β-Glucan (Oat; Medium Viscosity)

Fermentation of β-glucan fractions from barley [average molecular mass (MM), of 243, 172, and 137 kDa] and oats (average MM of 230 and 150 kDa) by the human faecal microbiota was investigated. Fractions were supplemented to pH-controlled anaerobic batch culture fermenters inoculated with human faecal samples from three donors, in triplicate, for each substrate. Microbiota changes were monitored by fluorescent in situ hybridization; groups enumerated were: Bifidobacterium genus, Bacteroides and Prevotella group, Clostridium histolyticum subgroup, Ruminococcus-Eubacterium-Clostridium (REC) cluster, Lactobacillus-Enterococcus group, Atopobium cluster, and clostridial cluster IX. Short-chain fatty acids and lactic acid were measured by HPLC. The C. histolyticum subgroup increased significantly in all vessels and clostridial cluster IX maintained high populations with all fractions. The Bacteroides-Prevotella group increased with all but the 243-kDa barley and 230-kDa oat substrates. In general β-glucans displayed no apparent prebiotic potential. The SCFA profile (51 : 32 : 17; acetate : propionate : butyrate) was considered propionate-rich. In a further study a β-glucan oligosaccharide fraction was produced with a degree of polymerization of 3-4. This fraction was supplemented to small-scale faecal batch cultures and gave significant increases in the Lactobacillus-Enterococcus group; however, the prebiotic potential of this fraction was marginal compared with that of inulin.

Carbohydrate active enzymes are valuable tools in cereal processing to valorise underutilized side streams. By solubilizing hemicellulose and modifying the fibre structure, novel food products with increased nutritional value can be created. In this study, a novel GH5_34 subfamily arabinoxylanase from Herbinix hemicellulosilytica, HhXyn5A, was identified, produced and extensively characterized, for the intended exploitation in cereal processing to solubilize potential prebiotic fibres; arabinoxylo-oligosaccharides (AXOS). The purified two-domain HhXyn5A (catalytic domain and CBM6) demonstrated high storage stability, showed a melting temperature T_m of 61 °C and optimum reaction conditions were determined to 55°C and pH 6.5 on wheat arabinoxylan (WAX). HhXyn5A demonstrated activity on various commercial cereal arabinoxylans and produced prebiotic AXOS, while the sole catalytic domain of HhXyn5A did not demonstrate detectable activity. HhXyn5A demonstrated no side activity on oat β-glucan. In contrast to the commercially available homologue CtXyn5A, HhXyn5A gave a more specific HPAEC–PAD oligosaccharide product profile when using WAX and alkali extracted oat bran fibres as substrate. Results from multiple sequence alignment of GH5_34 enzymes, homology modelling of HhXyn5A and docking simulations with ligands XXXA³, XXXA³XX, and X⁵, concluded that the active site of HhXyl5A catalytic domain is highly conserved and can accommodate both shorter and longer AXOS ligands. However, significant structural dissimilarities between HhXyn5A and CtXyn5A in the binding cleft of CBM6, due to lack of important ligand interacting residues, is suggested to cause the observed differences in substrate specificity and product formation.

Potentially beneficial microorganisms in the human gut microbiota may be promoted through exploiting dietary fibre (DF) characteristics. Wheat flour cell walls (WCW) are compositionally diverse and architecturally complex. Arabinoxylan (AX), mixed-linkage (1,3), (1,4)-β-glucan (MLG) and cellulose are the major components of WCW. This study investigated fermentation of WCW, individual polysaccharide components (AX, MLG and cellulose), and a physical mixture of them (Mix), using human faecal inoculum. Interestingly, the substrate order of final gas production and the degradation patterns of each were comparable to previous porcine faecal fermentation results, pointing to the important influence of substrate properties on fermentation outcomes. However, the human faecal fermentation was faster, with higher amounts of propionate and less n-butyrate produced than from the corresponding substrate fermented by porcine faecal inoculum. Microbiota variation during polysaccharide degradation were determined, and associations were made with fermentation end-products: Coprococcus 3, Bifidobacterium and Lachnospiraceae ND3007 group were positively correlated with AX fermentation and acetate production, while Catenibacterium and Lachnospiraceae NK4A136 group appeared to contribute to MLG degradation and n-butyrate production. These findings suggest that the nature of DF primarily dictates the relative extent of fermentation, while the fermentation rate, end-product composition and microbial shifts are more linked with specific microbial activity of individual bacterial groups, dependent on the initial microbiota composition.

Plant cell walls as well as their component polysaccharides in foods can be utilized to alter and maintain a beneficial human gut microbiota, but it is not known whether the architecture of the cell wall influences the gut microbiota population. In this study, wheat flour cell walls (WCW) were isolated and compared with their major constituents - arabinoxylan (AX), mixed linkage (1,3)(1,4)-β-glucan (MLG) and cellulose - both separately and as a physical mixture of polysaccharides (Mix) equivalent in composition to WCW. These samples underwent in vitro fermentation with a faecal inoculum from pigs fed a diet free of cereals and soluble-fibre to avoid prior adaptation to substrates. During fermentation, samples were collected for DNA extraction and 16S rRNA gene amplicon sequencing. Bioinformatics analyses revealed that the microbial communities promoted during fermentation by AX, MLG, Mix and WCW were similar at the genus level, but differed from the microbiota observed for the cellulose substrate. Differences in proportions of propionate and butyrate end-products were associated with differences in the relative levels of genera. These findings show that, in this experiment, the microbes that flourished were able to utilize diverse WCW polysaccharides alone, in mixtures or in intact cell walls in a similar way, but that different fermentation end-products were associated with AX (propionate) or MLG (butyrate) polysaccharides.

A recycle-integrated reactor-separator system was studied experimentally and based on simulations for the acid-catalyzed depolymerization of oat beta-glucan polysaccharide. The aim was to produce oligosaccharides with degree of polymerization in a narrow range (DP = 15-30). The reactor was operated intermittently at 80°C. Batch chromatography with Sephadex G-25 size-exclusion gel was found suitable for the separation of product from reactants and impurities. Part of the reaction mixture was periodically withdrawn and fed to the separation column. Molar mass distributions in four chromatographic fractions (waste, recycle, product, impurities) were monitored with SEC-MALLS. Experiments with 4 h mean residence time showed that the reactor-separator achieved approximately 2.0 and 2.5 times higher yield and purity of target DP than a batch reactor. Dimensionless operating parameters and equipment design parameters were introduced for analyzing performance of intermittent reactor-separators. The simulations show that intermittent operation offers higher yield and product purity than continuous operation (CSTR and chromatographic separation) when mean residence time in the reactor is long. Continuous operation is better when productivity is maximized by using short mean residence time and low yield.

Background: Non-digestible carbohydrates are added to infant formula to mimic the effects of human milk oligosaccharide by acting as prebiotics and stimulating the immune system. Although not yet used in infant formulas, β-glucans are known to have beneficial health effects, and are therefore of potential interest for supplementation. Methods and results: We investigated the in vitro fermentation of native and endo-1,3(4)-β-glucanase-treated oat β-glucan using pooled fecal inocula of 2- and 8-week-old infants. While native oat β-glucan was not utilized, both inocula specifically utilized oat β-glucan oligomers containing β(1→4)-linkages formed upon enzyme treatment. The fermentation rate was highest in the fecal microbiota of 2-week-old infants, and correlated with a high lactate production. Fermentation of media supplemented with native and enzyme-treated oat β-glucans increased the relative abundance of Enterococcus and attenuated pro-inflammatory cytokine production (IL-1β, IL-6, TNFα) in immature dendritic cells. This attenuating effect was more pronounced after enzyme treatment. This attenuation might result from the enhanced ability of fermented oat β-glucan to stimulate Dectin-1 receptors. Conclusion: Our findings demonstrate that endo-1,3(4)-β-glucanase treatment enhances the fermentability of oat β-glucan and attenuates pro-inflammatory responses. Hence, this study shows that especially enzyme-treated oat β-glucans have a high potential for supplementation of infant formula.

Reactivity of the glycosidic bonds in oat β-glucan in homogeneous acid catalyzed hydrolysis in aqueous solution was found to depend on their position in the polysaccharide chain. The bonds near the chain ends have lower hydrolytic stability in presence of HCl than the bonds in the middle. Reaction kinetics was monitored by measuring the molar mass distribution (DP ~ 30…3500) and formation of short oligosaccharides (DP < 6) under mild conditions (T = 50°C…80°C, c_HCL = 0.05…0.25 M). Accuracy of the molar mass determination using analytical scale size-exclusion chromatography and multi-angle laser light scattering was improved by carrying out preparative scale size-exclusion separation as a pretreatment. It was found that monitoring the short-chain degradation products in addition to the molar mass distribution provides valuable information about the hydrolysis rate. The data were successfully correlated with a kinetic model that takes into account the non-random structure of oat β-glucan. The rate of cleavage of the terminal glycosidic bonds was found to be almost three times higher than at a distance of 20 or more glucose units from the chain end. The β-(1,3) bonds in oat β-glucan were found less stable against acid catalyzed hydrolysis than the β-(1,4) bonds.

Depletion flocculation is a well-known instability mechanism that can occur in oil-in-water emulsions when the concentration of non-adsorbed polysaccharide exceeds a certain level. This critical flocculation concentration depends on the molecular characteristics of the polysaccharide molecules, such as their molecular weight and hydrodynamic radius. In this study, a range of analytical methods (dynamic shear rheology, optical microscopy, and static light-scattering) were used to investigate the interaction between lipid droplets and polysaccharides (guar gum and β-glucans) of varying weight-average molecular weight and hydrodynamic radius, and concentration. The aim of this work was to see if the health benefits of soluble fibers like β-glucans could be explained by their influence on the structure and digestibility of lipid emulsions. The apparent viscosity of the emulsions increased with increasing polysaccharide concentration, molecular weight, and hydrodynamic radius. Droplet flocculation was observed in the emulsions only at certain polysaccharide concentrations, which was attributed to a depletion effect. In addition, the water-soluble components in oat flakes, flour, and bran were extracted using aqueous solutions, to examine their impact on emulsion stability and properties. Then, the rate and extent of lipolysis of a sunflower oil-in-water emulsion in the presence of these oat extracts were monitored using the pH-stat method. However, the inhibition of lipolysis was not linearly related to the viscosity of the oat solutions. The water-soluble extracts of β-glucan collected from oat flakes had a significant inhibitory effect on lipolysis. The results of this study increase our understanding of the possible mechanisms influencing the impact of oat constituents on lipid digestion. This work also highlights the importance of considering the molecular properties of polysaccharides, and not just their impact on solution viscosity.

Oat β-Glucans were studied for their immunological impact before and after enzymatic digestion in order to enhance the efficacy of oat β-Glucans for application in functional foods. Oat β-Glucan is reported to have minimal impact compared to its fungal counterpart in vitro. Digestion with endo-glucanase enhanced its efficacy towards stimulating MCP-1, RANTES, IL-8, and IL-4 production in human dendritic cells as compared to the nondigested β-Glucan. This effect resulted from an enhanced activation of the Dectin-1 receptor. Our data suggest that the immune-stimulation was dependent on the β-(1-3) linkages and the reduced particle size of digested β-Glucans. Thus, we show that enzymatic pre-digestion of dietary fibres such as oat β-Glucan enhances its impact on specific immune receptors. We also demonstrate that particle size and/or molecular weight of oat β-Glucans and exposure of specific binding sites for the receptors might be important tools for designing efficacious functional feed and food additives.

Β-glucan is one of the components that differentiate oats from other cereals and that contribute to the health-related value of oats. However, so far oats cannot easily be applied in bread-like products without loss of product quality. Here we have studied how the content and viscosity of oat β-glucan affect the technological properties of oat dough in both a gluten-free and a gluten-containing system. In both systems, increasing the β-glucan concentration resulted in an increase of dough stiffness and in a reduction of dough extensibility. β-glucan negatively impacted the elastic properties that additional wheat gluten conferred to oat dough. This effect was smaller for medium-viscosity β-glucan than for high-viscosity β-glucan. Interestingly, dough made from low β-glucan flour (<2%) had increased gas retention capacity. Overall, the impact of β-glucan on the properties of oat dough systems was governed by concentration and viscosity, with or without additional wheat gluten. Our findings indicate that β-glucan is a key component that determines the rheology of oat-based dough systems and, with that, the technological functionality of oat in dough systems.