of the Study: These findings highlighted that caution is required when supplementing ITFs to restore intestinal homeostasis in the context of dysbiosis resulting from broad‐spectrum antibiotics. [ABSTRACT FROM AUTHOR]

Background: Fructans are added to infant formula and adult nutritionals for their prebiotic effect. A method (AOAC 2016.14) was developed for their analysis which has already demonstrated excellent performance during single laboratory validation. Objective: To determine repeatability and reproducibility of the method through a collaborative study. Methods: Fourteen laboratories from 11 different countries enrolled for the study. Participants analyzed a practice sample, then 8 formula or adult nutritionals in blind duplicate. Results and any method modifications were reported to the study director. Results: Twelve laboratories provided results on time for reporting. Precision results for five samples met the requirements of the Standard Method Performance Requirements (SMPR 2014.002), with RSDr ranging from 3.60 to 4.25% and RSDR ranging from 5.90 to 11.7%. The practice sample also met the requirements of SMPR 2014.002, with RSDr and RSDR of 2.53% and 6.70% respectively. Precision results for three test samples did not fully meet the SMPR, with RSDr ranging from 2.27 to 7.65% and RSDR ranging from 12.8 to 15.1%. After review, the AOAC Stakeholder Panel for Infant Formula and Adult Nutritional Expert Review Panel (SPIFAN ERP) concluded that the data presented mostly met the SMPR and hence recommended that the method to be advanced for adoption as an AOAC Final Action method. Conclusions: The method described in AOAC 2016.14 is suitable for the determination of fructans in infant formula and adult nutritionals. [ABSTRACT FROM AUTHOR]

Inulin-type fructans are known to exert different effects on the fermentation profile depending on the average and range of the degree of polymerization (DP). Here, swine fecal cultures were used to investigate the prebiotic properties of native chicory inulin (NIN), extracted from the chicory root, and synthetic inulin (SIN), which has a narrower DP distribution than NIN. Both NIN and SIN showed prebiotic effects, but NIN exhibited a significant decrease in pH and increase in the production of propionate and butyrate compared to SIN. There were also differences in the production of succinate and lactate, the precursors of propionate and butyrate, and the relative abundance of associated genes. Furthermore, NIN induced the growth of certain species of Bifidobacterium and Lactobacillus more strongly than SIN. These results suggest that NIN and SIN exhibit different prebiotic properties due to differences in DP, and that NIN might be more beneficial to host health. The superior prebiotic properties of native chicory inulin (NIN) compared to those of synthetic inulin (SIN), as demonstrated by propionate and butyrate production. [ABSTRACT FROM AUTHOR]

Extreme climatic events (ECEs) such as droughts and heat waves affect ecosystem functioning and species turnover. This study investigated the effect of elevated CO2 on species' resilience to ECEs. Monoliths of intact soil and their plant communities from an upland grassland were exposed to 2050 climate scenarios with or without an ECE under ambient (390 ppm) or elevated (520 ppm) CO2. Ecophysiological traits of two perennial grasses (Dactylis glomerata and Holcus lanatus) were measured before, during, and after ECE. At similar soil water content, leaf elongation was greater under elevated CO2 for both species. The resilience of D. glomerata increased under enhanced CO2 (+60%) whereas H. lanatus mostly died during ECE. D. glomerata accumulated 30% more fructans, which were more highly polymerized, and 4-fold less sucrose than H. lanatus. The fructan concentration in leaf meristems was significantly increased under elevated CO2. Their relative abundance changed during the ECE, resulting in a more polymerized assemblage in H. lanatus and a more depolymerized assemblage in D. glomerata. The ratio of low degree of polymerization fructans to sucrose in leaf meristems was the best predictor of resilience across species. This study underlines the role of carbohydrate metabolism and the species-dependent effect of elevated CO2 on the resilience of grasses to ECE. [ABSTRACT FROM AUTHOR]

Traditional enzyme-based methods for measurement of fructan were designed to measure just inulin and branched-type (agave) fructans. The enzymes employed, namely exo-inulinase and endoinulinase, give incompletely hydrolysis of levan. Levan hydrolysis requires a third enzyme, endolevanase. This paper describes a method and commercial test kit (Megazyme Fructan Assay Kit) for the determination of all types of fructan (inulin, levan, and branched) in a variety of animal feeds and pet foods. The method has been validated in a single laboratory for analysis of pure inulin, agave fructan, levan, and a range of fructan containing samples. Quantification is based on complete hydrolysis of fructan to fructose and glucose by a mixture of exo-inulinase, endo-inulinase, and endolevanase, followed by measurement of these sugars using the PAHBAH reducing sugar method which gives the same color response with fructose and glucose. Before hydrolysis of fructan, interfering sucrose and starch in the sample are specifically hydrolyzed and removed by borohydride reduction. The single-laboratory validation (SLV) outlined in this document was performed on commercially available inulin (Raftiline) and agave fructan (Frutafit®), levan purified from Timothy grass, two grass samples, a sample of legume hay, two animal feeds and two barley flours, one of which (Barley MAX®) was genetically enriched in fructan through plant breeding. Parameters examined during the validation included working range, target selectivity, recovery, LOD, LOQ, trueness (bias), precision (repeatability and intermediate precision), robustness, and stability. The method is robust, quick, and simple. [ABSTRACT FROM AUTHOR]

Asparagus (Asparagus officinalis L.) accumulates inulin- and inulin neoseries-type fructans. Fructose released by the hydrolysis of fructans is an energy source for emerging asparagus spears. Plant fructans are hydrolyzed by fructan exohydrolases (FEHs), whose presence in asparagus has not yet been fully characterized. Here, we describe for the first time the purification and characterization of an FEH from asparagus, and the functional analysis of its gene. The purified enzyme was predicted to exist as a dimer (approximately 130 kDa) consisting of two polypeptides with a molecular mass of approximately 68 kDa. N-terminal sequences of the purified enzyme were matched with the amino acid sequences of aoeh4a and aoeh4b cDNAs isolated from asparagus (cv. Gijnlim and Taihouwase). Native enzymes obtained from asparagus roots and recombinant enzymes produced by Pichia pastoris showed fructan 1-exohydrolase (1-FEH) activity via the hydrolysis of inulin-type fructan. Unlike other 1-FEHs, these enzymes showed minimal hydrolysis of 1-kestose but efficiently hydrolyzed neokestose. Therefore, the enzyme was termed 6G&1-FEH. Gene expression studies in asparagus roots showed that aoeh4 increased during root storage at 2 °C and spear harvesting. These findings suggest that 6G&1-FEH may be involved in fructan hydrolysis in asparagus roots to provide an energy source for emerging asparagus spears. [ABSTRACT FROM AUTHOR]

The article presents information related to fructans, multifunctional carbohydrates associated with stress responses in plants and microorganisms. It refers to the involvement of a fructan exohydrolase enzyme in degrading the neokestose-based fructans and central beta-2,6 linkage in neokestose has remained undetected.

In grassland plant communities, the ability of individual plants to regrow after defoliation is of crucial importance since it allows the restoration of active photosynthesis and plant growth. The aim of this study was to evaluate the effects of increasing defoliation intensity (0, 25, 65, 84, and 100% of removed leaf area) on sugar remobilization and N uptake, remobilization, and allocation in roots, adult leaves, and growing leaves of ryegrass over 2 days, using a 15N tracer technique. Increasing defoliation intensity decreased plant N uptake in a correlative way and increased plant N remobilization, but independently. The relative contribution of N stored before defoliation to leaf growth increased when defoliation intensity was severe. In most conditions, root N reserves also contributed to leaf regrowth, but much less than adult leaves and irrespective of defoliation intensity. A threshold of defoliation intensity (65% leaf area removal) was identified below which C (glucose, fructose, sucrose, fructans), and N (amino acids, soluble proteins) storage compounds were not recruited for regrowth. By contrast, nitrate content increased in elongating leaf bases above this threshold. Wounding associated with defoliation is thus not the predominant signal that triggers storage remobilization and controls the priority of resource allocation to leaf meristems. A framework integrating the sequential events leading to the refoliation of grasses is proposed on the basis of current knowledge and on the findings of the present work. [ABSTRACT FROM AUTHOR]

The sessile lifestyle of higher plants is accompanied by their remarkable ability to tolerate unfavorable environmental conditions. This is because, during evolution, plants developed a sophisticated repertoire of molecular and metabolic reactions to cope with changing biotic and abiotic challenges. In particular, the abiotic factors light intensity and ambient temperature are characterized by altering their amplitude within comparably short periods of time and are causative for onset of dynamic plant responses. These rapid responses in plants are also classified as 'acclimation reactions' which differ, due to their reversibility and duration, from non-reversible 'adaptation reactions'. In this review, we demonstrate the remarkable importance of stress-induced changes in carbohydrate homeostasis of plants exposed to high light or low temperatures. These changes represent a co-ordinated process comprising modifications of (i) the concentrations of selected sugars; (ii) starch turnover; (iii) intracellular sugar compartmentation; and (iv) corresponding gene expression patterns. The critical importance of these individual processes has been underlined in the recent past by the analyses of a large number of mutant plants. The outcome of these analyses raised our understanding of acclimation processes in plants per se but might even become instrumental to develop new concepts for directed breeding approaches with the aim to increase abiotic stress tolerance of crop species, which in most cases have high stress sensitivity. The latter direction of plant research is of special importance since abiotic stress stimuli strongly impact on crop productivity and are expected to become even more pronounced because of human activities which alter environmental conditions rapidly. [ABSTRACT FROM AUTHOR]

The article focuses on a study regarding the determination of fructans in infant formula and adult nutritionals with the help of high performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD). It mentions that blank corrections are required in the lowest fructan concentration and states that diluted sample are treated with a mixture of sucrase and α-glucanases to hydrolyze sucrose and α-glucans. It also mentions that Orafti P95 hydrolyzed inulin ingredient.

The article presents a study that determines the fructans in infant, adult, and pediatric nutritional formulas. It offers overview of the principle, apparatus and materials, and the preparation of standards and solutions in the study. It also outlines the result of the study as well as the calculations, validation protocol and accuracy of the findings.

A method for fructan analysis designed to comply with AOAC Standard Method Performance Requirements (SMPR®) 2014.002 is described. It is closely related to existing methods for fructan analysis, including AOAC 997.08 and 999.03, as well as a method previously published by Cuany et al. This new method achieves LOQ of 0.03% fructan on a ready-to-feed (RTF) basis with mean recoveries ranging from 93 to 108% in the presence of up to 9% sucrose (even at the 0.03% level of fructan). Repeatability ranged from 1.09 to 3.67%. Intermediate precision ranged from 2.46 to 6.79%. Sample preparation for quantitative analysis is simplified compared to some of the existing methodologies. The method incorporates a qualitative profile analysis to determine fructan size category. This allows assignment of appropriate correction factors without independent knowledge of fructan type. [ABSTRACT FROM AUTHOR]

Background and Aims Improving crops requires better linking of traits and metabolic processes to whole plant performance. In this paper, we present CN-Wheat, a comprehensive and mechanistic model of carbon (C) and nitrogen (N) metabolism within wheat culms after anthesis. Methods The culm is described by modules that represent the roots, photosynthetic organs and grains. Each of them includes structural, storage and mobile materials. Fluxes of C and N among modules occur through a common pool and through transpiration flow. Metabolite variations are represented by differential equations that depend on the physiological processes occurring in each module. A challenging aspect of CN-Wheat lies in the regulation of these processes by metabolite concentrations and the environment perceived by organs. Key Results CN-Wheat simulates the distribution of C and N into wheat culms in relation to photosynthesis, N uptake, metabolite turnover, root exudation and tissue death. Regulation of physiological activities by local concentrations of metabolites appears to be a valuable feature for understanding how the behaviour of the whole plant can emerge from local rules. Conclusions The originality of CN-Wheat is that it proposes an integrated view of plant functioning based on a mechanistic approach. The formalization of each process can be further refined in the future as knowledge progresses. This approach is expected to strengthen our capacity to understand plant responses to their environment and investigate plant traits adapted to changes in agronomical practices or environmental conditions. A companion paper will evaluate the model. [ABSTRACT FROM AUTHOR]

Background and Aims Simulating resource allocation in crops requires an integrated view of plant functioning and the formalization of interactions between carbon (C) and nitrogen (N) metabolisms. This study evaluates the functional–structural model CN-Wheat developed for winter wheat after anthesis. Methods In CN-Wheat the acquisition and allocation of resources between photosynthetic organs, roots and grains are emergent properties of sink and source activities and transfers of mobile metabolites. CN-Wheat was calibrated for field plants under three N fertilizations at anthesis. Model parameters were taken from the literature or calibrated on the experimental data. Key Results The model was able to predict the temporal variations and the distribution of resources in the culm. Thus, CN-Wheat accurately predicted the post-anthesis kinetics of dry masses and N content of photosynthetic organs and grains in response to N fertilization. In our simulations, when soil nitrates were non-limiting, N in grains was ultimately determined by availability of C for root activity. Dry matter accumulation in grains was mostly affected by photosynthetic organ lifespan, which was regulated by protein turnover and C-regulated root activity. Conclusions The present study illustrates that the hypotheses implemented in the model were able to predict realistic dynamics and spatial patterns of C and N. CN-Wheat provided insights into the interplay of C and N metabolism and how the depletion of mobile metabolites due to grain filling ultimately results in the cessation of resource capture. This enabled us to identify processes that limit grain mass and protein content and are potential targets for plant breeding. [ABSTRACT FROM AUTHOR]

Bacterial extracellular fructans, known as levans, have potential applications in food, pharmaceutical and cosmetic industries and high fructan producing strains could contribute into the cost reduction and more extensive commercial usage of them. An acetic acid bacteria ( AAB) isolate P1464 was obtained from the Microbial Strain Collection of Institute of Microbiology and Biotechnology, University of Latvia and identified as Gluconobacter nephelii by DNA- DNA hybridization and the formation of extracellular fructans by this strain was confirmed. Isolated extracellular fructose polymers were characterized using FT- IR spectroscopy and the structural features of fructan appeared as similar to the reference sample of bacterial levan. Molecular mass estimates showed that the isolated G. nephelii P1464 fructose polymer has a relatively small molecular weight (Mw 1122·939 ± 153·453 kDa) and a sizeable polydispersity (Mw/Mn = 21·57 ± 1·60), as compared with other AAB, which could promote their physiological activity, including the prebiotic effects. Obtained at different cultivation conditions characteristics of fructan production, including the biotechnological indices such as the productivity (Qp) and yield (Yp/s) ranging from 0·774 to 1·244 g l−1 h and from 0·181 to 0·436 g g−1, respectively, confirmed, that G. nephelii P1464 could be used as promising strain for commercial production of levan. Significance and Impact of the Study Bacterial fructans, known as levans, have extensive options for practical usage, however, actually limited due to high production costs. Therefore, the searches for efficient producer strains should be an urgent task to reduce costs. This study is the first report on the formation of fructans by a novel strain of acetic acid bacteria ( AAB) Gluconobacter nephelii P1464. Characteristics obtained at different cultivation conditions confirmed the operation of a competitive and perspective producer strain. Isolated extracellular fructans are characterized by a lower molecular weight as compared with other AAB which could promote their physiological activity, including the prebiotic effects. [ABSTRACT FROM AUTHOR]

Aims: To determine if Butyrivibrio fibrisolvens strain 3071 is able to use fructose polymers for growth and to identify the enzymes involved in their digestion. Methods and Results: Strain 3071 utilized 97, 89, 85 and 60% of sucrose, timothy grass fructan, inulin oligosaccharides and inulin, respectively, in the growth medium. A cell extract from timothy grass fructan-grown bacteria was used for identification of fructanolytic enzymes by anion exchange chromatography, gel filtration, zymography and thin-layer chromatography. The bacterium synthesizes a specific endolevanase and a nonspecific β-fructofuranosidase. Both enzymes occurred in two forms differing in molecular weight. The β-fructofuranosidase was not able to digest long-chain inulin or timothy grass fructan, but degraded inulin oligosaccharides and sucrose. Addition of 1,4-dithioerythritol to an enzyme solution did not affect the activity of endolevanase(s), but increased the ability of β-fructofuranosidase to digest sucrose. The digestion of timothy grass fructan by endolevanase(s) was described by Michaelis-Menten kinetics in which Km = 2.82 g-l and Vmax = 4.01 μmoles reducing sugar equivalents × mg-1-1 × min-1. Conclusion: Strain 3071 synthesizes enzymes enabling it to use grass fructans for growth. Significance and Impact of the Study: Butyrivibrio fibrisolvens strain 3071 can be considered a member of the rumen fructanolytic guild. [ABSTRACT FROM AUTHOR]

Background and Aims: Extreme climatic events such as severe droughts are expected to increase with climate change and to limit grassland perennity. The present study aimed to characterize the adaptive responses by which temperate herbaceous grassland species resist, survive and recover from a severe drought and to explore the relationships between plant resource use and drought resistance strategies. Methods: Monocultures of six native perennial species from upland grasslands and one Mediterranean drought-resistant cultivar were compared under semi-controlled and non-limiting rooting depth conditions. Above- and below-ground traits were measured under irrigation in spring and during drought in summer (50 d of withholding water) in order to characterize resource use and drought resistance strategies. Plants were then rehydrated and assessed for survival (after 15 d) and recovery (after 1 year). Key Results: Dehydration avoidance through water uptake was associated with species that had deep roots (>1·2 m) and high root mass (>4 kg m-3). Cell membrane stability ensuring dehydration tolerance of roots and meristems was positively correlated with fructan content and negatively correlated with sucrose content. Species that survived and recovered best combined high resource acquisition in spring (leaf elongation rate >9 mm d-1 and rooting depth >1·2 m) with both high dehydration avoidance and tolerance strategies. Conclusions: Most of the native forage species, dominant in upland grassland, were able to survive and recover from extreme drought, but with various time lags. Overall the results suggest that the wide range of interspecific functional strategies for coping with drought may enhance the resilience of upland grassland plant communities under extreme drought events. [ABSTRACT FROM AUTHOR]

Background and Aims: Prebiotic inulin-type fructans are widely consumed in the diet and may have contrasting effects in Crohn's disease by stimulating gut microbiota and/or by generating functional gastrointestinal symptoms. The aim of this study was to measure fructan and oligofructose intakes in patients with active and inactive Crohn's disease compared with healthy controls. Methods: Patients with active Crohn's disease (n = 98), inactive Crohn's (n = 99) and healthy controls (n = 106) were recruited to a case-control study. Dietary intake of inulin-type fructans was measured using a specific food frequency questionnaire and was compared between the three groups and between patients with different disease phenotypes (Montreal classification). Associations between intakes and disease activity (Harvey-Bradshaw Index, HBI) were also undertaken. Results: Patients with active Crohn's disease had lower fructan intakes (median 2.9 g/d, interquartile range [IQR] 1.8) than those with inactive Crohn's (3.6 g/d, 2.1, p = 0.036) or controls (3.9 g/d, 2.1, p = 0.003) and lower oligofructose intakes (2.8 g/d, 1.8) than those with inactive Crohn's (3.5 g/d, 2.2, p = 0.048) or controls (3.8 g/d, 2.1, p = 0.003). There were no differences in intakes related to disease site or behaviour. There were negative correlations between HBI well-being score and fructan intake (ρ = -0.154, p = 0.03) and oligofructose intake (ρ = -0.156, p = 0.028) and for the HBI abdominal pain score and fructan (ρ = -0.164, p = 0.021) and oligofructose intake (ρ = -0.157, p = 0.027). Conclusions: Patients with active Crohn's disease consume lower quantities of fructans and oligofructose than their inactive counterparts and healthy controls. The impact of lower intakes of prebiotic fructans on gut microbiota is unknown and warrants further research. [ABSTRACT FROM AUTHOR]

Background and Aims Chrysolaena obovata, an Asteraceae of the Brazilian Cerrado, presents seasonal growth, marked by senescence of aerial organs in winter and subsequent regrowth at the end of this season. The underground reserve organs, the rhizophores, accumulate inulin-type fructans, which are known to confer tolerance to drought and low temperature. Fructans and fructan-metabolizing enzymes show a characteristic spatial and temporal distribution in the rhizophores during the developmental cycle. Previous studies have shown correlations between abscisic acid (ABA) or indole acetic acid (IAA), fructans, dormancy and tolerance to drought and cold, but the signalling mechanism for the beginning of dormancy and sprouting in this species is still unknown. Methods Adult plants were sampled from the field across phenological phases including dormancy, sprouting and vegetative growth. Endogenous concentrations of ABA and IAA were determined by GC-MS-SIM (gas chromatography-mass spectrometry-selected ion monitoring), and measurements were made of fructan content and composition, and enzyme activities. The relative expression of corresponding genes during dormancy and sprouting were also determined. Key Results Plants showed a high fructan 1-exohydrolase (EC 3.2.1.153) activity and expression during sprouting in proximal segments of the rhizophores, indicating mobilization of fructan reserves, when ABA concentrations were relatively low and precipitation and temperature were at their minimum values. Concomitantly, higher IAA concentrations were consistent with the role of this regulator in promoting cell elongation and plant growth. With high rates of precipitation and high temperatures in summer, the fructan-synthesizing enzyme sucrose:sucrose 1-fructosyltransferase (EC 2.4.1.99) showed higher activity and expression in distal segments of the rhizophores, which decreased over the course of the vegetative stage when ABA concentrations were higher, possibly signalling the entry into dormancy. Conclusions The results show that fructan metabolism correlates well with endogenous hormone concentrations and environmental changes, suggesting that the co-ordinated action of carbohydrate metabolism and hormone synthesis enables C. obovata to survive unfavourable field conditions. Endogenous hormone concentrations seem to be related to regulation of fructan metabolism and to the transition between phenophases, signalling for energy storage, reserve mobilization and accumulation of oligosaccharides as osmolytes. [ABSTRACT FROM AUTHOR]

Fructan supplementation of a commercially available canned cat food was evaluated using senior (≥ 9 yr) cats to assess nitrogen (N) partitioning in excreta and stool metabolite and microbiota concentrations. Oligofructose (OF) or SynergyC (OF+IN) were added to the diet individually at 1% (dry weight basis). Cats were acclimated to the control diet for 7 d and then were randomly assigned to 1 of 3 treatment groups for 21 d (n = 6). Feces and urine were collected on d 22 through 28. No differences were observed in food intake; fecal output, DM percentage, score, pH, or short- or branched-chain fatty acids, fecal and urinary ammonia output, urinary felinine concentrations, or N retention. Supplemental OF+IN tended to decrease N digestibility (P = 0.102) and Bifidobacteria spp. (P = 0.073) and decrease fecal indole (P < 0.05), tyramine (P < 0.05), and Escherichia coli (P < 0.05) concentrations. Both fructan-supplemented treatments decreased (P < 0.05) fecal histamine concentrations. The tendency to a lower apparent N digestibility was likely due to increased colonic microbial protein synthesis of fructan-supplemented cats. Fructan supplementation may benefit senior cats as it modulates stool odor-forming compounds and decreases some protein catabolites and pathogenic gut microbiota concentrations without affecting N retention. [ABSTRACT FROM AUTHOR]

Aims The goals of this study were to determine if β-acid from hops ( Humulus lupulus L.) could be used to control fructan fermentation by equine hindgut micro-organisms, and to verify the antimicrobial mode of action on Streptococcus bovis, which has been implicated in fructan fermentation, hindgut acidosis and pasture-associated laminitis (PAL) in the horse. Methods and Results Suspensions of uncultivated equine faecal micro-organisms produced fermentation acids when inulin (model fructan) was the substrate, but β-acid (i.e. lupulone) concentrations ≥9 ppm inhibited lactate production and mitigated the decrease in pH. Inulin-fermenting Strep. bovis was isolated from the β-acid-free suspensions after enrichment with inulin. The isolates were sensitive to β-acid, which decreased the viable number of streptococci in faecal suspensions, as well as growth, lactate production and the intracellular potassium of Strep. bovis in pure culture. Conclusions These results are consistent with the hypothesis that hops β-acid prevented the growth of fructan-fermenting equine faecal bacteria, and that the mechanism of action was dissipation of the intracellular potassium of Strep. bovis. Significance and Impact of the Study Bacterial hindgut fermentation of grass fructans has been linked to PAL and other metabolic disorders in horses. Hops β-acid is a potential phytochemical intervention to decrease the growth of bacteria responsible for PAL. [ABSTRACT FROM AUTHOR]

Background and Aims Cold is a major constraint for cereal cultivation under temperate climates. Winter-hardy plants interpret seasonal changes and can acquire the ability to resist sub-zero temperatures. This cold acclimation process is associated with physiological, biochemical and molecular alterations in cereals. Brachypodium distachyon is considered a powerful model system to study the response of temperate cereals to adverse environmental conditions. To date, little is known about the cold acclimation and freezing tolerance capacities of Brachypodium. The main objective of this study was to evaluate the cold hardiness of seven diploid Brachypodium accessions. Methods An integrated approach, involving monitoring of phenological indicators along with expression profiling of the major vernalization regulator VRN1 orthologue, was followed. In parallel, soluble sugars and proline contents were determined along with expression profiles of two COR genes in plants exposed to low temperatures. Finally, whole-plant freezing tests were performed to evaluate the freezing tolerance capacity of Brachypodium. Key Results Cold treatment accelerated the transition from the vegetative to the reproductive phase in all diploid Brachypodium accessions tested. In addition, low temperature exposure triggered the gradual accumulation of BradiVRN1 transcripts in all accessions tested. These accessions exhibited a clear cold acclimation response by progressively accumulating proline, sugars and COR gene transcripts. However, whole-plant freezing tests revealed that these seven diploid accessions only have a limited capacity to develop freezing tolerance when compared with winter varieties of temperate cereals such as wheat and barley. Furthermore, little difference in terms of survival was observed among the accessions tested despite their previous classification as either spring or winter genotypes. Conclusions This study is the first to characterize the freezing tolerance capacities of B. distachyon and provides strong evidence that some diploid accessions such as Bd21 have a facultative growth habit. [ABSTRACT FROM PUBLISHER]

Dietary macronutrients affect the composition of the gut microbiota, and prebiotics are used to improve and maintain a healthy gut. The impact of prebiotics on dominant gut bacteria other than bifidobacteria, however, is under-researched. Here, we report carbohydrate utilisation patterns for representative butyrate-producing anaerobes, belonging to the Gram-positive Firmicutes families Lachnospiraceae and Ruminococcaceae, by comparison with selected Bacteroides and Bifidobacterium species. Growth assessments using anaerobic Hungate tubes and a new rapid microtitre plate assay were generally in good agreement. The Bacteroides strains tested showed some growth on basal medium with no added carbohydrates, utilising peptides in the growth medium. The butyrate-producing strains exhibited different growth profiles on the substrates, which included starch, inulin, fructooligosaccharides ( FOS), galactooligosaccharides ( GOS) and xylooligosaccharides ( XOS). Eleven were able to grow on short-chain FOS, but this number decreased as the chain length of the fructan substrates increased. Long-chain inulin was utilised by Roseburia inulinivorans, but by none of the Bifidobacterium species examined here. XOS was a more selective growth substrate than FOS, with only six of the 11 Firmicutes strains able to use XOS for growth. These results illustrate the selectivity of different prebiotics and help to explain why some are butyrogenic. [ABSTRACT FROM AUTHOR]

Although fructans play a crucial role in wheat kernel development, their metabolism during kernel maturation is far from being understood. In this study, all major fructan-metabolizing enzymes together with fructan content, fructan degree of polymerization and the presence of fructan oligosaccharides were examined in developing wheat kernels (Triticum aestivum L. var. Homeros) from anthesis until maturity. Fructan accumulation occurred mainly in the first 2 weeks after anthesis, and a maximal fructan concentration of 2.5 ± 0.3 mg fructan per kernel was reached at 16 days after anthesis (DAA). Fructan synthesis was catalyzed by 1-SST (sucrose:sucrose 1-fructosyltransferase) and 6-SFT (sucrose:fructan 6-fructosyltransferase), and to a lesser extent by 1-FFT (fructan:fructan 1-fructosyltransferase). Despite the presence of 6G-kestotriose in wheat kernel extracts, the measured 6G-FFT (fructan:fructan 6G-fructosyltransferase) activity levels were low. During kernel filling, which lasted from 2 to 6 weeks after anthesis, kernel fructan content decreased from 2.5 ± 0.3 to 1.31 ± 0.12 mg fructan per kernel (42 DAA) and the average fructan degree of polymerization decreased from 7.3 ± 0.4 (14 DAA) to 4.4 ± 0.1 (42 DAA). FEH (fructan exohydrolase) reached maximal activity between 20 and 28 DAA. No fructan-metabolizing enzyme activities were registered during the final phase of kernel maturation, and fructan content and structure remained unchanged. This study provides insight into the complex metabolism of fructans during wheat kernel development and relates fructan turnover to the general phases of kernel development. [ABSTRACT FROM PUBLISHER]

Fructans are the major component of temporary carbon reserve in the stem of temperate cereals, which is used for grain filling. Three families of fructosyltransferases are directly involved in fructan synthesis in the vacuole of Triticum aestivum. The regulatory network of the fructan synthetic pathway is largely unknown. Recently, a sucrose-upregulated wheat MYB transcription factor (TaMYB13-1) was shown to be capable of activating the promoter activities of sucrose:sucrose 1-fructosyltransferase (1-SST) and sucrose:fructan 6-fructosyltransferase (6-SFT) in transient transactivation assays. This work investigated TaMYB13-1 target genes and their influence on fructan synthesis in transgenic wheat. TaMYB13-1 overexpression resulted in upregulation of all three families of fructosyltransferases including fructan:fructan 1-fructosyltransferase (1-FFT). A γ-vacuolar processing enzyme (γ-VPE1), potentially involved in processing the maturation of fructosyltransferases in the vacuole, was also upregulated by TaMYB13-1 overexpression. Multiple TaMYB13 DNA-binding motifs were identified in the Ta1-FFT1 and Taγ-VPE1 promoters and were bound strongly by TaMYB13-1. The expression profiles of these target genes and TaMYB13-1 were highly correlated in recombinant inbred lines and during stem development as well as the transgenic and non-transgenic wheat dataset, further supporting a direct regulation of these genes by TaMYB13-1. TaMYB13-1 overexpression in wheat led to enhanced fructan accumulation in the leaves and stems and also increased spike weight and grain weight per spike in transgenic plants under water-limited conditions. These data suggest that TaMYB13-1 plays an important role in coordinated upregulation of genes necessary for fructan synthesis and can be used as a molecular tool to improve the high fructan trait. [ABSTRACT FROM AUTHOR]

In Exp. 1, 80 growing pigs (27.1 ± 0.7 kg) were used in a 42-d experiment to evaluate the effect of levan-type fructan on growth performance, digestibility, blood profile, and fecal microbiota. Pigs were randomly allocated to 1 of 4 treatments, according to initial BW and gender, with 5 replicate pens per treatment and 2 bar-rows and 2 gilts per pen. Treatments were com-soybean meal-based diets supplemented with 0%, 0.05%, 0.10%, or 0.20% levan-type fructan. Average daily gain and G:F increased (quadratic, P < 0.05), as dietary levan-type fructan increased from 0 to 0.2%. Similarly, the apparent total tract digestibility of N and GE increased (quadrat-ic, P < 0.05), as dietary supplementation of levan-type fructan increased. Dietary levan-type fructan supplemen-tation increased fecal Lactobacillus counts linearly (P < 0.05). In Exp. 2, 20 individually housed barrows (26.2 ± 0.6 kg) were used to evaluate immune responses after an Escherichia coli lipopolysaccharide (LPS) challenge. Pigs were fed com-soybean meal-based diets supple-mented with 0% or 0.10% levan-type fructan for 42 d. At d 42, 5 pigs from each treatment were injected with E. coli LPS (0.01% of BW) and the other 5 pigs with sterile saline solution, resulting in a 2 x 2 factorial arrangement of treatments. Blood was taken 0, 2, 4, 6, and 8 h after challenge. Challenge with LPS decreased blood lym-phocyte percentage and had an interactive effect with levan-type fructan inclusion at 4, 6, and 8 h (P < 0.01). Levan-type fructan supplementation increased (P < 0.05) white blood cells at 6 and 8 h, and increased (P < 0.05) lymphocyte percentage at 8 h after the challenge. Lipopolysaccharide injection increased (P < 0.05) rectal temperature at 2 and 4 h, and had an interactive effect (P < 0.05) with levan-type fructan supplementation at 4 h after the challenge. At 2, 4, 6, and 8 h, serum Cortisol, tumor necrosis factor-a, and IL-6 concentration increased (P < 0.05) by LPS challenge, and there was an interac-tive effect between LPS challenge and the inclusion of levan-type fructan (P < 0.05). Dietary levan-type fructan supplementation decreased serum Cortisol concentration at 8 h, tumor necrosis factor-a concentration at 4, 6, and 8 h, and IL-6 concentration at 6 and 8 h after the challenge (P < 0.05). In conclusion, dietary supplementation with 0.10% levan-type fructan can improve growth perfor-mance, digestibility, and fecal Lactobacillus counts, and has a beneficial effect on the immune response during an inflammatory challenge in growing pigs. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the effect of dietary fructan supplementation on growth performance, nutrient digestibility, meat quality, fecal microbial flora, and fecal noxious gas emission in finishing pigs. A total of 96 finishing pigs [(Yorkshire x Landrace) x Duroc] with an average BW of 73.1 ± 2.5 kg were used in a 6-wk study. Pigs were randomly allotted to 1 of 3 dietary treatments: 1) CON, basal diet, 2) CON + 1% fructan (FC1), and 3) CON + 2% fructan (FC2) with 8 replicate pens per treatment and 2 barrows and 2 gilts per pen. During the overall study, pigs fed the fructan supplementation diets had a greater (P < 0.05) ADG, G:F, the apparent total tract digestibility (ATTD) of DM and GE than pigs fed the CON diet. The levels of fructan supplementation did not affect growth performance and ATTD of DM, N, and GE. Fecal E. coli concentrations in the fructan treatments were lower (P < 0.001), while fecal Lactobacillus concentrations in the fructan treatments were greater (P < 0.001) compared with the CON treatment. Fecal ammonia and hydrogen sulfide emissions were lower (P < 0.05) in d 3, 5, and 7 in the fructan treatments, and total mercaptans emission was lower (P < 0.05) on d 3 and 5 compared with the CON treatment. In conclusion, fructan supplementation improved growth performance and ATTD of DM and GE, improved the fecal microbial balance, and inhibited the fecal E. coli. Furthermore, fructan may decrease fecal noxious gas emissions by finishing pigs. [ABSTRACT FROM AUTHOR]

A total of 720 1-d-old male Ross broilers (BW of 48.0 ± 0.3 g) were used to evaluate the effects of dietary levan fructan supplementation on growth performance, meat quality, relative organ weight, cecal microflora, and excreta noxious gas emission in broilers. This experiment lasted 31 d. Broilers were randomly allotted to 1 of 3 dietary treatments: 1) CON, basal diet, 2) CON + 0.25% fructan (FCl), and 3) CON + 0.50% fructan (FC2). Each treatment contained 16 pens with 15 chicks per pen. Broilers on levan fructan supplementation treatments (FCl + FC2) had a lower (P = 0.005 for d 15 to 31 ) ADFI and greater (P = 0.005 for d 15 to 31 and P = 0.022 for d 1 to 31 ) G:F than those on the CON. A decreased (P = 0.031) relative spleen weight was observed with levan fructan supplementation treatments compared with the CON. Cecal E. coli and C. perfringens concentrations in levan fructan treatments were decreased, while cecal Lactobacillus, as well as Bifidobacteria, concentrations in levan fructan treatments were increased compared with the CON. However, excreta NH3 concentrations were decreased (P=0.013 ) in levan fructan treatments compared with the CON. In conclusion, fructan supplementation improved later stage growth performance, increased cecal Lactobacillus and Bifidobacteria concentrations, and decreased cecal E. coli and C. perfringens concentrations, as well as excreta NH3, concentrations, in broilers. [ABSTRACT FROM AUTHOR]

Background and Aims There are many unresolved issues concerning the biochemistry of fructan biosynthesis. The aim of this paper is to address some of these by means of modelling mathematically the biochemical processes. Methods A model has been constructed for the step-by-step synthesis of fructan polymers. This is run until a steady state is achieved for which a polymer distribution is predicted. It is shown how qualitatively different distributions can be obtained. Key Results It is demonstrated how a set of experimental results on polymer distribution can by simulated by a simple parameter adjustments. Conclusions Mathematical modelling of fructan biosynthesis can provide a useful tool for helping elucidate the details of the biosynthetic processes. [ABSTRACT FROM PUBLISHER]

Recent in vitro, in vivo, and theoretical experiments strongly suggest that sugar-(like) molecules counteract oxidative stress by acting as genuine reactive oxygen species (ROS) scavengers. A concept was proposed to include the vacuole as a part of the cellular antioxidant network. According to this view, sugars and sugar-like vacuolar compounds work in concert with vacuolar phenolic compounds and the ‘classic’ cytosolic antioxidant mechanisms. Among the biologically relevant ROS (H2O2, O2·–, and ·OH), hydroxyl radicals are the most reactive and dangerous species since there are no enzymatic systems known to neutralize them in any living beings. Therefore, it is important to study in more detail the radical reactions between ·OH and different biomolecules, including sugars. Here, Fenton reactions were used to compare the ·OH-scavenging capacities of a range of natural vacuolar compounds to establish relationships between antioxidant capacity and chemical structure and to unravel the mechanisms of ·OH–carbohydrate reactions. The in vitro work on the ·OH-scavenging capacity of sugars and phenolic compounds revealed a correlation between structure and ·OH-scavenging capacity. The number and position of the C=C type of linkages in phenolic compounds greatly influence antioxidant properties. Importantly, the splitting of disaccharides and oligosaccharides emerged as a predominant outcome of the ·OH–carbohydrate interaction. Moreover, non-enzymatic synthesis of new fructan oligosaccharides was found starting from 1-kestotriose. Based on these and previous findings, a working model is proposed describing the putative radical reactions involving fructans and secondary metabolites at the inner side of the tonoplast and in the vacuolar lumen. [ABSTRACT FROM AUTHOR]

Agave tequilana fructans are the source of fermentable sugars for the production of tequila. Fructans are processed by acid hydrolysis or by cooking in ovens at high temperature. Enzymatic hydrolysis is considered an alternative for the bioconversion of fructans. We previously described the isolation of Aspergillus niger CH-A-2010, an indigenous strain that produces extracellular inulinases. Here we evaluated the potential application of A. niger CH-A-2010 inulinases for the bioconversion of A. tequilana fructans, and its impact on the production of ethanol. Inulinases were analyzed by Western blotting and thin layer chromatography. Optimal pH and temperature conditions for inulinase activity were determined. The efficiency of A. niger CH-A-2010 inulinases was compared with commercial enzymes and with acid hydrolysis. The hydrolysates obtained were subsequently fermented by Saccharomyces cerevisiae to determine the efficiency of ethanol production. Results indicate that A. niger CH-A-2010 predominantly produces an exo-inulinase activity. Optimal inulinase activity occurred at pH 5.0 and 50 °C. Hydrolysis of raw agave juice by CH-A-2010 inulinases yielded 33.5 g/l reducing sugars, compared with 27.3 g/l by Fructozyme (Novozymes Corp, Bagsværd, Denmark) and 29.4 g/l by acid hydrolysis. After fermentation of hydrolysates, we observed that the conversion efficiency of sugars into ethanol was 97.5 % of the theoretical ethanol yield for enzymatically degraded agave juice, compared to 83.8 % for acid-hydrolyzed juice. These observations indicate that fructans from raw Agave tequilana juice can be efficiently hydrolyzed by using A. niger CH-A-2010 inulinases, and that this procedure impacts positively on the production of ethanol. [ABSTRACT FROM AUTHOR]

A total of 150 weanling pigs [(Yorkshire × Landrace) × Duroc] with an average BW of 7.22 ± 0.80 kg (21 d of age) were used in a 28-d trial to determine the effects of dietary fructan and mannan oligosaccharides on growth performance, nutrient digestibility, blood profile, and diarrhea score in weanling pigs. Pigs were allotted randomly to 1 of 5 dietary treatments: 1) negative control (NC), basal diet; 2) positive control (PC), NC + 0.01% apramycin (165 mg/kg); 3) NC + 0.1% fructan (FC); 4) NC + 0.1% mannan oligosaccharide source (MO); and 5) NC + 0.05% fructan + 0.05% mannan oligosaccharide source (FM). There were 3 replications per treatment with 10 pigs per pen (5 barrows and 5 gilts). From d 0 to 14, ADG and ADFI of pigs fed the PC, MO, and FM diets were greater (P < 0.05) than pigs fed the NC diet. From d 15 to 28, there were no differences (P > 0.05) in ADG, ADFI, and G:F. During the overall period (d 0 to 28), pigs fed the MO diet had a greater ADG than pigs fed the NC diet (P < 0.05). Pigs fed the PC and MO diets increased ADFI (P < 0.05) compared with pigs fed the NC diet. However, no differences were detected among dietary treatments in G:F during the overall experimental period. On d 14, the apparent total tract digestibility (ATTD) of DM and N in pigs fed the PC, MO, and FM diets was greater (P < 0.05) than pigs fed the NC diet. The ATTD of DM increased (P < 0.05) in pigs fed the MO and FM diets compared with pigs fed the FC diet. However, at the end of the experiment, pigs fed the FM diet had a greater (P < 0.05) ATTD of DM compared with pigs fed the NC diet. Additionally, there were no differences in IgG, red blood cells, white blood cells, and lymphocyte counts among dietary treatments on d 0, 14, or 28. The diarrhea score in pigs fed the MO diet was reduced (P < 0.05) compared with pigs fed the NC diet. In conclusion, mannan oligosaccharides have a beneficial effect on growth performance and nutrient digestibility in weanling pigs. Furthermore, mannan oligosaccharides can decrease diarrhea score in weanling pigs. [ABSTRACT FROM AUTHOR]

The article presents a study which evaluates ameliorating effects of short-chain inulin-like fructans (SIF) with differing degrees of polymerization (DP) on the trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats. It states that the study monitored fecal myeloperoxidase (MPO) activity and IgA concentration. It concludes that DP4 significantly reduced colonic inflammation accompanied with higher concentrations of short-chain fatty acids.

The main storage compounds in Lolium perenne are fructans with prevailing β(2–6) linkages. A cDNA library of L. perenne was screened using Poa secunda sucrose:fructan 6-fructosyltransferase (6-SFT) as a probe. A full-length Lp6-SFT clone was isolated as shown by heterologous expression in Pichia pastoris. High levels of Lp6-SFT transcription were found in the growth zone of elongating leaves and in mature leaf sheaths where fructans are synthesized. Upon fructan synthesis induction, Lp6-SFT transcription was high in mature leaf blades but with no concomitant accumulation of fructans. In vitro studies with the recombinant Lp6-SFT protein showed that both 1-kestotriose and 6G-kestotriose acted as fructosyl acceptors, producing 1- and 6-kestotetraose (bifurcose) and 6G,6-kestotetraose, respectively. Interestingly, bifurcose formation ceased and 6G,6-kestotetraose was formed instead, when recombinant fructan:fructan 6G-fructosyltransferase (6G-FFT) of L. perenne was introduced in the enzyme assay with sucrose and 1-kestotriose as substrates. The remarkable absence of bifurcose in L. perenne tissues might be explained by a higher affinity of 6G-FFT, as compared with 6-SFT, for 1-kestotriose, which is the first fructan formed. Surprisingly, recombinant 6-SFT from Hordeum vulgare, a plant devoid of fructans with internal glucosyl residues, also produced 6G,6-kestotetraose from sucrose and 6G-kestotriose. In the presence of recombinant L. perenne 6G-FFT, it produced 6G,6-kestotetraose from 1-kestotriose and sucrose, like L. perenne 6-SFT. Thus, we demonstrate that the two 6-SFTs have close catalytic properties and that the distinct fructans formed in L. perenne and H. vulgare can be explained by the presence of 6G-FFT activity in L. perenne and its absence in H. vulgare. [ABSTRACT FROM PUBLISHER]

Twelve young adult (1.7 ± 0.1 yr) male cats were used in a replicated 3 × 3 Latin square design to determine the effects of fiber type on nutrient digestibility, fermentative end products, and fecal microbial populations. Three diets containing 4% cellulose, fructooligosaccharides (FOS), or pectin were evaluated. Feces were scored based on the 5-point system: 1 being hard, dry pellets, and 5 being watery liquid that can be poured. No differences were observed (P> 0.100) in intake of DM, OM, CP, or acid-hydrolyzed fat; DM or OM digestibility; or fecal pH, DM%, output on an as-is or DM basis, or concentrations of histamine or phenylalanine. Crude protein and fat digestibility decreased (P = 0.079 and 0.001, respectively) in response to supplementation with pectin compared with cellulose. Both FOS and pectin supplementation resulted in increased fecal scores (P < 0.001) and concentrations of ammonia (P = 0.003) and 4-methyl phenol (P = 0.003). Fecal indole concentrations increased (P = 0.049) when cats were supplemented with FOS. Fecal acetate (P = 0.030), propionate (P = 0.035), and total short-chain fatty acid (P = 0.016) concentrations increased in pectin-supplemented cats. Fecal butyrate (P = 0.010), isobutyrate (P = 0.011), isovalerate (P = 0.012), valerate (P = 0.026), and total branched-chain fatty acids + valerate (P = 0.008) concentrations increased with supplementation of FOS and pectin. Fecal cadaverine (P < 0.001) and tryptamine (P < 0.001) concentrations increased with supplementation of FOS and pectin. Fecal tyramine concentrations decreased (P = 0.039) in FOS-supplemented cats, whereas spermidine concentrations increased (P < 0.001) in pectin-supplemented cats. Whereas fecal concentrations of putrescine (P < 0.001) and total biogenic amines (P < 0.001) increased with FOS and pectin, the concentrations of these compounds were increased (P < 0.001) in cats supplemented with pectin. Fecal Bifido bacterium spp. concentrations increased (P = 0.006) and Escherichia coli concentrations decreased (P < 0.001) in FOS-supplemented cats. Fecal concentrations of Clostridium perfringens (P < 0.001), E. coli (P < 0.001), and Lactobacillus spp. (P = 0.030) also increased in pectin-supplemented cats. In addition to increasing populations of protein-fermenting microbiota, pectin increased production of fermentative end products associated with carbohydrate compared with protein fermentation. Pectin and FOS may be useful fiber sources in promoting intestinal health of the cat. [ABSTRACT FROM AUTHOR]

The article presents a study on the stimulation of the growth of exogenously administered Lactobacillus plantarum No.14 (LP14) by inulin-type fructans in the gastrointestinal tract of the mouse. The study examined the effects of the inulin-type fructans known as fructo-oligosaccharide (FOS). Moreover, the LP14 that was localized to the luminal contents has been showed by histological analysis following fluorescein-stained LP14 administration.

Homopolysaccharide (glucan and fructan) synthesis from sucrose by sucrase enzymes in lactic acid bacteria (LAB) has been well studied in the genera Leuconostoc, Streptococcus and Lactobacillus. This study aimed to identify and characterize genes encoding glucansucrase/glucosyltransferase (GTF) and fructansucrases/fructosyltransferase (FTF) enzymes from genomic DNA of ‘rare’ Indonesian exopolysaccharide-producing LAB. From a total of 63 exopolysaccharide-producing LAB isolates obtained from foods, beverages and environmental samples, 18 isolates showing the most slimy and mucoid colony morphologies on sucrose were chosen for further study. By comparing bacterial growth on De Man, Rogosa and Sharpe (MRS)-sucrose with that on MRS-raffinose, and using the results of a previous PCR screening study with degenerate primer pairs targeting the conserved catalytic domain of GTFs, various strains were identified as producers of fructan (13), of glucan only (five) or as potential producers of both glucan and fructan (nine). Here, we report the characteristics of three gtf genes and one ftf gene obtained from Weissella confusa strains MBF8-1 and MBF8-2. Strain MBF8-1 harbored two putative gtf genes with high sequence similarity to GTFB of Lactobacillus reuteri 121 and GTF180 of L. reuteri 180, respectively. Strain MBF8-2 possessed single gtf and ftf genes with high sequence similarity to GTFKg3 of Lactobacillus fermentum Kg3 and DSRWC of Weissella cibaria, and FTF levansucrase of L. reuteri 121, respectively. [ABSTRACT FROM AUTHOR]

Five ileal-cannulated adult dogs were utilized in a 5 x 5 Latin square design to determine the effects of fructan type and concentration on nutrient digestibility, stool metabolite concentrations, and fecal microbiota. Five diets were evaluated that contained cellulose alone or with inulin or short-chain fructooligosaccharides (scFOS) each at 0.2 or 0.4% of the diet. Dogs were fed 175 g of their assigned diet twice daily. Chromic oxide served as a digestibility marker. Nutrient digestibility; ileal and fecal pH and ammonia concentrations; ileal IgA concentrations; and fecal short- and branched-chain fatty acid concentrations, microbiota, and concentrations of phenol, indole, and biogenic amines were measured. No differences were observed in ileal pH or ammonia or fecal concentrations of indole or valerate. Ileal DM, OM, and CP digestibility coefficients; total tract DM and OM digestibility coefficients; and fecal concentrations of phenylethylamine increased linearly (P < 0.05), and fecal concentrations of phenol decreased linearly (P < 0.05) with inulin supplementation. Fecal concentrations of acetate, propionate, and total short-chain fatty acids decreased quadratically (P < 0.05) with inulin supplementation. Ileal DM, OM, and CP digestibility coefficients increased linearly (P < 0.05), and fecal phenol concentration decreased linearly (P < 0.05) with scFOS supplementation. Total tract DM and OM digestibility coefficients as well as fecal butyrate and isobutyrate concentrations increased quadratically (P < 0.05) with scFOS supplementation. Although a greater level of inclusion is needed to modify gut microbiota populations, low-level inclusion of inulin or scFOS is effective in modifying key nutritional outcomes in the dog. [ABSTRACT FROM AUTHOR]

Invertases cleave sucrose in glucose and fructose, using water as an acceptor. Fructosyltransferases catalyse the transfer of a fructosyl residue between sucrose and/or fructan molecules. Plant fructosyltransferases (FTs) evolved from vacuolar invertases by small mutational changes, leading to differences in substrate specificity. The S-type of enzymes (invertases, sucrose:sucrose 1-fructosyltransferases or 1-SSTs, and sucrose:fructan 6-fructosyltransferases or 6-SFTs) prefer sucrose as the donor substrate while F-type enzymes (fructan:fructan 1-fructosyltransferases or 1-FFTs and fructan:fructan 6G-fructosyltransferases or 6G-FFTs) preferentially use fructan as the donor substrate. Recently, a functional Asp/Arg or Asp/Lys couple in the Hypervariable Loop (HVL) was suggested to be essential to keep Asp in a favourable orientation for binding sucrose as the donor substrate in S-type enzymes. However, the F-type enzyme 1-FFT of Triticum aestivum (Ta1-FFT) also contains the Asp/Arg couple in the HVL, although it prefers fructan as the donor substrate. In this paper, mutagenesis studies on Ta1-FFT are presented. In Ta1-SST, Tyr282 (the Asp281 homologue) seems to be essential in creating a tight H-bond Network (HBN) in which the Arg-residue of the Asp/Arg couple is held in a fixed position. This tight HBN is disrupted in Ta1-FFT, leading to a more flexible Arg-residue and a dysfunctional Asp/Arg couple. A single D281Y mutation in Ta1-FFT restored the tight HBN and introduced typical S-type characteristics. Conclusively, in wheat FTs Asp281 (and its homologues) is involved in donor substrate specificity. [ABSTRACT FROM PUBLISHER]

Variation in the structures of plant fructans and their degree of polymerization (DP) can be explained as the result of diverse combinations of fructosyltransferases (FTs) with different properties. Although FT genes have been isolated in a range of plant species, sucrose:fructan 6-fructosyltransferase (6-SFT) cDNAs have only been functionally characterized in a few species such as wheat. A novel FT cDNA possessing 6-SFT activity has been identified and characterized from the temperate forage grass, timothy (Phleum pratense L.). The cDNA of an FT homolog, PpFT1, was isolated from cold-acclimated timothy. A recombinant PpFT1 protein expressed in Pichia pastoris showed 6-SFT/sucrose:sucrose 1-fructosyltransferase (1-SST) activity and produced linear β(2,6)-linked levans from sucrose with higher DPs than present in graminans formed in vitro by wheat 6-SFT (Wft1). PpFT1 and Wft1 showed remarkably different acceptor substrate specificities: PpFT1 had high affinity for 6-kestotriose to produce levans and low affinity for 1-kestotriose, whereas Wft1 preferentially used 1-kestotriose as an acceptor. The affinity of the PpFT1 recombinant enzyme for sucrose as a substrate was lower than that of the Wft1 recombinant enzyme. It is also confirmed that timothy seedlings had elevated levels of PpFT1 transcripts during the accumulation of fructans under high sucrose and cold conditions. Our results suggest that PpFT1 is a novel cDNA with unique enzymatic properties that differ from those of previously cloned plant 6-SFTs, and is involved in the synthesis of highly polymerized levans in timothy. [ABSTRACT FROM PUBLISHER]

In nature, no single plant completes its life cycle without encountering environmental stress. When plant cells surpass stress threshold stimuli, chemically reactive oxygen species (ROS) are generated that can cause oxidative damage or act as signals. Plants have developed numerous ROS-scavenging systems to minimize the cytotoxic effects of ROS. The role of sucrosyl oligosaccharides (SOS), including fructans and the raffinose family oligosaccharides (RFOs), is well established during stress physiology. They are believed to act as important membrane protectors in planta. So far a putative role for sucrose and SOS during oxidative stress has largely been neglected, as has the contribution of the vacuolar compartment. Recent studies suggest a link between SOS and oxidative defence and/or scavenging. SOS might be involved in stabilizing membrane-associated peroxidases and NADPH oxidases, and SOS-derived radicals might fulfil an intermediate role in oxido-reduction reactions taking place in the vicinity of membranes. Here, these emerging features are discussed and perspectives for future research are provided. [ABSTRACT FROM PUBLISHER]

Fructosyltransferases (FTs) synthesize fructans, fructose polymers accumulating in economically important cool-season grasses and cereals. FTs might be crucial for plant survival under stress conditions in species in which fructans represent the major form of reserve carbohydrate, such as perennial ryegrass (Lolium perenne). Two FT types can be distinguished: those using sucrose (S-type enzymes: sucrose:sucrose 1-fructosyltransferase [1-SST], sucrose:fructan 6-fructosyltransferase) and those using fructans (F-type enzymes: fructan:fructan I -fructosyltransferase [1-FFT], fructan:fructan 6G-fructosyltransferase [6G-FFT]) as preferential donor substrate. Here, we report, to our knowledge for the first time, the transformation of an F-type enzyme (6G- FFT/1-FFT) into an S-type enzyme (1-SST) using perem-tial ryegrass 6G-FFT/1-FFT (Lp6G-FFT/1-FFT) and 1-SST (Lpl-SST) as model enzymes. This transformation was accomplished by mutating three amino acids (N340D, W343R, and S415N) in the vicinity of the active site of Lp6C-FFT/1-FFT. In addition, effects of each amino acid mutation alone or in combination have been studied. Our results strongly suggest that the amino acid at position 343 (tryptophan or arginine) can greatly determine the donor substrate characteristics by influencing the position of the amino acid at position 340. Moreover, the presence of arginine-343 negatively affects the formation of neofructan-type linkages. The results are compared with recent findings on donor substrate selectivity within the group of plant cell wall invertases and fructan exohydrolases. Taken together, these insights contribute to our knowledge of structure/function relationships within plant family 32 glycosyl hydrolases and open the way to the production of tailor-made fructans on a larger scale. [ABSTRACT FROM AUTHOR]

During studies on fructan degradation in the rumen, a Treponema-like bacterium able to utilize Timothy grass fructan, commercial inulin and sucrose as the sole carbon source was recovered from sheep rumen. At least two different fructanolytic enzymes were identified in cell-free extracts of the isolated bacterium. Characterization of the strain by a polyphasic approach indicated that it can be regarded as a representative of a new bacterial species within the genus Treponema. Electron microscopy showed that the bacterium exhibited all of the features typical of spirochetes. The helical cells measured 5.4–11.5 μm × 0.42–0.51 μm and possessed up to seven regular coils. The bacterium utilized various plant mono- and disaccharides as fermentable substrates. Formate, acetate and ethanol in a molar ratio of 16 : 10 : 1 were the end products of glucose fermentation. The major cellular fatty acids were C13:0, C14:0, C14:1, C15:0, C15:1 and C16:0. The nearly complete 16S rRNA gene sequence was obtained, and phylogenetic analysis of the 16S rRNA gene showed the highest similarity to rumen Treponema strain CA. We propose the name Treponema zioleckii sp. nov. for this novel rumen spirochete with strain kT as the type strain. [ABSTRACT FROM AUTHOR]