Arabinoxylan Oligosaccharides Research Articles

Arabinoxylan source and xylanase specificity influence the production of oligosaccharides with prebiotic potential

Cereal arabinoxylans (AXs) are complex polysaccharides in terms of their pattern of arabinose and ferulic acid substitutions, which influence their properties in structural and nutritional applications. We have evaluated the influence of the molecular structure of three AXs from wheat and rye with distinct substitutions on the activity of β-xylanases from different glycosyl hydrolase families (GH 5_34, 8, 10 and 11). The arabinose and ferulic acid substitutions influence the accessibility of the xylanases, resulting in specific profiles of arabinoxylan-oligosaccharides (AXOS). The GH10 xylanase from Aspergillus aculeatus (AcXyn10A) and GH11 from Thermomyces lanuginosus (TlXyn11) showed the highest activity, producing larger amounts of small oligosaccharides in shorter time. The GH8 xylanase from Bacillus sp. (BXyn8) produced linear xylooligosaccharides and was most restricted by arabinose substitution, whereas GH5_34 from Gonapodya prolifera (GpXyn5_34) required arabinose substitution and produced longer (A)XOS substituted on the reducing end. The complementary substrate specificity of BXyn8 and GpXyn5_34 revealed how arabinoses were distributed along the xylan backbones. This study demonstrates that AX source and xylanase specificity influence the production of oligosaccharides with specific structures, which in turn impacts the growth of specific bacteria (Bacteroides ovatus and Bifidobacterium adolescentis) and the production of beneficial metabolites (short-chain fatty acids).

Profiling of cool-season forage arabinoxylans via a validated HPAEC-PAD method.

Cool-season pasture grasses contain arabinoxylans (AX) as their major cell wall hemicellulosic polysaccharide. AX structural differences may influence enzymatic degradability, but this relationship has not been fully explored in the AX from the vegetative tissues of cool-season forages, primarily because only limited AX structural characterization has been performed in pasture grasses. Structural profiling of forage AX is a necessary foundation for future work assessing enzymatic degradability and may also be useful for assessing forage quality and suitability for ruminant feed. The main objective of this study was to optimize and validate a high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) method for the simultaneous quantification of 10 endoxylanase-released xylooligosaccharides (XOS) and arabinoxylan oligosaccharides (AXOS) in cool-season forage cell wall material. The following analytical parameters were determined or optimized: chromatographic separation and retention time (RT), internal standard suitability, working concentration range (CR), limit of detection (LOD), limit of quantification (LOQ), relative response factor (RRF), and quadratic calibration curves. The developed method was used to profile the AX structure of four cool-season grasses commonly grown in pastures (timothy, Phleum pratense L.; perennial ryegrass, Lolium perenne L.; tall fescue, Schedonorus arundinaceus (Schreb.) Dumort.; and Kentucky bluegrass, Poa pratensis L.). In addition, the cell wall monosaccharide and ester-linked hydroxycinnamic acid contents were determined for each grass. The developed method revealed unique structural aspects of the AX structure of these forage grass samples that complemented the results of the cell wall monosaccharide analysis. For example, xylotriose, representing an unsubstituted portion of the AX polysaccharide backbone, was the most abundantly-released oligosaccharide in all the species. Perennial rye samples tended to have greater amounts of released oligosaccharides compared to the other species. This method is ideally suited to monitor structural changes of AX in forages as a result of plant breeding, pasture management, and fermentation of plant material.

The Effect of Hydrolyzed and Fermented Arabinoxylan-Oligo Saccharides (AXOS) Intake on the Middle-Term Gut Microbiome Modulation and Its Metabolic Answer.

Although fermentation and hydrolyzation are well-known processes to improve the bioavailability of nutrients and enable the fortification with dietary fibers, the effect of such pre-treatments on the prebiotic features of arabinoxylan-oligosaccharides (AXOS) had not been explored. The middle-term in vitro simulation through the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) demonstrated that the feeding with different formulations (namely oat bran, rye bran and wheat bran) containing hydrolyzed AXOS fermented by lactic acid bacteria significantly increased the synthesis of short-chain fatty acids (SCFA) by colon microbiota, with hydrolyzed and fermented rye bran displaying the highest effect. After two weeks from the interruption of intake, SCFA concentrations significantly decreased but remained still significantly higher compared to the original condition. The microbiome was also affected, with a significant abundance increase in Lactobacillaceae taxon after feeding with all fermented and hydrolyzed formulates. Hydrolyzed and fermented rye bran showed the highest changes. The fungal community, even if it had a lower variety compared to bacteria, was also modulated after feeding with AXOS formulations, with an increase in Candida relative abundance and a decrease in Issatchenkia. On the contrary, the intake of non-hydrolyzed and non-fermented wheat bran did not produce relevant changes of relative abundances. After two weeks from intake interruption (wash out period) such changes were mitigated, and the gut microbiome modulated again to a final structure that was more like the original condition. This finding suggests that hydrolyzed AXOS fermented by lactic acid bacteria could have a more powerful prebiotic effect compared to non-hydrolyzed and non-fermented wheat bran, shaping the colon microbiome and its metabolic answer. However, the intake should be continuous to assure persistent effects. Opening a window into the ecological evolutions and plausible underlying mechanisms, the findings reinforce the perspective to explore more in depth the use of hydrolyzed and fermented AXOS as additional ingredient for bread fortification.

Exogenous enzymes, meal size, and meal frequency: effect on ileal and total tract digestibility of carbohydrates, and energy and fiber degradation in growing pigs fed a wheat-barley grain-based high-fiber diet.

When conducting a digestibility trial, pigs are usually fed only twice a day with a restricted feed intake which is not representative of the feeding conditions in a commercial farm. This study aimed to determine the effects of meal size and frequency, and exogenous enzymes (xylanase and phytase) on the digestibility of a high-fiber diet using porcine in vivo and in vitro approaches. Pigs (n = 6) were fitted with a T cannula, and each received all treatments using a 6 × 6 Latin square experimental design. The diets were supplemented (Enz) or not with a combination of xylanase and phytase and distributed into three feeding programs: one received two meals per day that met three times the maintenance energy requirement (2M), one received the same quantity of feed in eight meals (8M), and another received an amount that met five times the maintenance energy requirements in eight meals (8M+). For in vitro experiment, the degradability of fiber with or without xylanase supplementation only was determined. Enzyme supplementation increased apparent ileal digestibility (AID) of dry matter, starch, and degradation of insoluble non-starch polysaccharides (I-NSP) in all in vivo treatments (P < 0.05). The 2M compared with 8M increased the AID of starch and total tract digestibility of organic matter and I-NSP (P < 0.05). Enzyme supplementation decreased the content of insoluble arabinoxylan (P < 0.05) and increased arabinoxylan oligosaccharides (P < 0.05) in the in vivo ileal digesta and in vitro incubation. The results of this study confirm degradation by xylanase of the fiber fraction at the ileal level, which resulted in less fermentation of fiber in the large intestine. However, number and size of meals had little influence on feed digestibility. The consequences of shifting fiber fermentation more towards the upper part of the gastrointestinal tract need further investigation. The in vitro model provided a confirmation of the action of xylanase on the degradation of non-starch polysaccharides.

Purification of wild type and recombinant xylanases and evaluation of their arabinoxylan hydrolysis ability

Arabinoxylan hydrolyzing enzymes are very rich and diversified groups, among these, endo-1,4-β-xylanase is the most important group which catalyzes the cleavage of xylan main skeleton to release arabinoxylan oligosaccharides such as D-xylose, xylobiose, L-arabinose, xylotetraose, xylopentose.  This group of enzyme was efficiently produced from natural microorganism including bacteria, yeast and fungi and recombinant strains. In this study, we have purified wild-type and recombinant xylanase to compare the difference in hydrolysis products of these two enzymes. Using sephadex G-100 and DEAE-sephadex column chromatography, we have purified xylanase from wild-type Aspergillus niger DSM1957 with molecular weight of 25 kDa, specific activity of 3240 IU/mL, the purity increases 1.68 folds compare to crude extract, and a recovery rate of 36%. The recombinant enzyme from Pichia pastoris GS115/pPXlnA was purified by ProbondTM affinity chromatography column, with a molecular weight of 36 kDa; the purity increased 3.2 folds than the crude extract, and with the recovery efficiency of 21.1%. The activity of commercial wild-type xylanase was better than that of our recombinant and wild-type enzymes. Our wild-type xylanase could hydrolyze arabinoxylan into L-arabinose in 50 mM CH3COONa pH 5.0. Our results showed that xylanases purified from different sources were stable and highly specific to xylan. Xylanase has potential application in the production of high quality biotechnological products.

Reclaiming lost nutrition

Food Science and TechnologyVolume 35, Issue 2 p. 32-33 FeaturesFree Access Reclaiming lost nutrition First published: 15 June 2021 https://doi.org/10.1002/fsat.3502_9.xAboutFiguresReferencesRelatedInformationPDFSections Food waste's untapped potential Circular production model Extracting better nutrition The prebiotic opportunity What's next?ReferencesPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessClose modalShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Rich Troyer describes Comet Bio's innovative extraction process for upcycling crop leftovers into high-quality, nutritious ingredients. Forty percent of the food produced around the globe currently goes to waste according to the Natural Resources Defense Council1. At the same time, nutrition gaps and food insecurity continue to persist, even in the United States, where more than 30% of the population is considered either obese or overweight2. This paradox of our food system producing more waste than ever, yet still not providing adequate nutrition, demonstrates the need for innovation. Comet Bio is working to address both challenges by extracting nutrition from food system waste. Its proprietary upcycling technology produces novel ingredients with significant nutritious potential, while simultaneously addressing critical environmental challenges. Food waste's untapped potential Food waste has been an increasingly trending topic for the last decades, especially as the world's population continues to grow. One of the most logical ways to prevent potential food supply shortages is to address the food already being produced and going to waste. The USDA's (United States Department of Agriculture) Loss-Adjusted Food Availability data series shows that the food wasted per person per day in the US in 2012 contained over 1,200 calories, 33g of protein, 5.9g of dietary fiber and significant amounts of other under-consumed nutrients, including calcium and potassium. These nutrients could go far in helping fill in nutrition gaps for many Americans3. For example, USDA data shows that US women fell short of meeting the recommended daily levels of dietary fiber by an average of 8.9g per day in 2012. The lost 5.9g of dietary fiber represents two-thirds of this gap. There is a lot of untapped potential in upcycling food waste for human consumption, especially given that most of the byproducts from manufacturing are fed to animals. It is a missed nutritional opportunity for humans because many of these byproducts have high nutritional value, which leads to economic value. The 1.3bn tons of food wasted each year represent not only valuable nutrition but also a $2.6tn economic value4. Circular production model Wasted food begins where our food supply chain begins – farming and harvesting. Comet Bio's founder, Andrew Richard, has a long history of working with crop leftovers and wood waste. Early on, he recognised the opportunity in upcycling these resources. Unlike some manufacturing systems that use a ‘take, make, waste’ model, upcycling solidifies a circular economy using a ‘take, make, remake and restore’ model. It takes materials that would otherwise go to waste and harvests them to develop an entirely new product, closing the loop on the food system. Comet Bio's innovative extraction process upcycles crop leftovers – such as straw, leaves and shells – into high-quality ingredients (Figure 1). We partner with farmers to understand where our crop leftovers are coming from and how they are being grown and collected. Not only does this help ensure we are sourcing our upcycled materials from organic and non-GMO crops, but it enables us to mitigate any mis-steps that could occur when purchasing farm leftovers. This partnership also benefits farmers by allowing them to profit from their whole harvest while ensuring the highest quality for our ingredients. Figure 1Open in figure viewerPowerPoint Upcycling production process With this approach, the company not only closes the loop on the food system but also reduce its carbon footprint. According to an independent well-to-wheel analysis, Comet Bio's dextrose production process emits over 60% fewer greenhouse gases than traditional manufacturing. Extracting better nutrition Comet Bio continues to research ways to use its technology to supply consumers with not only more sustainable but also healthier ingredients. For example, despite its prebiotic benefits, the plant fibre extract, arabinoxylan, has not been widely available for use due to inefficient extraction. Arabinoxylan is an FDA (Food and Drug Administration) recognised dietary fiber and can be procured from a variety of sources of hemicellulose, from spent brewer's grain to wheat stalks. Clinical research shows that consuming as little as 2.2g of agrabinoxylan per day promotes the growth of bifidobacteria in the gut, which is less than half the level required for other prebiotic ingredients5. Arabinoxylan is also clinically proven to support immune health and help maintain healthy blood glucose levels6. Comet Bio has utilised its extraction technology to make arabinoxylan available in an isolated, purified and fully soluble form. The proprietary process uses water and pressure to extract the arabinoxylan and purify it into a fully soluble powder (Figure 2). Figure 2Open in figure viewerPowerPoint Nutrient extraction process Recent development work has established how to adjust the operating conditions to obtain exact product specifications. There is a molecular weight range within arabinoxylan that can be used to fine tune the plant fibre to deliver different health benefits. There are ways of adjusting the process to produce different specs of arabinoxylan, such as a lighter colour product or a more potent immunity product. Arrabina prebiotic fiber and a sample showing its use in a chocolate bar The spec for the arabinoxylan prebiotic dietary fibre product (Arrabina) can be incorporated into a wide range of applications. In addition to being fully soluble, it has a low viscosity, remains stable at low pH levels and is extremely resistant to high temperatures. Arrabina's high functionality means it will not gel or impact texture when added as a supplement to protein powders or as a fiber boost for nutrition bars, baked goods and ready-to-drink beverages. It also works well in chocolate confectionary and can be added to help reduce sugar content. The prebiotic opportunity Demand for prebiotics has been growing steadily in recent years, resulting in strong growth for the category and a number of major suppliers looking to expand production or enter the market. According to Nutrition Business Journal (NBJ) data, the US digestive health market has nearly tripled in size within the last decade and is predicted to reach $5.7bn by 2024. Prebiotic sales currently make up a small portion of the overall digestive health market but are showing strong growth, having doubled every year since 2016 and surpassing $425m in sales in 20207. In addition, awareness of prebiotics has grown to 81%, with 35% of supplement users taking prebiotics at some level, according to Ingredient Transparency Center (ITC) Insights’ 2020 Consumer Survey8. Unfortunately, many popular prebiotic fiber options require high inclusion levels to be effective. For example, for inulin, a commonly used prebiotic fiber derived from chicory root, 5g of material per day or more is needed9. These higher inclusion rates render many prebiotic supplement, food and beverage formats impractical. Many popular prebiotic fiber options, including inulin, are also oligosaccharides, a type of fiber that the popular low-FODMAP diet (fermentable oligosaccharides, disaccharides, monosaccharides and polyols - short-chain carbohydrates that the small intestine absorbs poorly) cautions individuals to avoid due to tolerability concerns. Comet Bio's arabinoxylan prebiotic fiber is hemicellulose polysaccharide fiber. Its longer chain polysaccharide structure makes it better tolerated by the gut compared to oligosaccharides. Results from a recent clinical trial reveal that consumers can take up to 15g per day of Arrabina with no negative gut or bowel reaction10. At the same time, Arrabina is potent with an inclusion rate as low as 3g per day, making it easier to incorporate into existing food and beverage products. What's next? Comet Bio is reclaiming lost nutrition from food system waste and we are not alone. In 2020, the company was selected by Anheuser-Busch InBev, the world's largest brewer, to join its 100+ Accelerator to pilot their saved grains in our upcycling process. The Upcycled Food Association (UFA), of which we are members, now has over 65 mission-driven companies, each tackling the problem in slightly different ways. Consumers are recognising the opportunity too. According to UFA, 95% of consumers want to reduce food waste. Future Market Insights estimates that upcycled food could be worth $46bn and experience a 5% annual growth over the next decade. These collective efforts, including innovative approaches and industry partnerships, can give rise to a future with less waste and more nutrition in our food system. Rich Troyer, CEO of Comet Bio, Schaumburg, IL, USA Rich has a combination of corporate development, operations and finance expertise in industrial biotechnology. He was previously the Chief Business Officer at Coskata and before that he was a Managing Director at The Blackstone Group, responsible for investments in biofuels and bio-chemicals companies. Rich received his M.B.A. in Finance and Marketing from the Wharton School of the University of Pennsylvania, and a B.B.A. degree in accounting from Baylor University. Email rtroyer@comet-bio.com Web comet-bio.com/ References 1Gunders, Dana. "Wasted: How America is losing up to 40 percent of its food from farm to fork tolandfill."Natural Resources Defense Council26 (2012): 1-26. 2J. Eric Oliver, Taeku Lee; Public Opinion and the Politics of Obesity in America.JHealthPolitPolicyLaw1 October 2005; 30 (5): 923–954. doi:https://doi.org/10.1215/03616878-30-5-923 3Krittanawong, Chayakrit. "The appeal of alt: Understanding consumer demand for label-friendlyalternative products–report."Meta(2020). 4Juma,Calestous. The new harvest: agricultural innovation in Africa. Oxford University Press,2015. 5Cloetenset al. 2010. Tolerance of arabinoxylan-oligosaccharides and their prebiotic activity inhealthy subjects: a randomised, placebo-controlled crossoverstudy. British Journal of Nutrition, 103, 703- 713. 6Kjølbæk, L., Benítez-Páez, A., Pulgar, E. M., Brahe, L. K., Liebisch, G., Matysik, S., . . . Sanz, Y.(2019). Arabinoxylan oligosaccharides and polyunsaturated fatty acid effects on gut microbiotaand metabolic markers in overweight individuals with signs of metabolic syndrome: Arandomized cross-over trial. Clinical Nutrition. doi:https://doi.org/10.1016/j.clnu.2019.01.012 7https://store.newhope.com/products/april-2019-market-overview?_pos=1&_sid=2a7c210b5&_ss=r 8https://trusttransparency.com/insights/ 9Carlson JL, Erickson JM, Lloyd BB, Slavin JL. Health Effects and Sources of Prebiotic Dietary Fiber.Curr Dev Nutr. 2018 Jan 29;2(3):nzy005. doi: 10.1093/cdn/nzy005. PMID: 30019028; PMCID:PMC6041804. 10https://comet-bio.com/comet-bio-announces-positive-clinical-trial-results-for-its-prebiotic-arrabina/ Volume35, Issue2June 2021Pages 32-33 FiguresReferencesRelatedInformation RecommendedTHE USE OF SELECTED LACTIC ACID BACTERIA STARTER CULTURES FOR IMPROVED THAI SAUSAGE FERMENTATIONWIRAMSRI SRIPHOCHANART, WANWISA SKOLPAP, Journal of Food Processing and PreservationNisin Z-Producing Lactococcus lactis Subsp. Lactis GYl32 Isolated from BozaGozde Koral, Yasin Tuncer, Journal of Food Processing and PreservationEvaluation of lactic acid bacteria and component change during fermentation of Ishizuchi‐kurochaMasanori Horie, Atsumi Tada, Naoaki Kanamoto, Takahisa Tamai, Naohiro Fukuda, Sakiko Sugino, Takahito Toyotome, Yosuke Tabei, Journal of Food Processing and PreservationRedistributing surplus foodFood Science and TechnologyInnovation with purposeFood Science and Technology

The influence of xylanase on the fermentability, digestibility, and physicochemical properties of insoluble corn-based fiber along the gastrointestinal tract of growing pigs.

In theory, supplementing xylanase in corn-based swine diets should improve nutrient and energy digestibility and fiber fermentability, but its efficacy is inconsistent. The experimental objective was to investigate the impact of xylanase on energy and nutrient digestibility, digesta viscosity, and fermentation when pigs are fed a diet high in insoluble fiber (>20% neutral detergent fiber; NDF) and given a 46-d dietary adaptation period. A total of 3 replicates of 20 growing gilts were blocked by initial body weight, individually housed, and assigned to 1 of 4 dietary treatments: a low-fiber control (LF) with 7.5% NDF, a 30% corn bran high-fiber control (HF; 21.9% NDF), HF + 100 mg xylanase/kg (HF + XY [Econase XT 25P; AB Vista, Marlborough, UK]) providing 16,000 birch xylan units/kg; and HF + 50 mg arabinoxylan-oligosaccharide (AXOS) product/kg (HF + AX [XOS 35A; Shandong Longlive Biotechnology, Shandong, China]) providing AXOS with 3–7 degrees of polymerization. Gilts were allowed ad libitum access to fed for 36-d. On d 36, pigs were housed in metabolism crates for a 10-d period, limit fed, and feces were collected. On d 46, pigs were euthanized and ileal, cecal, and colonic digesta were collected. Data were analyzed as a linear mixed model with block and replication as random effects, and treatment as a fixed effect. Compared with LF, HF reduced the apparent ileal digestibility (AID), apparent cecal digestibility (ACED), apparent colonic digestibility (ACOD), and apparent total tract digestibility (ATTD) of dry matter (DM), gross energy (GE), crude protein (CP), acid detergent fiber (ADF), NDF, and hemicellulose (P < 0.01). Relative to HF, HF + XY improved the AID of GE, CP, and NDF (P < 0.05), and improved the ACED, ACOD, and ATTD of DM, GE, CP, NDF, ADF, and hemicellulose (P < 0.05). Among treatments, pigs fed HF had increased hindgut DM disappearance (P = 0.031). Relative to HF, HF + XY improved cecal disappearance of DM (162 vs. 98 g; P = 0.008) and NDF (44 vs. 13 g; P < 0.01). Pigs fed xylanase had a greater proportion of acetate in cecal digesta and butyrate in colonic digesta among treatments (P < 0.05). Compared with LF, HF increased ileal, cecal, and colonic viscosity, but HF + XY decreased ileal viscosity compared with HF (P < 0.001). In conclusion, increased insoluble corn-based fiber decreases digestibility, reduces cecal fermentation, and increases digesta viscosity, but supplementing xylanase partially mitigated that effect.

Xylanase modulates the microbiota of ileal mucosa and digesta of pigs fed corn-based arabinoxylans likely through both a stimbiotic and prebiotic mechanism.

The experimental objective was to characterize the impact of insoluble corn-based fiber, xylanase, and an arabinoxylan-oligosaccharide on ileal digesta and mucosa microbiome of pigs. Three replicates of 20 gilts were blocked by initial body weight, individually-housed, and assigned to 1 of 4 dietary treatments: a low-fiber control (LF), a 30% corn bran high-fiber control (HF), HF+100 mg/kg xylanase (HF+XY), and HF+50 mg/kg arabinoxylan oligosaccharide (HF+AX). Gilts were fed their respective treatments for 46 days. On day 46, pigs were euthanized and ileal digesta and mucosa were collected. The V4 region of the 16S rRNA was amplified and sequenced, generating a total of 2,413,572 and 1,739,013 high-quality sequences from the digesta and mucosa, respectively. Sequences were classified into 1,538 mucosa and 2,495 digesta operational taxonomic units (OTU). Hidden-state predictions of 25 enzymes were made using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States 2 (PICRUST2). Compared to LF, HF increased Erysipelotrichaceae_UCG-002, and Turicibacter in the digesta, Lachnospiraceae_unclassified in the mucosa, and decreased Actinobacillus in both (Q<0.05). Relative to HF, HF+XY increased 19 and 14 of the 100 most abundant OTUs characterized from digesta and mucosa, respectively (Q<0.05). Notably, HF+XY increased the OTU_23_Faecalibacterium by nearly 6 log2-fold change, compared to HF. Relative to HF, HF+XY increased genera Bifidobacterium, and Lactobacillus, and decreased Streptococcus and Turicibacter in digesta (Q<0.05), and increased Bifidobacterium and decreased Escherichia-Shigella in the mucosa (Q<0.05). Compared to HF, HF+AX increased 5 and 6 of the 100 most abundant OTUs characterized from digesta and mucosa, respectively, (Q<0.05), but HF+AX did not modulate similar taxa as HF+XY. The PICRUST2 predictions revealed HF+XY increased gene-predictions for enzymes associated with arabinoxylan degradation and xylose metabolism in the digesta, and increased enzymes related to short-chain fatty acid production in the mucosa. Collectively, these data suggest xylanase elicits a stimbiotic and prebiotic mechanism.

Effects of cereal fibers on short-chain fatty acids in healthy subjects and patients: a meta-analysis of randomized clinical trials.

Short-chain fatty acids (SCFAs) are involved in the regulation of a wide array of diseases. However, the effect of cereal dietary fibers on SCFA production remains unclear. We reviewed relevant clinical studies between 1950 and 2021 and aimed to evaluate the effect of cereal fiber consumption on SCFA production in healthy subjects and patients. PubMed, Web of Science, and the Cochrane Library databases were used for systematically searching published relevant trials with adults and a minimum intervention duration of 2 weeks. The effect size was estimated using standardized mean difference (SMD) and 95% confidence interval (CI). Of the 555 identified studies, 14 intervention groups involving 205 participants aged between 20 and 69 years are eligible. The results of meta-analysis revealed that cereal fiber supplementation significantly increased acetate [SMD: 0.86, 95% CI (0.46, 1.25), p < 0.0001], propionate [SMD: 0.48, 95% CI: (0.15, 0.81), p = 0.004], butyrate [SMD: 0.61, 95% CI: (0.20, 1.01), p = 0.003], and total SCFA [SMD, 0.96, 95% CI: (0.54, 1.39), p < 0.00001] concentrations. Subgroup analysis suggested that a long intervention duration (>4 weeks) significantly promoted acetate and propionate production, whereas a short intervention duration (≤4 weeks) significantly facilitated butyrate production. Cereal fiber supplementation had a more significant impact on overweight and obese subjects with body mass index (BMI) >29 kg m-2 than on individuals with BMI ≤29 kg m-2. Furthermore, we found that cereal fibers and wheat/rye arabinoxylan oligosaccharides, rather than wheat bran fibers, barley fibers, and barley β-glucan, could significantly elevate the SCFA concentration. Overall, our meta-analysis demonstrated that cereal fiber supplementation is helpful in increasing the SCFA concentration, which provided strong proof for the beneficial role of cereal fibers.

Microbial enterotypes beyond genus level: Bacteroides species as a predictive biomarker for weight change upon controlled intervention with arabinoxylan oligosaccharides in overweight subjects

ABSTRACT Recent studies indicate that microbial enterotypes may influence the beneficial effects of wholegrain enriched diets including bodyweight regulation. In a 4-week intervention trial, overweight subjects were randomized to consume either arabinoxylan-oligosaccharides (AXOS) (10.4 g/d) from wheat bran or polyunsaturated fatty acids (PUFA) (3.6 g/d). In the present study, we have stratified the subjects participating in the intervention (n = 29) according to the baseline Prevotella-to-Bacteroides (P/B) ratios through a post-hoc analysis and applied a linear mixed model analysis to identify the influence of this P/B ratio on the differences in weight changes in the intervention arms. Following AXOS consumption (n = 15), the high P/B group showed no bodyweight changes [−0.14 kg (95% CI: −0.67; 0.38, p = .59)], while the low P/B group gained 0.65 kg (95% CI: 0.16; 1.14, p = .009). Consequently, a difference of −0.79 kg was found between P/B groups (95% CI: −1.51; −0.08, p = .030). No differences were found between P/B groups following PUFA consumption (0.61 kg, 95% CI: −0.13; 1.35, p = .10). Among the Bacteroides species, B. cellulosilyticus relative abundance exhibited the highest positive rank correlation (Kendall’s tau = 0.51, FDR p = .070) with 4-week weight change on AXOS, and such association was further supported by using supervised classification methods (Random Forest). We outlined several carbohydrate-active enzyme (CAZy) genes involved in xylan-binding and degradation to be enriched in B. cellulosilyticus genomes, as well as multiple accessory genes, suggesting a supreme AXOS-derived glycan scavenging role of such species. This post-hoc analysis, ensuring species and strain demarcation at the human gut microbiota, permitted to uncover the predictive role of Bacteroides species over P/B enterotype in weight gain during a fiber-based intervention. The results of this pilot trial pave the way for future assessments on fiber fermentation outputs from Bacteroides species affecting lipid metabolism in the host and with direct impact on adiposity, thus helping to design personalized interventions.

Improving the digestibility of cereal fractions of wheat, maize, and rice by a carbohydrase complex rich in xylanases and arabinofuranosidases: an in vitro digestion study.

Cereal co-products rich in dietary fibres are increasingly used in animal feed. The high fibre content decreases the digestibility and reduces the nutrient and energy availability, resulting in lower nutritive value. Therefore, this study investigated the ability of two carbohydrase complexes to solubilize cell-wall polysaccharides, in particular arabinoxylan (AX), from different cereal fractions of wheat, maize, and rice using an in vitro digestion model of the pig gastric and small intestinal digestive system. The first complex (NSPase 1) was rich in cell-wall-degrading enzymes, whereas the second complex (NSPase 2) was additionally enriched with xylanases and arabinofuranosidases. The extent of solubilization of insoluble cell-wall polysaccharides after in vitro digestion was evaluated with gas-liquid chromatography and an enzymatic fingerprint of the AX oligosaccharides was obtained with high-performance anion-exchange chromatography with pulsed amperometric detection. The addition of carbohydrase increased the digestibility of dry matter and solubilized AX in particular, with the greatest effect in wheat fractions and less effect in maize and rice fractions. The solubilization of AX (expressed as xylose release) ranged from 6% to 41%, and there was an increased effect when enriching with xylanases and arabinofuranosidases in wheat aleurone and bran of 19% and 14% respectively. The enzymatic fingerprint of AX oligosaccharides revealed several non-final hydrolysis products of the enzymes applied, indicating that the hydrolysis of AX was not completed during in vitro digestion. These results indicate that the addition of a carbohydrase complex can introduce structural alterations under in vitro digestion conditions, and that enrichment with additional xylanases and arabinofuranosidases can boost this effect in wheat, maize, and rice. © 2020 Society of Chemical Industry.

Immunostimulatory Action of High-Content Active Arabinoxylan in Rice Bran.

Immunostimulatory activity comprises specific and nonspecific immune responses stimulated by internal and external factors. Arabinoxylan is well known for its immunostimulatory activity in vivo and in vitro, although the biological activities of arabinoxylan oligosaccharides depend on their structural features. In this study, we aimed to evaluate in vitro and in vivo the immunostimulatory activity of high-content active arabinoxylan (HCAA) obtained from rice bran through bioconversion by microorganisms and acid hydrolysis. Three microorganisms, Penicillium rocheforti, Aspergillus oryzae, and Pleurotus osteatus, and three different acid concentrations of hydrochloric acid (5, 10, and 20%) and acetic acid (25, 50, and 75%) were used for producing HCAA. HPLC analysis of arabinose and xylose content revealed that fermentation with P. rocheforti followed by hydrolysis with 5% hydrochloric acid was the most efficient to produce HCAA. GPC analysis of HCAA indicates that HCAA is a complex of various forms of saccharides and shows an average molecular weight of 625. Further, in vitro evaluation disclosed that exposure to HCAA (10–200 μg/mL) increased cell viability in mice splenic cells and RAW 264.7 cells. Additionally, exposure of mice to oral administration of HCAA (100 mg/kg) for 4–7 days increased lymphokine-activated killer (LAK)- and macrophage-mediated cytotoxic activity in cancer cells (YAC-1). Furthermore, in vitro exposure to HCAA and oral administrations in mice revealed increased interferon-γ (IFN-γ) and interleukin-10 (IL-10) protein expression through western blot analysis in RAW 264.7 cells and isolated splenic cells. Our results suggest that HCAA developed by bioconversion and acid hydrolysis may enhance immune responses in vivo and in vitro.

Relative abundance of the Prevotella genus within the human gut microbiota of elderly volunteers determines the inter-individual responses to dietary supplementation with wheat bran arabinoxylan-oligosaccharides

BackgroundThe human colon is colonised by a dense microbial community whose species composition and metabolism are linked to health and disease. The main energy sources for colonic bacteria are dietary polysaccharides and oligosaccharides. These play a major role in modulating gut microbial composition and metabolism, which in turn can impact on health outcomes.ResultsWe investigated the influence of wheat bran arabinoxylan oligosaccharides (AXOS) and maltodextrin supplements in modulating the composition of the colonic microbiota and metabolites in healthy adults over the age of 60. Male and female volunteers, (n = 21, mean BMI 25.2 ± 0.7 kg/m2) participated in the double-blind, cross over supplement study. Faecal samples were collected for analysis of microbiota, short chain fatty acids levels and calprotectin. Blood samples were collected to measure glucose, cholesterol and triglycerides levels. There was no change in these markers nor in calprotectin levels in response to the supplements. Both supplements were well-tolerated by the volunteers. Microbiota analysis across the whole volunteer cohort revealed a significant increase in the proportional abundance of faecal Bifidobacterium species (P ≤ 0.01) in response to AXOS, but not maltodextrin, supplementation. There was considerable inter-individual variation in the other bacterial taxa that responded, with a clear stratification of volunteers as either Prevotella-plus (n = 8; > 0.1% proportional abundance) or Prevotella-minus (n = 13; ≤0.1% proportional abundance) subjects founded on baseline sample profiles. There was a significant increase in the proportional abundance of both faecal Bifidobacterium (P ≤ 0.01) and Prevotella species (P ≤ 0.01) in Prevotella-plus volunteers during AXOS supplementation, while Prevotella and Bacteroides relative abundances showed an inverse relationship. Proportional abundance of 26 OTUs, including bifidobacteria and Anaerostipes hadrus, differed significantly between baseline samples of Prevotella-plus compared to Prevotella-minus individuals.ConclusionsThe wheat bran AXOS supplementation was bifidogenic and resulted in changes in human gut microbiota composition that depended on the initial microbiota profile, specifically the presence or absence of Prevotella spp. as a major component of the microbiota. Our data therefore suggest that initial profiling of individuals through gut microbiota analysis should be considered important when contemplating nutritional interventions that rely on prebiotics.Trial registrationClinical trial registration number: NCT02693782. Registered 29 February 2016 - Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT02693782?term=NCT02693782&rank=1

Colon-delivered short-chain fatty acids attenuate the cortisol response to psychosocial stress in healthy men: a randomized, placebo-controlled trial.

Short-chain fatty acids (SCFAs) are products of microbial fermentation of dietary fiber in the colon and may mediate microbiota-gut-brain communication. However, their role in modulating psychobiological processes that underlie the development of stress- and anxiety-related disorders is not mechanistically studied in humans. In this triple-blind, randomized, placebo-controlled intervention trial, we examine in a parallel group design the effects of 1-week colonic SCFA-mixture delivery in doses equivalent to fermentation of 10 g or 20 g of arabinoxylan oligosaccharides on responses to psychosocial stress and fear tasks in 66 healthy men. We demonstrate that low and high doses of SCFAs significantly attenuate the cortisol response to psychosocial stress compared to placebo. Both doses of SCFAs increase serum SCFA levels and this increase in circulating SCFAs co-varies significantly with the attenuation of the cortisol response to psychosocial stress. Colonic SCFA delivery does not modulate fecal SCFA concentrations, serum brain-derived neurotrophic factor, cortisol awakening response, fear learning and extinction, or subjective mood ratings. These results demonstrate that colon-delivered SCFAs modulate hypothalamic-pituitary-adrenal axis reactivity to psychosocial stress, thereby supporting their hypothesized role in microbiota-gut-brain communication.

Purification, Identification, and Characterization of an Endo-1,4-β-Xylanase from Wheat Malt.

In this study, an endo-1,4-β-xylanase was purified from wheat malt following the procedures of ammonium sulfate precipitation, cation-exchange chromatography, and two-step anion-exchange chromatography. The purified endo-1,4-β-xylanase had a specific activity of 3.94 u/mg, demonstrating a weight average molecular weight (Mw) of approximately 58,000 Da. After LC–MS/MS (Liquid chromatography-tandem mass spectrometry) identification, the purified enzyme had the highest matching degree with a GH10 (Glycoside Hydrolase 10) domain-containing protein from wheat, there were 23 match peptides with a score above the threshold and the prot-cover was 45.5%. The resulting purified enzyme was used to investigate its degradation ability on high viscosity wheat-derived water-extractable arabinoxylan (WEAX). Degradation experiments confirmed that the purified enzyme was a true endo-acting enzyme, which could degrade large WEAX into smaller WEAX. The average degree of polymerization (avDP) and the viscosity of WEAX decreased with the increasing reaction time. The enzyme could degrade a small amount of WEAX into arabinoxylan-oligosaccharides (AXOS) with a degree of polymerization of 2–6, but no monosaccharide was produced. The degradation occurred rapidly in the first 3.5 h and decreased with the further prolongation of reaction time.

Single or combined effects of dietary arabinoxylan-oligosaccharide and inulin on growth performance, gut microbiota, and immune response in Pacific white shrimp Litopenaeus vannamei

The single and synergistic effects of dietary arabinoxylan-oligosaccharide (AXOS) and inulin at different doses (2, 4, and 8 mg/g diet) on survival, growth performance, gut microbiota, and immune response in Pacific white shrimp Litopenaeus vannamei were assessed. Singular application of either inulin or AXOS at doses of 4 mg/g diet showed the most stimulatory effects on the growth rate and gene expression levels of chitinase, cathepsin L, chymotrypsin, and extracellular signal-regulated kinase (ERK) in each single prebiotic feeding trial. Compared with single prebiotic treatments, simultaneous application of AXOS and inulin at 4 mg/g diet enhanced remarkably the growth parameters of shrimp and the expression of related genes (chitinase, cathepsin L, chymotrypsin, ERK, myeloid differentiation factor 88, and phenoloxidase) after 8-week feeding (P < 0.05). Additionally, gut microbiota analysis indicated the dietary supplementation with combined prebiotics increased significantly the bacterial community richness and relative abundance of Bacillus, Pseudomonas, Bacteriovorax, and Lactobacillus, and reduced the abundance of Vibrio, Rhodococcus, and Photobacterium in the digestive tract of L. vannamei. Compared with the single prebiotic treatment and the control, combined prebiotics supplementation boosted notably the survival rate and expression levels of immune-related genes in shrimp infected with Vibrio alginolyticus or white spot syndrome virus. Therefore, simultaneous application of AXOS and inulin shall have a great potential of dietary supplement in the culture of L. vannamei.

Arabinoxylan-oligosaccharides kick-start arabinoxylan digestion in the aging broiler

While arabinoxylans (AX), an important dietary fiber fraction of wheat-based broiler diets, are known for exerting antinutritional effects in the gastrointestinal (GI) tract of broilers, the prebiotic potential of arabinoxylan-oligosaccharides (AXOS) is also well-documented. However, inconsistent performance responses as well as the effectiveness of low amounts of AXOS used in diets of previously conducted experiments put into question the classical prebiotic route being the sole mode of action of AXOS. The objective of this study was to investigate the effects of dietary AXOS addition on the rate of AX digestion in the gastrointestinal tract of broilers as a function of broiler age to gain more insight into the mode of action of these oligosaccharides. A feeding trial was performed on 480 one-day-old chicks (Ross 308) receiving a wheat-based diet supplemented with or without 0.50% AXOS, containing no endoxylanases. Digesta samples from ileum and caeca and fecal samples were analyzed for AX content, AX digestibility, intestinal viscosity, and microbial AX-degrading enzyme activities at 6 different ages (day 5, 10, 15, 21, 28, 35). Chicks fed from hatching with 0.50% AXOS demonstrated a higher ileal viscosity (P < 0.05). Also higher levels of AX solubilization and fermentation compared to control birds at 10 D were observed. This was noted by the higher total tract AX digestibility of water-extractable AX (WE-AX) and total AX (TOT-AX) at this age (P < 0.05). Although no significant difference in AX-degrading enzyme activities was observed among the dietary treatments, AXOS supplementation in young broilers was shown to stimulate or “kick-start” dietary AX digestion, thereby speeding up the development of a fiber-fermenting microbiome in the young broiler. This stimulation effect of AXOS could enable greater functional value to be extracted from dietary fiber in broiler feeds.

A Glycoside Hydrolase Family 62 A-L-Arabinofuranosidase from Trichoderma Reesei and Its Applicable Potential during Mashing.

Arabinoxylan is the second most abundant component in the endosperm cell wall of barley and it has been shown to have negative effects on the viscosity and filtration rate of wort and beer. In this study, a glycoside hydrolase (GH) family 62 α-L-arabinofuranosidase (AFase), termed as TrAbf62A, was purified from the culture filtrate of Trichoderma reesei CICC 41495 by a combined chromatographic method. The preferred substrates of the purified TrAbf62A were soluble, highly substituted arabinoxylan oligosaccharides and polymers, similar to the type found in barley grain. TrAbf62A exhibited activity towards oligomeric and polymeric arabinoxylans, as well as colorimetric arabinose-based substrates, thus meeting the criteria to be classified as a type B AFase. TrAbf62A released mainly arabinose and xylose from soluble wheat arabinoxylan, thus indicating a dual lytic enzyme activity. Supplementation of TrAbf62A during mashing, with a loading of 12 mU/g malt, resulted in a 36.3% decrease in arabinoxylan polymer content, a 5.6% reduction in viscosity, and finally, a 22.1% increase in filtration rate. These results revealed that TrAbf62A has a high potential value in improving lautering performance during mashing.

13C-DOSY-TOSY NMR Correlation for In Situ Analysis of Structure, Size Distribution, and Dynamics of Prebiotic Oligosaccharides

Arabinoxylan oligosaccharides (AXOS) are a complex mixture of cereal derived, water-soluble prebiotics, obtained by enzymatic hydrolysis of arabinoxylan, a group of dietary fibers exerting numerous nutritional and health-beneficial effects. Such complex biomolecular mixtures are notoriously difficult to characterize without initial physical fractionation. Here we present the in situ analysis of AXOS using a variety of state-of-the-art sensitivity-enhanced 13C-DOSY methods, enabling virtual separation and identification of the components. Three dimensional correlation plots displaying 13C diffusivity (DOSY: Diffusion Ordered SpectroscopY), relaxation parameters (TOSY: raTe of relaxation Ordered SpectrscopY), and chemical shift offer a unique way to elucidate the composition of mixtures. We have demonstrated this multifaceted 13C probed correlation strategy in standard mixtures of aliphatic and aromatic compounds, before implementing it on AXOS. These 3D-DOSY-TOSY plots in combination with 2D-NMR correlation experiments offer unprecedented clarity for assigning chemical functions, molecular size distribution, and dynamics of oligosaccharide mixtures.

Effect of wheat bran derived prebiotic supplementation on gastrointestinal transit, gut microbiota, and metabolic health: a randomized controlled trial in healthy adults with a slow gut transit

ABSTRACT Acute intake of the wheat bran extract Arabinoxylan-Oligosaccharide (AXOS) modulates the gut microbiota, improves stool characteristics and postprandial glycemia in healthy humans. Yet, little is known on how long-term AXOS intake influences gastrointestinal (GI) functioning, gut microbiota, and metabolic health. In this randomized, placebo-controlled, double-blind study, we evaluated the effects of AXOS intake on GI function and metabolic health in adults with slow GI transit without constipation. Forty-eight normoglycemic adults were included with whole-gut transit time (WGTT) of >35 h receiving either 15 g/day AXOS or placebo (maltodextrin) for 12-wks. The primary outcome was WGTT, and secondary outcomes included stool parameters, gut permeability, short-chain fatty acids (SCFA), microbiota composition, energy expenditure, substrate oxidation, glucose, insulin, lipids, gut hormones, and adipose tissue (AT) function. WGTT was unchanged, but stool consistency softened after AXOS. 12-wks of AXOS intake significantly changed the microbiota by increasing Bifidobacterium and decreasing microbial alpha-diversity. With a good classification accuracy, overall microbiota composition classified responders with decreased WGTT after AXOS. The incretin hormone Glucagon-like protein 1 was reduced after AXOS compared to placebo. Energy expenditure, plasma metabolites, AT parameters, SCFA, and gut permeability were unchanged. In conclusion, intake of wheat bran extract increases fecal Bifidobacterium and softens stool consistency without major effects on energy metabolism in healthy humans with a slow GI transit. We show that overall gut microbiota classified responders with decreased WGTT after AXOS highlighting that GI transit and change thereof were associated with gut microbiota independent of Bifidobacterium. NCT02491125