Beech Wood Sawdust Research Articles

A Preliminary Experimental Study on Biodegradation of 3D-Printed Samples from Biomass–Fungi Composite Materials

Products made from petroleum-derived plastic materials are linked to many environmental problems, such as greenhouse gas emissions and plastic pollution. It is desirable to manufacture products from environmentally friendly materials instead of petroleum-based plastic materials. Products made from biomass–fungi composite materials are biodegradable and can be utilized for packaging, construction, and furniture. In biomass–fungi composite materials, biomass particles (derived from agricultural wastes) serve as the substrate, and the fungal hyphae network binds the biomass particles together. There are many reported studies on the 3D printing of biomass–fungi composite materials. However, there are no reported studies on the biodegradation of 3D-printed samples from biomass–fungi composite materials. In this study, two types of biomass materials were used to prepare printable mixture hemp hurd and beechwood sawdust. The fungi strain used was Trametes versicolor. Extrusion based 3D printing was used to print samples. 3D-printed samples were left for five days to allow fungi to grow. The samples were then dried in an oven for 4 h at 120 °C to kill all the fungi in the samples. The samples were buried in the soil using a mesh bag and kept in an environmental chamber at 25 °C with a relative humidity of 48%. The weight of these samples was measured every week over a period of three months. During the testing period, the hemp hurd test samples lost about 33% of their original weight, whereas the beechwood sawdust samples lost about 30% of their original weight. The SEM (scanning electron microscope) micrographs showed the presence of zygospores in the test samples, providing evidence of biodegradation of the test samples in the soils. Additionally, the difference in peak intensity between the control samples and test samples (for both hemp hurd and beechwood sawdust) showed additional evidence of biodegradation of the test samples in the soils.

Molecular identification of Shiitake (Lentinula edodes), analysis and production of beta-glucan using beech wood sawdust waste

Lentinula edodes has the ability to grow and produce bioactive compounds on industrial by-products. This study aimed to produce B-glucan of cell wall Shiitake on Beechwood Sawdust (BWS) through a two-step procedure, which included fermentation and B-glucan extraction and purification. Shiitake mushrooms are cultivated by solid-state fermentation (SSF) using the Jamas method to increase the purity of B-glucan. The fermented substrate was first separated and then hydrolyzed by sodium hydroxide (NaOH) (10 M, 1 M), followed by acid hydrolysis extraction. The structure and purity of B-glucan were confirmed by FTIR, NMR, and AFM spectroscopy. The fungus used was molecularly identified by the 18 s rRNA method. Shiitake mushroom was produced by SSF using BWS and high purity β-glucan was extracted from the produced polysaccharide in the amount of 67.33 mg/g. FTIR, NMR, and AFM analyses proved the production of beta-glucan, and based on molecular identification, it was determined that the mushroom used was Lentinula edodes. The results obtained show that SSF is a valuable technology for the production of biomass and polysaccharides by utilizing the strain of L. edodes. To the best of our knowledge, the yield reported is the highest by the strain of L. edodes using SSF.

Isolation of Highly Crystalline Cellulose via Combined Pretreatment/Fractionation and Extraction Procedures within a Biorefinery Concept.

Sustainable production of bio-based materials and chemicals requires integrated approaches which utilize all fractions of lignocellulosic biomass. In this work, highly crystalline cellulose was isolated via combined pretreatment/fractionation and extraction processes from beechwood sawdust. The proposed approach was based on the selective recovery of hemicellulose components in the first step, followed by enhanced delignification in the second step, permitting the efficient recovery of the remaining cellulose via bleaching in the final step. Hydrothermal pretreatment under tailored conditions in neat water or dilute acid resulted in almost complete hemicellulose removal (80-96 wt %) in the liquid fraction. In the second step, the formed surface lignin was isolated via mild extraction while enhanced removal of both native/structural and surface lignin (71 wt %) was achieved by applying the organosolv treatment using dilute sulfuric acid as catalyst. Dilute sulfuric acid pretreatment followed by acid catalyzed organosolv pretreatment proved to be the most efficient combined approach, leading to 80 wt % hemicellulose removal as xylose monomer, and 71 wt % delignification. High crystallinity cellulose (<88%), with an overall cellulose recovery of 68-91 wt % based on native cellulose in parent biomass was isolated in the last step via bleaching of all pretreated biomass solids. The proposed integrated biorefinery procedures that aim to whole "waste" biomass valorization, replacing fossil resources, with the use of green solvents (water, ethanol) at relatively mild temperature/pressure conditions, are in line with the scope of several United Nations Sustainable Development Goals, such as UN SDG 8, 11, 12, and 13.

Conversion of beechwood organosolv lignin via fast pyrolysis and in situ catalytic upgrading towards aromatic and phenolic-rich bio-oil

Lignin, an abundant renewable biopolymer found in plant cell walls, is enriched in phenolic units within its complex molecular structure. Unlocking its potential as alternative feedstock in (bio)refining has posed a long-standing challenge, even though it holds immense promise for replacing fossil-derived phenolic and aromatic compounds. This study focuses on fast pyrolysis as effective thermochemical depolymerization method of lignin, coupled with the in situ catalytic upgrading aiming to produce valuable bio-oil enriched in dealkoxylated (alkyl)phenolic and aromatic compounds. Lignin was isolated via the organosolv process from beechwood sawdust (hardwood biomass). Various acidic aluminosilicate catalysts (e.g., zeolites, such as ZSM-5, Beta and USY, and amorphous silica alumina) were applied, having different Si/Al ratio, porous and acidic properties. Fast pyrolysis experiments were conducted on a fixed-bed bench-scale reactor at two distinct temperatures (500 and 600 °C), employing different contact times and lignin-to-catalyst ratios. Non-catalytic pyrolysis experiments revealed that higher temperature, significantly influences bio-oil’s composition and yield, resulting in the conversion of initially formed alkoxy-phenols to alkyl-phenolic compounds, reaching a 47% relative concentration at 600 °C, while also yielding high amount of bio-oil up to 43 wt%. Among the catalysts tested, zeolite ZSM-5 (Si/Al=40) proved to be the most efficient, shifting the chemical profile of bio-oil from phenolic to aromatic (mainly BTX) with relative concentration of 57%, owing to its unique microporous structure and acidity. Depending on the catalyst type, a balance between BTX monomer aromatics and naphthalenes was observed. Lignin, as well as the obtained products (bio-oil, non-condensable gases, char/coke-on-catalyst) were thoroughly characterized using various analytical techniques. The catalytic upgrading results were associated with the physicochemical properties of the catalysts, providing valuable insights into the underlying reaction mechanisms.

Experimental evidence that lignin-modifying enzymes are essential for degrading plant cell wall lignin by Pleurotus ostreatus using CRISPR/Cas9.

Lignin-modifying enzymes (LMEs), which include laccases (Lacs), manganese peroxidases (MnPs), versatile peroxidases (VPs), and lignin peroxidases (LiPs), have been considered key factors in lignin degradation by white-rot fungi because they oxidize lignin model compounds and depolymerize synthetic lignin in vitro. However, it remains unclear whether these enzymes are essential/important in the actual degradation of natural lignin in plant cell walls. To address this long-standing issue, we examined the lignin-degrading abilities of multiple mnp/vp/lac mutants of Pleurotus ostreatus. One vp2/vp3/mnp3/mnp6 quadruple-gene mutant was generated from a monokaryotic wild-type strain PC9 using plasmid-based CRISPR/Cas9. Also, two vp2/vp3/mnp2/mnp3/mnp6, two vp2/vp3/mnp3/mnp6/lac2 quintuple-gene mutants, and two vp2/vp3/mnp2/mnp3/mnp6/lac2 sextuple-gene mutants were generated. The lignin-degrading abilities of the sextuple and vp2/vp3/mnp2/mnp3/mnp6 quintuple-gene mutants on the Beech wood sawdust medium reduced drastically, but not so much for those of the vp2/vp3/mnp3/mnp6/lac2 mutants and the quadruple mutant strain. The sextuple-gene mutants also barely degraded lignin in Japanese Cedar wood sawdust and milled rice straw. Thus, this study presented evidence that the LMEs, especially MnPs and VPs, play a crucial role in the degradation of natural lignin by P. ostreatus for the first time.

Physiological function of hydrophobin Vmh3 in lignin degradation by white-rot fungus Pleurotus ostreatus.

Hydrophobins are small-secreted proteins comprising both hydrophobic and hydrophilic parts, that can self-assemble into an amphiphilic film at the air-liquid interface. More than 20 hydrophobin genes have been estimated in the white-rot fungus Pleurotus ostreatus. In our previous studies, three hydrophobin genes were shown to be predominantly expressed under ligninolytic conditions, and only vmh3 was downregulated in both the delignification-deficient mutant Δgat1 and Δhir1 strains. Here, we focused on the function of the hydrophobin Vmh3 to clarify its physiological role in lignin degradation. When the hyphae were observed by transmission electron microscopy, deletion of vmh3 resulted in the disappearance of black aggregates at the interface between the cell wall and outer environment. Deletion of vmh3 resulted in reduced hydrophobicity when 0.2% sodium dodecyl sulfate was dropped onto the mycelial surface. These results suggest that Vmh3 functions on the cell surface and plays a major role in mycelial hydrophobization. Furthermore, the Δvmh3 strain showed a marked delay in lignin degradation on beech wood sawdust medium, while the production of lignin-modifying enzymes was not reduced. This study demonstrated, for the first time, the possible effect of hydrophobin on lignin degradation by a white-rot fungus.

The lignin-degrading abilities of Gelatoporia subvermispora gat1 and pex1 mutants generated via CRISPR/Cas9.

White-rot fungi efficiently degrade wood lignin; however, the mechanisms involved remain largely unknown. Recently, a forward genetics approach to identify several genes in Pleurotus ostreatus (Agaricales) in which mutations cause defects in wood lignin degradation was used. For example, pex1 encodes a peroxisome biogenesis factor and gat1 encodes a putative Agaricomycetes-specific DNA-binding transcription factor. In this study, we examined the effects of single-gene mutations in pex1 or gat1 on wood lignin degradation in another white-rot fungus, Gelatoporia (Ceriporiopsis) subvermispora (Polyporales), to investigate conserved and derived degradation mechanisms in white-rot fungi. G. subvermispora pex1 and gat1 single-gene mutant strains were generated from a monokaryotic wild-type strain, FP-90031-Sp/1, using plasmid-based CRISPR/Cas9. As in P. ostreatus, Gsgat1 mutants were nearly unable to degrade lignin sourced from beech wood sawdust medium (BWS), while Gspex1 mutants exhibited a delay in lignin degradation. We also found that the transcripts of lignin-modifying enzyme-encoding genes, mnp4, mnp5, mnp6, mnp7, and mnp11, which predominantly accumulate in FP-90031-Sp/1 cultured with BWS, were greatly downregulated in Gsgat1 mutants. Taken together, the results suggest that Gat1 may be a conserved regulator of the ligninolytic system of white-rot fungi and that the contribution of peroxisomes to the ligninolytic system may differ among species.

Mild isolation and characterization of surface lignin from hydrothermally pretreated lignocellulosic forestry and agro-industrial waste biomass

Within the biorefining concept, the lignocellulosic biomass wastes can serve as feedstock to produce value-added chemicals and fuels. The sustainable valorization of biomass is based on integrated processes that aim to the utilization of “whole biomass”. To this end, the first step of the efficient biomass pretreatment towards selective isolation of cellulose, hemicellulose and lignin, is of paramount importance. Hydrothermal pretreatment of biomass in neat water is a green and well-established process that enhances hemicellulose removal and recovery in the liquid fraction, affording a cellulose and lignin enriched solid which can be enzymatically hydrolyzed to glucose. A simple and efficient method to facilitate the hydrolysis is the prior extraction of the formed “surface” lignin which hinders the action of enzymes. Within this context, in this work, forestry (beechwood sawdust) and agro-industrial (vine prunings, apricot kernel shells) wastes were hydrothermally pretreated in neat water (at 220 °C, 15 min) and the formed surface lignin was extracted via soxhlet or reflux methods under mild conditions, with the use of green solvents (ethanol, acetone). The delignification degree of the pretreated biomass was in the range of 35–67 wt%, depending on the solvent and biomass feedstock. The recovered surface lignins were of high purity and exhibited similar physicochemical and structural characteristics with the organosolv lignin. The fast pyrolysis of surface lignins afforded bio-oils enriched in alkoxy-phenols and oxy-aromatics. The biomass solids recovered after lignin extraction are enriched in cellulose (70%) and exhibit high relative crystallinity and improved textural properties, thus increasing their potential valorization.

Flow-through reductive catalytic fractionation of beech wood sawdust

The use of flow-through (FT) systems in the lignin-first approach can improve the proficiency of the process. Herein, the reductive catalytic fractionation of beech wood sawdust was conducted in an FT system using 35Ni/NDC pellets.

Examination and optimization of lignocellulolytic activity of Stereum gausapatum F28 on beechwood sawdust supplemented with molasses stillage

This study provides a detailed analysis of the lignocellulolytic activity of a new isolate Stereum gausapatum F28, a Serbian autochthonous fungi, on beechwood sawdust supplemented with cheap waste, sugar beet molasses stillage. Advanced multiple response optimization techniques were applied to improve ligninolytic and reduce hydrolytic activity as a requirement for potential biorefinery use. The applied techniques were supposed to select cultivation conditions that would give manganese peroxidase and laccase activities above 0.84 and 0.12 U g-1 substrate, respectively, and cellulase and xylanase activities below 1.12 and 1.4 U g-1 substrate. The optimal cultivation conditions that met the set requirements included molasses stillage concentration of 10 %, substrate moisture content of 53 %, incubation temperature of 23.5 ?C, and pH 5.2. The research showed that the addition of molasses stillage had a positive effect on enzyme production and that the optimal stillage concentration differed depending on the enzyme type (for laccase it was &lt;5 %, manganese peroxidase ?12 %, cellulase ?21 % and xylanase ?16 %), which should be taken into consideration when optimizing the desired process.

Overexpressing Pleurotus ostreatus rho1b results in transcriptional upregulation of the putative cellulolytic enzyme-encoding genes observed in ccl1 disruptants.

The transcriptional expression pattern of lignocellulolytic enzyme-encoding genes in white-rot fungi differs depending on the culture conditions. Recently, it was shown that 13 putative cellulolytic enzyme-encoding genes were significantly upregulated in most Pleurotus ostreatus ligninolysis-deficient mutant strains on beech wood sawdust medium. However, the mechanisms by which this transcriptional shift is triggered remain unknown. In this study, we identified one mechanism. Our previous study implied that histone H3 N-dimethylation at lysine 4 level possibly affects the shift; therefore, we analysed the expression pattern in the disruptants of P. ostreatus ccl1, which encodes a putative component of the COMPASS complex mediating the methylation. The results showed upregulation of 5 of the 13 cellulolytic enzyme-encoding genes. We also found that rho1b, encoding a putative GTPase regulating signal transduction pathways, was upregulated in the ccl1 disruptants and ligninolysis-deficient strains. Upregulation of at least three of the five cellulolytic enzyme-encoding genes was observed in rho1b-overexpressing strains but not in ccl1/rho1b double-gene disruptants, during the 20-day culture period. These results suggest that Rho1b may be involved in the upregulation of cellulolytic enzyme-encoding genes observed in the ccl1 disruptants. Furthermore, we suggest that Mpk1b, a putative Agaricomycetes-specific mitogen-activated protein kinase, functions downstream of Rho1b.

Application of the Natural Cellulosic Supports for the Treatment of the Industrial Effluents

Wastewater treatment is one of the most important concerns of the research studies in the field of environmental protection. The current study relates to the removal of Crystal violet dye from aqueous solutions through adsorption experiments onto natural cellulosic supports.&#x0D; Various waste materials that have in their composition lignocellulose are adsorbent supports with low costs for the removal of heavy metal ions and dyes from aqueous solutions.&#x0D; The use of chemically modified sawdust to obtain an inexpensive adsorbent for removing the Crystal violet dye (CV) from synthetic aqueous solutions has been presented in this research paper. The influence of some important parameters such as: initial dye concentration, adsorbent dose, contact time and temperature on adsorption efficiency for this dye removal have been studied. The Freundlich, Langmuir and Temkin mathematical models have been used to evaluate the data of Crystal violet adsorption from aqueous media using wood sawdust.&#x0D; The research study shows that beech wood sawdust (Fagus sylvatica), as an agricultural waste which is readily available and relatively inexpensive has the potential to be an applicable alternative adsorbent for Crystal violet dye removal from industrial effluents.&#x0D;

Catalytic fast pyrolysis of beech wood lignin isolated by different biomass (pre)treatment processes: Organosolv, hydrothermal and enzymatic hydrolysis

Lignin is one of the three main structural components of lignocellulosic biomass and is considered as the most abundant source of aromatic and phenolic compounds. Lignin is produced as side-stream in the pulp/paper industry or as residue in the production of second-generation bioethanol. More recently, novel biomass fractionation processes in biorefineries have been developed aiming at the production of high quality/purity lignin towards its more efficient down-stream catalytic conversion to chemicals, monomers, and fuels. Within this context, in this work, we studied the thermal (non-catalytic) and catalytic fast pyrolysis on a Py/GC–MS system and a fixed-bed reactor unit of three types of lignin, all originating from the same biomass (beech wood sawdust) but with different isolation processes: organosolv lignin derived by the organosolv pretreatment/fractionation of biomass, surface extracted lignin derived by the mild Soxhlet extraction (with ethanol or acetone) from the hydrothermally (HT) in pure water pretreated biomass, and the enzymatic hydrolysis lignin derived as a lignin-rich solid residue from the enzymatic hydrolysis of the HT pretreated biomass. Conventional microporous ZSM-5 and mesoporous ZSM-5 zeolites (with intracrystal mesopores, ˜ 9 nm) were used as catalysts in the pyrolysis experiments. Both zeolites were very active in converting the initially produced via thermal pyrolysis methoxy-substituted phenols, benzenes and benzaldehydes mainly towards BTX mono-aromatics, such as 1,3-dimethylbenzene/p- and o-xylenes, toluene and trimethylbenzenes, as well as polycyclic aromatic hydrocarbons (PAHs, mainly naphthalenes). The mesoporous ZSM-5 induced higher dealkoxylation reactivity compared to the microporous ZSM-5 leading to higher concentration of BTX aromatics without a consequent increase of PAHs. The pronounced dealkoxylation/aromatization reactivity of both ZSM-5 zeolites resulted to lower yields (but highly aromatic) organic bio-oils (e.g. from 32−35 wt.% to 15−22 wt.%) compared to thermal pyrolysis, increased non-condensable gases (mainly CO, CO2, methane, ethylene and propylene) and formation of relatively low amounts of coke on catalysts (e.g. 3−5 wt.% on lignin) in addition to thermal pyrolysis char. The observed moderate variations in the characteristics of the three types of beech wood lignin, i.e. molecular weight (GPC), S/G ratio, inter-unit linkages (2D HSQC NMR) and elemental composition, did not induce a systematic/substantial differentiation in the thermal and catalytic fast pyrolysis product yields and composition.

Preparation of Polyurethane Adhesive from Wood Sawdust polyol: Application of Response Surface Methodology for Optimization of Catalyst and Glycerol

Response surface methodology (RSM) was applied to optimize catalyst and glycerol values as suitable additives in polyurethane adhesive production and determine the appropriate gel time. Polyurethane adhesive was prepared using polyol, two types of diisocyanate (PMDI and TDI), catalyst, and glycerol with equal NCO/OH ratio. Polyol was produced using the liquefaction process of Beechwood sawdust with ethylene carbonate solvent at 130°C for 120 minutes under atmospheric pressure. Fourier transform infrared spectroscopy (FTIR) results revealed the existence of the hydroxyl groups in the liquefied wood and confirmed that the liquefied wood sawdust is a source of polyols and a potential chemical stock for the synthesis of polyurethane. Also, the adhesive bonding strength was evaluated by lap shear strength. The result of optimization by response surface methodology showed that catalyst and glycerol values were 0.11% and 1.56% for PU adhesive with PMDI, and catalyst and glycerol values of 0.34% and 3.12%for PU adhesive with TDI were appropriate.

Utilization of various lignocellulosic substrates for Pleurotus ostreatus mushroom cultivation in the manufacturing of polycaprolactone (PCL)-based biocomposite films

The possibility of using beech wood sawdust and hazelnut husk waste generated during the cultivation of Pleurotus ostreatus mushrooms as a filler in polycaprolactone (PCL)-based biocomposite films was investigated. Chemical and physical properties of the PCL biocomposites were determined. The beech sawdust and hazelnut husk were exposed to degradation for 73 d and 78 d, respectively, in slightly acidic environments at a relative humidity of 75%. The degraded materials caused the holocellulose and lignin contents in the PCL biocomposites to decrease, while the cellulose and α-cellulose contents increased. In general, as the lignocellulosic waste content increased, the tensile strength (TS) and elongation at break (EatB) values decreased and the tensile modulus (TM) and water absorption (WA) values increased. It was determined that the PCL biocomposite with the degraded beech sawdust absorbed more water than the composite with the undegraded beech sawdust. On the other hand, the PCL biocomposite with the degraded hazelnut husk absorbed less water than the composite with the raw hazelnut husk.

Comparative transcriptional analyses of Pleurotus ostreatus mutants on beech wood and rice straw shed light on substrate-biased gene regulation.

Distinct wood degraders occupying their preferred habitats have biased enzyme repertoires that are well fitted to their colonized substrates. Pleurotus ostreatus, commonly found on wood, has evolved its own enzyme-producing traits. In our previous study, transcriptional shifts in several P. ostreatus delignification-defective mutants, including Δhir1 and Δgat1 strains, were analyzed, which revealed the downregulation of ligninolytic genes and the upregulation of cellulolytic and xylanolytic genes when compared to their parental strain 20b on beech wood sawdust medium (BWS). In this study, rice straw (RS) was used as an alternative substrate to examine the transcriptional responses of P. ostreatus to distinct substrates. The vp1 gene and a cupredoxin-encoding gene were significantly upregulated in the 20b strain on RS compared with that on BWS, reflecting their distinct regulation patterns. The overall expression level of genes encoding glucuronidases was also higher on RS than on BWS, showing a good correlation with the substrate composition. Transcriptional alterations in the mutants (Δhir1 or Δgat1 versus 20b strain) on RS were similar to those on BWS, and the extracellular lignocellulose-degrading enzyme activities and lignin-degrading ability of the mutants on RS were consistent with the transcriptional alterations of the corresponding enzyme-encoding genes. However, transcripts of specific genes encoding enzymes belonging to the same CAZyme family exhibited distinct alteration patterns in the mutant strains grown on RS compared to those grown on BWS. These findings provide new insights into the molecular mechanisms underlying the transcriptional regulation of lignocellulolytic genes in P. ostreatus.Key Points• P. ostreatus expressed variable enzymatic repertoire-related genes in response to distinct substrates.• A demand to upregulate the cellulolytic genes seems to be present in ligninolysis-deficient mutants.• The regulation of some specific genes probably driven by the demand is dependent on the substrate.

Predtretman lignocelulozne biomase pomoću autohtonih gljiva Srbije

This research examined the potential use of isolated Serbian autochthonous fungi in lignocellulosic biomass pretreatment. Among 12 isolated fungi, the isolates identified as Trametes hirsuta F13 and Stereum gausapatum F28 stood out as ligninolytic enzyme producers and were selected for potential use in the pretreatment of a waste lignocellulosic biomass. An isolate identified as Myrmaecium fulvopruinatum F14 showed high hydrolytic activity, but negligible ligninolytic activity, and it was selected as a potential producer of important industrial hydrolytic enzymes. Further, the breakdown of lignocellulosic waste, beechwood sawdust, by T. hirsuta F13 and S. gausapatum F28 was examined. Both isolates efficiently degraded biomass, but T. hirsuta F13 exhibited greater selectivity (selectivity coefficient of 1.7) than S. gausapatum F28 (1.1). The isolate F13 was considered a better candidate for the pretreatment, and it was selected for further analysis which involved the use of molasses stillage as a supplement to improve the pretreatment.

Targeted disruption of hir1 alters the transcriptional expression pattern of putative lignocellulolytic genes in the white-rot fungus Pleurotus ostreatus

Pleurotus ostreatus is frequently used in molecular genetics and genomic studies on white-rot fungi because various molecular genetic tools and relatively well-annotated genome databases are available. To explore the molecular mechanisms underlying wood lignin degradation by P. ostreatus, we performed mutational analysis of a newly isolated mutant UVRM28 that exhibits decreased lignin-degrading ability on the beech wood sawdust medium. We identified that a mutation in the hir1 gene encoding a putative histone chaperone, which probably plays an important role in DNA replication-independent nucleosome assembly, is responsible for the mutant phenotype. The expression pattern of ligninolytic genes was altered in hir1 disruptants. The most highly expressed gene vp2 was significantly inactivated, whereas the expression of vp1 was remarkably upregulated (300–400 fold) at the transcription level. Conversely, many cellulolytic and xylanolytic genes were upregulated in hir1 disruptants. Chromatin immunoprecipitation analysis suggested that the histone modification status was altered in the 5ʹ-upstream regions of some of the up- and down-regulated lignocellulolytic genes in hir1 disruptants compared with that in the 20b strain. Hence, our data provide new insights into the regulatory mechanisms of lignocellulolytic genes in P. ostreatus.

Influence of drying mode and feed per tooth rate on the fine dust creation in pine and beech sawing on a mini sash gang saw

The experimental results of the study focused on the effect of drying processes of modified air drying and warm air-steam mixture drying of pine and beech wood on the size of sawdust particles created in cutting using PRW-15M sash gang saw, are presented in the paper. Particle size analysis of dry sawdust was performed using two methods—sieving method and laser diffraction analysis. The results showed that the drying process did not affect the general particle size distribution of the sawdust, but the content of very fine dust particles in beech wood sawdust is many times higher when the beech wood was dried in the warm air-steam mixture drying process and sawn at a feed per tooth of 0.105 mm.

Improvement of lignocellulosic biomass conversion by optimization of fungal ligninolytic enzyme activity and molasses stillage supplementation

Fungal treatment of lignocellulose is an ecologically and economically acceptable method. However, it needs improvement to increase the hydrolysis rate. A novel combination of multiple response optimization of ligninolytic activity of Trametes hirsuta F13 and supplementation of the lignocellulosic substrate (beechwood sawdust) with sugar beet molasses stillage was employed to improve and manage the desired type of fungal ligninolytic activity leading to a significant enhancement of biomass saccharification. Obtained optimal cultivation conditions (molasses stillage concentration, 13%; substrate moisture, 63%; and temperature, 25 °C) provided the desired combination of laccase and manganese-dependent peroxidase activity, and maintained the activities longer and at higher values (51.5 ± 3.5 U/L and 91 ± 4.24 U/L, respectively) than the conditions without molasses stillage (21.5 ± 2.12 U/L and 31.5 ± 4.9 U/L, respectively). Enzyme hydrolysis of the substrate treated for 7 days in the presence of molasses stillage released 20.54 ± 0.80 mg/mL fermentable sugars, and 63.58 ± 1.47 mg/mL from 18-days treated substrate, while sugar concentrations released from the substrates pretreated without the supplementation were 16.84 ± 0.30 mg/mL and 53.63 ± 2.16 mg/mL, respectively. The proposed new approach of the lignocellulose supplementation with sugar beet molasses stillage and manipulation and improvement of fungal ligninolytic activity proved to be a promising solution for the enhancement of lignocellulose bioconversion.