Two Subgroups within the GH43_36 α-l-Arabinofuranosidase Subfamily Hydrolyze Arabinosyl from Either Mono-or Disubstituted Xylosyl Units in Wheat Arabinoxylan

Fungal arabinofuranosidases (ABFs) catalyze the hydrolysis of arabinosyl substituents (Ara) and are key in the interplay with other glycosyl hydrolases to saccharify arabinoxylans (AXs). Most characterized ABFs belong to GH51 and GH62 and are known to hydrolyze the linkage of α-(1→2)-Ara and α-(1→3)-Ara in monosubstituted xylosyl residues (Xyl) (ABF-m2,3). Nevertheless, in AX a substantial number of Xyls have two Aras (i.e., disubstituted), which are unaffected by ABFs from GH51 and GH62. To date, only two fungal enzymes have been identified (in GH43_36) that specifically release the α-(1→3)-Ara from disubstituted Xyls (ABF-d3). In our research, phylogenetic analysis of available GH43_36 sequences revealed two major clades (GH43_36a and GH43_36b) with an expected substrate specificity difference. The characterized fungal ABF-d3 enzymes aligned with GH43_36a, including the GH43_36 from Humicola insolens (HiABF43_36a). Hereto, the first fungal GH43_36b (from Talaromyces pinophilus) was cloned, purified, and characterized (TpABF43_36b). Surprisingly, TpABF43_36b was found to be active as ABF-m2,3, albeit with a relatively low rate compared to other ABFs tested, and showed minor xylanase activity. Novel specificities were also discovered for the HiABF43_36a, as it also released α-(1→2)-Ara from a disubstitution on the non-reducing end of an arabinoxylooligosaccharide (AXOS), and it was active to a lesser extent as an ABF-m2,3 towards AXOS when the Ara was on the second xylosyl from the non-reducing end. In essence, this work adds new insights into the biorefinery of agricultural residues.     

Arabinoxylan and Pectin Metabolism in Crohn’s Disease Microbiota: An In Silico Study

Inflammatory bowel disease is a chronic disorder including ulcerative colitis and Crohn’s disease (CD). Gut dysbiosis is often associated with CD, and metagenomics allows a better understanding of the microbial communities involved. The objective of this study was to reconstruct in silico carbohydrate metabolic capabilities from metagenome-assembled genomes (MAGs) obtained from healthy and CD individuals. This computational method was developed as a mean to aid rationally designed prebiotic interventions to rebalance CD dysbiosis, with a focus on metabolism of emergent prebiotics derived from arabinoxylan and pectin. Up to 1196 and 1577 MAGs were recovered from CD and healthy people, respectively. MAGs of Akkermansia muciniphila, Barnesiella viscericola DSM 18177 and Paraprevotella xylaniphila YIT 11841 showed a wide range of unique and specific enzymes acting on arabinoxylan and pectin. These glycosidases were also found in MAGs recovered from CD patients. Interestingly, these arabinoxylan and pectin degraders are predicted to exhibit metabolic interactions with other gut microbes reduced in CD. Thus, administration of arabinoxylan and pectin may ameliorate dysbiosis in CD by promoting species with key metabolic functions, capable of cross-feeding other beneficial species. These computational methods may be of special interest for the rational design of prebiotic ingredients targeting at CD.     

Effects of genotype and harvest year on content and composition of dietary fibre in rye (Secale cereale L) grain

The aim of this study was to measure the variation in dietary fibre (DF) content and composition among different rye varieties grown in the same location in three successive years and to estimate the contributions of genotype and harvest year to this variation. The study included grain from 19 different varieties of rye, of which seven varieties were analysed in all three years. The content of total DF in all samples ranged from 147 to 209 g kg^?1 dry matter (dm), with 34–66 g kg^?1 being water‐extractable (WE) DF and 108–159 g kg^?1 being water‐unextractable (WUE) DF. The main DF component was arabinoxylan (AX) with a content of 80–121 g kg^?1 dm, of which 26–41 g kg^?1 dm was WE AX. The ratio of arabinose to xylose (ara/xyl) of total AX ranged from 0.59 to 0.69. The content of β‐glucan was 13–22 g kg^?1 dm and that of fructan was 45–64 g kg^?1 dm. Analysis of the data from seven varieties and three harvest years showed a highly significant influence of harvest year on most DF components. Significant genotype effects were found for the contents of total DF, total AX, β‐glucan and fructan and for the ara/xyl ratio of total and WUE AX. Estimation of variance components showed that yearly variations in the contents of total DF, total AX, WUE DF, WUE AX and β‐glucan were generally higher (27–55% of total variance) than genotype effects (14–19%). The ara/xyl ratio of total and WUE AX showed a high influence of genotype (46 and 43% respectively). Non‐adapted rye varieties with small kernel size had the highest contents of total DF and total AX. The content of β‐glucan was positively correlated with kernel weight (r = 0.68), whereas no overall correlation was found between kernel size and the content of DF. As a conclusion, the variation in DF content and composition in this study was significantly influenced by both harvest year and rye genotype. © 2002 Society of Chemical Industry     

麦麸中生理活性物质研究与应用进展

阿拉伯木聚糖、植酸、阿魏酸是麦麸中重要的生理活性物质,具有抗肿瘤、抗氧化等功能,加上其很强的饱水性、稳定性,使它在食品添加剂和功能食品开发上具有广泛的应用前景。不同的分离提取方法对这些生理活性物质的得率、结构影响明显,并影响其功用,本文综述了麦麸阿拉伯木聚糖、植酸、阿魏酸的提取方法及其应用情况。     

In vitro fermentation of cereal dietary fibre carbohydrates by probiotic and intestinal bacteria

A range of probiotic and other intestinal bacteria were examined for their ability to ferment the dietary fibre carbohydrates β‐glucan, xylan, xylo‐oligosaccharides (XOS) and arabinoxylan. β‐Glucan was fermented by Bacteroides spp and Clostridium beijerinckii but was not fermented by lactobacilli, bifidobacteria, enterococci or Escherichia coli. Unsubstituted xylan was not fermented by any of the probiotic bacteria examined. However, many Bifidobacterium species and Lactobacillus brevis were able to grow to high yields using XOS. XOS were also efficiently fermented by some Bacteroides isolates but not by E coli, enterococci, Clostridium difficile, Clostridium perfringens or by the majority of intestinal Lactobacillus species examined. Bifidobacterium longum strains were able to grow well using arabinoxylan as the sole carbon source. These organisms hydrolysed and fermented the arabinosyl residues from arabinoxylan but did not substantially utilise the xylan backbone of the polysaccharide. Arabinoxylan was not fermented by lactobacilli, enterococci, E coli, C perfringens or C difficile and has potential to be an applicable carbohydrate to complement probiotic Bif longum strains in synbiotic combinations. © 2002 Society of Chemical Industry     

Improvement of the Characteristics of Steamed Bread by Supplementation of Recombinant alpha-L-arabinofuranosidase Containing xylan-binding domain

Current study investigated the potential to increase the activity of α-L-arabinofuranosidase (TmAra) from Thermotoga marotima on insoluble arabinoxylan (IAX) through fusion to a family 6 xylan-binding domain (CtXBD6) from Clostridium thermocellum. CtXBD6 was fused to the carboxyl terminus of TmAra, and active forms of both TmAra and TmAraCtXBD6 were produced in E. coli and purified to homogeneity for biochemical characterization. TmAraCtXBD6 had similar temperature and pH profiles to TmAra with a decrease in the thermostability and pH stability at pH 7.0–8.2. It exhibited stronger IAX-binding and hydrolytic activity than TmAra. TmAra and TmAraCtXBD6-treated IAX were added to wheat flour to investigate the rheological and textural properties of the finished product. IAX alone had significant increase in dough development time (DDT) and dough stability (DS) and decrease in the softening degree (SD), crumb firmness (CF) and specific volume (SV). On adding TmAra and TmAraCtXBD6 to treat IAX texture, SV and DS improved but decreased DDT, SD, and CF. TmAraCtXBD6 reduced CF and increased SV, and had higher springiness, cohesiveness, and crumb chewiness than TmAra. These results demonstrated that the fusion of CtXBD6 into arabinofuranosidase enables the increase in catalytic performance on insoluble AX substrates, which can be used as more efficient biocatalysts that can improve the quality of AX-rich products.     

Kinetic analysis of bile salt passage across a dialysis membrane in the presence of cereal soluble dietary fibre polymers

The kinetics of passage of a model bile salt and complete porcine bile across a dialysis membrane, in the presence and absence of two cereal-derived soluble dietary fibre polysaccharides, were studied as a model for passage across the unstirred water layer that lines the small intestine. A first-order kinetic analysis allowed rate coefficients to be derived which quantified the effectiveness of barley mixed linkage β-glucan and wheat arabinoxylan in retarding the transport of bile. For both, a model bile salt and complete porcine bile, rate coefficients decreased with both concentration and viscosity. A combination of viscosity and molecular interaction effects is suggested to control the effect of the two polysaccharides on the transport of bile.     

Changes in Molecular Characteristics of Cereal Carbohydrates after Processing and Digestion

Different extraction, purification and digestion methods were used to investigate the molecular properties of carbohydrates in arabinoxylan and β-glucan concentrates, dietary fiber (DF) rich breads and ileum content of bread fed pigs. The breads studied were: a low DF wheat bread (WF), whole meal rye bread (GR), rye bread with kernels (RK), wheat bread supplemented with wheat arabinoxylan concentrate (AX) and wheat bread supplemented with oat β-glucan concentrate (BG). The weight average molecular weight (M_w) of extractable carbohydrates in β-glucan concentrate decreased eight-fold after inclusion in the BG bread when exposed to in vitro digestion, while the M_w of purified extractable carbohydrates in AX bread was reduced two-fold, and remained almost unaffected until reaching the terminal ileum of pigs. Similarly, the M_w of purified extractable carbohydrates in GR and RK bread was not significantly changed in the ileum. The AX bread resulted in the highest concentration of dissolved arabinoxylan in the ileum among all the breads that caused a substantial increased in ileal AX viscosity. Nevertheless, for none of the breads, the M_w of extractable carbohydrates was related neither to the bread extract nor ileal viscosity.     

阿拉伯木聚糖与壳聚糖复凝聚物的制备及表征

研究了不同因素对壳聚糖-阿拉伯木聚糖复凝聚的影响,通过测定平衡相的浊度和复凝聚相收率确定玉米阿拉伯木聚糖（CAX）和壳聚糖（CS）间复合物形成的条件。采用红外光谱、热重分析、扫描电子显微镜和流变学特性对复凝聚物进行表征。研究结果表明,在CAX/CS配比为9:1,体系pH值为4.0,总固形物浓度为3%（m/m）,室温下反应10 min,最大复凝聚相产率达76.04%。红外光谱和热重分析结果表明,CAX/CS复凝聚物是通过CS中的-NH3+和CAX中-COO-之间的相互吸引形成的。用SEM扫描成像显示复凝聚物具有规则且分布均匀的多孔网络结构。CAX/CS复凝聚物粘弹特性取决于发生复凝聚的pH值。在pH值3.5和4.0时,复凝聚物为液体粘弹行为;在pH值5.0时,复凝聚物为固体粘弹性行为。pH值5.0下形成的复凝聚物的G’最高,说明除静电作用外,其他因素也可能对CAX/CS复凝聚物粘弹特性起重要作用。CAX/CS复凝聚物可以作为包封营养素和功能性因子的微囊化壁材。     

In vitro fermentation of rye carbohydrates including arabinoxylans of different structure

Freeze‐dried ileal effluent from cannulated pigs fed rye bread diets based on either whole rye or one of three rye milling fractions enriched in pericarp/testa, aleurone and endosperm was incubated in an in vitro fermentation system with faecal inocula from pigs fed a whole grain rye diet. Three and 48 hours of in vitro degradation of arabinoxylans were comparable to the caecal and faecal degradation respectively. The productions of short‐chain fatty acids were highest from the ileum samples of the diets high in fermentable carbohydrates (whole rye, aleurone and endosperm). However, fermentation of the pericarp/testa ileum sample produced half the amount of short‐chain fatty acids resulting from the other ileum samples despite the fact that the pericarp/testa carbohydrates (CHO) were not fermented at all. Treating the pericarp/testa with alkali prior to the incubation increased fermentability considerably (from 0 to 49% of total arabinoxylans) and suggested that, at least in lignified tissue, alkali‐labile cross‐links were more important determinants of arabinoxylan (AX) degradability than AX structure itself. Oxidative delignification of the pericarp/testa ileal effluent increased fermentability too, but to a much lower degree, which indicates that the type and extent of links between polymers and lignin rather than the amount of lignin are important in determining degradability. © 2000 Society of Chemical Industry