Spatial relationships between polymers in Sitka spruce: proton spin-diffusion studies

The spatial arrangement of polymers in Sitka spruce (Picea sitchensis) was investigated by NMR proton spin-diffusion studies, supplemented by deuterium-exchange experiments monitored by FTIR spectroscopy. The FTIR spectra of earlywood sections after vapour-phase exchange with deuterium oxide showed that 43% of the hydroxyl groups were accessible to deuteration. This value is lower than predicted in the absence of aggregation of cellulose microfibrils into larger units, but greater than the predicted level of deuteration if 3.5-nm microfibrils surrounded by hemicellulose sheaths were aggregated into 4×4 arrays without space for deuterium oxide to penetrate between the microfibrils. The rate of proton spin diffusion between lignin and cellulose was consistent with the presence of microfibril arrays with approximately these dimensions and with lignin located outside them, in both earlywood and latewood. Proton spin-diffusion data for hemicelluloses were complicated by difficulties in assigning signals to glucomannans and xylans, but there was evidence for the spatial association of one group of hemicelluloses, including acetylated glucomannans, with cellulose surfaces, while another group of hemicelluloses was in spatial proximity to lignin. These data are consistent with a number of nanoscale models for the Sitka spruce cell wall, including a model in which glucomannans are associated with microfibril surfaces within the aggregate and water can penetrate partially between these surfaces, and one in which all non-cellulosic polymers and water are excluded from the interior of each microfibril aggregate

Structural studies of a fucogalactoxyloglucan from pinus radiata primary cell walls : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University

1. The changes in carbohydrate composition of elongating Pinus radiata primary cell walls were investigated. In the hemicellulose B extracts, a large increase in the percentage of non-starch, non-cellulosic, glucose was found to occur on cessation of cell-wall elongation. 2. By fractionation of the hemicellulose B extracts, with a variety of methods involving precipitation from an aqueous solution, a xyloglucan was purified. This xyloglucan was the major hemicellulose of the Pinus radiata hypocotyl cell wall. 3. Characterisation studies on the xyloglucan involved: quantitative analysis of the monosaccharides derived by nitric acid/urea hydrolysis; identification of the partial hydrolysis products derived by trifluoroacetic acid hydrolysis; quantitation of the sugar linkages using methylation by the Hakomori method; and analysis of the anomeric configuration of component sugars using chromium trioxide oxidation. 4. From the results a tentative structure has been suggested for the xyloglucan, consisting of a backbone of B-D-gluco-pyranose residues linked together by 1-4 glycosidic bonds, and with sidechains of single xylose residues linked through C-6 of the glucose units. Galacto and fuco-1,2- galacto sidechains are attached to some of the xylose residues, probably through the C-2 of the xylose

Deep Eutectic Solvent aqueous solutions as efficient media for the solubilization of hardwood xylans

This work contributes to the development of integrated lignocellulosic-based biorefineries by the pioneering exploitation of hardwood xylans by solubilization and extraction in deep eutectic solvents (DES). DES formed by choline chloride and urea or acetic acid were initially evaluated as solvents for commercial xylan as a model compound. The effects of temperature, molar ratio, and concentration of the DES aqueous solutions were evaluated and optimized by using a response surface methodology. The results obtained demonstrated the potential of these solvents, with 328.23 g L-1 of xylan solubilization using 66.7 wt % DES in water at 80 °C. Furthermore, xylans could be recovered by precipitation from the DES aqueous media in yields above 90 %. The detailed characterization of the xylans recovered after solubilization in aqueous DES demonstrated that 4-O-methyl groups were eliminated from the 4-O-methylglucuronic acids moieties and uronic acids (15 %) were cleaved from the xylan backbone during this process. The similar Mw values of both pristine and recovered xylans confirmed the success of the reported procedure. DES recovery in four additional extraction cycles was also demonstrated. Finally, the successful extraction of xylans from Eucalyptus globulus wood by using aqueous solutions of DES was demonstrated.publishe

Active Site Mapping of Xylan-Deconstructing Enzymes with Arabinoxylan Oligosaccharides Produced by Automated Glycan Assembly

Xylan-degrading enzymes are crucial for the deconstruction of hemicellulosic biomass, making the hydrolysis products available for various industrial applications such as the production of biofuel. To determine the substrate specificities of these enzymes, we prepared a collection of complex xylan oligosaccharides by automated glycan assembly. Seven differentially protected building blocks provided the basis for the modular assembly of 2-substituted, 3-substituted, and 2-/3-substituted arabino- and glucuronoxylan oligosaccharides. Elongation of the xylan backbone relied on iterative additions of C4-fluorenylmethoxylcarbonyl (Fmoc) protected xylose building blocks to a linker-functionalized resin. Arabinofuranose and glucuronic acid residues have been selectively attached to the backbone using fully orthogonal 2-(methyl)naphthyl (Nap) and 2-(azidomethyl)benzoyl (Azmb) protecting groups at the C2 and C3 hydroxyls of the xylose building blocks. The arabinoxylan oligosaccharides are excellent tools to map the active site of glycosyl hydrolases involved in xylan deconstruction. The substrate specificities of several xylanases and arabinofuranosidases were determined by analyzing the digestion products after incubation of the oligosaccharides with glycosyl hydrolases.Fil: Senf, Deborah. Max Planck Institut für Kolloid und Grenzflächenforschung; Alemania. Freie Universität; AlemaniaFil: Ruprecht, Colin. Max Planck Institut für Kolloid und Grenzflächenforschung; AlemaniaFil: de Kruijff, Goswinus H. M.. Max Planck Institut für Kolloid und Grenzflächenforschung; Alemania. Freie Universität; Alemania. University Mainz. Institute of Institute of Organic Chemistry, Johannes Gutenberg; AlemaniaFil: Simonetti, Sebastián Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Max Planck Institut für Kolloid und Grenzflächenforschung; AlemaniaFil: Schuhmacher, Frank. Max Planck Institut für Kolloid und Grenzflächenforschung; Alemania. Freie Universität; AlemaniaFil: Seeberger, Peter H.. Max Planck Institut für Kolloid und Grenzflächenforschung; Alemania. Freie Universität; AlemaniaFil: Pfrengle, Fabian. Max Planck Institut für Kolloid und Grenzflächenforschung; Alemania. Freie Universität; Alemani

Morphology and swelling of thin films of dialcohol xylan

Polysaccharides are excellent network formers and are often processed into films from water solutions. Despite being hydrophilic polysaccharides, the typical xylans liberated from wood are sparsely soluble in water. We have previously suggested that an additional piece to the solubilization puzzle is modification of the xylan backbone via oxidative cleavage of the saccharide ring. Here, we demonstrate the influence of the degree of modification, i.e., degree of oxidation (DO) on xylan solubilization and consequent film formation and stability. Oxidized and reduced wood xylans (i.e., dialcohol xylans) with the highest DO (77 %) within the series exhibited the smallest hydrodynamic diameter (dh) of 60 nm in dimethylsulfoxide (DMSO). We transferred the modified xylans into films credit to their established solubility and then quantified the film water interactions. Dialcohol xylans with intermediate DOs (42 and 63 %) did not form continuous films. The films swelled slightly when subjected to humidity. However, the film with the highest DO demonstrated a significant moisture uptake that depended on the film mass and was not observed with the other modified grades or with unmodified xylan

Extraction and structural study of hemicellulose B from tall fescue (Festuca arundinacea) : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Chemistry at Massey University

1.1 Hemicellulose The structural carbohydrates are mixtures of polysaccharides which together with lignin, constitute the cell wall. Usually they are divided into three fractions: pectic substances, hemicellulose and cellulose.1 Pectins, widely distributed in land plants are characterised by a main chain of 1 – 4 linked galacturonic acid units. They may also contain rhamnose, galactose, xylose, arabinose and fucose in varying amounts. These sugars form part the main chain in some pectins and branch chains in others.2 The pectic triad consists of galacturonan, araban and galactan. Pectic substances occur without exception in all higher plants. It is possible that pectic substances may serve as protective agents for natural rubber particles and may be important food reserve for the plant.3 They are found most abundantly in the primary cell walls and in the intercellular layer.4 [From Introduction

Cellulose-hemicellulose networks as target for in planta modification of the properties of natural fibres

Plant cell wall polysaccharides are predominant components of fibres. Natural fibres have a wide range of industrial applications, such as in paper and textile industries. Furthermore, their demand for use as bio-composites in building and automotive applications is also increasing. For the various applications, a gain of control over fibre characteristics is important. Inherent fibre characteristics are largely determined by the ratio and interactions of cellulose and hemicelluloses. Two main strategies for bioengineering fibre properties are reviewed: (i) modifying the cellulose/hemicellulose ratio (by biosynthesis or biodegradation of specific polysaccharides), and (ii) interference with cellulosehemicellulose interactions using carbohydrate-binding modules. These in planta approaches may have the potential of complementing the currently used surface modification approaches for modifying fibre characteristics

Fungi isolated from Miscanthus and sugarcane: biomass conversion, fungal enzymes, and hydrolysis of plant cell wall polymers.

BackgroundBiofuel use is one of many means of addressing global change caused by anthropogenic release of fossil fuel carbon dioxide into Earth's atmosphere. To make a meaningful reduction in fossil fuel use, bioethanol must be produced from the entire plant rather than only its starch or sugars. Enzymes produced by fungi constitute a significant percentage of the cost of bioethanol production from non-starch (i.e., lignocellulosic) components of energy crops and agricultural residues. We, and others, have reasoned that fungi that naturally deconstruct plant walls may provide the best enzymes for bioconversion of energy crops.ResultsPreviously, we have reported on the isolation of 106 fungi from decaying leaves of Miscanthus and sugarcane (Appl Environ Microbiol 77:5490-504, 2011). Here, we thoroughly analyze 30 of these fungi including those most often found on decaying leaves and stems of these plants, as well as four fungi chosen because they are well-studied for their plant cell wall deconstructing enzymes, for wood decay, or for genetic regulation of plant cell wall deconstruction. We extend our analysis to assess not only their ability over an 8-week period to bioconvert Miscanthus cell walls but also their ability to secrete total protein, to secrete enzymes with the activities of xylanases, exocellulases, endocellulases, and beta-glucosidases, and to remove specific parts of Miscanthus cell walls, that is, glucan, xylan, arabinan, and lignin.ConclusionThis study of fungi that bioconvert energy crops is significant because 30 fungi were studied, because the fungi were isolated from decaying energy grasses, because enzyme activity and removal of plant cell wall components were recorded in addition to biomass conversion, and because the study period was 2 months. Each of these factors make our study the most thorough to date, and we discovered fungi that are significantly superior on all counts to the most widely used, industrial bioconversion fungus, Trichoderma reesei. Many of the best fungi that we found are in taxonomic groups that have not been exploited for industrial bioconversion and the cultures are available from the Centraalbureau voor Schimmelcultures in Utrecht, Netherlands, for all to use

Xylanase production by Aspergillus niger ANL 301 using agro - wastes

Xylanase production by wild-type Aspergillus niger ANL301, newly isolated from wood-waste, was monitored at 24 h intervals for a period 168 h in media containing different carbon sources. The carbon sources were oat-spelt xylan (Fluka) and three agro-wastes (sawdust, sugarcane pulp and wheat bran). Highest xylanase activity of 6.47 units/mL was obtained at 96 h in media containing wheat bran as sole carbon source. Maximum activity value for the media containing sugarcane pulp was 0.95 units/mL obtained also at 96 h. Sawdust and oat spelt xylan gave the peak enzyme activities of 0.65 and 0.80 units/mL respectively at 120 h. High protein yield was obtained in media containing the agro-wastes, with wheat bran giving the highest value of 1.14 mg/mL at 96 h. The maximum specific xylanase activities were 3.86, 3.37, 5.69, and 9.36 units/ mg protein for sawdust, sugarcane pulp, wheat bran and oat spelt xylan, respectively. Out of the three agro-wastes used in this study, wheat bran holds greatest promise for low cost production of the xylanase enzyme

Xylanase production by Penicillium chrysogenum (PCL501) fermented on cellulosic wastes

Xylanase production by Penicillium chrysogenum PCL501, newly isolated from wood-wastes, was monitored at 24 h intervals for a period 168 h in media containing four different carbon sources (oatspelt xylan, wheat bran, sawdust, and sugarcane pulp). The highest xylanase activity of 6.47 Units mL-1 was obtained at 96 h in media containing wheat bran whereas media containing sugarcane pulp gave a peak value of 1.39 Units mL-1 at 144 h. Sawdust and xylan gave a peak xylanase activity of 1.35 and 0.79 Units mL-1 respectively at 120 h. Maximum protein released in xylan-containing media was 0.38 mg mL-1. Higher protein yield was obtained in media containing the agro-wastes, with wheat bran giving the highest value of 1.14 mg mL-1. The maximum specific xylanase activities were 2.59, 8.52, 16.06, and 9.36 Units mg Protein -1 for sawdust, sugarcane pulp, wheat bran and xylan respectively. Out of the three agro-wastes used in this study, wheat bran holds the greatest promise for cost-effective production of the xylanase enzyme. The carbon source is the highest inducer of the enzyme in the fungus