Effect of Lentinus sajor-caju on the chemical composition and antioxidant activity of highland barley straw under solid-state fermentation.

Introduction: The efficient utilization of straw resources as animal feed has gained considerable attention. The objective of this study was to evaluate whether Lentinus sajor-caju treatment alters the chemical composition and antioxidant activity of highland barley straw and enhances its functional value as a ruminant feed. Methods: The chemical composition, antioxidant capacity, and metabolomic profile of highland barley straw were determined after 21 days of solid-state fermentation with L. sajor-caju at 25°C. The differential metabolites between fermented and unfermented highland barley straw were identified by LC-MS and the relationship between the identified metabolites and antioxidant capacity was elucidated. Results: The results showed that, compared with untreated highland barley straw, the crude protein and ether extract contents were higher (51.55 and 76.43%, respectively) in highland barley straw after 21 days of incubation with L. sajor-caju, whereas the hemicellulose, cellulose, and acid detergent lignin contents were lower (2.48, 25.08, and 45%, respectively). The total antioxidant capacity was significantly higher in L. sajor-caju-treated than in untreated highland barley straw. In total, 600 differential metabolites (301 upregulated and 299 downregulated) were identified between L. sajor-caju-fermented and unfermented highland barley straw. Correlation analysis results showed that Fe2+ scavenging and total phenolic content were strongly correlated with total antioxidant capacity. Meanwhile, the differential flavonoid metabolites between fermented and unfermented highland barley straw were primarily associated with antioxidant activity, with kaempferol 3-xylosylglucoside, isoginkgetin, and rhoifolin being the most representative. Conclusion: Thus, this study demonstrates that L. sajor-caju could enhance the functional value of highland barley straw, showing the potential of L. sajor-caju for improving the utilization of agricultural straws in ruminants.

Synergistic adsorption and photocatalysis study of TiO2 and activated carbon composite.

The discharge of organic pollutants by the textile and dyeing industries presents an escalating threat to aquatic environments, necessitating the development of effective remediation strategies. This study introduces the utilization of graphite-like structured activated carbon (AC), derived from highland barley straw-a biomass unique to the Plateau regions of China, including Tibet, Qinghai, and Gansu-as a support material for the TiO2 catalyst. TiO2/AC composites with different TiO2 loadings were synthesized by ultrasonic impregnation. The TiO2/AC composites were found to be polycrystalline materials composed of anatase and rutile phases. The TiO2 nanoparticles are well-dispersed over the surface of the AC. The photocatalytic activity of these composites was evaluated through their capacity to degrade a methylene blue (MB) solution upon irradiation. It was observed that the inclusion of TiO2 increases the number of adsorption sites and active sites for methylene blue, with the photocatalytic activity being notably higher at a 3-wt% TiO2 loading, achieving a remarkable 99.6 % degradation efficiency for 100 mg/L MB within 100 min. The experimental kinetic data for the photocatalytic process follow the pseudo-first-order kinetic model. Furthermore, TiO2/AC retains high photocatalytic activity after five reaction cycles. This research provides valuable insights into the application of biomass-derived materials for the purification of water, offering a sustainable solution to both pollution and agricultural waste challenges in Plateau areas of China.

Solid-State Fermentation with White Rot Fungi (Pleurotus Species) Improves the Chemical Composition of Highland Barley Straw as a Ruminant Feed and Enhances In Vitro Rumen Digestibility.

Lignin degradation is important for enhancing the digestibility and improving the nutritive quality of ruminant feeds. White rot fungi are well known for their bioconversion of lignocellulosic biomass. The objective of this paper was to evaluate whether Lentinus sajor-caju, Pleurotus ostreatus, Phyllotopsis rhodophylla, Pleurotus djamor, Pleurotus eryngii, and Pleurotus citrinopileatus treatments altered the chemical compositions of highland barley straw constituents and enhanced their nutritional value as a ruminant feed. All white rot fungi significantly increased the relative crude protein (CP), ethyl ether extract (EE), starch, soluble protein (SP), and non-protein nitrogen (NPN) contents but decreased the ash, neutral detergent fiber (NDF), acid detergent fiber (ADF), acid detergent lignin (ADL), and acid detergent insoluble protein (ADFIP) contents. In addition, L. sajor-caju treatment increased (p < 0.001) the levels of PA, PB2, PB3, CA, CB1, CB2, and CNSC, but reduced (p < 0.001) the PC and CC in the solid-state fermentation of highland barley straw. Maximum ligninlysis (50.19%) was optimally produced in the presence of 1.53% glucose and 2.29% urea at 22.72 ℃. The in vitro dry matter digestibility and total volatile fatty acid concentrations of fermented highland barley straw, as well as the fermentability, were optimized and improved with L. sajor-caju, which degraded the lignocellulose and improved the nutritional value of highland barley straw as a ruminant feed.

Hydrogen Production from Barley Straw and Miscanthus by the Hyperthermophilic Bacterium, Cadicellulosirupter bescii.

This work aimed to evaluate the feasibility of biohydrogen production from Barley Straw and Miscanthus. The primary obstacle in plant biomass decomposition is the recalcitrance of the biomass itself. Plant cell walls consist of cellulose, hemicellulose, and lignin, which make the plant robust to decomposition. However, the hyperthermophilic bacterium, Caldicellulosiruptor bescii, can efficiently utilize lignocellulosic feedstocks (Barley Straw and Miscanthus) for energy production, and C. bescii can now be metabolically engineered or isolated to produce more hydrogen and other biochemicals. In the present study, two strains, C. bescii JWCB001 (wild-type) and JWCB018 (ΔpyrFA Δldh ΔcbeI), were tested for their ability to increase hydrogen production from Barley Straw and Miscanthus. The JWCB018 resulted in a redirection of carbon and electron (carried by NADH) flow from lactate production to acetate and hydrogen production. JWCB018 produced ~54% and 63% more acetate and hydrogen from Barley Straw, respectively than its wild-type counterpart, JWCB001. Also, 25% more hydrogen from Miscanthus was obtained by the JWCB018 strain with 33% more acetate relative to JWCB001. It was supported that the engineered C. bescii, such as the JWCB018, can be a parental strain to get more hydrogen and other biochemicals from various biomass.

Mechanistic insights into the digestion of complex dietary fibre by the rumen microbiota using combinatorial high-resolution glycomics and transcriptomic analyses.

There is a knowledge gap regarding the factors that impede the ruminal digestion of plant cell walls or if rumen microbiota possess the functional activities to overcome these constraints. Innovative experimental methods were adopted to provide a high-resolution understanding of plant cell wall chemistries, identify higher-order structures that resist microbial digestion, and determine how they interact with the functional activities of the rumen microbiota. We characterized the total tract indigestible residue (TTIR) from cattle fed a low-quality straw diet using two comparative glycomic approaches: ELISA-based glycome profiling and total cell wall glycosidic linkage analysis. We successfully detected numerous and diverse cell wall glycan epitopes in barley straw (BS) and TTIR and determined their relative abundance pre- and post-total tract digestion. Of these, xyloglucans and heteroxylans were of higher abundance in TTIR. To determine if the rumen microbiota can further saccharify the residual plant polysaccharides within TTIR, rumen microbiota from cattle fed a diet containing BS were incubated with BS and TTIR ex vivo in batch cultures. Transcripts coding for carbohydrate-active enzymes (CAZymes) were identified and characterized for their contribution to cell wall digestion based on glycomic analyses, comparative gene expression profiles, and associated CAZyme families. High-resolution phylogenetic fingerprinting of these sequences encoded CAZymes with activities predicted to cleave the primary linkages within heteroxylan and arabinan. This experimental platform provides unprecedented precision in the understanding of forage structure and digestibility, which can be extended to other feed-host systems and inform next-generation solutions to improve the performance of ruminants fed low-quality forages.

In Vitro Assessment of the Prebiotic Potential of Xylooligosaccharides from Barley Straw.

Barley straw was subjected to hydrothermal pretreatment (steam explosion) processing to evaluate its potential as a raw material to produce xylooligosaccharides (XOS) suitable for use as a prebiotic. The steam explosion pretreatment generated a liquid fraction containing solubilised hemicellulose. This fraction was purified using gel permeation chromatography to obtain a fraction rich in XOS DP2-DP6. The sample was characterised through analytical techniques such as HPAEC-PAD, FTIR and MALDI-TOF-MS. The prebiotic activity was evaluated using in vitro fermentation in human faecal cultures through the quantification of short-chain fatty acid (SCFA) and lactate production, the evolution of the pH and the consumption of carbon sources. The total SCFA production at the end of fermentation (30 h) was 90.1 mM. Positive significant differences between the amount of XOS from barley straw and fructooligosaccharides after incubation were observed.

Influence of cavitation, pelleting, extrusion and torrefaction petreatments on anaerobic biodegradability of barley straw and vine shoots.

When dealing with lignocellulosic biomass in anaerobic digestion, a pretreatment stage is always required to open the structure of the material, facilitating its degradation. Numerous methods have been developed to pretreat lignocellulosic biomass. Four of them: cavitation, pelleting, extrusion and torrefaction have been comparatively studied in this paper as ways to improve the production of methane by anaerobic digestion of two different feedstocks: barley straw and vine shoots. Additionally, how the selected pretreatments and the nature of the feedstock influence the formation of individual volatile fatty acids was examined. Cavitation was revealed as the most efficient pretreatment, increasing 240% and 360% the methane production for barley straw and vine shoots, respectively, although in absolute terms, barley straw has higher production rate and yield than vine shoots. Torrefaction carried out at 180 °C increased methane production, 81% for straw and 25% for vine shoots, while the process at higher temperatures (220 °C) negatively affected biogas production from both feedstocks. Finally, volatile fatty acids accumulation seems to neutralize any potential positive effects of densification pretreatments.

Complete Substitution of Clover Hay and its Impact on Performance of Barki Sheep in Arid and Semi-arid Regions.

BACKGROUND AND OBJECTIVE: Barley straw is a farm by product that may be useful in animal feeding but its use is limited due to its low nutritional value and high fiber content. Therefore, this study aimed to improving the nutritive value of barley straw by bacterial treatment with Condensed Molasses Soluble (CMS) in the ration of Barki lambs. MATERIALS AND METHODS: In Experiment 1, 12 Barki rams, which were divided into 3 equal groups (4 each), were given rations which consisted of 50% CFM+50%, either clover hay (C) or bacterial treated barley straw (T1) or T1+ CMS (T2). In Experiment 2, 10 Barki lambs were divided into 2 groups, one (C) and T2 as descripted in Experiment 1. Data analyzed by using one way ANOVA model. RESULTS: Digestibility of Dry Matter (DM), Organic Matter (OM), Crude Fiber (CF) and Ether Extract (EE) were insignificantly affected among all rations, while Digestible Crude Protein (DCP) recorded a higher value in T2 compared to T1. Changes in Total Digestible Nutrients (TDN) values were insignificant. Values of DCP were higher for T2 compared to C and T1. The Body Weight Gain (BWG) and Feed Conversion Ratio (FCR) of growing lambs didn't statistically differ among the groups. CONCLUSION: Finally, complete replacement of clover hay by treated barley straw plus CMS may be considered good feeding strategies in the feeding of Barki lambs in Arid and semi-arid regions.

Characteristics and unique sources of polycyclic aromatic hydrocarbons and nitro-polycyclic aromatic hydrocarbons in PM2.5 at a highland background site in northwestern China☆.

Polycyclic aromatic hydrocarbons (PAHs) and nitro-PAHs (NPAHs) in PM2.5 were first observed at a background site (Yuzhong site: YZ site) in the northwestern highlands of China in five seasonal campaigns. Compared with major northwestern cities, PAHs and NPAHs at the YZ site were at a lower level but showed consistent seasonal differences. The PAH and NPAH concentrations peaked in the winter campaigns, which were 36.11 ± 6.54 ng/m3 and 418.11 ± 123.55 pg/m3, respectively, in winter campaign 1 and 28.97 ± 10.07 ng/m3 and 226.89 ± 133.54 pg/m3, respectively, in winter campaign 2. These values were approximately a dozen times larger those in other campaigns. The diagnostic ratios indicate that vehicle emissions were the primary source of the PAHs throughout the five campaigns, and coal and biomass combustion also contributed during the winter, summer, and fall campaigns. Among NPAHs, 2-nitrofluoranthene and 2-nitropyrene were generated through OH radical-initiated reactions during atmospheric transport, while 1-nitropyrene came from combustion sources. There is an observation worth pondering, which is that the ratio between pyrene and fluoranthene increased abnormally in the spring and fall campaigns, which is presumably caused by the burning of Tibetan barley straw in the northwestern highlands. The backward trajectories over Tibetan areas in Qinghai and southwestern Gansu are consistent with this hypothesis. In addition, this study reported for the first time that the burning of Tibetan barley straw has become a seasonal contributor to air pollution in northwestern China and is participating in the atmospheric transport of air pollutants driven by the monsoon in East Asia, which urgently requires further research.

Effect of supplements on feed utilization of Tigray Highland sheep fed a basal diet of barley straw.

The feeding experiment was conducted at Ofla district, southern Tigray, North Ethiopia, to investigate feed utilization, diet digestibility, and its economic feasibility under different supplementation options. Twenty-four yearling intact growing lambs were used in randomized complete block design with three treatments and eight replications. The treatment diets were T1 (molasses 15% + wheat bran 48% + cotton seed cake 35%), T2 (maize grain 20% + wheat bran 43% + noug seed cake 35%) and T3 (maize grain 20% + wheat bran 33% + dried brewers' grain 45%). The diet was formulated as iso-nitrogenous and each animal has taken daily nutrient requirements according to their body weight. The collected data were analyzed using SAS (2007) statistical software. Economic analysis was done using standard partial budget analysis guidelines of CIMMIT (1988). The crude protein contents of the three treatment rations were comparable across treatments. There was a higher total dry matter intake recorded in animals fed on T3 as compared to the other treatments. The barley straw intake was higher and significant (P < 0.05) for the animals' group in T3 than that of T1 and T2. Animals fed with T3 had significantly (P < 0.05) higher organic matter intake as compared to those fed with T1 and T2. However, there was no significant (P > 0.05) differences in crude protein intake between treatments. The apparent digestibility of dry matter, organic matter, and crude protein had no significant differences (P > 0.05) between treatments. Similarly, the apparent digestibility of neutral detergent fiber and acid detergent fiber was not significantly different (P > 0.05) among T1, T2, and T3. In general, animals that fed on T3 achieved better feed utilization and economic return as compared to the others.

Effective pretreatment of lignocellulosic co-substrates using barley straw-adapted microbial consortia to enhanced biomethanation by anaerobic digestion.

Microbial pretreatments have been identified as a compatible and sustainable process with anaerobic digestion compared to energy-intensive physicochemical pretreatments. In this study, barley straw and hay co-substrate was pretreated with a microaerobic barley straw-adapted microbial (BSAM) consortium prior to anaerobic digestion. The improved digestibility was investigated through 16S rRNA gene sequencing, microbial counts and C:N ratios. BSAM pretreatment resulted in 15.2 L kg-1 TS of methane yield after 35 days, almost 40 times more than the control. The methane content in total biogas produced were 58% (v/v) and 10% (v/v) in BSAM and control, respectively. This research demonstrated that BSAM-based pretreatment significantly increased the digestibility and surface area of the lignocellulosic material and considerably enhanced biomethanation. This study generates new potential bio-research opportunities in the emerging field of lignocellulosic anaerobic digestion-biorefineries.

Feasibility of Barley Straw Fibers as Reinforcement in Fully Biobased Polyethylene Composites: Macro and Micro Mechanics of the Flexural Strength.

Awareness on deforestation, forest degradation, and its impact on biodiversity and global warming, is giving rise to the use of alternative fiber sources in replacement of wood feedstock for some applications such as composite materials and energy production. In this category, barley straw is an important agricultural crop, due to its abundance and availability. In the current investigation, the residue was submitted to thermomechanical process for fiber extraction and individualization. The high content of holocellulose combined with their relatively high aspect ratio inspires the potential use of these fibers as reinforcement in plastic composites. Therefore, fully biobased composites were fabricated using barley fibers and a biobased polyethylene (BioPE) as polymer matrix. BioPE is completely biobased and 100% recyclable. As for material performance, the flexural properties of the materials were studied. A good dispersion of the reinforcement inside the plastic was achieved contributing to the elevate increments in the flexural strength. At a 45 wt.% of reinforcement, an increment in the flexural strength of about 147% was attained. The mean contribution of the fibers to the flexural strength was assessed by means of a fiber flexural strength factor, reaching a value of 91.4. The micromechanical analysis allowed the prediction of the intrinsic flexural strength of the fibers, arriving up to around 700 MPa, and coupling factors between 0.18 and 0.19, which are in line with other natural fiber composites. Overall, the investigation brightness on the potential use of barley straw residues as reinforcement in fully biobased polymer composites.

Use of barley straw as a support for the production of conidiospores of Trichoderma harzianum.

In this work was to evaluate the conidiospores production of Trichoderma harzianum using barley straw as substrate. Four growth conditions were used; washed and unwashed barley straw and washed and unwashed barley straw supplemented with mineral salts. The highest spore production was observed when washed barley straw supplemented with mineral salts with 1.56 × 1010 conidiospores/gram of dry matter (gdm) at 216 h of cultivation was used. The effect of substrate moisture on spore production was studied, three initial moisture levels of the substrate were tested and it was observed that a humidity of 80 % of the substrate improves the production of conidiospores reaching a concentration of 2.35 × 1010 conidiospores/gdm at 136 h. Finally, conidiospores viability was evaluated for 12 months by keeping them on the conidia and substrate, and viability of 71 % of the conidiospores was observed, so this maintenance method is an excellent means of conserving the conidiospores viability.

Enzymatic hydrolysis of barley straw for biofuel industry using a novel strain of Trametes villosa from Paranaense rainforest.

Agricultural practices generate lignocellulosic waste that can be bioconverted by fungi to generate value-added products such as biofuels. In this context, fungal enzymes are presented as an alternative for their use in the hydrolysis of cellulose to sugars that can be fermented to ethanol. The aim of this work was to characterize LBM 033 strain and to analyze its efficiency in the hydrolysis of cellulosic substrates, including barley straw. LBM 033 strain was identified as Trametes villosa by molecular techniques, through the use of the ITS and rbp2 markers and the construction of phylogenetic trees. The cell-free supernatant of T. villosa LBM 033 showed high titers of hydrolytic enzymatic activities, necessary for the hydrolysis of the holocellulosic substrates, hydrolyzing pure cellulose to cellobiose and glucose and also degraded the polysaccharides contained in barley straw to short soluble oligosaccharides. These results indicate that macro fungi from tropical soil environments, such as T. villosa LBM 033 can be a valuable resource for in-house, cost effective production of enzymes that can be applied in the hydrolysis stage, which could reduce the total cost of bioethanol production.

Biobased Composites from Biobased-Polyethylene and Barley Thermomechanical Fibers: Micromechanics of Composites.

The cultivation of cereals like rye, barley, oats, or wheat generates large quantities of agroforestry residues, which reaches values of around 2066 million metric tons/year. Barley straw alone represents 53%. In this work, barley straw is recommended for the production of composite materials in order to add value to this agricultural waste. First of all, thermomechanical (TMP) fibers from barley straw are produced and later used to reinforce bio-polyethylene (BioPE) matrix. TMP barley fibers were chemically and morphologically characterized. Later, composites with optimal amounts of coupling agent and fiber content ranging from 15 to 45 wt % were prepared. The mechanical results showed the strengthening and stiffening capacity of the TMP barley fibers. Finally, a micromechanical analysis is applied to evaluate the quality of the interface and to distinguish how the interface and the fiber morphology contributes to the final properties of these composite materials.

Image Analysis of Sewage Sludge and Barley Straw as Biological Materials Composted under Different Conditions.

Composting is one of the most important methods of sewage sludge management. This paper describes the methods of computer image analysis used for objective comparison of the appearance of composted materials under diverse conditions in terms of size and thermal insulation of the composting chambers. The research material was a mixture of sewage sludge and barley straw. The composting process was performed under strictly controlled laboratory conditions, using 10 composting chambers with five different volumes. In half of them additional thermal insulation was used, while in the others no insulation was applied. A proper composting process run was observed only in the three chambers with the largest volume and with additional thermal insulation. The images of the materials were subjected to a wide analysis, wherein the values of 17 parameters regarding color and texture were estimated. Significant differences were observed in the appearances between materials obtained during the properly running composting processes and those obtained in the chambers of insufficient size and thermal insulation. The values of the considered parameters determined for images of the composted material under normal and abnormal conditions were significantly different from each other. Thus, these parameters may be used as indicators of a correctly conducted composting process. In the cases of 15 parameters, the values of these differences exceeded 10%, and in the cases of 10 parameters 50%, while in the cases of three parameters as much as 100%.

Bioconversion of barley straw lignin into biodiesel using Rhodococcus sp. YHY01.

Rhodococcus sp. YHY01 was studied to utilize various lignin derived aromatic compounds. It was able to utilize p-coumaric acid, cresol, and 2,6 dimethoxyphenol and resulted in biomass production i.e. 0.38 g dcw/L, 0.25 g dcw/L and 0.1 g dcw/L, and lipid accumulation i.e. 49%, 40%, 30%, respectively. The half maximal inhibitory concentration (IC50) value for p-coumaric acid (13.4 mM), cresol (7.9 mM), and 2,6 dimethoxyphenol (3.4 mM) was analyzed. Dimethyl sulfoxide (DMSO) solubilized barley straw lignin fraction was used as a carbon source for Rhodococcus sp. YHY01 and resulted in 0.130 g dcw/L with 39% w/w lipid accumulation. Major fatty acids were palmitic acid (C16:0) 51.87%, palmitoleic acid (C16:l) 14.90%, and oleic acid (C18:1) 13.76%, respectively. Properties of biodiesel produced from barley straw lignin were as iodine value (IV) 27.25, cetane number (CN) 65.57, cold filter plugging point (CFPP) 14.36, viscosity (υ) 3.81, and density (ρ) 0.86.

Valorization of egg shell as a detoxifying and buffering agent for efficient polymalic acid production by Aureobasidium pullulans NRRL Y-2311-1 from barley straw hydrolysate.

Stepwise formulation of a versatile and cost-effective medium based on barley straw hydrolysate and egg shell for efficient polymalic acid production by A. pullulans NRRL Y-2311-1 was carried out for the first time. The strain did not grow and produce polymalic acid when dilute acid pretreated barley straw hydrolysate (total fermentable sugars: 94.60 g/L; furfural: 1.01 g/L; hydroxymethylfurfural: 0.55 g/L; acetic acid: 5.06 g/L) was directly used in medium formulation without detoxification (e.g. charcoal pretreatment). When CaCO3 in the medium formulation was substituted with egg shell powder, efficient production of polymalic acid was achieved without a detoxification step. Utilization of 40 g/L of egg shell powder led to 43.54 g polymalic acid production per L with the productivity of 0.30 g/L/h and yield of 0.48 g/g. The bioprocess strategy used in this study can also be utilized for mass production of several other industrially important microbial organic acids and biomaterials.

Establishment of optimal barley straw biochar application conditions for rice cultivation in a paddy field.

This study was conducted to establish the optimal application conditions of barley straw biochar (BC) for rice cultivation and to determine the effects of combined application of BC and inorganic fertilizer (IF) on rice cultivation in a paddy field. Based on the characteristics of rice growth in pot-based experiments, the selected optimal application conditions of BC were application of 20 ton ha-1 at 14 days before rice transplanting. The effects of BC application on rice cultivation in a paddy field when using those conditions were then evaluated. Each treatment was separated by a control (Cn), IF, BC, and combined BC + IF treatments, respectively. The rice yields in the BC + IF treatment were 38.6, 21.7, and 24.5% greater than those in the Cn, IF, and BC treatments, respectively. In addition, yield components of rice were significantly improved in the BC + IF treatment relative to the other treatments. Following rice harvest, soil status was improved, showing greater soil aggregation stability, decreased bulk density, and increased porosity in the BC-treated areas compared to those in the Cn- and IF-treated areas. At the time of rice harvesting, soil chemical properties such as pH, EC, SOC, TN, Avail. P2O5, and CEC in the BC-treated areas were improved over those in other areas. The results of this study indicate that using BC as a soil amendment is effective at improving rice cultivation and can benefit the soil environment.

Saccharification efficiencies of multi-enzyme complexes produced by aerobic fungi.

In the present study, we have characterized high molecular weight multi-enzyme complexes in two commercial enzymes produced by Trichoderma reesei (Spezyme CP) and Penicillium funiculosum (Accellerase XC). We successfully identified 146-1000 kDa complexes using Blue native polyacrylamide gel electrophoresis (BN-PAGE) to fractionate the protein profile in both preparations. Identified complexes dissociated into lower molecular weight constituents when loaded on SDS PAGE. Unfolding of the secondary structure of multi-enzyme complexes with trimethylamine (pH >10) suggested that they were not a result of unspecific protein aggregation. Cellulase (CMCase) profiles of extracts of BN-PAGE fractionated protein bands confirmed cellulase activity within the multi-enzyme complexes. A microassay was used to identify protein bands that promoted high levels of glucose release from barley straw. Those with high saccharification yield were subjected to LC-MS analysis to identify the principal enzymatic activities responsible. The results suggest that secretion of proteins by aerobic fungi leads to the formation of high molecular weight multi-enzyme complexes that display activity against carboxymethyl cellulose and barley straw.