Modification of high-lignin kraft pulps with laccase. Part 2. Xylanase-enhanced strength benefits

The effects of xylanase pretreatment of high lignin content softwood (SW) kraft pulp on subsequent pulp treatment with laccase in combination with gallic acid were investigated. Although xylanase pretreatment was ineffective in enhancing the laccase-facilitated biografting of gallic acid to kraft fibers, it was beneficial for subsequent treatment with laccase exclusively. Treating pulp fibers with xylanase followed by laccase provided a collective 25% and 46% increase in dry and wet tensile strength properties, respectively.     

Grafting of softwood kraft pulps fibers with fatty acids under cold plasma conditions

Cold plasma treatment is used to modify the cellulosic fibers for a variety of applications. The grafting of softwood unbleached (UBP) and bleached (BP) kraft pulp fibers has been performed under the action of cold plasma discharges, using different kinds of fatty acids. The grafted samples are characterized by FTIR spectroscopy, X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), termogravimetry (TG-DTG) and X-ray diffraction (XRD). All these methods confirm the morphological and structural changes after plasma treatment which determines the modification in cellulosic fiber properties. The active centers created within the cellulose chains by plasma treatment were used to initiate grafting reactions with fatty acids. Such modification is useful to enhance the fibers properties such as softness and to change hydrophilic/hydrophobic balance.     

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.     

The pattern of cell wall deterioration in lignocellulose fibers throughout enzymatic cellulose hydrolysis

Cell wall deterioration throughout enzymatic hydrolysis of cellulosic biomass is greatly affected by the chemical composition and the ultrastructure of the fiber cell wall. The resulting pattern of cell wall deterioration will reveal information on cellulose activity throughout enzymatic hydrolysis. This study investigates the progression and morphological changes in lignocellulose fibers throughout enzymatic hydrolysis, using (transmission electron microscopy) TEM and field emission scanning electron microscopy (FE‐SEM). Softwood thermo‐mechanical pulp (STMP) and softwood bleached kraft pulp (SBKP), lignocellulose substrates containing almost all the original fiber composition, and with lignin and some hemicellulose removed, respectively, was compared for morphology changes throughout hydrolysis. The difference of conversion between STMP and SBKP after 48 h of enzymatic hydrolysis is 11 and 88%, respectively. TEM images revealed an even fiber cell wall cross section density, with uneven middle lamella coverage in STMP fibers. SKBP fibers exhibited some spaces between cell wall and lamella layers due to the removal of lignin and some hemicellulose. After 1 h hydrolysis in SBKP fibers, there were more changes in the fiber cross‐sectional area than after 10 h hydrolysis in STMP fibers. Cell wall degradation was uneven, and originated in accessible cellulose throughout the fiber cell wall. FE‐SEM images illustrated more morphology changes in SBKP fibers than STMP fibers. Enzymatic action of STMP fiber resulted in a smoother fiber surface, along with fiber peeling and the formation of ribbon‐disjunction layers. SBKP fibers exhibited structural changes such as fiber erosion, fiber cutting, and fiber splitting throughout enzymatic hydrolysis. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

Kraft Pulp Bleaching and Delignification by Dikaryons and Monokaryons of Trametes versicolor

The ability of 10 dikaryotic and 20 monokaryotic strains of Trametes (Coriolus) versicolor to bleach and delignify hardwood and softwood kraft pulps was assessed. A dikaryon (52P) and two of its mating-compatible monokaryons (52J and 52D) derived via protoplasting were compared. All three regularly bleached hardwood kraft pulp more than 20 brightness points (International Standards Organization) in 5 days and softwood kraft pulp the same amount in 12 days. Delignification (kappa number reduction) by the dikaryon and the monokaryons was similar, but the growth of the monokaryons was slower. Insoluble dark pigments were commonly found in the mycelium, medium, and pulp of the dikaryon only. Laccase and manganese peroxidase (MnP) but not lignin peroxidase activities were secreted during bleaching by all three strains. Their laccase and MnP isozyme patterns were compared on native gels. No segregation of isozyme bands between the monokaryons was found. Hardwood kraft pulp appeared to adsorb several laccase isozyme bands. One MnP isozyme (pI, 3.2) was secreted in the presence of pulp by all three strains, but a second (pI, 4.9) was produced only by 52P. A lower level of soluble MnP activity in one monokaryon (52D) was associated with reduced bleaching ability and a lower level of methanol production. Since monokaryon 52J bleached pulp better than its parent dikaryon 52P, especially per unit of biomass, this genetically simpler monokaryon will be the preferred subject for further genetic manipulation and improvement of fungal pulp biological bleaching.     

Nitrification, denitrification, and dechlorination in bleached kraft pulp mill wastewater

This study deals with combining the biologi cal removal of organic halogens with the removal of nitrogen from bleached kraft pulp mill wastewater in fluidized-bed reactors under nitrifying and denitrifying conditions. Untreated and biotreated bleached kraft pulp mill wastewaters had no detrimental effect on nitrification or denitrification. The nitrifying biofilm reactor, pregrown on synthetic inorganic feed with ammonia, removed without a lag phase adsorbable organic halogens [7.2 mg Cl (g biomass volatile solids)⁻¹day⁻¹] from bleached kraft pulp mill wastewater and selected chlorophenols from synthetic wastewater. Electron microscopical examination of the biofilm showed that bacteria, morphologically similar to the nitrifying species Nitrosomonas or Nitrobacter, and Nitrosospira were dominant. The denitrifying fluidized-bed reactor, pregrown on nitrate and methanol, denitrified without a lag phase bleached kraft pulp mill wastewater. Under denitrifying conditions, 35% of the total organic carbon content of untreated bleached kraft pulp mill waste water was removed. The reducing power delivered by untreated bleached kraft pulp mill wastewater for denitrification was 2 mmol electrons/mmol carbon mineralized. Dechlorination under denitrifying conditions was negligible. Received: 21 November 1996 / Received revision: 27 January 1997 / Accepted: 1 February 1997     

Chemical force microscopy of cellulosic fibers

Atomic force microscopy with chemically modified cantilever tips (chemical force microscopy) was used to study the pull-off forces (adhesion forces) on cellulose model surfaces and bleached softwood kraft pulp fibers in aqueous media. It was found that for the –COOH terminated tips, the adhesion forces are dependent on pH, whereas for the –CH₃ and –OH terminated tips adhesion is not strongly affected by pH. Comparison between the cellulose model surfaces and cellulosic fibers under our experimental conditions reveal that surface roughness does not affect adhesion strongly. X-ray photoelectron spectroscopy (XPS) and Fourier Transformed Infrared (FTIR) spectroscopy reveal that both substrate surfaces have homogeneous chemical composition. The results show that chemical force microscopy can be used for the chemical characterization of cellulose surfaces at a nano-level.     

Effect of reaction conditions on the properties and behavior of wood cellulose nanocrystal suspensions

Sulfuric acid hydrolysis of native cellulose fibers produces stable suspensions of cellulose nanocrystals. Above a critical concentration, the suspensions spontaneously form an anisotropic chiral nematic liquid crystal phase. We have examined the effect of reaction time and acid-to-pulp ratio on nanocrystal and suspension properties for hydrolyzed black spruce acid sulfite pulp. Longer hydrolysis times produced shorter, less polydisperse black spruce cellulose nanocrystals and slightly increased the critical concentration for anisotropic phase formation. Increased acid-to-pulp ratio reduced the dimensions of the nanocrystals thus produced; the critical concentration was increased and the biphasic range became narrower. A suspension made from a bleached kraft eucalyptus pulp gave very similar properties to the softwood nanocrystal suspension when prepared under similar hydrolysis conditions.     

Effect of Residual Lignin Type and Amount on Bleaching of Kraft Pulp by Trametes versicolor

The white rot fungus Trametes (Coriolus) versicolor can delignify and brighten unbleached hardwood kraft pulp within a few days, but softwood kraft pulps require longer treatment. To determine the contributions of higher residual lignin contents (kappa numbers) and structural differences in lignins to the recalcitrance of softwood kraft pulps to biobleaching, we tested softwood and hardwood pulps cooked to the same kappa numbers, 26 and 12. A low-lignin-content (overcooked) softwood pulp resisted delignification by T. versicolor, but a high-lignin-content (lightly cooked) hardwood pulp was delignified at the same rate as a normal softwood pulp. Thus, the longer time taken by T. versicolor to brighten softwood kraft pulp than hardwood pulp results from the higher residual lignin content of the softwood pulp; possible differences in the structures of the residual lignins are important only when the lignin becomes highly condensed. Under the conditions used in this study, when an improved fungal inoculum was used, six different softwood pulps were all substantially brightened by T. versicolor. Softwood pulps whose lignin contents were decreased by extended modified continuous cooking or oxygen delignification to kappa numbers as low as 15 were delignified by T. versicolor at the same rate as normal softwood pulp. More intensive O₂ delignification, like overcooking, decreased the susceptibility of the residual lignin in the pulps to degradation by T. versicolor.     

Treatment of recycled kraft pulps with Trichoderma reesei hemicellulases and cellulases

Effects of recycling ECF-bleached softwood kraft pulp on pulp properties were evaluated in the laboratory. The tensile strength, fiber flexibility and WRV lost during drying of the pulp were recovered by refining between the cycles which, however, resulted in deteriorated drainage properties. The recycled pulps were treated with purified Trichoderma reesei cellulases and hemicellulases and the changes in fiber properties due to enzymatic treatments were characterized. The endoglucanases (EG I and EG II) significantly improved pulp drainage already at low dosage levels, and EG II was found to be more effective at a given level of carbohydrate solubilization. Combining hemicellulases with the endoglucanase treatments increased the positive effects of the endoglucanases on pulp drainage. However, as a result of the endoglucanase treatments a slight loss in strength was observed. Combining mannanase with endoglucanase treatments appeared to increase this negative effect, whereas the impact of xylanase was not significant. Although the drainage properties of the pulps could be improved by selected enzymes, the water retention capacity of the dried hornified fibers could not be recovered by any of the enzymes tested.     

Modification of high lignin content kraft pulps with laccase to improve paper strength properties. 1. Laccase treatment in the presence of gallic acid

Laccase was reacted with gallic acid in the presence of a high-kappa (91) kraft pulp. The result was a modified pulp with 34%, 20%, and 72% improvements in burst, tensile, and wet tensile strength compared to untreated control samples. Fully bleached pulps were not responsive to the laccase treatment, indicating lignin was the major target for the fiber modification. The results indicate that the strength increases were a combined effect of improvements of hydrogen bonding between fibers and creation of phenoxy radical cross-links within the sheet.     

Water transmission barrier properties of biodegradable films based on cellulosic whiskers and xylan

The reinforcement of natural biopolymers with cellulosic whiskers has been shown to be beneficial for physical strength properties including xylan films. This study examines the water transmission properties of xylan films reinforced with cellulosic whiskers prepared from kraft pulp hydrolyzed with sulfuric acid. Measurements of water transmission rate (WVTR) were accomplished by a modification of wet cup method described by ASTM E 96-95. The results showed that films prepared by xylan reinforced by 10% sulfonated whiskers exhibited a 74% reduction in specific water transmission properties with respect to xylan film and a 362% improvement with respect to xylan films reinforced with 10% softwood kraft fibers.     

The effects of fungi pre-treatment of poplar chips on the kraft fiber properties

Poplar chips were pre-treated by Trametes versicolor for 1, 2 and 3 weeks. Pre-treated chips, after washing, have been air dried for kraft pulping to achieve pulp kappa number of about 20. Pulp samples have been analyzed by Bauer Mc Nett, Kajaani analyzer and SEM. The results indicated that fungi pre-treatment of chips can degrade lignin and carbohydrates and affect kraft pulping and fiber characteristics. Higher chemical charge in pulping, lower fine and higher long fiber fraction were observed in pre-treated pulp samples in comparison with others. Fiber length, cross sectional area, width, cell wall thickness and volume index were increased by increasing pre-treatment time while fine length, fiber coarseness and curl have been reduced. Based on the study findings, with respect to higher fiber length, lower fine, and lower fiber curl and coarseness, 2-weeks pre-treatment of chips was recommended to produce acceptable overall fiber properties in kraft pulping.     

Characteristics of antimicrobial fibers prepared with wood periodate oxycellulose

A kind of antimicrobial fibers, a composite of Chitosan-Dialdehyde Cellulose (C-DAC) fibers, was prepared by using commercially elemental chlorine-free (ECF) bleached kraft softwood cellulose fibers oxidized by periodate and then further grafted by chitosan with different molecular weights. The characteristics of the C-DAC fibers and physical properties as well as antimicrobial activities of the handsheet made of C-DAC fibers were measured. The results show that the dry and rewet tensile indices, antimicrobial activity against Staphylococcus aureus and Escherichia coli, can be improved significantly as the molecular weight of grafted chitosan on the dialdehyde cellulose fibers decreases.     

A comparison of enzyme-aided bleaching of softwood paper pulp using combinations of xylanase, mannanase and α-galactosidase

Enzymatic pretreatment of softwood kraft pulp was investigated using xylanase A (XylA) from Neocallimastix patriciarum in combination with mannanase and α-galactosidase. Mannanase A (ManA) from Pseudomonas fluorescens subsp. cellulosa and ManA from Clostridium thermocellum, both family 26 glycosyl hydrolases, are structurally diverse and exhibit different pH and temperature optima. Although neither mannanase was effective in pretreating softwood pulp alone, both enzymes were able to enhance the production of reducing sugar and the reduction of single-stage bleached κ number when used with the xylanase. Sequential incubations with XylA and P. fluorescens ManA produced the largest final κ number reduction in comparison to control pretreated pulp. The release of galactose from softwood pulp by α-galactosidase A (AgaA) from P. fluorescens was enhanced by the presence of ManA from the same microorganism, and a single pretreatment with these enzymes, in combination with XylA, gave the most effective κ number reduction using a single incubation. Results indicated that mixtures of hemicellulase activities can be chosen to enhance pulp bleachability. Received: 16 August 1999 / Received revision: 13 December 1999 / Accepted: 15 December 1999     

Cellulase-assisted refining of chemical pulps: Impact of enzymatic charge and refining intensity on energy consumption and pulp quality

Paper production requires fibres to be refined, meaning mechanically treated to present sufficient bonding potential. As it is a highly energy consuming stage, various charges of cellulase as a pre-treatment were investigated to reduce the energy consumption and improve paper properties. The enzyme was added during pulp slushing and conditions of treatment were chosen to be compatible with an industrial application. Results obtained after low consistency disc refining of a softwood bleached kraft pulp were compared at a given drainage index. Enzymatically-treated samples showed a better development of fibrillation leading to a stronger paper. Moreover, fibre swelling was significantly improved. The impact of cellulase charge added to the pulp was identified. However, for all cellulase charges tested, fibre intrinsic resistance was lowered. Consequently fibres became shorter and tear index dropped. A solution was found by applying a milder treatment consisting in reducing refining intensity. This was a way to limit the intense fibre cutting at high refining levels. Moreover, by treating pulp with cellulase, it became possible to reduce refining intensity by 33%, keeping breaking length similar to control.     

Prebleaching of kraft pulp with full-length and truncated forms of a thermostable modular xylanase from Rhodothermus marinus

Full-length and truncated forms of a modular thermostable xylanase (EC 3.2.1.8., glycoside hydrolase family 10) were used in bleaching sequences of hardwood and softwood kraft pulps. Enzymatic treatment led to brightness gains of all pulps but the result depended on the pulp source. The presence of the additional domains in the full-length enzyme (including carbohydrate-binding modules) did not improve the bleaching process. No significant change in viscosity was seen after enzyme treatments indicating an unaffected pulp fibre length.     

The assessment of chromophores in bleached cellulosic pulps employing UV-Raman spectroscopy

UV-Resonance Raman (UV-RR) coupled with UV-visible Diffuse Reflectance (UV-vis DR) spectroscopy was applied to a solid-state study of chromophores in Eucalyptus globulus kraft cellulosic pulps bleached by chlorine dioxide and hydrogen peroxide. The UV-RR spectra were acquired at 325 nm laser beam excitation, which was shown to be appropriate for selective analysis of chromophore structures in polysaccharides. The proposed approach allowed the monitoring of chromophores in pulps and to track the extent of polysaccharide oxidation. However, precaution was suggested while performing a quantitative analysis of chromophores at the characteristic band of ∼1600 cm⁻¹ because of charge transfer complexes (CTCs) that exist in the pulp. These CTCs can affect the intensity of the aforementioned band by diminishing the conjugate state in the chromophore moieties. The amount of carbonyl and carboxyl groups in polysaccharides correlated with the intensity of the band at 1093 cm⁻¹. The analysis of UV-RR spectra revealed xylan as an important source of chromophores in eucalypt kraft pulp.     

New approach for upgrading pulp & paper quality: Mild potassium permanganate treatment of already bleached pulps

The present work introduces mild – room temperature – potassium permanganate treatment of cellulosic materials, namely already bleached pulps. Such treatment represents a new approach for upgrading pulp and paper quality, which is lacking in the literature. Potassium permanganate was investigated as a purifying and mild oxidizing agent for commercial already bleached softwood and bagasse pulps. It was found that treatment of the bleached beaten pulps, with 0.25–2% KMnO₄ (based on pulp weight), led to significant improvement in paper properties. The strength (breaking length) increased greatly and the brightness increased significantly due to treatment. The improvements were related to the degree of polymerization, and to the alphacellulose content of pulps.Moreover, potassium permanganate serves as a disinfectant and deodorizer. Thus treatment of bleached pulps with KMnO₄ is a promising remedy for the side effects which pulps suffer, during transportation and storage, before papermaking.     

Biological bleaching of hardwood kraft pulp using Trametes (Coriolus) versicolor immobilized in polyurethane foam

Summary Incubation of hardwood kraft pulp (HWKP) in agitated aerated cultures of the white-rot fungus Trametes versicolor increases pulp brightness and decreases its residual lignin content. A consequence of this biobleaching with whole cultures is that the resulting pulp also contains fungal biomass (up to ca. 10% (w/w)). In this report culture conditions for the immobilization of T. versicolor on polyurethane foam and bleaching of HWKP with the immobilized fungus are described. The major advantage of using immobilized fungus to bleach HWKP is that the fungal biomass can be separated from the pulp after treatment, resulting in a biologically bleached pulp free of fungal mycelium. From an analysis of pulp samples bleached with free and foam-immobilized mycelium, we conclude that fungal biomass in pulp treated with free mycelium accounts for up to 25% of the reduction in pulp viscosity (indication of cellulose chain length) whereas the zero span breaking length (indication of fibre strength) is not significantly affected by the presence of the fungus. Immobilization of the fungus on polyurethane foam also allows the repeated use of the same fungal biomass to bleach successive batches of pulp, either immediately or after storage at 4°C. Offprint requests to: I. D. ReidIssued as NRCC no. 30975