Non-cellulosic Polysaccharides Research Articles

Acid hydrolysis of O-acetyl-galactoglucomannan

Acid hydrolysis of O-acetyl-galactoglucomannan (GGM), the main non-cellulosic polysaccharide in coniferous wood, was studied at 90 °C in the pH range of 0.5–2.0 using different acids (hydrochloric acid, sulfuric acid, trifluoroacetic acid, oxalic acid, and formic acid) as homogeneous catalysts and cation exchange resins (Smopex 101 and Amberlyst 15) as heterogeneous catalysts. The hydrolysis rate was exclusively dependent on the pH of the solution, but independent of the concentration and type of acid. Analysis of the formation rates of oligomers with DP 2–5 revealed that GGM hydrolysis occurs rather randomly. The final products were mannose, galactose, and glucose. In the beginning of the reaction, the formation of galactose was slightly faster than those of other sugars, since galactose is a side substituent to the glucose–mannose main chain of GGM. However, with longer reaction times, the formation rates of sugar monomers were correlated with their initial abundances in the polysaccharide. Autocatalytic kinetics was observed in the presence of the heterogeneous catalysts: an induction time was observed, after which the reaction proceeded with the same rates as with homogeneous acids. The autocatalytic effect is probably explained by the increase of the hydrolysis rate constant, as the GGM chain length decreases and the molecule becomes more accessible for the active sites on the outer surface of the catalyst.

Designing Cell Walls for Improved Bioenergy Production

Dwindling supply and unpredictable price fluctuations of fossil fuels have necessitated a renewed worldwide search for alternative fuels. Plant cell walls, rich in cellulose and hemicellulose, are actively pursued for their use in production of second-generation biofuels. It has been envisioned that degradation of these carbohydrate polymers to their sugar monomer subunits (saccharification) will provide necessary substrates for their yeast-mediated fermentation to ethanol. But the major component of plant cell walls, cellulose, is not easily amenable to this type of deconstruction and is tightly interlinked with hemicelluloses and lignin, making cell walls highly recalcitrant to saccharification.

Equine pre-caecal and total tract digestibility of individual carbohydrate fractions and their effect on caecal pH response

The working hypothesis was that a minor postprandial caecal pH decline would affect apparent total tract digestibility (ATTD) of the fibre fraction in horses and, hence, that soluble fibre would amplify fermentation and consequently increase ATTD of fibre. This study was a 4 × 4 Latin Square design with a sequence of 17 days adaptation to the ration followed by 8 sampling days. The feed rations consisted of only timothy hay (Group H), hay plus molassed sugar beet pulp combined with either whole oats (Group OB) or barley (Group BB) and hay plus loose chaff based concentrate (Group M). Four horses fitted with permanent caecal cannulas and collection harnesses were used. A pH electrode with logger was inserted through the cannula and caecal pH was recorded at 1 min intervals for 8 h. The mobile nylon bag technique was used to quantify pre-caecal loss (PCL) of individual feedstuffs. Fibre was analysed as dietary fibre (DF), non-starch polysaccharides, soluble non-cellulosic polysaccharides (S-NCP), insoluble non-cellulosic polysaccharide (I-NCP) and neutral detergent fibre. The ATTD of the S-NCP fraction was above 0.8, which was 60% higher than for the I-NCP fraction. The PCL of starch were 0.98 (oats) and 0.75 (barley). The BB diet lowered (p < 0.001) postprandial caecal pH more than the other diets and a significant correlation was found between the lowest pH and ATTD of I-NCP (r = −0.66; p = 0.005). In conclusion, this study successfully measured the in vivo digestibility of individual fibre fractions and found that S-NCP was more digestible than the I-NCP, and that a single meal of unprocessed barley was sufficient to decrease caecal pH to such an extent that the fibre digestibility of the whole diet was negatively affected.

Engineering of plants with improved properties as biofuels feedstocks by vessel-specific complementation of xylan biosynthesis mutants

BackgroundCost-efficient generation of second-generation biofuels requires plant biomass that can easily be degraded into sugars and further fermented into fuels. However, lignocellulosic biomass is inherently recalcitrant toward deconstruction technologies due to the abundant lignin and cross-linked hemicelluloses. Furthermore, lignocellulosic biomass has a high content of pentoses, which are more difficult to ferment into fuels than hexoses. Engineered plants with decreased amounts of xylan in their secondary walls have the potential to render plant biomass a more desirable feedstock for biofuel production.ResultsXylan is the major non-cellulosic polysaccharide in secondary cell walls, and the xylan deficient irregular xylem (irx) mutants irx7, irx8 and irx9 exhibit severe dwarf growth phenotypes. The main reason for the growth phenotype appears to be xylem vessel collapse and the resulting impaired transport of water and nutrients. We developed a xylan-engineering approach to reintroduce xylan biosynthesis specifically into the xylem vessels in the Arabidopsis irx7, irx8 and irx9 mutant backgrounds by driving the expression of the respective glycosyltransferases with the vessel-specific promoters of the VND6 and VND7 transcription factor genes. The growth phenotype, stem breaking strength, and irx morphology was recovered to varying degrees. Some of the plants even exhibited increased stem strength compared to the wild type. We obtained Arabidopsis plants with up to 23% reduction in xylose levels and 18% reduction in lignin content compared to wild-type plants, while exhibiting wild-type growth patterns and morphology, as well as normal xylem vessels. These plants showed a 42% increase in saccharification yield after hot water pretreatment. The VND7 promoter yielded a more complete complementation of the irx phenotype than the VND6 promoter.ConclusionsSpatial and temporal deposition of xylan in the secondary cell wall of Arabidopsis can be manipulated by using the promoter regions of vessel-specific genes to express xylan biosynthetic genes. The expression of xylan specifically in the xylem vessels is sufficient to complement the irx phenotype of xylan deficient mutants, while maintaining low overall amounts of xylan and lignin in the cell wall. This engineering approach has the potential to yield bioenergy crop plants that are more easily deconstructed and fermented into biofuels.

Trans‐Golgi Network—An Intersection of Trafficking Cell Wall ComponentsF

The cell wall, a crucial cell compartment, is composed of a network of polysaccharides and proteins, providing structural support and protection from external stimuli. While the cell wall structure and biosynthesis have been extensively studied, very little is known about the transport of polysaccharides and other components into the developing cell wall. This review focuses on endomembrane trafficking pathways involved in cell wall deposition. Cellulose synthase complexes are assembled in the Golgi, and are transported in vesicles to the plasma membrane. Non-cellulosic polysaccharides are synthesized in the Golgi apparatus, whereas cellulose is produced by enzyme complexes at the plasma membrane. Polysaccharides and enzymes that are involved in cell wall modification and assembly are transported by distinct vesicle types to their destinations; however, the precise mechanisms involved in selection, sorting and delivery remain to be identified. The endomembrane system orchestrates the delivery of Golgi-derived and possibly endocytic vesicles carrying cell wall and cell membrane components to the newly-formed cell plate. However, the nature of these vesicles, their membrane compositions, and the timing of their delivery are largely unknown. Emerging technologies such as chemical genomics and proteomics are promising avenues to gain insight into the trafficking of cell wall components.

Analysis of non-cellulosic polysaccharides helps to reveal the history of thick organic surface layers on calcareous Alpine soils

Background and aimsWe investigated the potential of non-cellulosic polysaccharides (NCP) as biomarkers to identify the plant types that dominate present and past litter input into organic surface covers on calcareous Alpine soils and to reveal historic vegetation changes.MethodsAt two sites in the Alps, NCP monomers were quantified in different organs of site-dominating plants, the Oa horizon of four Folic Leptosols, and different sections of thick organic surface layers of four Folic Histosols on calcareous bedrock.ResultsThe dominating plant types at our study sites differ markedly in their NCP composition and (galactose + mannose)/(arabinose + xylose) [GM/AX] ratio (grasses and sedges: 0.2; dicots Fagus and Vaccinium: 0.2–0.6; conifers Abies, Picea, Pinus: 0.7–2.4; mosses: 5). For all except one soil, the NCP signature of the uppermost Oa horizon reflects the present vegetation. For all Histosol O horizons, NCP signatures indicate a dominance of conifer litter throughout their development (up to 1,500 years). Different NCP and GM/AX depth profiles reflect specific patterns of O layer genesis.ConclusionsNCP and GM/AX depth profiles in organic surface covers of soils provide important information about dominating litter sources in the past and can be valuable tools to reveal historic vegetation and/or land use changes.

Chemical and physicochemical characterisation of various horse feed ingredients

There is an increasing need for identifying energy dense fibre rich feed ingredients, because starch has shown to cause detrimental health problems in sports horses. This study aimed at evaluating feeds considered to be suitable for horses by use of comprehensive carbohydrate analytical methods. Eighteen feeds of diverse botanical origin ranging from apple pulp, root crops, cereal grains and roughages were analysed for dry matter, ash, crude protein, fat, carbohydrates and lignin. Carbohydrates were analysed for sugar, starch, fructans, oligosaccharides and non-starch polysaccharides (NSP) using enzymatic-chemical methods and lignin by gravimetry. The results for total dietary fibre (DF) were compared with conventional crude fibre (CF) and neutral detergent fibre (aNDFom) methods. The physicochemical properties were quantified based on swelling and water binding capacity (WBC). Between 755g and 970g/kg DM was detected. The highest total sugar content was measured in root crops with 803g/kg DM in raw sugar beet followed by 581g/kg DM in carrots. The starch content ranged between 343g/kg DM and 754g/kg DM in cereal grains. The DF method detected (P≤0.04) more NSP as compared to the aNDFom and CF methods. The greatest difference between the DF and aNDFom methods was found in root crops and apple pulp in which the soluble non-cellulosic polysaccharides (S-NCP) fraction made up 350–581g/kg of total NSP. The physicochemical properties were compared to fibre content and were associated to WBC (P=0.04). The correlation ranged from 0.52 to 0.53 for WBC and from 0.03 to 0.16 for swelling. The S-NCP fraction is lost in the aNDFom and CF methods due to solubilisation, which explains the higher recovery of total NSP by the DF method. This illustrates that a feed's potential as a fibre source may be underestimated depending on the analytical method. Quantifying the soluble NCP fraction is beneficial as it has been shown to have health beneficial properties and contributes to the total energy supply. These results suggest that the DF method should be used when evaluating feeds for horses.

Chemical Characterization and Water Holding Capacity of Fibre-rich Feedstuffs Used for Pigs in Vietnam.

During two years, four samples per year were collected in Vietnam from rice bran, cassava residue, brewer’s grain, tofu residue, soybean meal, coconut cake, sweet potato vines and water spinach for chemical analysis and assessment of water holding capacity (WHC). The selected feedstuffs represent fibre-rich plant sources and agro-industry co-products commonly used in pig feeding in Vietnam. The content (g/kg DM) of crude protein (CP), ether extract (EE) and non-starch polysaccharides (NSP) varied between feedstuffs and ranged from 21 to 506 for CP, from 14 to 118 for EE and from 197 to 572 for NSP. Cassava residue had a high starch content of 563 g/kg DM, while sweet potato vines, water spinach, coconut cake and soybean meal had a high content of sugars (63–71 g/kg DM). The content of individual neutral sugars varied between feed ingredients, with the highest content of arabinose, galactose and glucose in tofu residue, the highest content of xylose in brewer’s grain and the highest content of mannose in coconut cake. The content of uronic acid was high for cassava residue, tofu residue, sweet potato vines and water spinach (57–88 g/kg DM). The content of soluble non-cellulosic polysaccharides (S-NCP) was positively correlated (r2 = 0.82) to the WHC. The content (g/kg DM) of CP, NDF, neutral sugars, total NSP, total NCP, S-NCP and total dietary fibre in tofu residue, water spinach and coconut cake varied (p<0.05) between years. In conclusion, diet formulation to pigs can be improved if the variation in chemical composition of the fibre fraction and in WHC between potential feed ingredients is taken into account.

Hot-water extraction and characterization of spruce bark non-cellulosic polysaccharides

Abstract Non-cellulosic polysaccharides (NCP) from bark offer large potential as a class of natural raw materials for functional materials development and production of biochemicals. We have elaborated a process for sequential extraction of NCP from industrial Norway spruce bark using an accelerated solvent extraction (ASE) with water at 100 to 160°C. Carbohydrates, Klason lignin and ash content as well as size-exclusion chromatography (SEC) analyses were performed for all hot-water extracts. NCP were mainly composed of glucose, arabinose and galacturonic acid units which revealed the presence of starch, arabinose-rich hemicelluloses and pectins. In total, the industrial bark of Norway spruce contained up to 20% of NCP which were extracted with pressurized hot water. NCP were mainly extractable at 140°C and started to undergo degradation at higher temperature.

Golgi-Mediated Synthesis and Secretion of Matrix Polysaccharides of the Primary Cell Wall of Higher Plants

The Golgi apparatus of eukaryotic cells is known for its central role in the processing, sorting, and transport of proteins to intra- and extra-cellular compartments. In plants, it has the additional task of assembling and exporting the non-cellulosic polysaccharides of the cell wall matrix including pectin and hemicelluloses, which are important for plant development and protection. In this review, we focus on the biosynthesis of complex polysaccharides of the primary cell wall of eudicotyledonous plants. We present and discuss the compartmental organization of the Golgi stacks with regards to complex polysaccharide assembly and secretion using immuno-electron microscopy and specific antibodies recognizing various sugar epitopes. We also discuss the significance of the recently identified Golgi-localized glycosyltransferases responsible for the biosynthesis of xyloglucan (XyG) and pectin.

Extraction and chemical characterization of Norway spruce inner and outer bark

Abstract Possible chemical utilization of bark requires appropriate knowledge of its composition. Extraction of valuable components before burning is an interesting option for utilization of bark. Here, Norway spruce inner and outer bark were extracted separately with a successive series of solvents of increasing polarity and the extracts, as well as the residues, were analyzed to obtain an overall picture of the bark composition. The lipophilic extractives contained the same major components as found in wood. Inner bark contained over 10% of stilbene glucosides with piceatannol (astringenin) as the main stilbene. Tannins of the proanthocyanidin type were extracted with hot water. Further extraction with pressurized hot water at 140°C or 160°C yielded 11-14% of non-cellulosic polysaccharides, on original bark basis, with pectic polysaccharides built up of arabinose, galacturonic acid and rhamnose dominating. Inner bark contained two times more cellulose than outer bark, but the opposite was true for lignin, determined as Klason “lignin”. Among the potentially valuable components, stilbene glucosides could be extracted with water even at low temperatures, while tannins could be extracted with hot water in a second step. The pectic polysaccharides are also of potential interest and should be studied further. The amount and true chemical character of lignin is also not yet fully elucidated.

Localization of Cell Wall Polysaccharides in Normal and Compression Wood of Radiata Pine: Relationships with Lignification and Microfibril Orientation

The distribution of noncellulosic polysaccharides in cell walls of tracheids and xylem parenchyma cells in normal and compression wood of Pinus radiata, was examined to determine the relationships with lignification and cellulose microfibril orientation. Using fluorescence microscopy combined with immunocytochemistry, monoclonal antibodies were used to detect xyloglucan (LM15), β(1,4)-galactan (LM5), heteroxylan (LM10 and LM11), and galactoglucomannan (LM21 and LM22). Lignin and crystalline cellulose were localized on the same sections used for immunocytochemistry by autofluorescence and polarized light microscopy, respectively. Changes in the distribution of noncellulosic polysaccharides between normal and compression wood were associated with changes in lignin distribution. Increased lignification of compression wood secondary walls was associated with novel deposition of β(1,4)-galactan and with reduced amounts of xylan and mannan in the outer S2 (S2L) region of tracheids. Xylan and mannan were detected in all lignified xylem cell types (tracheids, ray tracheids, and thick-walled ray parenchyma) but were not detected in unlignified cell types (thin-walled ray parenchyma and resin canal parenchyma). Mannan was absent from the highly lignified compound middle lamella, but xylan occurred throughout the cell walls of tracheids. Using colocalization measurements, we confirmed that polysaccharides containing galactose, mannose, and xylose have consistent correlations with lignification. Low or unsubstituted xylans were localized in cell wall layers characterized by transverse cellulose microfibril orientation in both normal and compression wood tracheids. Our results support the theory that the assembly of wood cell walls, including lignification and microfibril orientation, may be mediated by changes in the amount and distribution of noncellulosic polysaccharides.

Microsphere valorization of forestry derived hydrolysates

A value-adding approach to the material utilization of non-cellulosic polysaccharides (NCPs) released from the lignocellulosic feedstock was realized via the formulation of renewable microspheres from wood hydrolysates using a purposely elaborated all edible water-in-oil emulsion technique. Four compositionally different hemicellulose rich wood hydrolysates were recovered from process waters in pulping and other hydrothermal treatments of hardwood and softwood. Multivariate screening designs were employed allowing for the identification and quantitation of significant process parameters and interaction effects governing the conversion of hydrolysates into small, smooth and well-defined microspheres with narrow size dispersity.

Biosynthesis of non-cellulosic polysaccharides of plant cell walls

Enzymes that make the polymer backbones of plant cell wall polysaccharides have proven to be recalcitrant to biochemical purification. Availability of mutational genetics and genomic tools paved the way for rapid progress in identifying genes encoding various cell wall glycan synthases. Mutational genetics, the primary tool used in unraveling cellulose biosynthesis, was ineffective in assigning function to any of the hemicellulosic, polymerizing glycan synthases. A combination of comparative genomics and functional expression in a heterologous system allowed identification of various cellulose synthase-like (Csl) sequences as being involved in the formation of β-1,4-mannan, β-1,4-glucan, and mixed-linked glucan. A number of xylose-deficient mutants have led to a variety of genes, none of which thus far possesses the motifs known to be conserved among polymerizing β-glycan synthases. Except for xylan synthase, which appears to be an agglomerate of proteins just like cellulose synthase, Golgi glycan synthases already identified suggest that the catalytic polypeptide by itself is sufficient for enzyme activity, most likely as a homodimer. Several of the Csl genes remain to be assigned a function. The possibility of the involvement of various Csl genes in making more than one product remains.

Mannan synthase activity in the CSLD family

Cellulose Synthase Like (CSL) proteins are a group of plant glycosyltransferases that are predicted to synthesize β-1,4-linked polysaccharide backbones. CSLC, CSLF and CSLH families have been confirmed to synthesize xyloglucan and mixed linkage β-glucan, while CSLA family proteins have been shown to synthesize mannans. The polysaccharide products of the five remaining CSL families have not been determined. Five CSLD genes have been identified in Arabidopsis thaliana and a role in cell wall biosynthesis has been demonstrated by reverse genetics. We have extended past research by producing a series of double and triple Arabidopsis mutants and gathered evidence that CSLD2, CSLD3 and CSLD5 are involved in mannan synthesis and that their products are necessary for the transition between early developmental stages in Arabidopsis. Moreover, our data revealed a complex interaction between the three glycosyltransferases and brought new evidence regarding the formation of non-cellulosic polysaccharides through multimeric complexes.

Flame retarded composite panels from sugar beet residues

Sugar beet residues and various additives were used to prepare panels by forming the semi-dry or wet mixtures and pressing the composites at a temperature of 150 °C and pressures of 100–5410 kPa for up to 105 min. The highest panel density and second highest thickness were observed when a combination of calcium hydroxide and boric acid were used as additives using the semi-dry procedure. SEM images revealed that at pressures over 1000 kPa the cell wall structure of sugar beet was completely unrecognizable. The FTIR results indicated that the non-cellulosic polysaccharides contributed significantly to the properties of the panels by acting as adhesives. The best flame retardant parameters were also obtained with the calcium hydroxide/boric acid formulation. In comparison to composites prepared from recycled paper and mixtures of sugar beet shreds with recycled paper, higher density panels were prepared at lower pressure from sugar beet sources.

Characterization of kenaf phloem fibers in relation to stem growth

Microscopic study on phloem fibers in kenaf stem was carried out to examine the developmental process of phloem fiber and localization of noncellulosic polysaccharides in the cell walls. Histochemical observation showed that layered phloem fiber bundles developed external to the vascular cambium as stems grew after planting, and the number of layered bundles was higher in the bottom region of the stem regardless of growth stage. Based on immunocytochemical observations, cells making up the phloem fiber bundles had thick secondary cell walls containing mixed-linkage, (1→3, 1→4)-β-d-glucan as a component of hemicellulose. Other polysaccharides present in cell walls, namely xyloglucan, rhamnogalacturonan, and methyl-esterified homogalacturonan, were localized only, in the primary cell walls and/or in the junction between contiguous cells. In addition, mechanical evaluation on isolated kenaf fibers showed that the fiber strength varied by stem maturity and region: both higher tensile strength and initial Young's modulus were recorded in fibers obtained from the middle region of stems harvested 4–6 months after planting.

Cell Wall Modifications in Maize Pulvini in Response to Gravitational Stress

Changes in cell wall polysaccharides, transcript abundance, metabolite profiles, and hormone concentrations were monitored in the upper and lower regions of maize (Zea mays) pulvini in response to gravistimulation, during which maize plants placed in a horizontal position returned to the vertical orientation. Heteroxylan levels increased in the lower regions of the pulvini, together with lignin, but xyloglucans and heteromannan contents decreased. The degree of substitution of heteroxylan with arabinofuranosyl residues decreased in the lower pulvini, which exhibited increased mechanical strength as the plants returned to the vertical position. Few or no changes in noncellulosic wall polysaccharides could be detected on the upper side of the pulvinus, and crystalline cellulose content remained essentially constant in both the upper and lower pulvinus. Microarray analyses showed that spatial and temporal changes in transcript profiles were consistent with the changes in wall composition that were observed in the lower regions of the pulvinus. In addition, the microarray analyses indicated that metabolic pathways leading to the biosynthesis of phytohormones were differentially activated in the upper and lower regions of the pulvinus in response to gravistimulation. Metabolite profiles and measured hormone concentrations were consistent with the microarray data, insofar as auxin, physiologically active gibberellic acid, and metabolites potentially involved in lignin biosynthesis increased in the elongating cells of the lower pulvinus.

Comparison of Cell Wall Polysaccharide Hydrolysis by a Dilute Acid/Enzymatic Saccharification Process and Rumen Microorganisms

Evaluation of biomass crops for breeding or pricing purposes requires an assay that predicts performance in the bioenergy conversion process. Cell wall polysaccharide hydrolysis was compared for a dilute sulfuric acid pretreatment at 121°C followed with cellulase hydrolysis for 72 h conversion assay (CONV) with in vitro rumen microflora incubation for 72 h (RUMEN) for a set of maize (Zea mays L.) stover samples with a wide range in cell wall composition. Residual polysaccharides from the assays were analyzed for sugar components and extent of hydrolysis calculated. Cell wall polysaccharide hydrolysis was different for all sugar components between the CONV and RUMEN assays. The CONV assay hydrolyzed xylose-, arabinose-, galactose-, and uronic acid-containing polysaccharides to a greater degree than did the RUMEN assay, whereas the RUMEN assay was more effective at hydrolyzing glucose- and mannose-containing polysaccharides. Greater hydrolysis of hemicelluloses and pectins by CONV can be attributed to the acid hydrolysis mechanism of the CONV assay for noncellulosic polysaccharides, whereas the RUMEN assay was dependent on enzymatic hydrolysis. While CONV and RUMEN hydrolysis were correlated for most polysaccharide components, the greatest correlation was only r = 0.70 for glucose-containing polysaccharides. Linear correlations and multiple regressions indicated that polysaccharide hydrolysis by the RUMEN assay was negatively associated with lignin concentration and ferulate ether cross linking as expected. Corresponding correlations and regressions for CONV were less consistent and occasionally positive. Use of rumen microbial hydrolysis to characterize biomass performance in a conversion process may have some limited usefulness for genetic evaluations, but such assays would be unreliable for biomass pricing.

Antioxidant Activities of Soluble Dietary Fiber Extracted from Brewers’ Spent Grain

Brewers’ spent grain (BSG) is the most common by-product of the brewing industry, contains about 16.8% cellulose, 28.4% non-cellulosic polysaccharides and 27.8% lignin, which is a good raw material for industrial production of soluble dietary fiber (SDF).The antioxidant activity of crude dietary fiber (CDF) and grade dietary fiber (GDF) isolated from BSG was investigated in vitro. The scavenging hydroxyl radical capability of CDF and GDF was respectively reached to15.4% and 69.2% at the concentration of 0.1mg /mL. DF concentration from 0.02mg /mL to 0.1m g /mL was used. The scavenging capability of CDF and GDF to superoxide anion radical was increased from 10.68% and 12.33% to 35.78% and 78.39%. Both of the two DF showed some effect in scavenging superoxide radical and 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) radical with the concentration increase. The result showed that BSG DF can be used as a good additive with some functionality.