Dissolution behavior of different celluloses

Celluloses from different origins were dissolved stepwise in N,N-dimethylacetamide/lithium chloride (9% v/w; DMAc/LiCl) with the aim to study the time course of the dissolution process, completeness of dissolution in the dissolved fractions, possible discrimination effects, and differences between the celluloses. Cellulosic pulps from both annual plants and different wood species were analyzed. The obtained fractions were subject to gel permeation chromatography (GPC) with multiple detection to monitor the development of molecular mass distribution (MMD), molecular mass, and recovered mass. The dissolution behavior of accompanying xylans was followed by quantitative analysis of the uronic acids by fluorescence labeling--GPC. The morphological changes at the remaining fibers in the stepwise dissolution were addressed by SEM. The time needed to dissolve completely the cellulosic pulp differed from species to species, mainly between pulps from annual plants and pulps from wood. Annual plants generally needed much longer to dissolve completely. In the beginning of the dissolution, the dissolved fractions of annual plants showed a distinct discrimination effect because they were enriched in hemicellulose. By contrast, wood pulps dissolve fast and without distinct changes in the MMD of the dissolved fractions over time. Bagasse pulp is an exception to the observation for annual plants and rather resembled the behavior of wood celluloses. Prolonged dissolution times, as often practiced in cellulose GPC, do not lead to any improvements regarding the determination of molecular mass, MMD, and recovered mass of injected sample, so that the dissolution times required for reliable GPC analysis can be significantly shortened, which will be important for biorefinery analytics with high numbers of samples.     

Enzymatic hydrolysis of biomass from wood

Current research and development in cellulosic ethanol production has been focused mainly on agricultural residues and dedicated energy crops such as corn stover and switchgrass; however, woody biomass remains a very important feedstock for ethanol production. The precise composition of hemicellulose in the wood is strongly dependent on the plant species, therefore different types of enzymes are needed based on hemicellulose complexity and type of pretreatment. In general, hardwood species have much lower recalcitrance to enzymes than softwood. For hardwood, xylanases, beta‐xylosidases and xyloglucanases are the main hemicellulases involved in degradation of the hemicellulose backbone, while for softwood the effect of mannanases and beta‐mannosidases is more relevant. Furthermore, there are different key accessory enzymes involved in removing the hemicellulosic fraction and increasing accessibility of cellulases to the cellulose fibres improving the hydrolysis process. A diversity of enzymatic cocktails has been tested using from low to high densities of biomass (2–20% total solids) and a broad range of results has been obtained. The performance of recently developed commercial cocktails on hardwoods and softwoods will enable a further step for the commercialization of fuel ethanol from wood.     

Direct Compression of Cellulose and Lignin Isolated by a New Catalytic Treatment

Tablet compression of softwood cellulose and lignin prepared by a new catalytic oxidation and acid precipitation method were investigated and compared with the established pharmaceutical direct compression excipients. Catalytic pretreated softwood cellulose (CPSC) and lignin (CPSL) were isolated from pine wood (Pinus sylvestris). The compaction studies were carried out with an instrumented eccentric tablet machine. The plasticity and elasticity of the materials under compression were evaluated using force-displacement treatment and by determining characteristic plasticity (PF) and elasticity (EF) factors. With all biomaterials studied, the PF under compression decreased exponentially as the compression force increased. The compression force applied in tablet compression did not significantly affect the elasticity of CPSC and microcrystalline cellulose (MCC) while the EF values for softwood lignins increased as compression force increased. CPSL was clearly a less plastically deforming and less compactable material than the two celluloses (CPSC and MCC) and hardwood lignin. CPSL presented deformation and compaction behaviour almost identical to that of lactose monohydrate. In conclusion, the direct tablet compression behaviour of native lignins and celluloses can greatly differ from each other depending on the source and isolation method used.     

Biological Pretreatment of Lignocellulosic Substrates for Enhanced Delignification and Enzymatic Digestibility

Sheer enormity of lignocellulosics makes them potential feedstock for biofuel production but, their conversion into fermentable sugars is a major hurdle. They have to be pretreated physically, chemically, or biologically to be used by fermenting organisms for production of ethanol. Each lignocellulosic substrate is a complex mix of cellulose, hemicellulose and lignin, bound in a matrix. While cellulose and hemicellulose yield fermentable sugars, lignin is the most recalcitrant polymer, consisting of phenyl-propanoid units. Many microorganisms in nature are able to attack and degrade lignin, thus making access to cellulose easy. Such organisms are abundantly found in forest leaf litter/composts and especially include the wood rotting fungi, actinomycetes and bacteria. These microorganisms possess enzyme systems to attack, depolymerize and degrade the polymers in lignocellulosic substrates. Current pretreatment research is targeted towards developing processes which are mild, economical and environment friendly facilitating subsequent saccharification of cellulose and its fermentation to ethanol. Besides being the critical step, pretreatment is also cost intensive. Biological treatments with white rot fungi and Streptomyces have been studied for delignification of pulp, increasing digestibility of lignocellulosics for animal feed and for bioremediation of paper mill effluents. Such lignocellulolytic organisms can prove extremely useful in production of bioethanol when used for removal of lignin from lignocellulosic substrate and also for cellulase production. Our studies on treatment of hardwood and softwood residues with Streptomyces griseus isolated from leaf litter showed that it enhanced the mild alkaline solubilisation of lignins and also produced high levels of the cellulase complex when growing on wood substrates. Lignin loss (Klason lignin) observed was 10.5 and 23.5% in case of soft wood and hard wood, respectively. Thus, biological pretreatment process for lignocellulosic substrate using lignolytic organisms such as actinomycetes and white rot fungi can be developed for facilitating efficient enzymatic digestibility of cellulose.     

Solid-state fermentation of wood residues by Streptomyces griseus B1, a soil isolate,and solubilization of lignins

The actinomycete strain Streptomyces griseus B₁ isolated from soil, when grown on cellulose powder as submerged culture produced high levels of all the three components i.e. filter paper lyase (FPase), CMCellulase and β-glucosidase of the cellulolytic enzyme system. FP activity and CMCellulase were present only extracellularly, while β-glucosidase was both intra- and extra-cellular. It produced highest FPase activity when grown on hardwood powder under submerged culture. It was unable to use lignin monomers (ferulic acid, vanillic acid and syringic acid) as carbon source. While growing on hardwood and softwood powders under solid-state conditions, it depleted them of cellulose (36.3 in the case of softwood and 14.4 in the case of hardwood). It also caused partial loss of lignin content in both the substrates by solubilizing them. These solubilized lignins could be recovered as acid precipitable polymeric lignins (APPL) from extracts of wood powders upon acidification. Extracts of inoculated wood powders yielded higher amounts of APPL than uninoculated controls. Also, the APPLs from Streptomyces-treated wood powders differed from control APPLs in their molecular weight distribution, as observed from their elution pattern using Sephadex G-100.     

Vibrational spectroscopy and X-ray diffraction methods to establish the differences between hardwood and softwood

FT-IR spectrometry and X-ray diffraction were applied to probe the differences between pulp fibers from Eucalyptus wood (hardwood) and Norway spruce wood (softwood). Wood processing was found to induce certain structural alterations within its components depending on the type of wood and the applied procedure. These differences were established by using techniques such as; spectral comparison of wood samples with those of individual component fractions, derivative spectroscopy, bands deconvolution, etc. FT-IR spectroscopy was shown to be an important tool that provided details about the structural characteristics of hardwood and softwood samples. Using second-derivative spectra and deconvolution processes small differences between spectra became apparent that allowed correlations to be made related to wood composition. In addition a correlation was established between the integral absorptions for the various bands and lignin content as well as the lignin/carbohydrate content. Relations between various spectral characteristics and the degree of crystallinity and sample composition were established.     

Predicting the most appropriate wood biomass for selected industrial applications: comparison of wood,pulping, and enzymatic treatments using fluorescent-tagged carbohydrate-binding modules

Background

Lignocellulosic biomass will progressively become the main source of carbon for a number of products as the Earth’s oil reservoirs disappear. Technology for conversion of wood fiber into bioproducts (wood biorefining) continues to flourish, and access to reliable methods for monitoring modification of such fibers is becoming an important issue. Recently, we developed a simple, rapid approach for detecting four different types of polymer on the surface of wood fibers. Named fluorescent-tagged carbohydrate-binding module (FTCM), this method is based on the fluorescence signal from carbohydrate-binding modules-based probes designed to recognize specific polymers such as crystalline cellulose, amorphous cellulose, xylan, and mannan.

Results

Here we used FTCM to characterize pulps made from softwood and hardwood that were prepared using Kraft or chemical-thermo-mechanical pulping. Comparison of chemical analysis (NREL protocol) and FTCM revealed that FTCM results were consistent with chemical analysis of the hemicellulose composition of both hardwood and softwood samples. Kraft pulping increased the difference between softwood and hardwood surface mannans, and increased xylan exposure. This suggests that Kraft pulping leads to exposure of xylan after removal of both lignin and mannan. Impact of enzyme cocktails from Trichoderma reesei (Celluclast 1.5L) and from Aspergillus sp. (Carezyme 1000L) was investigated by analysis of hydrolyzed sugars and by FTCM. Both enzymes preparations released cellobiose and glucose from pulps, with the cocktail from Trichoderma being the most efficient. Enzymatic treatments were not as effective at converting chemical-thermomechanical pulps to simple sugars, regardless of wood type. FTCM revealed that amorphous cellulose was the primary target of either enzyme preparation, which resulted in a higher proportion of crystalline cellulose on the surface after enzymatic treatment. FTCM confirmed that enzymes from Aspergillus had little impact on exposed hemicelluloses, but that enzymes from the more aggressive Trichoderma cocktail reduced hemicelluloses at the surface.

Conclusions

Overall, this study indicates that treatment with enzymes from Trichoderma is appropriate for generating crystalline cellulose at fiber surface. Applications such as nanocellulose or composites requiring chemical resistance would benefit from this enzymatic treatment. The milder enzyme mixture from Aspergillus allowed for removal of amorphous cellulose while preserving hemicelluloses at fiber surface, which makes this treatment appropriate for new paper products where surface chemical responsiveness is required.

     

Effect of solvent exchange on the supramolecular structure, the molecular mobility and the dissolution behavior of cellulose in LiCl/DMAc

We investigated the effect of solvent exchange on the supramolecular structure and the molecular mobility of the cellulose molecule to clarify the mechanism of the dissolution of cellulose in lithium chloride/N,N-dimethylacetamide (LiCl/DMAc). Among the celluloses that were solvent exchanged in different ways, the DMAc-treated celluloses dissolved most rapidly. Dissolution of the acetone-treated celluloses was much slower than the DMAc-treated ones, but considerably faster than the untreated one. Such differences in the dissolution behavior were well explained by the differences in the surface fractal dimension calculated from the small-angle X-ray scattering profiles and in the (1)H spin-lattice and spin-spin relaxation times estimated from the solid-state NMR spectroscopic measurements. Furthermore, it was suggested from the IR spectra and the (13)C spin-lattice relaxation times of cellulose that DMAc is adsorbed on the surface of cellulose even after vacuum-drying and affects the molecular mobility and hydrogen-bonding state of cellulose.     

The Spatial Distribution of Fungi on Decomposing Woody Litter in a Freshwater Stream,Western Ghats,India

We mapped filamentous fungal association with mechanically “hard” and “soft” woody litter naturally deposited in a stream of the Western Ghats of India. Using a durometer (rubber hardness tester), the toughness of surface of wood collected from stream was determined by considering durometer reading from 60–72 to 30–37 as hardwood and softwood, respectively. From each wood (1.5 cm diameter), two segments each of 3 cm length were excised and vertically cut into nine sections comprising eight marginal and one central section. From three stream locations, hardwood and softwood sections were assessed for the occurrence of lignicolous and Ingoldian fungi. A first set of wood sections was incubated in damp chambers up to 4 months with periodical screening (every 2 weeks) for lignicolous fungi. Another set was incubated in bubble chambers up to 72 h to ascertain colonization of Ingoldian fungi. In hardwood sections, 17 lignicolous fungi (ascomycetes, four; mitosporic fungi, 13; mean, 6.8; range, 6–8/section) and ten Ingoldian fungi (mean, 2; range, 0–4/section) comprising nine lignicolous (11.1–40.7%) and three Ingoldian (11.1–14.8%) fungi as core-group taxa were recovered. In softwood, ten lignicolous fungi (ascomycetes, 0; mitosporic fungi, ten; mean, 3.8; range, 2–5/section) and 26 Ingoldian fungi (mean, 8.1; range, 5–10/section) comprising six lignicolous (11.1–85.2%) and 12 Ingoldian (11.1–88.9%) fungi as core-group taxa were recovered. The ratio of lignicolous fungi/Ingoldian fungi was higher in hardwood than softwood (1.7 vs. 0.4). The spore output of Ingoldian fungi was higher in softwood (mean, 901 g⁻¹; range, 80–2546 g⁻¹) than hardwood (mean, 21 g⁻¹; range, 0–140 g⁻¹). The Shannon diversity of lignicolous fungi was higher in hardwood than softwood (3.604 vs. 2.665), whereas it was opposite for Ingoldian fungi (3.116 vs. 3.918). The overall fungal diversity was higher in softwood than hardwood (4.413 vs. 4.219). The range of Jaccard’s index of similarity among wood sections was higher in lignicolous fungi (8–71% and 13–75%) than Ingoldian fungi (0–50% and 8–55%) in hardwood and softwood. The rarefaction indices of expected number of taxa against hardwood sections revealed higher and persistent lignicolous fungi than the Ingoldian fungi, while the Ingoldian fungi were persistent in softwood sections, although they were lower than lignicolous fungi. Our study demonstrated the dominance of lignicolous fungi and Ingoldian fungi in hardwood and softwood, respectively.     

Rheological properties and molecular structure of tunicate cellulose in LiCl/1,3-dimethyl-2-imidazolidinone

Solution properties and molecular structure of tunicate cellulose (TC), an animal cellulose from Halocynthia roretzi, were investigated in terms of rheological and dilute solution properties. The solvent used is 8 wt % LiCl/1,3-dimethyl-2-imidazolidinone (DMI). A solution of dissolving pulp (DP), derived from plant, was also used for comparison. The weight-average molecular weight, Mw, and the limiting viscosity number, [eta], of the TC were evaluated to be 413 x 10(6) and 2645 mL/g, respectively. The TC solution showed the same concentration dependence of GN (GN=5.49 x 10(6)phiw(2.1)4 Pa; phiw: weight fraction of cellulose in solution; GN: plateau modulus) as the DP solution and, moreover, also as the solution of cotton linter (CC) in 8 wt % LiCl/N,N-dimethylacetamide (DMAc). This exponent of 2.1(4) indicates that network structure by entanglements was formed in these solutions. According to the theory of Fetters et al., moreover, such identity means that all of these celluloses have the identical chain structure though their biological origins are far different. On the other hand, the phiw-dependence of eta0-etas (eta0=zero shear rate viscosity of solution; etas=solvent viscosity) was different between the TC and the DP solution in the semidilute regime: the TC solution exhibited eta0-etas proportional, variant phiw(7.5) and the DP solution eta0-etas proportional, variant phiw4. According to the theory of Doi-Edwards, this exponent of 4 (the DP solution) indicates that the DP behaves as flexible polymers in the solution. In contrast, the dependence for the TC solution seems unexplainable on the basis of molecular theories. This difference probably signifies the difference in the relaxation process or mechanism in entanglement systems.     

Variation of cellulose microfibril angles in softwoods and hardwoods-a possible strategy of mechanical optimization

Position-resolved small-angle X-ray scattering was used to investigate the nanostructure of the wood cell wall in two softwood species (Norwegian spruce and Scots pine) and two hardwood species (pedunculate oak and copper beech). The tilt angle of the cellulose fibrils in the wood cell wall versus the longitudinal cell axis (microfibril angle) was systematically studied over a wide range of annual rings in each tree. The measured angles were correlated with the distance from the pith and the results were compared. The microfibril angle was found to decrease from pith to bark in all four trees, but was generally higher in the softwood than in the hardwood. In Norwegian spruce, the microfibril angles were higher in late wood than in early wood; in Scots pine the opposite was observed. In pedunculate oak and copper beech, low angles were found in the major part of the stem, except for the very first annual rings in pedunculate oak. The results are interpreted in terms of mechanical optimization. An attempt was made to give a quantitative estimation for the mechanical constraints imposed on a tree of given dimensions and to establish a model that could explain the general decrease of microfibril angles from pith to bark.     

Cellulose nanofibers prepared by TEMPO-mediated oxidation of native cellulose

Never-dried and once-dried hardwood celluloses were oxidized by a 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated system, and highly crystalline and individualized cellulose nanofibers, dispersed in water, were prepared by mechanical treatment of the oxidized cellulose/water slurries. When carboxylate contents formed from the primary hydroxyl groups of the celluloses reached approximately 1.5 mmol/g, the oxidized cellulose/water slurries were mostly converted to transparent and highly viscous dispersions by mechanical treatment. Transmission electron microscopic observation showed that the dispersions consisted of individualized cellulose nanofibers 3-4 nm in width and a few microns in length. No intrinsic differences between never-dried and once-dried celluloses were found for preparing the dispersion, as long as carboxylate contents in the TEMPO-oxidized celluloses reached approximately 1.5 mmol/g. Changes in viscosity of the dispersions during the mechanical treatment corresponded with those in the dispersed states of the cellulose nanofibers in water.     

A review of the production of ethanol from softwood

Ethanol produced from various lignocellulosic materials such as wood, agricultural and forest residues has the potential to be a valuable substitute for, or complement to, gasoline. One of the major resources in the Northern hemisphere is softwood. This paper reviews the current status of the technology for ethanol production from softwood, with focus on hemicellulose and cellulose hydrolysis, which is the major problem in the overall process. Other issues of importance, e.g. overall process configurations and process economics are also considered.     

Cellulose acetates from linters and sisal: correlation between synthesis conditions in DMAc/LiCl and product properties

We report the acetylation of celluloses from sisal (untreated and alkali treated) and cotton linters (alkali treated), under homogeneous solution conditions, using DMAc/LiCl as solvent system. Our target was to evaluate the effects of cellulose dissolution and reactions conditions on the product properties. The products were characterized in terms of degree of substitution (DS) by 1H NMR, and molar weight distribution (MWD) by size exclusion chromatography. Changes in the DS of the products were correlated with reaction conditions and solution properties. It was found that the dissolution of celluloses and degree of substitution of cellulose derivatives depends on a fine adjustment of the dissolution/derivatization conditions, as well as on the origin (sisal or linters) of celluloses.     

Universal fractionation of lignin–carbohydrate complexes (LCCs) from lignocellulosic biomass: an example using spruce wood

It is of both theoretical and practical importance to develop a universally applicable approach for the fractionation and sensitive lignin characterization of lignin–carbohydrate complexes (LCCs) from all types of lignocellulosic biomass, both natively and after various types of processing. In the present study, a previously reported fractionation approach that is applicable for eucalyptus (hardwood) and flax (non‐wood) was further improved by introducing an additional step of barium hydroxide precipitation to isolate the mannan‐enriched LCC (glucomannan‐lignin, GML), in order to suit softwood species as well. Spruce wood was used as the softwood sample. As indicated by the recovery yield and composition analysis, all of the lignin was recovered in three LCC fractions: a glucan‐enriched fraction (glucan‐lignin, GL), a mannan‐enriched fraction (GML) and a xylan‐enriched fraction (xylan‐lignin, XL). All of the LCCs had high molecular masses and were insoluble or barely soluble in a dioxane/water solution. Carbohydrate and lignin signals were observed in ¹H NMR, ¹³C CP‐MAS NMR and normal‐ or high‐sensitivity 2D HSQC NMR analyses. The carbohydrate and lignin constituents in each LCC fraction are therefore believed to be chemically bonded rather than physically mixed with one another. The three LCC fractions were found to be distinctly different from each other in terms of their lignin structures, as revealed by highly sensitive analyses by thioacidolysis‐GC, thioacidolysis‐SEC and pyrolysis‐GC.     

Distribution of carboxylate groups introduced into cotton linters by the TEMPO-mediated oxidation

The 2,2,6,6-tetramethylpiperidine-1-oxy radial (TEMPO)-mediated oxidation was applied to aqueous slurries of cotton linters. The water-insoluble fibrous fractions thus obtained in the yields of more than 78% were characterized by solid-state ¹³C-NMR, X-ray diffraction and scanning electron microscopic analyses for evaluation of distribution of carboxylate groups formed in the TEMPO-oxidized celluloses. The patterns of solid-state ¹³C-NMR spectra revealed that the oxidation occurred at the C6 primary hydroxyl groups of cellulose. X-ray diffraction and scanning electron microscopic analyses showed that such C6 oxidation took place at the surfaces of cellulose I crystallites without any oxidation at the C6 of inside cellulose I crystallites. Thus, carboxylate and aldehyde groups introduced into the TEMPO-oxidized celluloses are densely present on the surfaces of cellulose I crystallites. In addition, the obtained results revealed that the shoulder signal due to non-crystalline C6 carbons at about 63 ppm in solid-state ¹³C-NMR spectra of native celluloses is ascribed to those of surfaces of cellulose I crystallites or those of cellulose microfibrils.     

Influence of the supramolecular structure and physicochemical properties of cellulose on its dissolution in a lithium chloride/N,N-dimethylacetamide solvent system

The present work deals with the effects of structural variables of celluloses on their dissolution in the solvent system LiCl/N,N-dimethylacetamide, LiCl/DMAc. Celluloses from fast growing sources (sisal and linters), as well as microcrystalline cellulose (avicel PH-101) were studied. The following structural variables were investigated: index of crystallinity, I(c); crystallite size; polymer porosity; and degree of polymerization determined by viscosity, DPv. Mercerization of fibrous celluloses was found to decrease DPv, I(c), the specific surface area, and the ratio pore volume/radius. The relevance of the structural properties of cellulose to its dissolution is discussed. Rate constants and activation parameters of cellulose decrystallization, prior to its solubilization, have been determined under nonisothermal conditions. The kinetic parameters calculated showed that dissolution is accompanied with small, negative enthalpy and a large, negative entropy of activation.     

Influence of the cycle number in processing of cellulose from waste paper on its ability to hydrolysis by cellulases

Hydrolytic ability of laboratory enzyme preparations from fungus of the Penicillium genus was investigated using kraft pulp from nonbleached softwood and bleached hardwood cellulose as substrates. The enzyme preparations were shown to efficiently hydrolyze both softwood and hardwood cellulose. The yields of glucose and reducing sugars were 24–36 g/l and 27–37 g/l from 100 g/l of dry substrate in 48 h, respectively, and depended on the number of substrate grinding cycles.     

Solute exclusion from cellulose in packed columns: experimental investigation and pore volume measurements

A packed column approach was used in this investigation to determine pore volume and surface area distributions of several celluloses. Specific surface areas for Avicel PH 102, Solka Floc BW 300, and two size fractions of corn cobs that have been pretreated to remove lignin and hemicellulose were measured using this technique. In addition to measuring pore volume and specific surface area, the molecular diameters of several PEGs (polyethylene glycol) were estimated using viscosity measurements. Also, the influence of cellulose particle size, molecular diameter of PEGs, and PEG solution velocity on dispersion and tailing were investigated. Molecular diameter estimates from this investigation were 30%-35% lower than those reported in the literature. This discrepancy is due to earlier investigators using an inappropriate relationship for estimating molecular diameter from viscosity measurements. The precision of the column approach to solute exclusion was higher than that obtained by investigators using a batch approach. Dispersion increased with increasing particle size. Tailing of the elution curve was increased with increasing solute molecular diameter and elution rate. For a cellulase with a molecular diameter of 5.1 nm, estimated specific surface area ranged from 7.2 to 10.5 m(2) g(-1).