Delignification of wood pulp by a thermostable xylanase from Bacillus stearothermophilus strain T-6

During the bleaching of wood pulp for the paper industry, large amounts of chlorinated aromatic compounds are produced and released into the environment. These compounds are extremely toxic and are a major source of pollution. The paper and pulp industry is seeking for alternative methods for bleaching pulp. One such method involves the use of hemicellulases to release the colored lignohemicellulose. We have isolated and characterized several thermophilic bacteria which produce xylanases. One such strain, T-6, produced high levels of extracellular xylanase, free of cellulase and proteinase activities. Strain T-6 was classified as a strain of Bacillus stearothermophilus and was able to grow on defined medium containing xylose, methionine and asparagine at 65 °C. Xylanase activity was induced by either xylose or xylan; no activity was detected with other carbon sources, such as glycerol, acetate, lactose, glucose, maltose, fructose, mannose, galactose or sucrose. Xylanase constitutive mutants were obtained following mutagenesis and detection on p-nitrophenol -d-xylopyranoside containing agar plates. Xylanase T-6 was produced on large scale, and was purified and concentrated by a single adsorption-desorption step from a cation exchanger. The overall purification yield of a 1000 liter fermentation was 45%, resulting in a 98% pure enzyme. Xylanase T-6 was shown to partially remove lignin from unbleached pulp at 65 °C and pH 9.0, without loss in pulp viscosity. The enzyme-treated pulp was used to make handsheets that had higher brightness than untreated pulp.     

The effects of wood decay type on the growth of bryophyte gametophytes

Deadwood is an important habitat for bryophytes in boreal and subalpine forests. The type of decay in wood (white, brown, and soft rot) caused by fungal colonizers has been revealed to affect bryophyte communities. However, little is known about the effects of decay type on the growth of bryophytes. We tested the effect of wood decay type on gametophyte growth for two common bryophyte species, Scapania bolanderi Austin and Pleurozium schreberi (Brid.) Mitt., which dominate the logs in subalpine coniferous forest on Mt. Ontake, in central Honshu, Japan. We used pot culture experiments in an open-sky nursery field. After eight months of cultivation, the growth of S. bolanderi was larger on brown rot wood than white rot wood, but the growth of P. schreberi was not. Mixed cultures of the two species also showed greater growth on brown rot wood. However, growth of S. bolanderi was significantly smaller than P. schreberi in mixed culture. These results suggest that brown rot wood enhances growth of S. bolanderi, but growth may be reduced under competition from P. schreberi. The results are in agreement with the field observation that brown rot wood has a positive association with S. bolanderi coverage on deadwood.     

Potential of xylanase from thermophilic Bacillus sp. XTR-10 in biobleaching of wood kraft pulp

An extracellular xylanase produced under optimal conditions by a thermophilic strain of Bacillus sp. XTR-10 was evaluated for its potential application in biobleaching of wood kraft pulp. Spectrophotometric analysis showed considerable release of lignin derived compounds and chromophoric material by the xylanase treated pulp samples. Xylanase was found to be effective in the liberation of reducing sugars in the pulp filtrates with increment in enzyme dose and reaction time. Eight hours pretreatment with 40 IU of xylanase/g of dry pulp resulted in 16.2% reduction of kappa number with 25.94% ISO increase in brightness as compared to the control. The same treatment slightly lowered the tensile strength and burst index, however. Enzyme pretreatment of the pulp saved 15% active chlorine charges in single step and 18.7% in multiple steps chemical bleaching with attainment of brightness at the level of the control. These results indicate the potential of enzymatic pretreatment of pulp for reduction in environmental discharge of hazardous waste from the pulp and paper industry.     

The effect of xylanase on lignocellulosic components during the bleaching of wood pulps

HPLC, SEM and XRD techniques have been proposed as methods for ascertaining the changes occurring in polysaccharides (cellulose and xylans) and fibres during the xylanase bleaching processes. TCF and ECF bleached pulps with and without enzyme pretreatment were analysed. The ratio of carbohydrates present in the pulp, observation of changes occurring in the surface of the fibres and the crystallinity and accessibility of the bleached fibres were determinated. These characteristics have been related with pulp properties. Xylan content decreased when pulp was bleached. Xylanase treatment substantially reduced the xylose content present in pulp, measured by HPLC after the hydrolysis method of the sample. Morphological changes in the fibres occurred when the enzymatic treatment was applied. Bleaching increased the crystallinity of the pulp and enzyme pretreatment also affected the crystallinity of cellulose fibres     

Freeze-thaw effects on metabolic enzymes in wood frog organs.

To determine whether episodes of natural freezing and thawing altered the metabolic makeup of wood frog (Rana sylvatica) organs, the maximal activities of 28 enzymes of intermediary metabolism were assessed in six organs (brain, heart, kidney, liver, skeletal muscle, gut) of control (5 degrees C acclimated), frozen (24 h at -3 degrees C), and thawed (24 h back at 5 degrees C) frogs. The enzymes assessed represented pathways including glycolysis, gluconeo-genesis, amino acid metabolism, fatty acid metabolism, the TCA cycle, and adenylate metabolism. Organ-specific responses seen included (a) the number of enzymes affected by freeze-thaw (1 in gut ranging to 17 in heart), (b) the magnitude and direction of response (most often enzyme activities decreased during freezing and rebounded with thawing but, liver showed freeze-specific increases in several enzymes), and (c) the response to freezing versus thawing (enzyme activities in gut and kidney changed during freezing, whereas most enzymes in skeletal muscle responded to thawing). Overall, the data show that freeze-thaw implements selected changes to the maximal activities of various enzymes of intermediary metabolism and that these may aid organ-specific responses that alter fuel use during freeze-thaw, support cryoprotectant metabolism, and aid organ endurance of freeze-induced ischemia.     

Xylanase prebleaching of fractions of Douglas-fir kraft pulp of different fibre length

A commercial xylanase from Trichoderma longibrachiatum was used to treat the fractions of Douglas-fir kraft pulp of different fibre length. Enzymatic prebleaching was followed by chelation and peroxide bleaching. An evaluation of both optical and physical properties of the distinct fractions was conducted. A difference in susceptibility of the fractions of different fibre length to xylanase prebleaching was observed. The bleach-boosting effect observed with all fractions appeared to be related to the high-molecular-mass UV-absorbing material solubilized during enzyme treatments. Xylanase treatments resulted in beneficial effects to handsheet density and roughness as well as to some of the strength properties. However, the response to the xylanase treatments exhibited by all fractions of different fibre length was not uniform, indicating that fiber composition played a key role in the effectiveness of xylanase treatments.     

Effects of surfactants on the enzymatic bleaching of kraft pulp by xylanase

A xylanase was purified from a commercial crude xylanase, Pulpzyme HC, and used for the bleaching of kraft pulp in the absence or in the presence of nonionic surfactants, Tween 20, Tween 80, and Igepal C930. The purified xylanase has a molecular weight of 23,500 as determined by a reducing SDS-PAGE. Tween 20 was most effective to enhance the efficiency of the enzymatic bleaching of kraft pulp by xylanase.     

Short-term effects of nitrogen availability on wood formation and fibre properties in hybrid poplar

The application of nitrogen-containing fertilisers is one approach used to increase growth rates and productivity of forest tree plantations. However, the effects of nitrogen fertilisation on wood properties have not been systematically assessed. The aim of this work was to document the impacts of nitrogen fertilisation on wood formation and secondary xylem fibre properties. We used three fertilisation treatments in which the level of ammonium nitrate was adjusted to 0, 1 and 10 mM in a complete nutrient solution applied daily over a period of 28 days in standardised greenhouse experiments with clonal material of Populus trichocarpa (Torr and Gray) × deltoides (Bartr. ex Marsh). We showed that there was a short-term and repeatable response in which xylem fibre morphology and secondary cell wall structure adapt to a shift in N availability. Under high-nitrogen exposure, xylem fibres were 17% wider and 18% shorter compared to the adequate nitrogen treatment. A very significant thickening of the fibre cell walls was also observed throughout the stem of trees receiving the high-N treatment. It appeared that cell wall structure was greatly affected by the high-N treatment as fibres developed a modified inner cell wall layer. Histological observations indicated that the internal cell wall layer was enriched in cellulose and chemical determinations showed that wood contained more holocellulose. Together, these results indicate that the response of poplar to nitrogen availability may involve marked effects on secondary xylem formation.     

Impact of xylanase pretreatment on peroxide bleaching stage of hemp pulp

Pretreatment of hemp pulp with xylanase was investigated. Unbleached hemp pulp was treated with commercial xylanase, and then bleached with hydrogen peroxide. Control pulp bleached with out xylanase was compared with xylanase bleached pulp. Application of xylanase was found to have a positive effect on followed peroxide stage in terms of low kappa number and high brightness of pulp.     

The effect of extracellular enzymes of wood-rotting fungi on wood

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The effect of hemicelluloses on wood pulp nanofibrillation and nanofiber network characteristics

Hemicelluloses as matrix substances showed an important role in nanofibrillation of wood pulp. Never-dried and once-dried pulps with different amounts of hemicelluloses were fibrillated using a grinding treatment. The degree of fibrillation was evaluated by scanning electron microscopy observation of the fibrillated pulps and light transmittance measurements of the fibrillated pulp/acrylic resin composites. With a one-pass grinding treatment, the once-dried pulp with higher hemicellulose content was fibrillated into 10-20 nm wide fibers as easily as the never-dried pulps, while the once-dried pulp with lower hemicellulose content was not fibrillated into uniform nanosized fibers. This result indicates that hemicelluloses act as inhibitors of the coalescence of microfibrils during drying and facilitate the nanofibrillation of once-dried pulp. Furthermore, hemicelluloses provide adhesion between nanofibers, contributing to reduction of thermal expansion and enhancement of mechanical properties in the composites.     

Effect of steam treatment on the properties of wood cell walls

Steam treatment is a hygrothermal method of potential industrial significance for improving the dimensional stability and durability of wood materials. The steaming results in different chemical and micromechanical changes in the nanostructured biocomposite that comprise a wood cell wall. In this study, spruce wood ( Picea abies Karst.) that had been subjected to high-temperature steaming up to 180 °C was examined, using imaging Fourier Transform Infrared (FT-IR) microscopy and nanoindentation to track changes in the chemical structure and the micromechanical properties of the secondary cell wall. Similar changes in the chemical components, due to the steam treatment, were found in earlywood and latewood. A progressive degradation of the carbonyl groups in the glucuronic acid unit of xylan and a loss of mannose units in the glucomannan backbone, that is, a degradation of glucomannan, together with a loss of the C═O group linked to the aromatic skeleton in lignin, was found. The development of the hygroscopic and micromechanical properties that occurred with an elevation in the steam temperature correlated well with this pattern of degradation in the constituents in the biocomposite matrix in the cell wall (hemicellulose and lignin).     

Comparative study on application of T.lanuginosus SSBP xylanase and commercial xylanase on biobleaching of non wood pulps

Biobleaching of three non wood kraft pulps (bagasse, rice straw and wheat straw) by Thermomyces lanuginosus SSBP xylanase and commercial xylanase (cartazyme sandoz), was studied in order to investigate their potential and effect on their various properties (reduction sugars, chlorine dioxide, kappa number, brightness and chromophores). In generally, xylanases released chromophores and reducing sugars and decreased kappa number of pulps. These samples gained over six brightness points over controls. Biobleaching of rice straw pulp with xylanase cartazyme (Sandoz) produced chlorine dioxide savings of up to 25% or 3.5-4 kg chlorine dioxide/ton pulp.     

The effects of climate change on the birch pollen season in Denmark

During the last two decades the climate inDenmark has become warmer and in climatescenarios (IPCC, 2001) it is foreseen that thetemperature will increase in the comingdecades. This predicted future increase intemperature will probably affect both theflowering of plants and the dispersion ofpollen in the air. In this study the alreadyobserved effects on the birch pollen season arestudied.Trend analyses of the birch pollen seasonfor two stations in Denmark more than 200 kmapart give similar results. In Copenhagen thereis a marked shift to an earlier season – itstarts about 14 days earlier in year 2000 thanin 1977, the peak-date is 17 days earlier andthe season-end is 9 days earlier. For Viborgthe trend to an earlier season is in generalthe same, but slightly smaller.During the same period there has also beena distinct rise in the annual-total amount ofbirch pollen, peak-values and days withconcentrations above zero.Rising mean temperatures during winter andspring can explain the calculated trends towardearlier pollen season. Models for estimation ofthe starting date based on Growing Degree Hours(GDH's) give very fine results with acorrelation coefficient around 0.90 and rmserror around 4.2 days.For annual-total there is a significantpositive correlation with the mean temperaturein the growing season the previous year.     

Synergistic effects of cellulolytic and pectinolytic enzymes in degrading sugar beet pulp

Three cellulases, one hemicellulase and three pectinases were used, separately or in binary and ternary combinations, to hydrolyze dried beet-pulp, a by-product of the sugar industry. By IE-HPLC the compositions and concentrations of the sugars released were determined. The results obtained by enzymatic saccharification were compared to those obtained by acid hydrolysis. The synergistic action of cellulolytic and pectinolytic enzymes in release of total monosaccharides, and of glucose, arabinose and galacturonic acid was also studied. The combination of cellulase, hemicellulase and pectinase, commercially available, was as effective in degrading the beet pulp as the acid hydrolysis. Pectinase appeared to be the most important enzyme, since by hydrolyzing the pectic surface of the lignocellulosic substrate, it favoured the degradation of cellulose and hemicellulose by the respective enzymes.     

酶在纸浆漂白中的应用

介绍了酶在纸浆漂白工业中的研究及应用概况,特别是木聚糖酶在纸浆漂白中的应用及其研究进展。     

Extracting value from Eucalyptus wood before kraft pulping: effects of hemicelluloses solubilization on pulp properties

Eucalyptus globulus wood samples were subjected to autohydrolysis for extracting hemicelluloses, and the resulting solids were assayed as substrates for kraft pulping and further Totally Chlorine Free (TCF) bleaching. The susceptibility of treated solids to kraft processing was assessed under selected experiments covering the optimum experimental range. In order to establish a basis for comparison, samples of untreated wood were also subjected to kraft delignification. The best kraft pulps obtained from autohydrolyzed solids were subjected to an optimized TCF bleaching sequence involving double alkaline oxygen and pressurized H₂O₂ processing, and characterized using standard methods. The suitability of the final product obtained by autohydrolysis-kraft delignification-TCF bleaching for specific purposes is discussed on the basis of the experimental results.     

Microbial impacts on plant-herbivore interactions: the indirect effects of a birch pathogen on a birch aphid

The role of indirect interactions in structuring communities is becoming increasingly recognised. Plant fungi can bring about changes in plant chemistry which may affect insect herbivores that share the same plant, and hence the two may interact indirectly. This study investigated the indirect effects of a fungal pathogen ( Marssonina betulae) of silver birch ( Betula pendula) on an aphid ( Euceraphis betulae), and the processes underpinning the interaction. There was a strong positive association between natural populations of the aphid and leaves bearing high fungal infection. In choice tests, significantly more aphids settled on leaves inoculated with the fungus than on asymptomatic leaves. Individual aphids reared on inoculated leaves were heavier, possessed longer hind tibiae and displayed enhanced embryo development compared with aphids reared on asymptomatic leaves; population growth rate was also positively correlated with fungal infection when groups of aphids were reared on inoculated branches. Changes in leaf chemistry were associated with fungal infection with inoculated leaves containing higher concentrations of free-amino acids. This may reflect a plant-initiated response to fungal attack in which free amino acids from the degradation of mesophyll cells are translocated out of infected leaves via the phloem. These changes in plant chemistry are similar to those occurring during leaf senescence, and are proposed as the mechanistic basis for the positive interaction between the fungus and aphid.     

The effect of acetyl groups on the hydrolysis of ryegrass cell walls by xylanase and cellulase from trichoderma koningii

Delignified ryegrass cell walls were effectively hydrolysed by a mixture of endo-1,4-β-glucanase and xylanase, but the rate and extent of hydrolysis was greater when the cellobiohydrolase part of the cellulase system was also present. Deacetylation of the xylan in the cell walls had a significant effect on the rate but not on the extent of hydrolysis of delignified cell walls. Deacetylation followed by endoglucanase-xylanase action resulted in a significant decrease in the proportion of xylose present in the residual cell walls. However, when cellobiohydrolase was acting in admixture with the endoglucanase-xylanase, it was the cellulose component of deacetylated cell walls that was preferentially hydrolysed. The proportion of galactose in the unhydrolysed fraction of the cell walls increased significantly after enzyme action by the cellobiohydrolase-endoglucanase-xylanase system.