Characteristics of degraded cellulose obtained from steam-exploded wheat straw

The isolation of cellulose from wheat straw was studied using a two-stage process based on steam explosion pre-treatment followed by alkaline peroxide post-treatment. Straw was steamed at 200 degrees C, 15 bar for 10 and 33 min, and 220 degrees C, 22 bar for 3, 5 and 8 min with a solid to liquid ratio of 2:1 (w/w) and 220 degrees C, 22 bar for 5 min with a solid to liquid ratio of 10:1, respectively. The steamed straw was washed with hot water to yield a solution rich in hemicelluloses-derived mono- and oligosaccharides and gave 61.3%, 60.2%, 66.2%, 63.1%, 60.3% and 61.3% of the straw residue, respectively. The washed fibre was delignified and bleached by 2% H2O2 at 50 degrees C for 5 h under pH 11.5, which yielded 34.9%, 32.6%, 40.0%, 36.9%, 30.9% and 36.1% (% dry wheat straw) of the cellulose preparation, respectively. The optimum cellulose yield (40.0%) was obtained when the steam explosion pre-treatment was performed at 220 degrees C, 22 bar for 3 min with a solid to liquid ratio of 2:1, in which the cellulose fraction obtained had a viscosity average degree of polymerisation of 587 and contained 14.6% hemicelluloses and 1.2% klason lignin. The steam explosion pre-treatment led to a significant loss in hemicelluloses and alkaline peroxide post-treatment resulted in substantial dissolution of lignin and an increase in cellulose crystallinity. The six isolated cellulose samples were further characterised by FT-IR and 13C-CP/MAS NMR spectroscopy and thermal analysis.     

Effects of rhamnolipid on the cellulase and xylanase in hydrolysis of wheat straw

The effects of biosurfactant rhamnolipid (RL) and chemical surfactant Triton X-100 on the production of cellulases and xylanase from Penicillium expansum (P. expansum) in untreated, acid- and alkali-pretreated wheat straw submerged fermentations were studied, and the influences on the activity and stability of Cellulase R-10 were also investigated. The results showed that RL and Triton X-100 enhanced the activities of cellulases and xylanase to different extents and the stimulatory effects of RL were superior to those of Triton X-100. During the peak enzyme production phase, RL (60 RE mg/l) increased cellulases activities by 25.5-102.9%, in which the raise of the same enzyme in acid-pretreated straw broths was the most. It was found that the reducing sugars by hydrolyzing wheat straw with Cellulase R-100 were not visibly increased after adding RL. However, it distinctly protected Cellulase R-10 from degradation or inactivation, keeping the reducing sugars yield at about 17%.     

Laccase detoxification of steam-exploded wheat straw for second generation bioethanol

In this work we compared the efficiency of a laccase treatment performed on steam-exploded wheat straw pretreated under soft conditions (water impregnation) or harsh conditions (impregnation with diluted acid). The effect of several enzymatic treatment parameters (pH, time of incubation, laccase origin and loading) was analysed. The results obtained indicated that severity conditions applied during steam explosion have an influence on the efficiency of detoxification. A reduction of the toxic effect of phenolic compounds by laccase polymerization of free phenols was demonstrated. Laccase treatment of steam-exploded wheat straw reduced sugar recovery after enzymatic hydrolysis, and it should be better performed after hydrolysis with cellulases. The fermentability of hydrolysates was greatly improved by the laccase treatment in all the samples. Our results demonstrate the action of phenolic compounds as fermentation inhibitors, and the advantages of a laccase treatment to increase the ethanol production from steam-exploded wheat straw.     

Kinetic analysis of enzymatic hydrolysis of crystalline cellulose by cellobiohydrolase using an amperometric biosensor

An amperometric biosensor for the detection of cellobiose has been introduced to study the kinetics of enzymatic hydrolysis of crystalline cellulose by cellobiohydrolase. By use of a sensor in which pyrroloquinoline quinone-dependent glucose dehydrogenase was immobilized on the surface of electrode, direct and continuous observation of the hydrolysis can be achieved even in a thick cellulose suspension. The steady-state rate of the hydrolysis increased with increasing concentrations of the enzyme to approach a saturation value and was proportional to the amount of the substrate. The experimental results can be explained well by the rate equations derived from a three-step mechanism consisting of the adsorption of the free enzyme onto the surface of the substrate, the reaction of the adsorbed enzyme with the substrate, and the liberation of the product. The catalytic constant of the adsorbed enzyme was determined to be 0.044+/-0.011s(-1).     

Superfine grinding of steam-exploded rice straw and its enzymatic hydrolysis

The combination of low severity steam explosion and superfine grinding has been studied with respect to side products generation and enzymatic hydrolysis efficiency. Chemical compositions, fiber characteristics and composed cells contents in the superfine ground product and the ground residue particles produced by superfine grinding were also studied. At the determined parameters using FJM-200 fluidized bed opposed jet mill, 78% superfine ground steam-exploded rice straw (SERS) products with the mean fiber length of 60 μm were obtained, the particles yield was 179% higher than that from the native rice straw (RS). Enzymatic hydrolysis, chemical composition, fiber characteristics and composed cells proportion of the superfine ground SERS product and the ground residue all show great differences. The difference in enzymatic hydrolysis and structural properties indicates that superfine grinding is a good way to fractionate SERS into easily bio-converted part and difficultly hydrolysed part.     

蒸汽爆破玉米秸秆酶解动力学

为了掌握蒸汽爆破玉米秸秆的酶解特性,研究了不同底物浓度、酶浓度、温度对反应速率的影响。运用米氏方程对酶解动力学过程进行拟合,结果表明,纤维素酶对该玉米秸秆的水解反应在反应前3 h符合一级反应,可用米氏方程对其进行拟合。在转速为120 r/min、酶浓度为1.2 FPU/mL、pH 5.0、温度为45 ℃时米氏常数Km为11.71 g/L,最大反应速率Vm为1.5 g/(L·h)。确立了包括底物浓度、酶浓度、温度在内的酶解动力学模型,该模型适合温度为30 ℃~50 ℃。     

Improving enzymatic hydrolysis of wheat straw using ionic liquid 1-ethyl-3-methyl imidazolium diethyl phosphate pretreatment

This study aims to establish a cellulose pretreatment process using ionic liquids (ILs) for efficient enzymatic hydrolysis. The IL 1-ethyl-3-methyl imidazolium diethyl phosphate ([EMIM]DEP) was selected in view of its low viscous and the potential of accelerating enzymatic hydrolysis, and it could be recyclable. The yield of reducing sugars from wheat straw pretreated with this IL at 130 °C for 30 min reached 54.8% after being enzymatically hydrolyzed for 12 h. Wheat straw regenerated were hydrolyzed more easily than that treated with water. The fermentability of the hydrolyzates, obtained after enzymatic saccharification of the regenerated wheat straw, was evaluated using Saccharomyces cerevisiae. This microbe could ferment glucose efficiently, and the ethanol production was 0.43 g/g glucose within 26 h. In conclusion, the IL [EMIM]DEP shows promise as pretreatment solvent for wheat straw, although its cost should be reduced and in-depth exploration of this subject is needed.     

Comparison of the rates of enzymatic hydrolysis of pretreated rice straw and bagasse with celluloses

The rates of enzymatic hydrolysis of pretreated rice straw and bagasse have been studied and compared with the hydrolysis rates of microcrystalline cellulose powder (MCCP) and Solka Floc. The effects of particle size reduction and enzyme loading on the rates of hydrolysis of rice straw and bagasse were also studied. It was found that the rates of hydrolysis of pretreated rice straw and bagasse are much higher than that of MCCP and Solka Floc. For both rice straw and bagasse, particle size reduction had very little effect in enhancing the rate of hydrolysis. Lignin present at <10% did not seem to hinder the accessibility of the enzyme to the cellulose surface. An enzyme loading > 40 Ug^?1 had no effect on the hydrolysis rate of rice straw or bagasse.     

Studies on biological decomposition of wheat straw

Summary The incorporation of undecomposed wheat straw in the soil along-with the micro-organisms favourably increased the yield of groundnut crop. An increase of 37 per cent in yield was recorded when wheat straw was inoculated withPenicillium digitatum and the C:P ratio was adjusted to 65. Inoculated treatments of narrower C:P ratio gave a higher yield than wider C:P ratio treatments inoculated with the same cultures. An increase in nitrogen uptake by groundnut plants was recorded due to incorporation of straw alongwith the micro-organisms in soil. The organic carbon and nitrogen content of the soil increased with all the treatments except control. The highest increase in organic carbon and nitrogen of the soil was observed with a treatment of wheat straw of 65 C:P ratio inoculated withS. coccosporum. The yield of wheat crop after groundnut was significantly more with several treatments than control plots. The highest increase of 79 per cent in grain yield of wheat was observed in the plots previouslq received with wheat straw of 200 C:P ratio.This paper is based on the data presented at IV Southern Regional Conference on Microbial Inoculants, held at Parbhani during 3–4 July 1978.     

Enzymatic hydrolysis of wheat straw: Kinetic analysis

Pretreated wheat straw was enzymatically hydrolyzed in 250-ml flasks using the culture filtrate of Trichoderma reesei QM-9414. The influence of the initial enzymatic activity in the liquid phase was studied. The unreacted core model was used to analyze the experimental data obtained at 40, 46 and 50°C. The model adequately describes the data for hydrolysis times lower than 10 h.     

Economic evaluation of enzymatic hydrolysis of phenol-pretreated wheat straw

Experimental data from the enzymatic hydrolysis of phenol-pretreated Swedish wheat straw have been used to evaluate the cost fractions of capital and utility, enzyme, and raw material. Two different raw material prices and varying enzyme prices have been used. The evaluation is based on an empirical model for the enzymatic hydrolysis and a computer program where utility and equipment, enzyme, and raw material prices can be varied. The optimal residence time for the enzymatic hydrolysis is in the range of 70-110 h. A fed-batch procedure with substrate concentrations higher than 10% oven-dried material (ODM) and enzyme concentrations in the range (6-10) . 10(6) FPU/ton ODM should be used.     

Reducing agents improve enzymatic hydrolysis of cellulosic substrates in the presence of pretreatment liquid

Enzymatic hydrolysis of pretreated lignocellulosic substrates has emerged as an interesting option to produce sugars that can be converted to liquid biofuels and other commodities using microbial biocatalysts. Lignocellulosic substrates are pretreated to make them more accessible to cellulolytic enzymes, but the pretreatment liquid partially inhibits subsequent enzymatic hydrolysis. The presence of pretreatment liquid from Norway spruce resulted in a 63% decrease in the enzymatic saccharification of Avicel compared to when the reaction was performed in a buffered aqueous solution. The addition of 15 mM of a reducing agent (hydrogen sulfite, dithionite, or dithiothreitol) to reaction mixtures with the pretreatment liquid resulted in up to 54% improvement of the saccharification efficiency. When the reducing agents were added to reaction mixtures without pretreatment liquid, there was a 13-39% decrease in saccharification efficiency. In the presence of pretreatment liquid, the addition of 15 mM dithionite to Avicel, α-cellulose or filter cake of pretreated spruce wood resulted in improvements between 25 and 33%. Positive effects (6-17%) of reducing agents were also observed in experiments with carboxymethyl cellulose and 2-hydroxyethyl cellulose. The approach to add reducing agents appears useful for facilitating the utilization of enzymes to convert cellulosic substrates in industrial processes.     

Solid state fermentation of a Mycelia Sterilia laccase using steam-exploded wheat straw

Mycelia Sterilia YY-5, an endophytic fungus isolated from Rhus Chinensis Mill, was used in SSF for laccase production using steam-exploded wheat straw (SEWS). The fermentation period of YY-5 in solid state fermentation (SSF) shortened to 4 days compared with 5 days of submerged liquid fermentation (SmF) and the maximum laccase activity was 678.1 IU g⁻¹ substrate. The steam-explosion intensity (Log₁₀ R ₀) of SEWS had a significant effect on the growth of YY-5 and laccase activity, since SEWS could provide enough carbon source for YY-5 and inducers for laccase. The optimum SSF conditions using SEWS with Log₁₀ R ₀ = 3.597 as substrate were: inoculating with liquid inocula, keeping the solid-to-liquid ratio (S/L) for 1:4 and cultivating at 26°C. Under the optimum fermentation condition the laccase activity of YY-5 reached 849.5 ± 42.5 IU g⁻¹ substrate. The enzyme composition analysis indicated that laccase was the dominant enzyme of YY-5. Assayed with SDS-PAGE and active PAGE electrophoresis, the molecular weight of YY-5 laccase was approximately 45 kDa.     

Dilute acid pretreatment, enzymatic saccharification and fermentation of wheat straw to ethanol

Wheat straw consists of 48.57 ± 0.30% cellulose and 27.70 ± 0.12% hemicellulose on dry solid (DS) basis and has the potential to serve as a low cost feedstock for production of ethanol. Dilute acid pretreatment at varied temperature and enzymatic saccharification were evaluated for conversion of wheat straw cellulose and hemicellulose to monomeric sugars. The maximum yield of monomeric sugars from wheat straw (7.83%, w/v, DS) by dilute H₂SO₄ (0.75%, v/v) pretreatment and enzymatic saccharification (45 °C, pH 5.0, 72 h) using cellulase, β-glucosidase, xylanase and esterase was 565 ± 10 mg/g. Under this condition, no measurable quantities of furfural and hydroxymethyl furfural were produced. The yield of ethanol (per litre) from acid pretreated enzyme saccharified wheat straw (78.3 g) hydrolyzate by recombinant Escherichia coli strain FBR5 was 19 ± 1 g with a yield of 0.24 g/g DS. Detoxification of the acid and enzyme treated wheat straw hydrolyzate by overliming reduced the fermentation time from 118 to 39 h in the case of separate hydrolysis and fermentation (35 °C, pH 6.5), and increased the ethanol yield from 13 ± 2 to 17 ± 0 g/l and decreased the fermentation time from 136 to 112 h in the case of simultaneous saccharification and fermentation (35 °C, pH 6.0).     

Isolation and characterization of cellulose nanofibrils from wheat straw using steam explosion coupled with high shear homogenization

Cellulose nanofibrils of diameter 10–50 nm were obtained from wheat straw using alkali steam explosion coupled with high shear homogenization. High shear results in shearing of the fiber agglomerates resulting in uniformly dispersed nanofibrils. The chemical composition of fibers at different stages were analyzed according to the ASTM standards and showed increase in α-cellulose content and decrease in lignin and hemicellulose. Structural analysis of steam exploded fibers was carried out by Fourier Transform Infrared (FT-IR) spectroscopy and X-ray diffraction (XRD). Thermal stability was higher for cellulose nanofibrils as compared to wheat straw and chemically treated fibers. The fiber diameter distribution was obtained using image analysis software. Characterization of the fibers by AFM, TEM, and SEM showed that fiber diameter decreases with treatment and final nanofibril size was 10–15 nm. FT-IR, XRD, and TGA studies confirmed the removal of hemicellulose and lignin during the chemical treatment process.     

Solid-state anaerobic digestion of spent wheat straw from horse stall

The spent wheat straw from horse stall bedding has lower cellulose and hemicellulose contents, but higher volatile fatty acid content than raw wheat straw. Biogas production from solid-state anaerobic digestion (SS-AD) of spent wheat straw and raw wheat straw was compared in this study. The SS-AD tests were conducted at 22% total solids (TS) content using inoculum from a liquid AD system at three feedstock-to-inoculum (F/I) ratios of 2.0, 4.0, and 6.0. Daily methane yields of spent wheat straw peaked 8 and 3 days earlier than those of raw wheat straw at F/I ratios of 2.0 and 4.0, respectively. The highest methane yield of 150.0 L/kg volatile solids (VS) was obtained from spent wheat straw at an F/I ratio of 4.0, which was 56.2% higher than that of raw wheat straw. The corresponding cellulose and hemicellulose degradation of spent wheat straw was 24.1% and 49.4% higher than those of raw wheat straw, respectively.     

Alkaline-assisted screw press pretreatment affecting enzymatic hydrolysis of wheat straw

Bioprocess and Biosystems Engineering - Screw press processing of biomass can be considered as a suitable mechanically based pretreatment for biofuel production since it disrupts the structure of...     

Modification of wheat straw lignin by solid state fermentation with white-rot fungi

The potential of crude enzyme extracts, obtained from solid state cultivation of four white-rot fungi (Trametes versicolor, Bjerkandera adusta, Ganoderma applanatum and Phlebia rufa), was exploited to modify wheat straw cell wall. At different fermentation times, manganese-dependent peroxidase (MnP), lignin peroxidase (LiP), laccase, carboxymethylcellulase (CMCase), avicelase, xylanase and feruloyl esterase activities were screened and the content of lignin as well as hydroxycinnamic acids in fermented straw were determined. All fungi secreted feruloyl esterase while LiP was only detected in crude extracts from B. adusta. Since no significant differences (P > 0.05) were observed in remaining lignin content of fermented straw, LiP activity was not a limiting factor of enzymatic lignin removal process. The levels of esterified hydroxycinnamic acids degradation were considerably higher than previous reports with lignocellulosic biomass. The data show that P. rufa, may be considered for more specific studies as higher ferulic and p-coumaric acids degradation was observed for earlier incubation times.     

Enhanced enzymatic hydrolysis of wheat straw by aqueous glycerol pretreatment

Considering the practical technology-economy of glycerol processing from oleochemicals industry, the ensuing work was proposed to further explore the atmospheric aqueous glycerol autocatalytic organosolv pretreatment (AAGAOP) to improve the enzymatic hydrolysis of lignocellulosic biomass. With the liquid-solid ratio of 20 g g(-1) at 220 degrees C for 3h, the AAGAOP enabled wheat straw to remove approximately 70% hemicelluloses and approximately 65% lignin, with approximately 98% cellulose retention. The pretreated fiber was achieved with approximately 90% of the enzymatic hydrolysis yield after 48 h. At oven-drying, dehydration was likely to cause the hornification of fiber, which was responsible for the low enzymatic hydrolysis of dried fiber. With SEM observations, the AAGAOP disrupted wheat straw into thin and fine fibrils, with a small average size and more surface area. The AAGAOP technique, as a novel strategy, enhanced the enzymatic hydrolysis of lignocellulosic biomass by removing the chemically compositional barrier and altering the physically structural impediment.     

Effect of antioxidant extraction on the enzymatic hydrolysis and bioethanol production of the extracted steam-exploded sugarcane bagasse

Ethyl acetate extraction (EAE) of the steam exploded sugarcane bagasse may be an effective and economic way to extract antioxidants as well as enhance the enzymatic hydrolysis and bioethanol yield from the extracted residue. Therefore, the effects of EAE on steam-exploded sugarcane bagasse (SESB) were studied. Under boiling solvent extraction (BSE), the efficiency of EAE for obtaining phenolics from SESB was approximately 20%. EA extracts obtained under BSE showed an H₂O₂ scavenging activity (210 μL) of 99%. The IC₅₀ values for 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power of BSE40 were 50.89 and 256.38 μg/mL, respectively, while those of vitamin C were 24 and 112 μg/mL, respectively. EAE improved the glucose yield by 30% but had no significant effect on the xylose yield during the enzymatic hydrolysis obtained using Celluclast 1.5L and Novozym 188. EAE also increased the ethanol yield by 8.78% by employing simultaneous saccharification and fermentation. The present study may be of great importance in industrial bioethanol production from steam-exploded biomass environmentally friendly and economically.