Production of biocatalysts on the basis of recombinant heterologous xylanase producer strains in the Penicillium verruculosum fungus: Their application in the hydrolysis of timber and wood processing industry wastes

The aim of this work was to create biocatalysts with an increased heterologous expression of endo-β-1,4-xylanase of P. canescens using recombinant P. verruculosum strains, to analyze the properties of new enzyme preparations, and to study the saccharifying activity of these preparations in the hydrolysis of plant raw materials, such as hogged aspen and detarred pine wood wastes of the timber and wood processing industries. The xylanase activity of the existing enzymatic preparations is insufficiently high to hydrolyze a xylan-rich biomass. The creation of increasingly xylanolytically active P. verruculosum-based recombinant strains containing homologous or heterologous genes of xylanase and mannanase is therefore a problem of great interest. Using the methods of genetic engineering, we obtained enzymatic preparations that are biocatalysts for the hydrolysis of plant raw material wastes of the sawmilling and wood processing industries and, according to the data of chromatographic fractionation, have compositions of 45–60% cellulase and 20–50% xylanase (which is optimal for the saccharifying of bagasse, along with aspen and pine wood). The originality of our technique lies in the creation of biocatalysts with predetermined properties, thus reducing appreciably the cost of enzyme preparation by eliminating the need to mix components of the carbohydrase complex for the hydrolysis of plant raw materials, e.g., aspen and pine wood.     

New aspects in cationization of lignocellulose materials. VII. Modification of spruce wood meal with quarternary ammonium groups

Trimethylammonium-2-hydroxypropyl (TMAHP) derivatives of spruce wood meal (SWM) and holocellulose of this specia were prepared by the reaction of wood meal with 3-chlor-2-hydroxy-propyltrimethylammoniumchloride (CHMAC) in alkaline medium. The TMAHP samples were fractionated and yields and exchange capacity (Q) of individual fractions were compared with beech and aspen fractions obtained under the same conditions. As it is evident from ¹³C-NMR spectroscopy and GPC analysis the water soluble fraction from TMAHP–SWM consists only of lignin–saccharide degradation products. The NaOH extracts of TMAHP–SWM and TMAHP–holocellulose as well as the water-soluble fraction from TMAHP–holocellulose are polymeric materials. From TMAHP–SWM only 3.1% of alkali-soluble material could be extracted, while from TMAHP–holocellulose 15.7% of water-soluble and 7.9% of alkali-soluble materials were obtained.     

Effect of Pretreatment of Poplar Wood Upon Enzymatic Saccharification

The wood of young poplar grown in short rotation coppices was used as a substrate for enzymatic saccharification. Several pretreatments of the wood, both physical and chemical, including delignification were applied to enhance the polysaccharide conversion into fermentable sugars. Comparing the yields obtained on a delignified material and on alkali treated material pointed out that lignin is not the major obstacle to saccharification. On the other hand, the swelling and dissolution effect of the potent cellulose solvent, N-methyl morpholine N-oxide, on wood brought about a nearly quantitative sugar recovery. This shows the importance of the ultrastructural organization of the plant cell wall over its enzymatic hydrolysis.     

Thermal,crystallization, and dynamic rheological behavior of wood particle/HDPE composites: Effect of removal of wood cell wall composition

This study investigated the effect of removal of wood cell wall composition on thermal, crystallization, and dynamic rheological behavior of the resulting high density polyethylene (HDPE) composites. Four types of wood particle (WP) with different compositions: native wood flour (WF), hemicellulose‐removed wood particle (HR), lignin‐removed wood particle (holocellulose, HC), and both hemicellulose and lignin removed particle (α‐cellulose, αC) were prepared and compounded with HDPE using extruder, both with and without maleated polyethylene (MAPE). Results show that removal of the hemicellulose improved the thermal stability of composites, while removal of the lignin facilitated thermal decomposition. WPs acted as nucleating agents and facilitated the process of crystallization, thereby increasing the crystallization temperature and degree of crystallinity. The crystallization nucleation and growth rate of αC and HR based composites without MAPE decreased, as compared with WF based one. Composite melts with and without MAPE exhibited a decreasing order of storage modulus, loss modulus, and complex viscosity as αC > WF > HR > HC and αC > HR > WF > HC, respectively. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40331.     

New aspects in cationization of lignocellulose materials. V. Modification of rotten aspen wood meal with quarternary ammonium groups

Holocelluloses prepared from clear and rotten aspen wood were gradually fractionated with 2.5% NH₄OH, 4.5% NaOH, and 17.5% NaOH, respectively. A higher yield of polysaccharides (24.4%) was obtained from the rotten sample in comparison with clear wood (20.9%). Trimethyl ammonium-2-hydroxypropyl (TMAHP) derivatives of rotten aspen were prepared by the reaction of wood with 3-chlor-2-hydroxypropyltrimethylammoniumchloride (CHMAC) in alkaline medium. The quantity of TMAHP—hemicelluloses (yield 14.1%) is only slightly lower in comparison with TMAHP—hemicelluloses (yield 15.8%) obtained by modification and subsequent extraction from clear aspen wood meal. The hemicelluloses isolated from the rotten aspen wood meal are contaminated with low molecular cellulose fraction, the degradation products of cellulose attacked by fungi. The lignin component of rotten wood is less intensively attacked by fungi than the polysaccharidic one.     

A new approach for the production of cellulose acetate: Acetylation of mechanical pulp with subsequent isolation of cellulose acetate by differential solubility

A heretofore uninvestigated approach to the production of cellulose acetate, the acetylation of whole wood pulp with subsequent isolation of the cellulose derivative by differential solubility, is described. The mechanical pulp used was produced by refining aspen wood chips with a disc refiner. Two conventional acetylation techniques, the fibrous and solution process, were employed to acetylato all components of the pulp. The cellulose acetate was isolated from the acetylated lignin and hemicellulose by dissolving in dichloromethane/methanol (9:1, v/v). The advantage of this new approach is that the high cost involved in using an extensively purified dissolving pulp are avoided. Both acetylation techniques yielded a product that was about 84% cellulose acetate. The remaining acetylated components were lignin and hemicellulose. The yield of cellulose acetate, based on the cellulose content of the original pulp and the product, was 75–80%.     

Softening of wood polymers induced by moisture studied by dynamic FTIR spectroscopy

The softening of the wood polymers is very important in the utilization of wood fiber products. To better to understand how the hemicelluloses interact with the other wood polymers in the fiber ultrastructure and contribute to the mechanical properties of the wood, their softening inside the cell wall of pulp fibers was studied by dynamic FTIR spectroscopy under humid conditions. The two hemicelluloses of spruce, glucomannan and xylan, exhibited different softening behaviors indicating a different organization of the two hemicelluloses inside the cell wall. Lignin in mechanical pulp showed a more viscoelastic behavior already under dry conditions than the polysaccharides of the cell wall. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 2032–2040, 2004     

The dependence of char and carbon yield on lignocellulosic precursor composition

A set of lignocellulosic precursors of widely varying composition was pyrolyzed under argon at 15°C/min to 500°C. Variation in the total mass and carbon yields among precursors was found to be due to variation in composition, i.e. distribution of the main organic components (lignin, holocellulose and extractives). Simple models for total mass and carbon yields of the precursors allowed determination by multiple regression analysis of the total mass and carbon yield of the components pyrolyzing in their native state in the lignocellulosic matrix. The component yields thus estimated were compared to results observed in studies of isolated components, such as reagent-grade cellulose. The char yield from lignin (53%) was found to be three times that of cellulose (18%) because of the higher initial carbon content in lignin (63 vs 44%) as well as the higher carbon yield (76 vs 40%).     

Enhancing exterior performance of clear coatings through photostabilization of wooden surfaces. Part 1: Treatment and characterization

The aim of this work was to produce a less photodegradation-prone substrate for clear coating by partially delignifying the surface cells of Pinus radiata boards to a depth of 2–3 mm while maintaining the integrity of the wood surface tissues in the delignified zone. To achieve this, several surface oxidative delignification treatments were trialled in the method development process and peracetic acid was chosen as the method for refinement and deployment. The treatment method was optimized to yield a significant degree of delignification compatible with the aim of producing a photostabilized yet intact wooden surface. A preweathering technique was also used as a second delignification method for producing photostabilized boards. Microscopic and chemical techniques were used to characterize the effects of chemical and preweathering treatments, which produced delignified surface envelopes 2–3 mm and 100 μm deep, respectively. Acetyl bromide lignin analyses, infrared spectroscopic analyses, and density changes of 30-μm-thick sections of peracetic-acid-treated samples, as a function of depth from the wood surface, suggested that a partial delignification had occurred that diminished with depth. Light and transmission electron microscopy provided evidence of delignification at the cellular level. In the surface layers of peracetic-acid-treated boards, all cell wall regions were delignified, with the middle lamella being the most severely affected. Lignin appeared to be completely removed from the cell corner middle lamella regions, but tracheids were still joined in other parts of the middle lamella. The S₁, S₂, and S₃ walls were also delignified. In subsurface layers, cell walls were only partially delignified and the tissues held their integrity. In contrast, in the preweathered boards, cell walls in the outermost layers were completely separated at the middle lamella from photodegradation. Preferential lignin removal over that of hemicelluloses was achieved via oxidative treatment of solid wood. The outcome of this was the successful fulfillment of our aim to produce partially delignified wooden surfaces that retained sufficient strength and aesthetic appearance and were suitable for application in wooden structures, provided that the necessary protection measures were carried out. Due to the narrow surface zone produced, machining of such surfaces would not be recommended as the most delignified zone would be removed first. In Part 2 of this series, the photostability of treated boards, which had been clear coated and weathered, will be considered.     

植物纤维原料及纸浆中的木素―碳水化合物复合体

造纸工业中所用的木质纤维原料主要是由纤维素、半纤维素和木素组成的。纤维聚合物之间存在相互作用。研究证明,在木材和纸浆中,部分木素和碳水化合物发生化学键联接形成木素―碳水化合物复合体结构。文章着重介绍了复合物中三种重要化学键的键型;结合前人的研究成果,综述了云杉木、杨木、麦草以及各种针叶木浆和阔叶木浆中的木素―碳水化合物复合体(LCC);此外,对LCC的形成机理和蒸煮过程中形成新LCC的可能性加以阐述。     

Structural changes of cellulose,wood, and paper under shear deformation and high pressure

Structural changes of wood and its components have been studied after shear deformation under high pressure (SDHP) at up to 6 GPa. Cellulose amorphization and chain depolymerization was observed. Approximately 30% of microcrystalline cellulose was soluble in water after a 360-degree twist of the Bridgman anvils. The water-insoluble part was re-crystallized into cellulose II lattice. Repeated treatment applied to the nondissolved part of the sample, the so-called cascade experiment, permits the dissolving of about 30% of the residual nondissolved material once again. Extraction with water, followed by 10% sodium hydroxide, allows almost complete dissolving of microcrystalline cellulose (98%). Water-soluble saccharides were studied by HPLC and ¹³C NMR. It was found that destruction of the wood lignin network needs more severe treatment conditions than cellulose destruction does. Lignin domains in wood act as “grinding stones” during cellulose destruction. Long-living lignin free radicals have been detected with EPR after SDHP. ¹³C NMR CP/MAS spin diffusion studies showed that SDHP leads to separation of wood components into different biopolymer domains, which turns the system toward thermo-dynamic equilibrium. SDHP does not permit achievement of initial compulsary compatibility of components in native wood. SDHP technique appears as a promising method for wood delignification and carbohydrate saccharification in the solid state without using harmful chemical reagents of solvents, which is important for technological safety and ecology. © 1994 John Wiley & Sons, Inc.     

Studies on the Mechanism of Formation of the Products of Liquefaction of Cellulose and Aspen Poplar Wood

Abstract

Studies are reported on the chemical nature and mechanism of formation of the components of the oil fraction obtained by the alkali-catalyzed liquefaction of aspen poplar wood and separately of cellulose. Along with the smaller molecular weight compounds reported earlier (5) evidence has been obtained for the presence of polymeric fractions involving ether linkages and which show a bimodal pattern of molecular weights. Using appropriate model substances, some evidence ha3 been obtained for the production and involvement of one- and two-carbon fragments as well as ketene-type intermediates in the liquefaction process. Examination of the role of pH during the conversion process suggests that the initially alkaline medium is required to degrade the cellulose to smaller fragments, including possible saccharinic acids, with a corresponding decrease in pH. Then at an appropriate pH (now acidic) and temperature the resulting degradation products are converted to components of the final oil.     

Wood Fiber Quality and Kraft Pulping Efficiencies of Trembling Aspen (Populus tremuloides Michx) Clones

Abstract

The natural variation in wood and pulp fiber quality of 15 aspen (Populus tremuloides Michx) clones, represented by 47 trees, was assessed from 4 different sites in British Columbia, Canada. Kraft pulping trials revealed substantial variation in the pulping efficiencies, illustrated by differences of 6% in total pulp yield, ～30% differences in H‐factor required to attain a target kappa of 21, and differences of up to 2 ISO brightness units in bleachability of kappa 21 pulp. Clearly, enormous variation exists in the natural stands of aspen, and presents some exciting opportunities for selecting clonal aspen for targeted end‐product applications. A comprehensive characterization of wood chemical composition, wood density, and fiber properties indicated that pulp yield is directly related to syringyl lignin monomer composition, and not inherent wood density, regardless of geographic locations, whereas pulp bleachability and viscosity appear to be associated with the inherent cell wall thickness of the starting wood resources (fiber coarseness). These findings suggest that geographic location imparts influences on wood fiber coarseness traits, while substantial genetic variability exists on all sites.     

New aspects in cationization of lignocellulose materials. VI. Modification of steam-exploded aspen wood chips with quarternary ammonium groups

Holocellulose prepared from steam-exploded aspen wood was gradually fractionated with 2.5% NH₄OH, 4.5% NaOH, and 17.5% NaOH, respectively. Some residual polysaccharides were extracted from the material in this way, but their yield was only about 5%. Trimethylammonium-2-hydroxypropyl (TMAHP)-derivatives, exploded aspen wood (EXAW), were prepared by the reaction with 3-chlor-2-hydroxypropyl trimethylammoniumchloride (CHMAC) in alkaline medium. The quantity of water and alkali extracted TMAHP–polysaccharides (yield 2.6%) was lower than the yields from modified healthy beech and aspen, as well as rotten aspen. On the other hand, we can obtain more modified cellulose-rich and lignin-rich material than from previously mentioned species. The steam-explosion process is suitable for increasing the accessibility of cellulose component in lignocellulose material for chemical modification.     

稻草秸秆综纤维素对阿司匹林片崩解性能的影响

以稻草秸秆为原料,经过SO₃微热爆预处理、木质素降解、漂白等工序制备出了稻草秸秆综纤维素,并将其用作片剂崩解剂进行了性能评价。参照《中国药典》(2015版)中的有关规定,检测了稻草秸秆综纤维素的各项指标;以阿司匹林为模型药物,以市售微晶纤维素为对照,测定了稻草秸秆综纤维素的崩解性能。结果表明,稻草秸秆综纤维素质量符合《中国药典》(2015版)中的相关规定,且崩解性明显优于市售微晶纤维素,可用作片剂崩解剂。在稻草秸秆综纤维素用量为3％~5%时即可达到微晶纤维素用量为15%~20%的崩解效果,这一效果的呈现与稻草秸秆综纤维素的形态结构以及含有半纤维素组分有着密切联系。     

The Xyloglucan Endotransglucosylase/Hydrolase Gene XTH22/TCH4 Regulates Plant Growth by Disrupting the Cell Wall Homeostasis in Arabidopsis under Boron Deficiency

TCH4 is a xyloglucan endotransglucosylase/hydrolase (XTH) family member. Extensive studies have shown that XTHs are very important in cell wall homeostasis for plant growth and development. Boron (B), as an essential micronutrient for plants, plays an essential role in the cross-linking of cell wall pectin. However, the effect of B on cell wall organization is unclear. This study aimed to explore the mechanism of plant adaption to B stress by investigating the role of TCH4 in cell wall homeostasis. We conducted both plate and hydroponic cultures of wild-type Col-0 and overexpression and gene knockout lines of XTH22/TCH4 to analyze the phenotype, components, and characteristics of the cell wall using immunofluorescence, atomic force microscopy (AFM), and transmission electron microscopy (TEM). B deficiency induces the expression of TCH4. The overexpression lines of TCH4 presented more sensitivity to B deficiency than the wild-type Col-0, while the knockout lines of TCH4 were more resistant to low B stress. Up-regulation of TCH4 influenced the ratio of chelator-soluble pectin to alkali-soluble pectin and decreased the degree of methylesterification of pectin under B-deficient conditions. Moreover, we found that B deficiency disturbed the arrangement of cellulose, enlarged the gap between cellulose microfibrils, and decreased the mechanical strength of the cell wall, leading to the formation of a thickened and deformed triangular region of the cell wall. These symptoms were more profound in the TCH4 overexpression lines. Consistently, compared with Col-0, the O₂⁻ and MDA contents in the TCH4 overexpression lines increased under B-deficient conditions. This study identified the B-deficiency-induced TCH4 gene, which regulates cell wall homeostasis to influence plant growth under B-deficient conditions.     

New aspects in cationization of lignocellulose materials. II. Distribution of functional groups in lignin,hemicellulose, and cellulose components

The distribution of functional groups in the three main components of beech after alkylation with 3-chlor-2-hydroxypropyltrimethylammoniumchloride (CHMAC) was studied. By fractional extraction of hemicellulose combined with delignification it was found that the three components were substituted to various extents. The exchanging capacity decreased in the following order: hemicellulose, lignin, and cellulose. The obtained degree of substitution is related to the accessibility and reactivity of the individual components in the wood. Hemicellulose, which is substituted the most, can be extracted from modified sawdust with water. This represents a new method of hemicellulose isolation from native plant materials.