cellulase and sulfuric-acid

Lignocellulosic biomass mainly consists of hemicellulose, lignin, and cellulose, which differently affect the enzymatic digestibility of cellulose. As for the typical representative for inert woody biomass, three components of cellulose were proposed conceptually for poplar sawdust, i.e., active cellulose, inert cellulose, and resistant cellulose. Dilute sulfuric acid pretreatment, hydrogen peroxide-sulfuric acid delignification, and sulfuric acid-assisted glycerol swelling were, respectively, proven to break the three obstacle mechanisms that affect the cellulase of poplar. The removal of key obstacles improved the cellulase digestibility of poplar enzyme-hydrolyzed residues by 188.7 %, and glucose yield increased from 34.6 % to 99.9 %. Therefore, a total of 39.5 g glucose was obtained from 100 g poplar sawdust by integrating the above three technologies. This work presented insight into and removed the key obstacles to enzymatic digestibility of poplar cellulose and developed an integrated technology to effectively convert full cellulose fraction to glucose from woody biomass.

Topics: Cellulase; Cellulose; Digestion; Glucose; Hydrolysis; Lignin; Populus; Technology

Exploiting bamboo shoot shells (BSS) as feedstocks for biorefining is a crucial scheme to advance the bioavailability of bamboo shoots. This work applied traditional dilute sulfuric acid pretreatment (DAP) to treat BSS and simultaneously prepared the solid-acid-catalyst by using BSS as carbon-based carriers. The biocatalysis of the prehydrolysate from DAP and enzymatic hydrolysis of pretreated BSS was subsequently performed to achieve efficient bioconversion of its carbohydrates. The results displayed that 0.1 g/L H

Topics: Acids; Carbohydrates; Catalysis; Cellulase; Hydrolysis; Lignin

Waterborne polyurethanes (WBPUs) have been proposed as ecofriendly elastomers with several applications in coatings and adhesives. WBPU's physicochemical properties can be enhanced by the addition of cellulose nanocrystals (CNCs). The way CNCs are isolated has a strong effect on their properties and can determine their role as reinforcement. In this work, CNCs produced using ancestral endoglucanase (EnCNCs) were used as reinforcement for WBPU and compared with CNC produced by sulfuric acid hydrolysis (AcCNC). The enzymatic method produced highly thermostable and crystalline CNCs. The addition of small contents of EnCNCs improved the thermomechanical stability and mechanical properties of WBPUs, even better than commercial AcCNCs. Besides, WBPU reinforced by adding EnCNCs was studied as a coating for paper materials, increasing its abrasion resistance and as electrospun nanocomposite mats where EnCNCs helped maintaining the morphology of the fibers.

Topics: Adhesives; Cellulase; Cellulose; Construction Materials; Green Chemistry Technology; Humans; Hydrolysis; Nanocomposites; Nanoparticles; Polyurethanes; Recombinant Proteins; Sulfuric Acids; Water

Traditional surfactants have been reported to enhance enzymatic saccharification of lignocellulose, however, it is important to transfer these findings to a system that uses a high-efficiency and low-toxicity natural surfactant instead. In this work, a novel hybrid method involving use of the natural surfactant (humic acid, HA) during mild acid (H

Topics: Cellulase; Hydrolysis; Lignin; Sulfuric Acids; Surface-Active Agents; Triticum

The synergism of cellulase (C), pectinase (P), and xylanase (X) for the saccharification of sweet potato residues (SPR) was investigated. The removal of starch from SPR was easily achieved by using amylase, but the cellulose conversion of de-starched SPR was relatively low, thus dilute H

Topics: Cellulase; Endo-1,4-beta Xylanases; Hydrolysis; Ipomoea batatas; Polygalacturonase; Sulfuric Acids

After chemical pretreatment, improved amenability of agrowaste biomass for enzymatic saccharification needs an understanding of the effect exerted by pretreatments on biomass for enzymatic deconstruction. In present studies, NaOH, NH

Topics: Ammonium Hydroxide; Biomass; Cellulase; Cellulose; Hydrolysis; Lignin; Microscopy, Fluorescence; Saccharum; Sodium Hydroxide; Sulfuric Acids

In this work, conditions for an enzymatic pretreatment prior to NCC isolation from cotton linter were assessed. Different cellulase doses and reaction times were studied within an experimental design and NCC were obtained. At optimal enzymatic conditions (20U, 2 h), a total yield greater than 80% was achieved and the necessary enzymatic treatment time was reduced 90%. Different intensities of enzymatic treatments led to proportional decreases in fiber length and viscosity and also were inversely proportional to the amount of released oligosaccharides. These differences within fibers lead to quantitative differences in NCC: increase in acid hydrolysis yield, reduction of NCC surface charge and crystallinity increase. Benefits produced by enzymatic treatments did not have influence over other NCC characteristics such as their sulfur content (≈1%), size (≈200 nm), zeta potential (≈-50 mV) or degree of polymerization (≈200). Evidence presented in this work would reduce the use of harsh sulfuric acid generating a cleaner stream of profitable oligosaccharides.

Topics: Cellulase; Cellulose; Oligosaccharides; Sulfuric Acids

Bio-refinery processes require use of the most suitable lignocellulosic biomass for enzymatic saccharification and microbial fermentation. Glucose yield from biomass solid fractions obtained after dilute sulfuric acid (1%) pretreatment (at 180 °C) was investigated using 14, 8, and 16 varieties of rice, wheat, and sorghum, respectively. Biomass solid fractions of each crop showed similar cellulose content. However, glucose yield after enzymatic hydrolysis (cellulase loading at 6.6 filter paper unit/g-biomass) was different among the varieties of each crop, indicating genotypic differences for rice, wheat, and sorghum. Nuclear magnetic resonance method revealed that the high residual level of lignin aromatic regions decreased glucose yield from solid fraction of sorghum.

Topics: Biomass; Cellulase; Fermentation; Glucose; Hydrolysis; Lignin; Oryza; Sorghum; Species Specificity; Sulfuric Acids; Triticum

In this study, commercial surfactants have been investigated at economically viable dosage to enhance the enzymatic saccharification of pretreated wheat straw at high solid loadings. Twenty one surfactants were evaluated with pilot scale pretreated wheat straw and mechanism of surfactant action has been elucidated. One surfactant has improved the saccharification of dilute acid wheat straw (DAWS) by 26.4% after 24h and 23.1% after 48h while, steam exploded wheat straw (SEWS) saccharification was increased by 51.2% after 24h and 36.4% after 48h at 10% solid loading. At 20% solid loading, about 31% increase in yield was obtained on DAWS and about 55% on SEWS after 48h. Further, lignin was isolated from pretreated wheat straws and characterized which revealed that SEWS derived lignin was more hydrophobic than DAWS lignin. This investigation suggests that surfactant supplementation during saccharification is an effective strategy to achieve higher saccharification yield.

Topics: Adsorption; Biomass; Bioreactors; Carbohydrate Metabolism; Cellulase; Enzyme Stability; Fermentation; Lignin; Pilot Projects; Proton Magnetic Resonance Spectroscopy; Reproducibility of Results; Spectroscopy, Fourier Transform Infrared; Steam; Substrate Specificity; Sulfuric Acids; Surface-Active Agents; Triticum; Waste Products

Dilute acid pre-extraction enhanced the mechanically refined poplar pulp substrates' enzymatic hydrolysis efficiency obviously. The results showed that the surface lignin distribution was changed significantly in residual wood chips and pulp substrates, and the surface lignin distribution showed important impact on the following enzymatic hydrolysis. Acid pre-extraction can lead to a redistribution of lignin in fiber cell walls, i.e., the lignin was degraded and migrated to fiber surface in the form of re-deposited lignin and pseudo-lignin. However, higher pre-extraction intensity was not desired due to the formation of redeposited lignin and pseudo-lignin. This study will help to reach a deeper understanding on the lignin distribution in the view of molecular and ultrastructure, and promote the development of a cost-efficient pretreatment strategy for biomass processing.

Topics: Biomass; Biotechnology; Cell Wall; Cellulase; Cellulose; Glucose; Hydrolysis; Lignin; Manganese; Photoelectron Spectroscopy; Populus; Sulfuric Acids; Wood

This study aims to propose a biorefinery pretreatment technology for the bioconversion of sugarcane bagasse (SB) into biofuels and N-fertilizers. Performance of diluted acid (DA), aqueous ammonia (AA), oxidate ammonolysis (OA) and the combined DA with AA or OA were compared in SB pretreatment by enzymatic hydrolysis, structural characterization and acetone-butanol-ethanol (ABE) fermentation. Results indicated that DA-OA pretreatment improves the digestibility of SB by sufficiently hydrolyzing hemicellulose into fermentable monosaccharides and oxidating lignin into soluble N-fertilizer with high nitrogen content (11.25%) and low C/N ratio (3.39). The enzymatic hydrolysates from DA-OA pretreated SB mainly composed of glucose was more suitable for the production of ABE solvents than the enzymatic hydrolysates from OA pretreated SB containing high ratio of xylose. The fermentation of enzymatic hydrolysates from DA-OA pretreated SB produced 12.12g/L ABE in 120h. These results suggested that SB could be utilized efficient, economic, and environmental by DA-OA pretreatment.

Topics: Acetone; Ammonia; Biofuels; Bioreactors; Butanols; Cellulase; Cellulose; Ethanol; Fermentation; Hydrolysis; Oxidation-Reduction; Saccharum; Solvents; Sulfuric Acids

Corncob is a potential feedstock in Thailand that can be used for fermentable sugar production through dilute sulfuric acid pretreatment and enzymatic hydrolysis. To recover high amounts of monomeric sugars from corncob, the sulfuric pretreatment conditions were optimized by using response surface methodology with three independent variables: sulfuric acid concentration, temperature, and time. The highest response of total sugars, 48.84 g/L, was found at 122.78°C, 4.65 min, and 2.82% (v/v) H2SO4. With these conditions, total sugars from the confirmation experiment were 46.29 g/L, with 5.51% error from the predicted value. The hydrolysate was used as a substrate for acetone-butanol-ethanol fermentation to evaluate its potential for microbial growth. The simultaneous saccharification and fermentation (SSF) showed that C. beijerinckii TISTR 1461 can generate acetone-butanol-ethanol products at 11.64 g/L (5.29 g/L acetone, 6.26 g/L butanol, and 0.09 g/L ethanol) instantly using sugars from the hydrolysed corncob with Novozymes 50013 cellulase enzyme without an overliming process.

Topics: Acetone; Butanols; Carbohydrates; Cellulase; Clostridium beijerinckii; Ethanol; Fermentation; Hydrolysis; Industrial Microbiology; Microscopy, Electron, Scanning; Particle Size; Sulfuric Acids; X-Ray Diffraction; Zea mays

This study reported a new solution of lignocellulose feedstock storage based on the distributed pretreatment concept. The dry dilute sulfuric acid pretreatment (DDAP) was conducted on corn stover feedstock, instead of ammonia fiber explosion pretreatment. Then the dry dilute acid pretreated corn stover was stored for three months during summer season with high temperature and humidity. No negative aspects were found on the physical property, composition, hydrolysis yield and ethanol fermentability of the long term stored pretreated corn stover, plus the additional merits including no chemicals recovery operation, anti-microbial contaminant environment from stronger acid and inhibitor contents, as well as the mild and slow hydrolysis in the storage. The new pretreatment method expanded the distributed pretreatment concept of feedstock storage with potential for practical application.

Topics: Cellulase; Ethanol; Fermentation; Hydrolysis; Lignin; Sulfuric Acids; Time Factors; Waste Products; Zea mays

A two-stage process was evaluated to increase sugar recovery. Firstly, corn stover was treated with dilute hydrochloric acid to recover the xylose, and then the residue was subjected to a wet-milling pretreatment. Dilute hydrochloric acid showed a high xylose recovery during the first stage. The optimal condition was 120°C and 40min for 0.7wt% dilute hydrochloric acid pretreatment followed by wet-milling pretreatment for 15min. The xylose and glucose yield were 81.0% and 64.0%, respectively, with a cellulase dosage at 3FPU/g of substrate. This two-stage process was effective on account of the removal of hemicelluloses in the first stage and the delamination of cell wall in the second stage, increasing the possibility of adsorption of cellulose to enzymes, and resulting in a high sugar recovery with a very low enzyme loading.

Topics: Biotechnology; Carbohydrates; Cellulase; Glucose; Hydrochloric Acid; Hydrolysis; Porosity; Sulfuric Acids; Temperature; Time Factors; Waste Products; Xylose; Zea mays

Soluble coffee, being one of the world's most popular consuming drinks, produces a considerable amount of spent coffee ground (SCG) along with its production. The SCG could function as a potential lignocellulosic feedstock for production of bioproducts. The objective of this study is to investigate the possible optimal condition of dilute acid hydrolysis (DAH) at high solids and mild temperature condition to release the reducing sugars from SCG. The optimal condition was found to be 5.3 % (w/w) sulfuric acid concentration and 118 min reaction time. Under the optimal condition, the mean yield of reducing sugars from enzymatic saccharification of defatted SCG acid hydrolysate was 563 mg/g. The SCG hydrolysate was then successfully applied to culture Lipomyces starkeyi for microbial oil fermentation without showing any inhibition. The results suggested that dilute acid hydrolysis followed by enzymatic saccharification has the great potential to convert SCG carbohydrates to reducing sugars. This study is useful for the further developing of biorefinery using SCG as feedstock at a large scale.

Topics: Analysis of Variance; Bacteria; Biomass; Carbohydrate Metabolism; Carbohydrates; Cellulase; Coffee; Fermentation; Hydrolysis; Models, Theoretical; Oils; Sulfuric Acids; Temperature; Waste Products

In this work, to elucidate why the acid-pretreated bamboo shows disappointingly low enzymatic digestibility comparing to the alkali-pretreated bamboo, residual lignins in acid-pretreated and kraft pulped bamboo were isolated and analyzed by adsorption isotherm to evaluate their extents of nonproductive enzyme adsorption. Meanwhile, physicochemical properties of the isolated lignins were analyzed and a relationship was established with non-productive adsorption. Results showed that the adsorption affinity and binding strength of cellulase on acid-pretreated bamboo lignin (MWLa) was significantly higher than that on residual lignin in pulped bamboo (MWLp). The maximum adsorption capacity of cellulase on MWLp was 129.49 mg/g lignin, which was lower than that on MWLa (160.25 mg/g lignin). When isolated lignins were added into the Avicel hydrolysis solution, the inhibitory effect on enzymatic hydrolysis efficiency of MWLa was found to be considerably stronger than that with MWLp. The cellulase adsorption on isolated lignins was correlated positively with hydrophobicity, phenolic hydroxyl group, and degree of condensation but negatively with surface charges and aliphatic hydroxyl group. These results suggest that the higher nonproductive cellulase adsorption and physicochemical properties of residual lignin in acid-pretreated bamboo may be responsible for its disappointingly low enzymatic digestibility.

Topics: Adsorption; Carbon-13 Magnetic Resonance Spectroscopy; Cellulase; Cellulose; Chemical Phenomena; Hydrolysis; Kinetics; Lignin; Paper; Poaceae; Sulfuric Acids; Temperature

The recalcitrance in grasses varies according to cell type and tissue. In this study, dilute acid pretreatment was performed on Miscanthus×giganteus internodes that include rind and pith regions which showing heterogeneous structural and chemical changes. Pretreatment on pith effectively hydrolyzed 73.33% hemicelluloses and separated cohesive cell walls from the compound middle lamella due to lignin migration. Lignin droplets with an average diameter of 49.5±29.3nm were concurrently coalesced on wall surface, that in turn exposed more microfibrils deep in walls to be enzymatically hydrolyzed reaching 82.55%. By contrast, the rind with a relatively intergrated cell structure was covered by larger lignin droplets (101.2±44.1nm) and filled with inaccessible microfibrils limiting enzymatic sacchrification (31.50%). Taken together, the cellulose digestibility of biomass was not majorly influenced by cellulose crystallinity, while it was strongly correlated with the positive effects of hemicelluloses degradation, lignin redistribution, cellulose exposure and loosening cell wall structure.

Topics: Biomass; Cell Wall; Cellulase; Cellulose; Hydrolysis; Lignin; Microscopy, Atomic Force; Microscopy, Electron, Transmission; Microscopy, Fluorescence; Poaceae; Sulfuric Acids

A two stage pretreatment approach for biomass is developed in the current work in which dilute acid (DA) pretreatment is followed by a solvent based pretreatment (N-methyl morpholine N oxide - NMMO). When the combined pretreatment (DAWNT) is applied to sugarcane bagasse and corn stover, the rates of hydrolysis and overall yields (>90%) are seen to dramatically improve and under certain conditions 48h can be taken off the time of hydrolysis with the additional NMMO step to reach similar conversions. DAWNT shows a 2-fold increase in characteristic rates and also fractionates different components of biomass - DA treatment removes the hemicellulose while the remaining cellulose is broken down by enzymatic hydrolysis after NMMO treatment to simple sugars. The remaining residual solid is high purity lignin. Future work will focus on developing a full scale economic analysis of DAWNT for use in biomass fractionation.

Topics: Biomass; Biotechnology; Cellulase; Cellulose; Cyclic N-Oxides; Glucans; Hydrolysis; Lignin; Morpholines; Saccharum; Solvents; Sulfuric Acids; Time Factors; Zea mays

Pretreatment of rice straw on pilot scale steam explosion has been attempted to achieve maximum sugar recovery. Three different reaction media viz. water, sulfuric acid and phosphoric acid (0.5%, w/w) were explored for pretreatment by varying operating temperature (160, 180 and 200°C) and reaction time (5 and 10min). Using water and 0.5% SA showed almost similar sugar recovery (∼87%) at 200 and 180°C respectively. However, detailed studies showed that the former caused higher production of oligomeric sugars (13.56g/L) than the later (3.34g/L). Monomeric sugar, followed the reverse trend (7.83 and 11.62g/L respectively). Higher oligomers have a pronounced effect in reducing enzymatic sugar yield as observed in case of water. Mass balance studies for water and SA assisted SE gave total saccharification yield as 81.8% and 77.1% respectively. However, techno-economical viability will have a trade-off between these advantages and disadvantages offered by the pretreatment medium.

Topics: Biotechnology; Carbohydrates; Cellulase; Hydrolysis; Oryza; Phosphoric Acids; Pilot Projects; Steam; Sulfuric Acids; Temperature; Waste Products

The structural and physicochemical characteristics are associated with resistance of plant cell walls to saccharification by enzymes. The effect of physicochemical properties on glucose yield of bagasse from different varieties of sugarcane at low and high enzyme dosages was investigated. The result showed that glucose yield at low enzyme dosage was positively linear correlated with the yield at high enzyme dosage, for both the untreated and pretreated materials. The pretreatment significantly increased the accessibility of substrates by enzyme due to the increase of internal and external surface area. Glucose yield also showed a linear correlation with dye adsorption. However, the increase in glucose yield as a result of pretreatment did not correlate with the increases in crystallinity index and decreases in degree of polymerization. The Principal Component Analysis of infrared data indicated that lignin was the main component that differentiated the varieties before and after pretreatment. These results suggested that the key differences in pretreatment responses among varieties could be mainly attributed to their differences in the internal and external surface area after pretreatment.

Topics: beta-Glucosidase; Biofuels; Biotechnology; Cellulase; Cellulose; Chemical Phenomena; Crystallization; Ethanol; Glucose; Hydrolysis; Polymerization; Saccharum; Spectroscopy, Fourier Transform Infrared; Sulfuric Acids

In this study, steam explosion pretreatment was performed in cotton stalks, leading to 5-6 folds enhancements on biomass enzymatic saccharification distinctive in Gossypium barbadense and Gossypium hirsutum species. Sequential 1% H2SO4 pretreatment could further increase biomass digestibility of the steam-exploded stalks, and also cause the highest sugar-ethanol conversion rates probably by releasing less inhibitor to yeast fermentation. By comparison, extremely high concentration alkali (16% NaOH) pretreatment with raw stalks resulted in the highest hexoses yields, but it had the lowest sugar-ethanol conversion rates. Characterization of wall polymer features indicated that biomass saccharification was enhanced with steam explosion by largely reducing cellulose DP and extracting hemicelluloses. It also showed that cellulose crystallinity and arabinose substitution degree of xylans were the major factors on biomass digestibility in cotton stalks. Hence, this study has provided the insights into cell wall modification and biomass process technology in cotton stalks and beyond.

Topics: Biomass; Biotechnology; Carbohydrate Metabolism; Cell Wall; Cellulase; Cellulose; Fermentation; Gossypium; Hexoses; Lignin; Polymerization; Sodium Hydroxide; Steam; Sulfuric Acids; Waste Products

This paper reports on processing options for the conversion of feedlot cattle manures into composite sugars for ethanol fermentation. Small-scale anaerobic digestion trials revealed that the process significantly reduces the content of glucan and xylan (ca. 70%) without effecting lignin. Moreover, anaerobic digestate (AD) fibres were poor substrates for cellulase (Cellic® CTec 2) saccharification, generating a maximum combined sugar yield of ca. 12% per original dry weight. Dilute acid pretreatment and enzyme saccharification of raw manures significantly improved total sugar recoveries, totalling 264 mg/g (79% theoretical). This was attained when manures were pretreated with 2.5% H2SO4 for 90 min at 121°C and saccharified with 50 FPU CTec 2/g glucan. Saccharomyces cerevisiae efficiently fermented crude hydrolysates within 6 h, yielding 7.3 g/L ethanol, representing glucose to ethanol conversion rate of 70%. With further developments (i.e., fermentation of xylose), this process could deliver greater yields, reinforcing its potential as a biofuel feedstock.

Topics: Anaerobiosis; Animals; Batch Cell Culture Techniques; Biofuels; Biomass; Biotechnology; Carbohydrates; Cattle; Cellulase; Ethanol; Fermentation; Glucose; Hydrolysis; Manure; Saccharomyces cerevisiae; Sulfuric Acids; Time Factors; Xylose

A novel integrated process to coproduce xylose, lignosulfonate and ethanol from wheat straw was investigated. Firstly, wheat straw was treated by dilute sulfuric acid and xylose was recovered from its hydrolyzate. Its optimal conditions were 1.0wt% sulfuric acid, 10% (w/v) wheat straw loading, 100°C, and 2h. Then the acid treated wheat straw was treated by sulfomethylation reagent and its hydrolyzate containing lignosulfonate was directly recovered. Its optimal conditions were 150°C, 15% (w/v) acid treated wheat straw loading, and 5h. Finally, the two-step treated wheat straw was converted to ethanol through enzymatic hydrolysis and microbial fermentation. Under optimal conditions, 1kg wheat straw could produce 0.225kg xylose with 95% purity, 4.16kg hydrolyzate of sulfomethylation treatment containing 5.5% lignosulfonate, 0.183kg ethanol and 0.05kg lignin residue. Compared to present technology, this process is a potential economically profitable wheat straw biorefinery.

Topics: Acids; Biotechnology; Carbohydrates; Cellulase; Cellulose; Ethanol; Fermentation; Glucose; Hydrolysis; Lignin; Saccharomyces cerevisiae; Sulfuric Acids; Temperature; Triticum; Xylose

Three strategies were presented to achieve high solids loading while maximizing carbohydrate conversion, which are fed-batch, splitting/thickening, and clarifier processes. Enzymatic hydrolysis was performed at water insoluble solids (WIS) of 15% using washed dilute-acid pretreated corn stover. The carbohydrate concentration increased from 31.8 to 99.3g/L when the insoluble solids content increased from 5% to 15% WIS, while the final carbohydrate conversion was decreased from 78.4% to 73.2%. For the fed-batch process, a carbohydrate conversion efficiency of 76.8% was achieved when solid was split into 60:20:20 ratio, with all enzymes added first. For the splitting/thickening process, a carbohydrate conversion of 76.5% was realized when the filtrate was recycled to simulate a steady-state process. Lastly, the clarifier process was evaluated and the highest carbohydrate conversion of 81.4% was achieved. All of these results suggests the possibility of enzymatic hydrolysis at high solids to make the overall conversion cost-competitive.

Topics: Acids; Carbohydrates; Cellulase; Enzyme Activation; Hydrolysis; Industrial Waste; Plant Components, Aerial; Sulfuric Acids; Viscosity; Zea mays

The aim of this study is to investigate the technical feasibility of converting macroalgae cellulosic residue (MCR) into bioethanol. An attempt was made to present a novel, environmental friendly and economical pretreatment process that enhances enzymatic conversion of MCR to sugars using Dowex (TM) Dr-G8 as catalyst. The optimum yield of glucose reached 99.8% under the optimal condition for solid acid pretreatment (10%, w/v biomass loading, 4%, w/v catalyst loading, 30min, 120°C) followed by enzymatic hydrolysis (45FPU/g of cellulase, 52CBU/g of β-glucosidase, 50°C, pH 4.8, 30h). The yield of sugar obtained was found more superior than conventional pretreatment process using H2SO4 and NaOH. Biomass loading for the subsequent simultaneous saccharification and fermentation (SSF) of the pretreated MCR was then optimized, giving an optimum bioethanol yield of 81.5%. The catalyst was separated and reused for six times, with only a slight drop in glucose yield.

Topics: beta-Glucosidase; Biofuels; Biomass; Catalysis; Cellulase; Cellulose; Ethanol; Fermentation; Glucose; Hydrolysis; Microscopy, Electron, Scanning; Resins, Synthetic; Saccharomyces cerevisiae; Seaweed; Sodium Hydroxide; Sulfuric Acids; Temperature

High-efficiency xylose utilization is one of the restrictive factors of bioethanol industrialization. However, xylonic acid (XA) as a new bio-based platform chemical can be produced by oxidation of xylose with microbial. So, an applicable technology of XA bioconversion was integrated into the process of bioethanol production. After corn stover was pretreated with acid-catalyzed steam-explosion, solid and liquid fractions were obtained. The liquid fraction, also named as acid-catalyzed steam-exploded corn stover (ASC) prehydrolyzate (mainly containing xylose), was catalyzed with Gluconobacter oxydans NL71 to prepare XA. After 72 h of bioconversion of concentrated ASC prehydrolyzate (containing 55.0 g/L of xylose), the XA concentration reached a peak value of 54.97 g/L, the sugar utilization ratio and XA yield were 94.08 and 95.45 %, respectively. The solid fraction was hydrolyzed to produce glucose with cellulase and then fermented with Saccharomyces cerevisiae NL22 to produce ethanol. After 18 h of fermentation of concentrated enzymatic hydrolyzate (containing 86.22 g/L of glucose), the ethanol concentration reached its highest value of 41.48 g/L, the sugar utilization ratio and ethanol yield were 98.72 and 95.25 %, respectively. The mass balance showed that 1 t ethanol and 1.3 t XA were produced from 7.8 t oven dry corn stover.

Topics: Biofuels; Catalysis; Cellulase; Ethanol; Fermentation; Hydrolysis; Saccharomyces cerevisiae; Steam; Sugar Acids; Sulfuric Acids; Waste Products; Zea mays

In this study, hydrothermal acid treatment for efficient recovery of sugar from Golenkinia sp. was investigated. The initial glucose and XMG (xylose, mannose, and galactose) contents of a prepared Golenkinia sp. solution (40g/L) were 15.05 and 5.24g/L, respectively. The microalgal cell walls were hydrolyzed, for sugar recovery, by enzymatic saccharification and/or hydrothermal acid treatment. Among the various hydrothermal acid treatment conditions, the most optimal were the 2.0% H2SO4 concentration at 150°C for 15min, under which the glucose- and XMG-extraction yields were 71.7% and 64.9%, respectively. By pH 4.8, 50°C enzymatic hydrolysis after optimal hydrothermal acid treatment, the glucose- and XMG-extraction yields were additionally increased by 8.3% and 0.8%, respectively. After hydrothermal acid treatment, the combination with the enzymatic hydrolysis process improved the total sugar yield of Golenkinia sp. to 75.4%.

Topics: beta-Glucosidase; Carbohydrates; Cell Fractionation; Cellulase; Chlorophyta; Ethanol; Hot Temperature; Hydrolysis; Microalgae; Sulfuric Acids; Water

In this work the introduction of a cellulase treatment prior to NCC isolation was assessed. NCC was produced using sulfuric acid at two different concentrations (62 and 64% wt.). The effect of pore size for filtration step was also assessed. The smaller acid dose leaded to yields up to 65-70% and average size up to 160 nm. It also produced crystals with reduced sulfur content (0.6-1%). Cellulase pretreatment influenced NCC characteristics, as it increased overall yield a 12%, increased average particle size around 35 nm and reduced NCC sulfur content up to a 0.8%. We found that different conditions of enzymatic treatments led to quantitative differences on their effects on NCC. Acetate buffer used for enzymatic treatments was found to counteract effects of acid. The evidence presented in this work suggested that pretreating fibers with this cellulase represents a very interesting option to partially replace chemicals on NCC isolation.

Topics: Cellulase; Cellulose; Crystallization; Gossypium; Hydrolysis; Nanoparticles; Particle Size; Plant Extracts; Sulfuric Acids

Aqueous solutions of tetrabutylphosphonium hydroxide have been evaluated as pretreatment media for rice husks, prior to sulphuric acid hydrolysis or cellulase enzymatic hydrolysis. Varying the water:tetrabutylphosphonium hydroxide ratio varied the rate of delignification, as well as silica, lignin and cellulose solubility. Pre-treatment with 60wt% hydroxide dissolved the rice husk and the regenerated material was thus heavily disrupted. Sulphuric acid hydrolysis of 60wt%-treated samples yielded the highest amount of glucose per gram of rice husk. Solutions with good lignin and silica solubility but only moderate to negligible cellulose solubility (10-40wt% hydroxide) were equally effective as pre-treatment media for both acid and enzymatic hydrolysis. However, pre-treatment with 60wt% hydroxide solutions was incompatible with downstream enzymatic hydrolysis. This was due to significant incorporation of phosphonium species in the regenerated biomass, which significantly inhibited the activity of the cellulase enzymes.

Topics: Acids; Biomass; Cellulase; Cellulose; Glucose; Hydrolysis; Lignin; Organophosphorus Compounds; Oryza; Solubility; Sulfuric Acids; Temperature; Water

The response and behavior of bamboo green, timber, and yellow of moso bamboo (Phyllostachys heterocycla) to three pretreatments, sulfite (SPORL), dilute acid (DA), and alkali (NaOH), were investigated and compared with varied chemical loadings at 180°C for 30 min with a 6.25:1 (v/w) liquor-to-bamboo ratio. All the pretreatments improved the enzymatic digestibility of bamboo substrates. Under the investigated conditions, the DA pretreatment achieved better enzymatic digestibility, but had lower sugar recovery yield, and formed more fermentation inhibitors. The results suggested that the SPORL pretreatment be able to generate more readily digestible bamboo substrate with higher sugar yield and fewer fermentation inhibitors than the corresponding DA pretreatment if hemicelluloses are sufficiently removed by adding more acid to bring down the pretreatment pH. Bamboo timber had higher sugar content and better enzymatic digestibility and therefore was a better feedstock for bioconversion than bamboo green and yellow.

Topics: Bambusa; beta-Glucosidase; Carbohydrate Metabolism; Cell Wall; Cellulase; Fermentation; Hydrolysis; Lignin; Sodium Hydroxide; Sulfites; Sulfuric Acids; Time Factors; Wood

The main objective of this study was to screen endogenous microorganisms grown on olive mill solid wastes (OMSW) with the potential to ferment pentoses and produce ethanol. Two yeasts were isolated and identified as Issatchenkia orientalis, and Pichia galeiformis/manshurica. The adaptation of the strains displayed a positive impact on the fermentation process. In terms of xylose utilization and ethanol production, all strains were able to utilize xylose and produce xylitol but no ethanol was detected. Separate hydrolysis and fermentation process on hydrolysate undergo detoxification, strain I. orientalis showed the best efficiency in producing of ethanol when supplemented with glucose. Using simultaneous saccharification and fermentation process following pretreatment of OMSW, the average ethanol yield was 3 g/100 g dry OMSW. Bioethanol production from OMSW is not economic despite the raw material is cheap.

Topics: beta-Glucosidase; Biofuels; Carbohydrate Metabolism; Cellulase; Charcoal; Ethanol; Fermentation; Glucose; Hydrolysis; Industrial Waste; Olea; Solid Waste; Sulfuric Acids; Xylose; Yeasts

This study evaluated a cost-effective approach for the conversion of rice straw into fermentable sugars. The composition of rice straw pretreated with 1 % sulfuric acid or 1 % sodium hydroxide solution was compared to rice straw with no chemical pretreatment. Enzymatic saccharification experiments on non-pretreated rice straw (NPRS), pretreated rice straw (PRS), and pretreated rice straw with acid hydrolysate (PRSAH) were conducted in a series of batch reactors. The results indicated that pretreating the rice straw with dilute acid and base increased the cellulose content from 38 % to over 50 %. During enzymatic saccharification, straight aliphatic cellulose was hydrolyzed before branched hemicellulose, and glucose was the major hydrolysis product. The glucose yield was 0.52 g glucose/g for NPRS and was comparable to the yields of 0.50 g glucose/g for PRS and 0.58 g glucose/g for PRSAH. The hydrolysis of rice straw to produce glucose can be described by a first-order reaction with a rate constant of 0.0550 d(-1) for NPRS, 0.0653 d(-1) for PRSAH, and 0.0654 d(-1) for PRS. Overall, the production of fermentable sugars from ground rice straw will be more cost effective if the straw is not pretreated with chemicals.

Topics: Acids; beta-Glucosidase; Biomass; Biotechnology; Carbohydrates; Cellulase; Cellulose; Enzymes; Fermentation; Glucose; Hydrolysis; Oryza; Polymers; Polysaccharides; Sodium Hydroxide; Sulfuric Acids; Temperature; Time Factors

The enzymatic cocktail of cellulases is one of the most costly inputs affecting the economic viability of the biochemical route for biomass conversion into biofuels and other chemicals. Here, the influence of liquid hot water, dilute acid, alkali, and combined acid/alkali pretreatments on sugarcane bagasse (SCB) used for cellulase production was investigated by means of spectroscopic and imaging techniques. Chemical composition and structural characteristics, such as crystallinity (determined by X-ray diffraction), functional groups (Fourier transform infrared spectroscopy), and microstructure (scanning electron microscopy), were used to correlate SCB pretreatments with enzymatic biosynthesis by a strain of the filamentous fungus Aspergillus niger under solid-state fermentation. The combined acid/alkali pretreatment resulted in a SCB with higher cellulose content (86.7%). However, the high crystallinity (74%) of the resulting biomass was detrimental to microbial uptake and enzyme production. SCB pretreated with liquid hot water yielded the highest filter paper cellulase (FPase), carboxymethyl cellulase (CMCase), and xylanase activities (0.4, 14.9, and 26.1 U g(-1), respectively). The results showed that a suitable pretreatment for SCB to be used as a substrate for cellulase production should avoid severe conditions in order to preserve amorphous cellulose and to enhance the physical properties that assist microbial access.

Topics: Aspergillus niger; Biofuels; Cellulase; Cellulose; Crystallization; Endo-1,4-beta Xylanases; Ethanol; Fermentation; Hot Temperature; Hydrolysis; Microscopy, Electron, Scanning; Saccharum; Sodium Hydroxide; Spectroscopy, Fourier Transform Infrared; Sulfuric Acids; Water; X-Ray Diffraction

Gracilaria sp., a red alga, was used as a feedstock for the production of bioethanol. Saccharification of Gracilaria sp. by sequential acid and enzyme hydrolysis in situ produced a high quality hydrolysate that ensured its fermentability to produce ethanol. The optimal saccharification process resulted in total 11.85g/L (59.26%) of glucose and galactose, Saccharomyces cerevisiae Wu-Y2 showed a good performance on co-fermentability of glucose and galactose released in the hydrolysate from Gracilaria sp. The final ethanol concentrations of 4.72g/L (0.48g/g sugar consumed; 94% conversion efficiency) and the ethanol productivity 4.93g/L/d were achieved. 1g of dry Gracilaria can be converted to 0.236g (23.6%) of bioethanol via the processes developed. Efficient alcohol production by immobilized S. cerevisiae Wu-Y2 in batch and repeated batch fermentation was also demonstrated. The findings of this study revealed that Gracilaria sp. can be a potential feedstock in biorefinery for ethanol production.

Topics: Biofuels; Biomass; Biotechnology; Carbohydrate Metabolism; Cells, Immobilized; Cellulase; Ethanol; Fermentation; Gracilaria; Hydrogen-Ion Concentration; Hydrolysis; Sulfuric Acids; Time Factors

In the present study, two commonly used catalysts in chemical pretreatment, sulfuric acid and sodium hydroxide, were tested to evaluate the effect of solid-to-liquid ratio on pretreatment and enzymatic hydrolysis. Solid to liquid ratio (S/L) was influential on sugars released with an increase in the S/L ratio between 0.03 and 0.2. Enzymatic digestibility of 0.25 M H2SO4 pretreated corncobs were released more sugars (415.12 mg/mL); whereas, corncobs pretreated with NaOH released 350.12 mg/mL of reducing sugars at S/L 0.05. Further, in comparison with NaOH pretreated corncobs, acid treated material substantially increased the accessibility and digestibility of cellulose during crude enzymatic hydrolysis (28.96 FPU) and released 398.95 mg/mL reducing sugars.

Topics: Biotechnology; Carbohydrates; Cellulase; Hydrolysis; Sodium Hydroxide; Sulfuric Acids; Zea mays

Two commonly used chemical pretreatment processes, sulphuric acid, and sodium hydroxide, were tested to provide comparative performance data. A connection between solid to liquid ratio (S/L) and sugars released was observed with an increase in S/L ratio between 0.02 and 0.2. Enzymatic digestibility of 1 M of NaOH-pretreated corncobs were released 210.7 mg ml(-1) of sugars. Further, compared with different concentrations of acid pretreatments at 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, and 0.5 M concentrations, sodium hydroxide pretreatment of corncob substantially increased accessibility and digestibility of cellulose. Another additional observation made was whole-cell and crude enzymatic hydrolysis of different concentrations of acid and NaOH (0.05, 0.1, 0.25 M)-treated materials released lower amount of sugars compared with the sugars released (310.9 mg ml(-1)) with whole-cell hydrolysis of 1 M of NaOH-treated corncobs. NaOH-pretreated corncobs contained higher content of sugars and which is more suitable for production of reducing sugars.

Topics: Alkalies; Biotechnology; Carbohydrates; Cellulase; Dose-Response Relationship, Drug; Hydrolysis; Phanerochaete; Sodium Hydroxide; Sulfuric Acids; Zea mays

Sorghum is one of the commercially feasible lignocellulosic biomass and has a great potential of being sustainable feedstock for renewable energy. As with any lignocellulosic biomass, sorghum also requires pretreatment which increases its susceptibility to hydrolysis by enzymes for generating sugars which can be further fermented to alcohol. In the present study, sorghum biomass was evaluated for deriving maximum fermentable sugars by optimizing various pretreatment parameters using statistical optimization methods. Pretreatment studies were done with H2SO4, followed by enzymatic saccharification. The efficiency of the process was evaluated on the basis of production of the total reducing sugars released during the process. Compositional analysis was done for native as well as pretreated biomass and compared. The biomass pretreated with the optimized conditions could yield 0.408 g of reducing sugars /g of pretreated biomass upon enzymatic hydrolysis. The cellulose content in the solid portion obtained after pretreatment using optimised conditions was found to be increased by 43.37% with lesser production of inhibitors in acid pretreated liquor.

Topics: Biomass; Carbohydrates; Cellulase; Fermentation; Hydrochloric Acid; Hydrogen-Ion Concentration; Hydrolysis; Nitric Acid; Plant Extracts; Plant Stems; Polysaccharides; Sorghum; Sulfuric Acids; Temperature; Xylose

The effects of dilute H2SO4 concentration, forage:sulfuric acid ratio, digestion time, and digestion temperature were evaluated to determine effects on ethanol yield of Triarrhena sacchariflora (Maxim.) Nakai. Twenty single factor experiments were conducted to evaluate H2SO4 concentration (0.5, 1.0, 1.5, 2.0, and 2.5%, w/w), forage:sulfuric acid ratio (1:6, 1:8, 1:10, 1:12, and 1:14, g/ml), digestion time (15, 30, 45, 60, and 90, min), digestion temperature (80, 100, 110, 120, and 125 °C) for 3 replicates of the 5 levels of each factor. Based on results of the single factor experiments, an incomplete factorial was designed to evaluate ethanol yield from the best combinations of single factors. Finally, the best combination was tested by enzymatic hydrolysis and fermentation experiment in selected combinations according to pretreatment results. Percentage cellulose, hemicellulose, and lignin contents of forage residue after pretreatment, and glucose and xylose concentrations of the filtrate were analyzed prior to enzymatic hydrolysis, and percentage crystallinity was observed in untreated grass and pretreated residue. In addition, the solid residues were then hydrolysed and fermented by cellulase and yeast, the concentrations of glucose and ethanol being monitored for 96 h. Results showed that the order of the effect of main effect factors was as follows: digestion temperature > dilute H2SO4 concentration > digestion time > forage:sulfuric acid ratio. The best process parameters evaluated were sulfuric acid concentration of 1.5%, forage:sulfuric acid ratio of 1:6, digestion time of 15 min, and digestion temperature of 120°C. With this combination of factors, 80% of the cellulose was hydrolysed in 96 h, and 78% converted to ethanol. The findings identified that hemicelluloses were the key deconstruction barrier for pretreatment of Triarrhena sacchariflora (Maxim.) Nakai for ethanol production. The results of this research provide evidence of appropriate combinations of processing factors for production of ethanol from Triarrhena sacchariflora (Maxim.) Nakai.

Topics: Bioreactors; Biotechnology; Cellulase; Dose-Response Relationship, Drug; Ethanol; Fermentation; Glucose; Hydrolysis; Kinetics; Lignin; Poaceae; Sulfuric Acids; Temperature; Xylose

Cellulose consists of amorphous and crystalline regions. It is the crystalline regions which may be exploited to produce nanocrystalline cellulose (NCC). In order to extract nanocrystalline cellulose from native cellulose, sulfuric acid hydrolysis is typically used. The amorphous regions of cellulose are hydrolyzed and degraded into soluble products while the crystalline regions remain intact. In an effort to make the NCC extraction process more feasible, a new process was developed to recover and utilize the hydrolyzed regions of cellulose as a byproduct. The acid hydrolyzed amorphous regions were separated and then recovered (regenerated) into solid particles. XRD data revealed that the recovered material is characteristic of cellulose II. Hydrolysis conditions were optimized to maximize the yield of the recovered material and at the same time produce NCC material. Preliminary experiments showed yield values of approximately 61% for the cellulose I crystalline portions and values of about 21.7% for the recovered material (cellulose II). Enzymatic hydrolysis experiments of the recovered material revealed high susceptibility to enzymatic hydrolysis which makes it a promising source for biofuels production.

Topics: Biofuels; Cellulase; Cellulose; Chemical Fractionation; Gossypium; Hydrolysis; Microscopy, Electron, Scanning; Nanostructures; Nanotechnology; Solubility; Sulfuric Acids; Trichoderma; X-Ray Diffraction

The acidic ionic liquid 1-H-3-methylimidazolium chloride can effectively pretreat yellow pine wood chips under mild conditions for enzymatic saccharification. Wood samples were treated at temperatures between 110 and 150°C for up to 5 h in the ionic liquid and three fractions collected; a cellulose rich fraction, lignin, and an aqueous fraction. This treatment caused the hemicellulose and the lignin to be degraded and dissolved from the cell walls of the pine wood. The lignin was depolymerized and subsequently dissolved in the ionic liquid. This process occurred more quickly at higher temperatures, although at the highest temperatures tested, significant cellulose degradation also occurred. The cellulose rich fraction was saccharified using cellulase from Trichoderma viride, with longer pretreatment times at 130°C resulting in higher glucose yields.

Topics: Biotechnology; Borates; Cellulase; Glucose; Imidazoles; Ionic Liquids; Lignin; Mannose; Pinus; Polysaccharides; Sulfuric Acids; Trichoderma

Cassava cellulose accounts for one quarter of cassava residues and its utilization is important for improving the efficiency and profit in commercial scale cassava ethanol industry. In this study, three scenarios of cassava cellulose utilization for ethanol production were experimentally tested under same conditions and equipment. Based on the experimental results, a rigorous flowsheet simulation model was established on Aspen plus platform and the cost of cellulase enzyme and steam energy in the three cases was calculated. The results show that the simultaneous co-saccharification of cassava starch/cellulose and ethanol fermentation process (Co-SSF) provided a cost effective option of cassava cellulose utilization for ethanol production, while the utilization of cassava cellulose from cassava ethanol fermentation residues was not economically sound. Comparing to the current fuel ethanol selling price, the Co-SSF process may provide an important choice for enhancing cassava ethanol production efficiency and profit in commercial scale.

Topics: Biotechnology; Carbohydrate Metabolism; Cellulase; Cellulose; Computer Simulation; Costs and Cost Analysis; Ethanol; Fermentation; Hydrolysis; Manihot; Saccharomyces cerevisiae; Steam; Sulfuric Acids

Solid wood, metal finnish: Instead of burning waste wood treated with chromated copper arsenite (CCA) or disposing of it in landfills, the CCA-treated wood can be used as a raw material for the production of chemicals. Catalytic or alkaline oxidation together with very mild sulfuric acid extraction produces an easily enzymatically hydrolyzable material. Usage as a raw material for the chemical industry in this manner demonstrates a sustainable and value-added waste management process.

Topics: Arsenates; beta-Glucosidase; Catalysis; Cellulase; Cellulose; Edetic Acid; Hazardous Waste; Hydrolysis; Oxalates; Oxidation-Reduction; Phenanthrolines; Recycling; Sulfuric Acids; Wood

This study investigates the potential for extracting sugars from the polysaccharides of Eastern redcedar. Pretreatment temperature, time, sulfuric acid loading, sodium bisulfite loading and impregnation time were varied using factorial treatment design experiments for identifying near optimal overall wood glucan-to-glucose yields during acid bisulfite pretreatments. The highest overall wood glucan-to-glucose yield of 87% was achieved when redcedar was impregnated with pretreatment liquor containing 3.75 g of sulfuric acid/100g of dry wood and 20 g of sodium bisulfite/100g of dry wood at 90 °C for 3h followed by increasing the temperature to 200 °C with a hold time of 10 min. Hemicellulose and lignin removal during pretreatments made the substrate amenable to enzymatic hydrolysis using 0.5 ml of Accelerase® 1500/g of glucan at 2% (w/w) solid loading. Preliminary mass balances showed 97% glucan recovery at pretreatment condition with 87% overall wood glucan-to-glucose yield and 59% delignification.

Topics: Biotechnology; Carbohydrate Metabolism; Cellulase; Glucans; Glucose; Hydrolysis; Juniperus; Sulfites; Sulfuric Acids; Surface-Active Agents; Temperature; Time Factors; Wood

Spent mushroom substrate (SMS) was pretreated with alkaline reagents including potassium hydroxide, lime and ammonia to enhance enzymatic saccharification. Under the best pretreatment conditions (1M KOH, 80 °C, 90 min; 1M lime, 80 °C, 120 min; 10 M ammonia, 70 °C, 120 min), the total reducing sugar (TRS) yield reached 258.6, 204.2 and 251.2 mg/g raw SMS, which were respectively 6.15, 4.86, and 5.98 times of untreated SMS. The effects of pretreatment by above alkaline reagents and sulfuric acid on the composition and structure of SMS were evaluated to provide comparative performance data. A new process, combined alkali and acid (CAA) pretreatment followed by enzymatic hydrolysis, was innovatively proposed to improve the cost-effectiveness and avoid environmental problems. The SMS residue after CAA pretreatment-enzymatic hydrolysis process was converted to biofertilizer with Pichia farinose FL7 and a cell density of 3.0×10(8) cfu/g in biomass was attained.

Topics: Agaricales; Alkalies; Biotechnology; Carbohydrates; Cellulase; Fertilizers; Hydrolysis; Hydroxides; Lignin; Oxidation-Reduction; Polysaccharides; Potassium Compounds; Recycling; Sulfuric Acids

Various pretreatments on Ceiba pentandra (L.) Gaertn. (kapok) fiber prior to enzymatic hydrolysis for sugar production were optimized in this study. The optimum conditions for water, acid, and alkaline pretreatments were 170°C for 45 min, 120°C for 45 min in 1.0% (v/v) H2SO4 solution and 120°C for 60 min in 2.0% (v/v) NaOH solution, respectively. Among the three pretreatments, the alkaline pretreatment achieved the highest total glucose yield (glucose yield calculated based on the untreated fiber) (38.5%), followed by the water (35.0%) and acid (32.8%) pretreatments. As a result, the relative effectiveness of the pretreatment methods for kapok fiber was verified as alkali>water>acid at the condition stated.

Topics: Alkalies; Biofuels; Biotechnology; Carbohydrate Metabolism; Carbohydrates; Ceiba; Cellulase; Ethanol; Hydrolysis; Sulfuric Acids; Water

The effects of subcritical water (SCW) and dilute acid pretreatments on the shedding bark of Melaleuca leucadendron (paper bark tree, PBT) biomass morphology, crystallinity index (CrI) and enzymatic saccharification were studied. The morphology of PBT bark was characterized by X-ray diffraction, scanning electron microscopy and Fourier transform infrared spectroscopy. SCW pretreatment mainly extracted amorphous parts of the biomass hence its CrI increased, partial decrystallization of cellulose and exposing of intact nanofibers of cellulose were observed for SCW pretreatment at 180°C. On the other hand, dilute acid pretreatment at 160°C exhibited a large decrease in CrI, an increase in surface area, a decrease in lignin content and decrystallization of cellulose as well as the peel-off and degradation of some nanofiber bundles. Dilute acid and SCW pretreatments of PBT biomass resulted in about 4.5 fold enhancement in glucose release relative to the untreated one.

Topics: beta-Glucosidase; Biomass; Carbohydrate Metabolism; Cellulase; Crystallization; Hydrolysis; Melaleuca; Plant Bark; Spectroscopy, Fourier Transform Infrared; Sulfuric Acids; Water; X-Ray Diffraction

In this work the pretreatment of sunflower stalks by dilute sulfuric acid is studied. Pretreatment temperature and the concentration of acid solution were selected as operation variables and modified according to a central rotatable composite experimental design. Based on previous studies pretreatment time was kept constant (5 min) while the variation range for temperature and acid concentration was centered at 175°C and 1.25% (w/v) respectively. Following pretreatment the insoluble solids were separated by filtration and further submitted to enzymatic hydrolysis, while liquid fractions were analyzed for sugars and inhibitors. Response surface methodology was applied to analyze results based on the combined severity of pretreatment experiments. Optimized results show that up to 33 g of glucose and xylose per 100g raw material (65% of the glucose and xylose present in the raw material) may be available for fermentation after pretreatment at 167°C and 1.3% sulfuric acid concentration.

Topics: Biotechnology; Carbohydrates; Cellulase; Fermentation; Glucose; Helianthus; Hydrolysis; Sulfuric Acids; Waste Products; Xylose

In this study, the effects of different acid catalysts and pretreatment factors on the hydrolysis of mixed softwood were investigated over a range of thermochemical pretreatments. Maleic, oxalic, and sulfuric acids were each used, under different pretreatment conditions. The most influential factor for fermentable sugar production in the dicarboxylic acid pretreatment of softwood was the pH. Reaction temperature was the next significant factor. However, during sulfuric acid pretreatment, fermentable sugar production was more dependent on reaction temperature, than time or pH. Enzymatic hydrolysis yields differed, depending on acid catalyst and pretreatment factor, regardless of lignin content in pretreated biomass. The highest enzymatic hydrolysis yield was found following maleic acid pretreatment, which reached 61.23%. The trend in enzymatic hydrolysis yields that were detected concomitantly with pretreatment condition or type of acid catalyst was closely related to the fermentable sugar production in the hydrolysate.

Topics: beta-Glucosidase; Biomass; Biotechnology; Carbohydrates; Catalysis; Cellulase; Cellulose; Fermentation; Hydrolysis; Oligosaccharides; Pinus; Sulfuric Acids; Wood

The pretreatment of lignocellulosic biomass is crucial for efficient subsequent enzymatic hydrolysis and ethanol fermentation. In this study, wet explosion (WEx) pretreatment was applied to cocksfoot grass and pretreatment conditions were tailored for maximizing the sugar yields using response surface methodology. The WEx process parameters studied were temperature (160-210 °C), retention time (5-20 min), and dilute sulfuric acid concentration (0.2-0.5 %). The pretreatment parameter set E, applying 210 °C for 5 min and 0.5 % dilute sulfuric acid, was found most suitable for achieving a high glucose release with low formation of by-products. Under these conditions, the cellulose and hemicellulose sugar recovery was 94 % and 70 %, respectively. The efficiency of the enzymatic hydrolysis of cellulose under these conditions was 91 %. On the other hand, the release of pentose sugars was higher when applying less severe pretreatment conditions C (160 °C, 5 min, 0.2 % dilute sulfuric acid). Therefore, the choice of the most suitable pretreatment conditions is depending on the main target product, i.e., hexose or pentose sugars.

Topics: Carbohydrates; Cellulase; Cellulose; Computer Simulation; Models, Chemical; Poaceae; Steam; Sulfuric Acids

This work explores the feasibility of recycling cellulase by electroultrafiltration (EUF), an ultrafiltration process enhanced by an electric field, to reduce the cost of enzymatic transformation of cellulose. The effect of electric field under different operating conditions (buffer concentration, acid treated wheat straw concentration, current and temperature) on flux during EUF was examined. The results showed that EUF was effective to reduce concentration polarization (CP) and enhance filtration flux in recycling cellulase. The flux improvement by the electric field could be strengthened at low buffer concentration (5 mM) and relatively low temperature (room temperature) and high current (150 mA). The flux for 2% (substrate concentration, w/v) lignocellulosic hydrolyzate increased by a factor of 4.4 at 836 V/m and room temperature, compared to that without electric field. This work shows that under appropriate operating conditions EUF can efficiently recycle cellulase from lignocellulosic hydrolyzate and thus substantially reduce hydrolysis cost.

Topics: Buffers; Cellulase; Electricity; Hydrolysis; Permeability; Recycling; Sulfuric Acids; Temperature; Trichoderma; Triticum; Ultrafiltration; Waste Products

Aiming at increasing the efficiency of transferring corn stover into sugars, a biological pretreatment was developed and investigated in this study. The protocol was characterized by the pretreatment with crude ligninolytic enzymes from Phanerochete chrysosporium and Coridus versicolor to break the lignin structure in corn stover, followed by a washing procedure to eliminate the inhibition of ligninolytic enzyme on cellulase. By a 2 d-pretreatment, sugar yield from corn stover hydrolysis could be increased by 50.2% (up to 323 mg/g) compared with that of the control. X-ray diffractometry and FT-IR analysis revealed that biological pretreatment could partially remove the lignin of corn stover, and consequently enhance the enzymatic hydrolysis efficiency of cellulose and hemeicellulose. In addition, the amount of microbial inhibitors, such as acetic acid and furfural, were much lower in biological pretreatment than that in acid pretreatment. This study provided a promising pretreatment method for biotransformation of corn stovers.

Topics: Carbohydrates; Cellulase; Cellulose; Hydrolysis; Laccase; Lignin; Sulfuric Acids; Waste Products; Zea mays

Saccharification of sugarcane bagasse pretreated at the pilot-scale with different processes (in combination with steam-explosion) was evaluated. Maximum glucan conversion with Celluclast 1.5L (15-25FPU/g glucan) was in the following order: glycerol/HCl>HCl>H2SO4>NaOH, with the glycerol system achieving ≈ 100% conversion. Surprisingly, the NaOH substrate achieved optimum saccharification with only 8 FPU/g glucan. Glucan conversions (3.6-6%) obtained with mixtures of endo-1,4-β-glucanase (EG) and β-glucosidase (βG) for the NaOH substrate were 2-6 times that of acid substrates. However, glucan conversions (15-60%) obtained with mixtures of cellobiohydrolase (CBH I) and βG on acidified glycerol substrate were 10-30% higher than those obtained for NaOH and acid substrates. The susceptibility of the substrates to enzymatic saccharification was explained by their physical and chemical attributes. Acidified glycerol pretreatment offers the opportunity to simplify the complexity of enzyme mixtures required for saccharification of lignocellulosics.

Topics: beta-Glucosidase; Biomass; Carbohydrate Metabolism; Cellulase; Cellulose; Hydrolysis; Microscopy, Electron, Scanning; Pilot Projects; Recombinant Proteins; Saccharum; Sodium Hydroxide; Solutions; Spectroscopy, Fourier Transform Infrared; Steam; Sulfuric Acids; X-Ray Diffraction

Pretreatment is an essential step in the enzymatic hydrolysis of biomass and subsequent production of bioethanol. The current study is focused on two different pretreatment methods of wheat straw using mild temperatures (100°C for 2h and RT for overnight). In one method, native substrate was treated with 1.5% (w/v) NaOH at two different above mentioned conditions followed by acid hydrolysis (0.75% (v/v) sulfuric acid at 100°C for 2h). In another method, the native substrate was initially treated with acid (0.75% (v/v) sulfuric acid at 100°C for 2h) followed by treatment with 1.5% (w/v) NaOH at two different above conditions. After the pretreatments, the residues were treated with Accellerase 1500 (26U/g) and maximum yield of glucose (65.2gL(-1)) were found with 0.75% sulfuric acid (100°C for 2h) followed by alkali (1.5% NaOH at 100°C for 2h). Fermentation of this hydrolyzate using Saccharomyces cerevisiae strain produced 24.4gL(-1) of ethanol with corresponding yield of 0.44g/g.

Topics: Biofuels; Biomass; Cellulase; Ethanol; Fermentation; Hydrolysis; Lignin; Saccharomyces cerevisiae; Sodium Hydroxide; Sulfuric Acids; Triticum

An extremely low acid (ELA) pretreatment using 0.06% (w/w) sulfuric acid at 170 °C for 15 min was employed to extract non-glucan components from Saccharina japonica, a brown macroalgae. Subsequent simultaneous saccharification and fermentation (SSF) was conducted using Saccharomyces cerevisiae DK 410362 and cellulase (15 FPU/g-glucan) and ß-glucosidase (70 pNPGU/g-glucan). Deionized water was used for making fermentation suspension. After the ELA pretreatment, a glucan content of 29.10% and an enzymatic digestibility of 83.96% was obtained for pretreated S. japonica. These values are 4.2- and 2.4-fold higher, respectively, than those of obtained with untreated S. japonica. In SSF, a bioethanol concentration of 6.65 g/L was obtained, corresponding to a glucose equivalent concentration of 13.01 g/L, which indicated an SSF yield of 67.41% based on the total available glucan of the pretreated S. japonica. The remaining separated liquid hydrolysate, which contains mannitol and alginate-derived oligosaccharides can be applied to other fermentations.

Topics: beta-Glucosidase; Biofuels; Cellulase; Chromatography, High Pressure Liquid; Ethanol; Fermentation; Glucans; Hydrolysis; Phaeophyceae; Saccharomyces cerevisiae; Sulfuric Acids; Temperature; Time Factors

Lodgepole wood chips were pretreated by sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL) at 25% solids loading and 180 °C for 20 min with sulfuric acid and sodium bisulfite charges of 2.2 and 8 wt/wt% on an oven-dry wood basis, respectively. The pretreated wood chips were disk-milled with pretreatment spent liquor and water, and the solid fraction was separated from the liquor stream. The liquor was neutralized and concentrated through vacuum evaporation. Quasi-simultaneous enzymatic saccharification of the cellulosic solids and combined fermentation with the concentrated liquor was conducted at up to 20% total solids loading. Fed-batching of the solids facilitated liquefaction and saccharification, as well as managing instantaneous inhibitor concentrations. At a commercial cellulase (CTec2) loading of only 9 FPU or 0.06 mL/g untreated wood, a maximum ethanol titer of 47.4 g/L was achieved, resulting in a calculated yield of 285 L/tonne of wood using Saccharomyces cerevisiae YRH400 at 35 °C and pH 5.5.

Topics: Biofuels; Cellulase; Ethanol; Fermentation; Pinus; Saccharomyces cerevisiae; Sulfites; Sulfuric Acids

Wheat straw fractionation by ethanol organosolv was studied as pretreatment for enzymatic cellulose hydrolysis. A parametric study focusing on temperature, reaction time, acid catalyst dose, solvent concentration, and particle size was performed to determine their influence on delignification, xylan hydrolysis, and enzymatic cellulose digestibility. Major process parameters were found to be temperature, ethanol concentration, and acid dose. Optimisation of the process towards enzymatic digestibility resulted in a maximum glucose yield of 86% without the use of a catalyst (lignin yield 84%, organosolv at 210 °C, 50% w/w aqueous EtOH). Using 30 mM H2SO4 as catalyst resulted in similar glucose and lignin yields at a lower temperature (190 °C, 60% w/w aqueous EtOH). Lowering the pretreatment temperature by using an acid catalyst substantially improved the yield of the hemicellulose derivatives xylose and furfural. A systematic approach in pretreatment optimisation is vital for development of efficient lignocellulosic biorefineries.

Topics: Biocatalysis; Biomass; Cellulase; Cellulose; Chemical Fractionation; Ethanol; Hydrogen-Ion Concentration; Lignin; Particle Size; Solvents; Sulfuric Acids; Temperature; Time Factors; Triticum; Waste Products

Degradation of cellulose by anaerobic microbial consortium is brought about either by an exocellular process or by secretion of extracellular enzymes. In this work, a novel route for synthesis of nanocellulose is described where in an anaerobic microbial consortium enriched for cellulase producers is used for hydrolysis. Microcrystalline cellulose derived from cotton fibers was subjected to controlled hydrolysis by the anaerobic microbial consortium and the resultant nanocellulose was purified by differential centrifugation technique. The nanocellulose had a bimodal size distribution (43±13 and 119±9 nm) as revealed by atomic force microscopy. A maximum nanocellulose yield of 12.3% was achieved in a span of 7 days. While the conventional process of nanocellulose preparation using 63.5% (w/w) sulfuric acid resulted in the formation of whisker shaped nanocellulose with surface modified by sulfation, controlled hydrolysis by anaerobic microbial consortium yielded spherical nanocellulose also referred to as nano crystalline cellulose (NCC) without any surface modification as evidenced from Fourier transform infrared spectroscopy. Also, it scores over chemo-mechanical production of nanofibrillated cellulose by consuming less energy due to enzyme (cellulase) assisted catalysis. This implies the scope for use of microbial prepared nanocellulose in drug delivery and bio-medical applications requiring bio-compatibility.

Topics: Bacteria, Anaerobic; Bacterial Proteins; Biocompatible Materials; Cellulase; Cellulose; Cotton Fiber; Drug Delivery Systems; Fungal Proteins; Hydrogen Bonding; Hydrolysis; Microbial Consortia; Microscopy, Atomic Force; Nanospheres; Particle Size; Pliability; Powders; Spectroscopy, Fourier Transform Infrared; Sulfuric Acids; Ultrafiltration

In this study, synergetic effect of mild acid and alkali with electron beam irradiation (EBI) on the enzymatic hydrolysis of a selected grass biomass was assessed. Biomass samples prepared by soaking with 1% H2SO4, or 1% NaOH, were exposed to 75 and 150 kGy of EBI. Water presoaked biomass was used as control. Hydrolysis of pretreated samples was carried out using cellulase (15 FPU/g biomass) for 120 h. Structural changes were studied by FTIR and XRD analyses. Reducing sugar and glucose yields from enzymatic hydrolysis were significantly higher in acid and alkali presoaked EBI exposed samples. Theoretical glucose yield showed 40% increase from control in alkali presoaked EBI exposed (150 kGy) samples. Removal of hemicellulose, decreased crystallinity and structural changes were major factors for the combined treatment effect favoring the hydrolysis.

Topics: Alkalies; Cellulase; Electrons; Enzyme Activation; Hydrolysis; Lignin; Plant Extracts; Poaceae; Polysaccharides; Radiation Dosage; Sodium Hydroxide; Sulfuric Acids

The purpose of this study was to fractionate switchgrass (SG) to obtain hemicellulose-, lignin-rich fractions and highly digestible pulp, using a clean fractionation (CF) approach. The main objective was to produce highest glucose yield in the enzymatic hydrolysis of pulp. Effects of processing factors such as time (10-50 min), temperature (120-160 °C), catalyst concentration (0.21-0.93% w/w sulfuric acid) and organic solvent mixture composition (7-43% w/w methyl isobutyl ketone) were evaluated. Response surface methodology and central composite design were used for process optimization and statistical analyses. High lignin (75-93%) and xylan (83-100%) removal from biomass were obtained, leaving solid pulp rich in glucan (78-94%). High enzymatic hydrolysis glucose yields (more than 90%) were obtained for selected optimal conditions. Pulp can be used for ethanol production while separated xylan and lignin fractions can be used as a feedstock for value-added products which suggests the applicability of clean fractionation technology in a biorefinery concept.

Topics: beta-Glucosidase; Cellulase; Chemical Fractionation; Computer Simulation; Glucose; Hydrolysis; Lignin; Models, Chemical; Plant Extracts; Poaceae; Sulfuric Acids

Dilute sulfuric acid pretreatment of oil palm empty fruit bunches (EFB) followed by the whole slurry fermentation of the pretreated EFB slurry was investigated. The optimized pretreatment conditions were at 1% (w/v) sulfuric acid with 3 min ramping to 190 °C in a microwave digester. Pretreated and washed EFB exhibited enzymatic digestibility of 88.5% of theoretical glucose yield after 48 h of hydrolysis. When the whole slurry of pretreated and neutralized EFB was used in simultaneous saccharification and fermentation (SSF) using cellulase and Saccharomyces cerevisiae, sulfuric acid-pretreated EFB resulted in 52.5% of theoretical ethanol yield based on total glucan in the untreated initial EFB after 72 h of SSF. When pretreated EFB slurry was treated with activated carbon before subjecting to SSF, the SSF furnished 87.5% ethanol yield based on the initial glucan content in untreated EFB (after 48 h of SSF).

Topics: Arecaceae; Biotechnology; Cellulase; Charcoal; Ethanol; Fermentation; Fruit; Glucose; Hydrolysis; Lignin; Microwaves; Saccharomyces cerevisiae; Sulfuric Acids

Conventional Alamo switchgrass and its transgenic counterparts with reduced/modified lignin were subjected to dilute sulfuric acid pretreatment for improved sugar production. At 150 °C, the effects of acid concentration (0.75%, 1%, 1.25%) and residence time (5, 10, 20, 30 min) on sugar productions in pretreatment and enzymatic hydrolysis were investigated, with the optimal pretreatment conditions determined for each switchgrass genotype based on total sugar yield and the amounts of sugar degradation products generated during the pretreatment. The results show that genetic engineering, although did not cause an appreciable lignin reduction, resulted in a substantial increase in the ratio of acid soluble lignin:acid insoluble lignin, which led to considerably increased sugar productions in both pretreatment and enzymatic hydrolysis. At an elevated threshold concentration of combined 5-hydroxyfuranmethal and furfural (2.0 g/L), the overall carbohydrate conversions of conventional switchgrass and its transgenic counterparts, 10/9-40 and 11/5-47, reached 75.9%, 82.6%, and 82.2%, respectively.

Topics: Carbohydrates; Cellulase; Panicum; Plants, Genetically Modified; Sulfuric Acids

Dilute acid pretreatment is a leading pretreatment technology for biomass to ethanol conversion due to the comparatively low chemical cost and effective hemicellulose solubilization. The conventional dilute acid pretreatment processes use relatively large quantities of sulfuric acid and require alkali for pH adjustment afterwards. Significant amounts of sulfate salts are generated as by-products, which have to be properly treated before disposal. Wastewater treatment is an expensive, yet indispensable part of commercial level biomass-to-ethanol plants. Therefore, reducing acid use to the lowest level possible would be of great interest to the emerging biomass-to-ethanol industry. In this study, a dilute acid pretreatment process was developed for the pretreatment of corn stover. The pretreatment was conducted at lower acid levels than the conventional process reported in the literature while using longer residence times. The study indicates that a 50% reduction in acid consumption can be achieved without compromising pretreatment efficiency when the pretreatment time was extended from 1-5 min to 15-20 min. To avoid undesirable sugar degradation and inhibitor generation, temperatures should be controlled below 170°C. When the sulfuric acid-to-lignocellulosic biomass ratio was kept at 0.025 g acid/g dry biomass, a cellulose-to-glucose conversion of 72.7% can be achieved at an enzyme loading of 0.016 g/g corn stover. It was also found that acid loading based on total solids (g acid/g dry biomass) governs the pretreatment efficiency rather than the acid concentration (g acid/g pretreatment liquid). While the acid loading on lignocellulosic biomass may be achieved through various combinations of solids loading and acid concentration in the pretreatment step, this work shows that it is unlikely to reduce acid use without undermining pretreatment efficiency simply by increasing the solid content in pretreatment reactors, therefore acid loading on biomass is indicated to be the key factor in effective dilute acid pretreatment.

Topics: Biomass; Carbohydrate Metabolism; Cellulase; Cellulose; Ethanol; Glycoside Hydrolases; Lignin; Polysaccharides; Sulfuric Acids; Temperature; Zea mays

Process simulation and lab trials were carried out to demonstrate and confirm the efficiency of the concept that recycling hydrolysate at low total solid enzymatic hydrolysis is one of the options to increase the sugar concentration without mixing problems. Higher sugar concentration can reduce the capital cost for fermentation and distillation because of smaller retention volume. Meanwhile, operation cost will also decrease for less operating volume and less energy required for distillation. With the computer simulation, time and efforts can be saved to achieve the steady state of recycling process, which is the scenario for industrial production. This paper, to the best of our knowledge, is the first paper discussing steady-state saccharification with recycling of the filtrate form enzymatic hydrolysis to increase sugar concentration. Recycled enzymes in the filtrate (15-30% of the original enzyme loading) resulted in 5-10% higher carbohydrate conversion compared to the case in which recycled enzymes were denatured. The recycled hydrolysate yielded 10% higher carbohydrate conversion compared to pure sugar simulated hydrolysate at the same enzyme loading, which indicated hydrolysis by-products could boost enzymatic hydrolysis. The high sugar concentration (pure sugar simulated) showed inhibition effect, since about 15% decrease in carbohydrate conversion was observed compared with the case with no sugar added. The overall effect of hydrolysate recycling at WinGEMS simulated steady-state conditions with 5% total solids was increasing the sugar concentration from 35 to 141 g/l, while the carbohydrate conversion was 2% higher for recycling at steady state (87%) compared with no recycling strategy (85%). Ten percent and 15% total solid processes were also evaluated in this study.

Topics: Biomass; Carbohydrates; Carbonates; Cellulase; Cellulose; Chromatography, High Pressure Liquid; Computer Simulation; Endo-1,4-beta Xylanases; Ethanol; Fermentation; Hydrolysis; Lignin; Recycling; Sulfides; Sulfuric Acids; Wood; Xylans

Pretreatment processes are key technologies for generating fermentable sugars based on lignocellulosic biomass. In this study, we developed a novel method for empty palm fruit bunch fiber (EPFBF) using sequential pretreatment with dilute acid and then alkali. Dilute sulfuric acid was used in the first step, which removed 90% of the hemicellulose and 32% of the lignin, but left most of the cellulose under the optimum pretreatment condition. Sodium hydroxide was then applied in the second step, which extracted lignin effectively with a 70% delignification yield, partially disrupting the ordered fibrils of the EPFBF and thus enhancing the enzyme digestibility of the cellulose. The sequentially pretreated biomass consisted of 82% cellulose, less than 1% hemicellulose, and 30% lignin content afterward. The pretreated biomasses morphologically revealed rough, porous, and irregularly ordered surfaces for enhancing enzyme digestibility. These results indicate that the sequentially acid/alkali-pretreated EPFBF could be broadly useful as a novel biomass.

Topics: Arecaceae; Biotechnology; Carbohydrate Metabolism; Cellulase; Dietary Fiber; Fruit; Hydrolysis; Lignin; Sodium Hydroxide; Sulfuric Acids

This study seeks to investigate the effects of biomass mixtures on overall sugar recovery from the combined processes of dilute acid pretreatment and enzymatic hydrolysis. Aspen, a hardwood species well suited to biochemical processing, was chosen as the model species for this study. Balsam, a high-lignin softwood species, and switchgrass, an herbaceous energy crop with high ash content, were chosen as adjuncts. A matrix of three different dilute acid pretreatment severities and three different enzyme loading levels was used to characterize interactions between pretreatment and enzymatic hydrolysis. No synergism or antagonism was observed for any of the feedstock mixtures. Maximum glucose yield was 70% of theoretical for switchgrass and maximum xylose yield was 99.7% of theoretical for aspen. Supplemental β-glucosidase increased glucose yield from enzymatic hydrolysis by an average of 15%. Total sugar recoveries for mixtures could be predicted to within 4% by linear interpolation of the pure species results.

Topics: Biomass; Biotechnology; Cellulase; Glucose; Hydrolysis; Panicum; Sulfuric Acids; Wood; Xylose

Pseudo-lignin, which can be broadly defined as aromatic material that yields a positive Klason lignin value and is not derived from native lignin, has been recently reported to form during the dilute acid pretreatment of poplar holocellulose. To investigate the chemistry of pseudo-lignin formation, GPC, FT-IR and 13C NMR were utilized to characterize pseudo-lignin extracted from dilute-acid pretreated α-cellulose and holocellulose. The results showed that pseudo-lignin consisting of carbonyl, carboxylic, aromatic and aliphatic structures was produced from dilute acid pretreated cellulose and hemicellulose. Pseudo-lignin extracted from holocellulose pretreated at different conditions had similar molecular weights (Mn∼1000 g/mol; Mw∼5000 g/mol) and structural features (carbonyl, carboxylic, aromatic and methoxy structures). These characterizations have provided the pseudo-lignin formation mechanisms during pretreatment. The presence and structure of pseudo-lignin is important since pseudo-lignin decreases the enzymatic conversion.

Topics: beta-Glucosidase; Biotechnology; Carbohydrate Metabolism; Cellulase; Cellulose; Glucose; Hydrolysis; Lignin; Molecular Weight; Populus; Spectroscopy, Fourier Transform Infrared; Sulfuric Acids; Time Factors

A lignocellulose pretreatment process was developed with formic acid delignification (FAD) followed by alkaline deformylation (AD), which was termed as Formiline process. In FAD, more than 80% of lignin and hemicellulose were removed, but cellulose formylation also happened. Formic acid concentration (FAC) was the most important factor affecting delignification and cellulose formylation. Increasing FAC could enhance degree of delignification but also increased cellulose formylation. The presence of formyl group could inhibit the enzymatic hydrolysis of cellulose; however, removing formyl group with a small loading of alkali well recovered cellulose digestibility. The spent liquor could be directly recycled for delignification thus significantly decreasing energy consumption in solvent recovery. The Formiline-pretreated substrates showed an excellent enzymatic digestibility and could be very well converted to ethanol by simultaneous saccharafication and fermentation (SSF). The final ethanol concentrations were 55.4 and 80.1g/L respectively at initial solid consistencies of 15% and 20%.

Topics: Alkalies; beta-Glucosidase; Biotechnology; Carbohydrate Metabolism; Cellulase; Cellulose; Chemical Fractionation; Ethanol; Fermentation; Formates; Lignin; Recycling; Saccharomyces cerevisiae; Saccharum; Sulfuric Acids; Time Factors; Water

Pretreatment of biomass for bioethanol production makes it necessary to use large amounts of water for removing inhibitors and neutralization. In order to reduce water usage, separate batches of corncobs were hydrolyzed in 1M NaOH and 0.05 M H(2)SO(4), respectively, and the hydrolysis products were mixed to achieve a pH of 7. This approach lowered water usage by 10-fold compared with neutralization by distilled and recycling wash water. Mixing of the pretreated biomasses (121°C, 20 min) increased release of reducing sugars during enzymatic hydrolysis with cellulases (38.49 FPU(IU)) produced by Phanerochaete chrysosporium NCIM 1106 by 2- and 15-fold compared with the sugars released from the unmixed NaOH- and H(2)SO(4)-treated corncobs, respectively. Enzymatic hydrolysis (EH, cell free extract) of the mixed material released 395.15 mg/ml of sugars during 48 h, slightly less than what was achieved by microbial hydrolysis (whole cell hydrolysis), 424.50mg after 120 h.

Topics: Carbohydrates; Cellulase; Phanerochaete; Plant Components, Aerial; Plant Extracts; Sodium Hydroxide; Sulfuric Acids; Water; Zea mays

Direct and efficient enzymatic synthesis of long-chain cellulose from cellobiose in its original form was successfully achieved via the combination of a surfactant-enveloped enzyme (SEE) and a protic acid in an aprotic organic solvent, lithium chloride/N,N-dimethylacetamide system. The SEE biocatalyst was prepared by protecting the surface of cellulase with the nonionic surfactant dioleyl-N-D-glucona-L-glutamate for keeping its enzymatic activity in nonaqueous media. Fourier transform infrared and nuclear magnetic resonance analyses elucidated the successful synthesis of cellulose, β-1,4-linked D-glucopyranose polymer, through the reverse hydrolysis of cellobiose. By using protic acid cocatalysts, a degree of polymerization of as-synthesized cellulose reached more than 120, in a ca. 26% conversion, which was 5 times higher than that obtained in an acid-free SEE system. A novel-concept biocatalysis, i.e., a protic acid-assisted SEE-mediated reaction, enables a facile, one-step chain elongation of carbohydrates without any activation via multistep organic chemistry, and can provide potential applications in the functional design of glycomaterials.

Topics: Biocatalysis; Cellobiose; Cellulase; Cellulose; Hydrolysis; Lithium Chloride; Magnetic Resonance Spectroscopy; Solvents; Spectroscopy, Fourier Transform Infrared; Sulfuric Acids; Surface-Active Agents; Water

Downregulated lignin transgenic black cottonwood (Populus trichocarpa) was used to elucidate the effect of lignin and xylan content on enzymatic saccharification. The lignin contents of three transgenic samples (4CL1-1, 4CL1-4, and CH8-1-4) were 19.3, 16.7, and 15.0 %, respectively, as compared with the wild type (21.3 %). The four pretreatments were dilute acid (0.1 % sulfuric acid, 185 °C, 30 min), green liquor (6 % total titratable alkali, 25 % sulfidity based on TTA, 185 °C, and 15 min.), autohydrolysis (185 °C, 30 min), and ozone delignification (25 °C, 30 min). Following the pretreatment, enzymatic saccharification was carried out using an enzyme charge of 5 FPU/g of substrates. The removal of lignin and hemicellulose varies with both the types of pretreatments and the lignin content of the transgenic trees. Due to the greatest removal of lignin, green liquor induced the highest sugar production and saccharification efficiency, followed by acid, ozone, and autohydrolysis in descending order. The results indicated that lignin is the main recalcitrance of biomass degradation. At a given lignin content, pretreatment with ozone delignification had lower saccharification efficiency than the other pretreatment methods due to higher xylan content.

Topics: Cellulase; Down-Regulation; Hydrogen-Ion Concentration; Hydrolysis; Lignin; Ozone; Plants, Genetically Modified; Polysaccharides; Populus; Sulfuric Acids; Wood

Agave americana is one of commonly grown agave species but currently less valuable because its large flower stalk cannot be used for producing alcoholic beverage. In the present study, the stalk was pretreated with dilute acid (DA), sulfite (SPORL), and sodium hydroxide (NaOH) to preliminarily assess its potential as feedstock for bioethanol production. The changes of cell wall components during the pretreatments, enzymatic digestibility of the pretreated stalks, and the adsorption of cellulases on the substrates were investigated. Results indicated that the pretreatments significantly improved the enzymatic digestibility of the agave stalk. SPORL pretreatment gave higher substrate and sugar yields, while NaOH pretreated stalk had better digestibility under the investigated conditions. The better hydrolysability of NaOH-pretreated stalk was attributed to low lignin and hemicellulose content and high affinity to cellulases.

Topics: Agave; Carbohydrate Metabolism; Cellulase; Cellulose; Hydrolysis; Sodium Hydroxide; Solid Waste; Sulfites; Sulfuric Acids; Time Factors; Xylans

Rising fuel prices and environmental issues have paved the way for the exploration of cellulosic ethanol. However, challenges involving substrate hydrolysis and cost-effectiveness still limit the efficient bioconversion and utilization of cellulosic ethanol. We aimed to evaluate a cheaper and abundantly available wild sugarcane variety, Saccharum spontaneum, as the raw substrate for bioconversion of ethanol by Pichia stipitis NCIM3498. Three different strategies for substrate hydrolysis using acid (dilute sulfuric acid) and alkali (dilute sodium hydroxide) and aqueous ammonia (AA) treatment followed by enzymatic hydrolysis were studied. A maximum of 631.5±3.25 mg/g sugars with 89.38% hydrolytic efficiency (HE) could be achieved after enzymatic hydrolysis of AA-pretreated S. spontaneum. All the substrate hydrolysates were evaluated for ethanol conversion in batches by P. stipitis. The microbial fermentation of released sugars into ethanol showed (g/g) 0.36±0.011, 0.384±0.022, 0.391±0.02, and 0.40±0.01 yield from detoxified acid hydrolysate and acid-, NaOH- and AA-pretreated substrate S. spontaneum enzymatic hydrolysates, respectively.

Topics: Alkalies; Aspergillus; Biofuels; Biotechnology; Cellulase; Ethanol; Hydrolysis; Pichia; Plant Weeds; Saccharum; Sulfuric Acids; Time Factors

Dilute sulfuric acid was used to pretreat coastal Bermuda grass at high temperature prior to enzymatic hydrolysis. After both pretreatment and enzymatic hydrolysis processes, the highest yield of total sugars (combined xylose and glucose) was 97% of the theoretical value. The prehydrolyzate liquor was analyzed for inhibitory compounds (furfural, hydroxymethylfurfural (HMF)) in order to assess potential risk for inhibition during the following fermentation. Accounting for the formation of the inhibitory compounds, a pretreatment with 1.2% acid at 140 °C for 30 min with a total sugar yield of 94% of the theoretical value may be more favorable for fermentation. From this study, it can be concluded that dilute sulfuric acid pretreatment can be successfully applied to coastal Bermuda grass to achieve high yields of monomeric glucose and xylose with acceptable levels of inhibitory compound formation.

Topics: Analysis of Variance; beta-Glucosidase; Biomass; Carbohydrates; Cellulase; Computer Simulation; Cynodon; Fermentation; Furaldehyde; Glucose; Hot Temperature; Hydrolysis; Lignin; Oxidation-Reduction; Seawater; Sulfuric Acids; Xylose

This study aimed to correlate the efficiency of enzymatic hydrolysis of the cellulose contained in a sugarcane bagasse sample pretreated with dilute H(2)SO(4) with the levels of independent variables such as initial content of solids and loadings of enzymes and surfactant (Tween 20), for two cellulolytic commercial preparations. The preparations, designated cellulase I and cellulase II, were characterized regarding the activities of total cellulases, endoglucanase, cellobiohydrolase, cellobiase, β-glucosidase, xylanase, and phenoloxidases (laccase, manganese and lignin peroxidases), as well as protein contents. Both extracts showed complete cellulolytic complexes and considerable activities of xylanases, without activities of phenoloxidases. For the enzymatic hydrolyses, two 2(3) central composite full factorial designs were employed to evaluate the effects caused by the initial content of solids (1.19-4.81%, w/w) and loadings of enzymes (1.9-38.1 FPU/g bagasse) and Tween 20 (0.0-0.1 g/g bagasse) on the cellulose digestibility. Within 24 h of enzymatic hydrolysis, all three independent variables influenced the conversion of cellulose by cellulase I. Using cellulase II, only enzyme and surfactant loadings showed significant effects on cellulose conversion. An additional experiment demonstrated the possibility of increasing the initial content of solids to values much higher than 4.81% (w/w) without compromising the efficiency of cellulose conversion, consequently improving the glucose concentration in the hydrolysate.

Topics: beta-Glucosidase; Cellulase; Cellulases; Cellulose; Conservation of Energy Resources; Conservation of Natural Resources; Ethanol; Hydrolysis; Polysorbates; Saccharum; Sulfuric Acids

The combined pretreatment of rice straw using dilute-acid and steam explosion followed by enzymatic hydrolysis was investigated and compared with acid-catalyzed steam explosion pretreatment. In addition to measuring the chemical composition, including glucan, xylan and lignin content, changes in rice straw features after pretreatment were investigated in terms of the straw's physical properties. These properties included crystallinity, surface area, mean particle size and scanning electron microscopy imagery. The effect of acid concentration on the acid-catalyzed steam explosion was studied in a range between 1% and 15% acid at 180°C for 2 min. We also investigated the influence of the residence time of the steam explosion in the combined pretreatment and the optimum conditions for the dilute-acid hydrolysis step in order to develop an integrated process for the dilute-acid and steam explosion. The optimum operational conditions for the first dilute-acid hydrolysis step were determined to be 165°C for 2 min with 2% H(2)SO(4) and for the second steam explosion step was to be carried out at 180°C for 20 min; this gave the most favorable combination in terms of an integrated process. We found that rice straw pretreated by the dilute-acid/steam explosions had a higher xylose yield, a lower level of inhibitor in the hydrolysate and a greater degree of enzymatic hydrolysis; this resulted in a 1.5-fold increase in the overall sugar yield when compared to the acid-catalyzed steam explosion.

Topics: Carbohydrates; Cellulase; Ethanol; Oryza; Plant Components, Aerial; Steam; Sulfuric Acids; Systems Integration

A low temperature alkali pretreatment method was proposed for improving the enzymatic hydrolysis efficiency of lignocellulosic biomass for ethanol production. The effects of the pretreatment on the composition, structure and enzymatic digestibility of sweet sorghum bagasse were investigated. The mechanisms involved in the digestibility improvement were discussed with regard to the major factors contributing to the biomass recalcitrance. The pretreatment caused slight glucan loss but significantly reduced the lignin and xylan contents of the bagasse. Changes in cellulose crystal structure occurred under certain treatment conditions. The pretreated bagasse exhibited greatly improved enzymatic digestibility, with 24-h glucan saccharification yield reaching as high as 98% using commercially available cellulase and β-glucosidase. The digestibility improvement was largely attributed to the disruption of the lignin-carbohydrate matrix. The bagasse from a brown midrib (BMR) mutant was more susceptible to the pretreatment than a non-BMR variety tested, and consequently gave higher efficiency of enzymatic hydrolysis.

Topics: beta-Galactosidase; Biofuels; Cellulase; Cellulose; Chromatography, High Pressure Liquid; Cold Temperature; Ethanol; Hydrolysis; Lignin; Sorghum; Sulfuric Acids; X-Ray Diffraction; Xylans

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%.

Topics: Carbohydrates; Cellulase; Fermentation; Glycolipids; Hydrolysis; Lignin; Penicillium; Sodium Hydroxide; Sulfuric Acids; Triticum; Waste Products

Thermo-mechanical extrusion pretreatment for lignocellulosic biomass was investigated using soybean hulls as the substrate. The enzyme cocktail used to hydrolyze pretreated soybean hulls to fermentable sugars was optimized using response surface methodology (RSM). Structural changes in substrate and sugar yields from thermo-mechanical processing were compared with two traditional pretreatment methods that utilized dilute acid (1% sulfuric acid) and alkali (1% sodium hydroxide). Extrusion processing parameters (barrel temperature, in-barrel moisture, screw speed) and processing aids (starch, ethylene glycol) were studied with respect to reducing sugar and glucose yields. The conditions resulting in the highest cellulose to glucose conversion (95%) were screw speed 350rpm, maximum barrel temperature 80°C and in-barrel moisture content 40%wb. Compared with untreated soybean hulls, glucose yield from enzymatic hydrolysis of soybean hulls increased by 69.6%, 128.7% and 132.2%, respectively, when pretreated with dilute acid, alkali and extrusion.

Topics: Alkalies; beta-Glucosidase; Biofuels; Cellulase; Ethylene Glycol; Fermentation; Glucose; Glycine max; Hot Temperature; Hydrolysis; Lignin; Sulfuric Acids

Spent mushroom substrate (SMS) was treated with dilute sulfuric acid followed by cellulase and xylanase treatment to produce hydrolysates that could be used as the basis for media for the production of value added products. A L9 (3(4)) orthogonal experiment was performed to optimize the acid treatment process. Pretreatment with 6% (w/w) dilute sulfuric acid at 120°C for 120 min provided the highest reducing sugar yield of 267.57 g/kg SMS. No furfural was detected in the hydrolysates. Exposure to 20PFU of cellulase and 200 XU of xylanase per gram of pretreated SMS at 40°C resulted in the release of 79.85 g/kg or reducing sugars per kg acid pretreated SMS. The dilute sulfuric acid could be recycled to process fresh SMS four times. SMS hydrolysates neutralized with ammonium hydroxide, sodium hydroxide, or calcium hydroxide could be used as the carbon source for cultivation of Lactococcus lactis subsp. lactis W28 and a cell density of 2.9×10(11)CFU/mL could be obtained. The results provide a foundation for the development of value-added products based on SMS.

Topics: Agaricales; Cellulase; Hydrolysis; Lactococcus lactis; Sulfuric Acids; Xylosidases

Native aspen (Populus tremuloides) was pretreated using sulfuric acid and sodium bisulfite (SPORL) and dilute sulfuric acid alone (DA). Simultaneous enzymatic saccharification and fermentation (SSF) was conducted at 18% solids using commercial enzymes with cellulase loadings ranging from 6 to 15 FPU/g glucan and Saccharomyces cerevisiae Y5. Compared with DA pretreatment, the SPORL pretreatment reduced the energy required for wood chip size-reduction, and reduced mixing energy of the resultant substrate for solid liquefaction. Approximately 60% more ethanol was produced from the solid SPORL substrate (211 L/ton wood at 59 g/L with SSF efficiency of 76%) than from the solid DA substrate (133 L/ton wood at 35 g/L with SSF efficiency 47%) at a cellulase loading of 10 FPU/g glucan after 120 h. When the cellulase loading was increased to 15 FPU/g glucan on the DA substrate, the ethanol yield still remained lower than the SPORL substrate at 10 FPU/g glucan.

Topics: Biofuels; Cellulase; Chromatography, Gas; Chromatography, Ion Exchange; Ethanol; Fermentation; Populus; Saccharomyces cerevisiae; Sulfites; Sulfuric Acids

Dacotah switchgrass was pretreated with sulfuric acid concentrations of 0.5, 1.0, and 2.0 wt.% at 140, 160, and 180 °C and with 1 and 3 wt.% sulfur dioxide at 180 °C over a range of times. Sulfur dioxide loadings of 0%, 1%, 3%, 5%, and 10%wt.% of dry biomass were also tested at 180 °C for 10 min. Sugar yields were tracked for pretreatment and subsequent enzymatic hydrolysis to identify conditions for the highest total sugar yields. Pretreatment with 1 wt.% dilute sulfuric acid at 140 °C for 40 min followed by enzymatic hydrolysis with 48.6 mg enzyme/g initial glucan in raw biomass resulted in ∼86% of theoretical yield for glucose and xylose combined. For sulfur dioxide pretreatment, the highest total sugar yield of about 87% occurred at 5% SO₂ for 10 min and 180 °C. However, xylose yields were higher at shorter times and glucose yields at longer times.

Topics: Biofuels; Carbohydrates; Cellulase; Hydrolysis; Panicum; Plant Stems; Sulfur Dioxide; Sulfuric Acids

Bioethanol is one of the alternatives of the conventional fossil fuel. In present study, effect of different carbon sources on the production of cellulolytic enzyme (CMCase) from Trichoderma reesei at different temperatures, duration and pH were investigated and conditions were optimized. Acid treated Kans grass (Saccharum sponteneum) was subjected to enzymatic hydrolysis to produce fermentable sugars which was then fermented to bioethanol using Saccharomyces cerevisiae. The maximum CMCase production was found to be 1.46 U mL(-1) at optimum condition (28°C, pH 5 and cellulose as carbon source). The cellulases and xylanase activity were found to be 1.12 FPU g(-1) and 6.63 U mL(-1), respectively. Maximum total sugar was found to be 69.08 mg/g dry biomass with 20 FPU g(-1) dry biomass of enzyme dosage under optimum condition. Similar results were obtained when it was treated with pure enzyme. Upon fermentation of enzymatic hydrolysate, the yield of ethanol was calculated to be 0.46 g g(-1).

Topics: Biofuels; Biomass; Biotechnology; Carbohydrates; Carbon; Cellulase; Ethanol; Fermentation; Hydrogen-Ion Concentration; Hydrolysis; Lignin; Poaceae; Saccharomyces cerevisiae; Sulfuric Acids; Temperature; Trichoderma; Xylose

A combination of a twin-screw extrusion and an acid-catalyzed hot water extraction process performed at a bench-scale was used to prepare high monomeric xylose hydrolysate for cellulosic production. The influences of the screw speed (30-150 rpm), barrel temperature (80-160 °C) and corresponding specific mechanical energy of the extruder on the structural properties of the pretreated rice straw, sugar concentration and conversion were investigated. The optimal condition for the extrusion step was determined to be 40 rpm with 3% H2SO4 at 120 °C; the optimal condition for the extraction step was determined to be 130 °C for 20 min. After the pretreatment at the optimal condition, 83.7% of the xylan was converted to monomeric xylose, and the concentration reached levels of 53.7 g/L. Finally, after the subsequent enzymatic hydrolysis, an 80% yield of the total saccharification was obtained.

Topics: Biotechnology; Cellulase; Cellulose; Ethanol; Glucose; Hydrolysis; Oryza; Sulfuric Acids; Temperature; Waste Products; Water; Xylose

A comparison between the classic Plackett-Burman design (PB) ANOVA analysis and a genetic algorithm (GA) approach to identify significant factors have been carried out. This comparison was made by applying both analyses to data obtained from the experimental results when optimizing both chemical and enzymatic hydrolysis of three lignocellulosic feedstocks (corn and wheat bran, and pine sawdust) by a PB experimental design. Depending on the kind of biomass and the hydrolysis being considered, different results were obtained. Interestingly, some interactions were found to be significant by the GA approach and allowed to identify significant factors, that otherwise, based only in the classic PB analysis, would have not been taken into account in a further optimization step. Improvements in the fitting of c.a. 80% were obtained when comparing the coefficient of determination (R2) computed for both methods.

Topics: Algorithms; Cellulase; Dietary Fiber; Hydrolysis; Lignin; Models, Statistical; Pinus; Sulfuric Acids; Trichoderma; Waste Products; Zea mays

A low level of phosphoric acid (1% w/w on dry bagasse basis, 160 degrees C and above, 10 min) was shown to effectively hydrolyze the hemicellulose in sugar cane bagasse into monomers with minimal side reactions and to serve as an effective pre-treatment for the enzymatic hydrolysis of cellulose. Up to 45% of the remaining water-insoluble solids (WIS) was digested to sugar monomers by a low concentration of Biocellulase W (0.5 filter paper unit/gWIS) supplemented with beta-glucosidase, although much higher levels of cellulase (100-fold) were required for complete hydrolysis. After neutralization and nutrient addition, phosphoric acid syrups of hemicellulose sugars were fermented by ethanologenic Escherichia coli LY160 without further purification. Fermentation of these syrups was preceded by a lag that increased with increased pre-treatment temperature. Further improvements in organisms and optimization of steam treatments may allow the co-fermentation of sugars derived from hemicellulose and cellulose, eliminating need for liquid-solid separation, sugar purification, and separate fermentations.

Topics: Biotechnology; Carbohydrates; Cellulase; Cellulose; Ethanol; Fungi; Hydrolysis; Lignin; Phosphoric Acids; Saccharum; Sulfuric Acids; Temperature; Time Factors; Xylose

This study investigated the effects of chemical pretreatment and disk-milling conditions on energy consumption for size-reduction and the efficiency of enzymatic cellulose saccharification of a softwood. Lodgepole pine wood chips produced from thinnings of a 100-year-old unmanaged forest were pretreated by hot-water, dilute-acid, and two SPORL processes (Sulfite Pretreatment to Overcome Recalcitrance of Lignocellulose) at acid charge on oven dry (od) wood of 0% and 2.21%. The pretreated wood chips were then milled using a laboratory disk mill under various solids-loadings and disk-plate gaps to produce substrates for enzymatic hydrolysis. We found that post-chemical-pretreatment size-reduction of forest biomass can decrease size-reduction energy consumption by 20-80% depending on the pretreatment applied under 20% solids-loading and a disk-plate gap of 0.76 mm in milling. SPORL with a sodium bisulfite charge of 8% and sulfuric acid charge of 2.21% on wood was the most effective in decreasing size-reduction energy consumption. Solids-loading had the most significant effect on disk-milling energy. When solids-loading was reduced from 30% to 3%, disk-milling energy could be decreased by more than a factor of 10 for wood chips pretreated by both SPORL and dilute-acid at an acid charge of 2.21%. The enzymatic hydrolysis glucose yields (EHGY) from the substrates produced by all pretreatments were independent of the solids-loading in milling, indicating that these energy savings in size-reduction can be realized without affecting EHGY. When wood chips were pretreated by SPORL with 2.21% acid charge, size-reduction energy consumption was decreased to less than 50 Wh/kg od wood at a practical solids-loading of approximately 10-20%, equivalent to that used in size-reduction of agriculture biomass, with excellent EHGY of about 370 g per kg od wood. Similar effects on size-reduction energy savings and excellent EHGY were also achieved when large disk-plate gaps (up to 1.52 mm studied) were applied in disk-milling of wood chips pretreated by SPORL with acid.

Topics: beta-Glucosidase; Biofuels; Biotechnology; Cellulase; Chromatography, Ion Exchange; Conservation of Energy Resources; Hydrolysis; Pinus; Sulfites; Sulfuric Acids; Wood

The production of fermentable sugars from rice hull was studied by dilute acid pretreatment and enzymatic saccharification. Rice hull (15%, w/v) was pretreated by 1% (v/v) sulfuric acid at high temperature (120 approximately 160 degrees C) for 15, 30, 45, and 60 min, respectively. The maximum sugar concentration from rice hull in the prehydrolysate was obtained at 140 degrees C for 30 min, but the enzymatic saccharification yield from the corresponding pretreated rice hull is not high. To another aspect, the maximum enzymatic saccharification yield was achieved at 160 degrees C for 60 min, while the recovery of fermentable sugars was the poorest. To take account of fermentable sugars from pretreatment and enzymatic saccharification, the maximum yield of sugars was obtained only when rice hull was treated at 140 degrees C for 30 min. Under this condition, 72.5% (w/w) of all sugars generated from the raw material can be recovered. The kinetic study on the enzymatic saccharification of dilute acid pretreated rice hull was also performed in this work by a modified Michaelis-Menten model and a diffusion-limited model. After calculation by a linear and a non-linear regression analysis, both models showed good relation with the experimental results.

Topics: Biotechnology; Carbohydrate Metabolism; Cellulase; Fermentation; Hydrolysis; Kinetics; Oryza; Regression Analysis; Sulfuric Acids; Temperature; Thermogravimetry; Time Factors; Waste Products

In this study, the effect and the optimum pretreatment condition of corncobs using low strength of H2SO4 were investigated, in which H2SO4 was used to improve the enzymatic digestibility of corncobs for saccharification without degradation of sugars released. The optimum pretreatment condition was found to be the addition of 0.5% (vol./vol.) H2SO4 and autoclaving at 122°C for 20 min. Under this condition, the structural integrity of corncob was altered to make cellulose microfibrils more accessible for cellulase enzymes, and the enzymatic digestion of corncobs could be significantly enhanced. A high yield of sugar, 80% (wt./wt.), could be obtained at a low enzyme dosage of 0.024 g enzymes/g cobs, when pretreated. As a result, the ethanol production was obviously improved by the pretreatment, i.e., the ethanol yield of 77% (wt./wt.) was obtained within 36 h in the SSF fermentation using Saccharomyces cerevisiae NBRC2114.

Topics: Cellulase; Cellulose; Ethanol; Microscopy, Electron, Scanning; Sulfuric Acids; Zea mays

Bovine serum albumin (BSA), Tween-20, and polyethylene glycol (PEG6000) were added to washed corn stover solids produced by ammonia fiber expansion (AFEX), ammonia recycled percolation (ARP), dilute sulfuric acid (DA), lime, controlled pH, and sulfur dioxide (SO(2)) pretreatments and to untreated corn stover (UT) and pure Avicel glucan prior to adding cellulase supplemented with beta-glucosidase at an activity ratio of 1:2/g and a moderate enzyme loading of 16.1 mg/g glucan in the raw corn stover. The additives were applied individually at 150, 300, and 600 mg/g glucan in the pretreated solids and in combinations of equal amounts of each that totaled 600 mg/g. The greatest increase in total sugar release was by Tween-20 with SO(2) pretreated solids followed by PEG6000 with ARP solids and Tween-20 with lime solids. The effectiveness of the additives was observed to depend on the type of sugars left in the solids, suggesting that it may be more beneficial to use the mixture of these additives to realize a high total sugar yield. In addition, little enhancement in sugar release was possible beyond a loading of 150 mg additives/g glucan for most pretreatments, and combinations did not improve sugar release much over use of additives alone for all except SO(2). Additives were also found to significantly increase concentrations of cellobiose and cellooligomers after 72 h of Avicel hydrolysis.

Topics: Ammonia; Animals; beta-Glucosidase; Calcium Sulfate; Cattle; Cellulase; Cellulose; Glucans; Glucose; Hydrogen-Ion Concentration; Hydrolysis; Lignin; Polyethylene Glycols; Polysorbates; Serum Albumin, Bovine; Sulfur Dioxide; Sulfuric Acids; Time Factors; Xylose; Zea mays

Rice straw has recently attracted interest in Japan as a potential source of raw material for ethanol production. Wet disk milling, a continuous pretreatment to enhance the enzymatic digestibility of rice straw, was compared with conventional ball milling and hot-compressed water treatment. Pretreated rice straw was evaluated by enzymatic hydrolysis using Acremonium cellulase and characterized by X-ray diffraction and scanning electron microscopy. Glucose and xylose yields by wet disk milling, ball milling, and hot-compressed water treatment were 78.5% and 41.5%, 89.4% and 54.3%, and 70.3% and 88.6%, respectively. Wet disk milling and hot-compressed water treatment increased sugar yields without decreasing their crystallinity. The feature size of the wet disk milled rice straw was similar to that of hot-compressed water-treated rice straw. The energy consumption of wet disk milling was lower than that of other pretreatments. Thus, wet disk milling is an economical, practical pretreatment for the enzymatic hydrolysis of lignocellulosic biomass, especially herbaceous biomass such as rice straw.

Topics: Acremonium; Alkalies; Cellulase; Enzyme Activation; Glucose; Oryza; Plant Components, Aerial; Sulfuric Acids; Xylose

Distillers' grains and corn fiber are the coproducts of the corn dry grind and wet milling industries, respectively. Availability of distillers' grains and corn fiber at the ethanol plant and their high levels of lignocellulosic material make these coproducts attractive feedstocks for conversion to ethanol. In this study, dilute sulfuric acid hydrolysis of these coproducts was investigated in a multistage scheme. After the completion of each pretreatment stage, the liquid substrate was separated and reused in the succeeding pretreatment stage with a fresh substrate. The substrate from each stage was also subjected to enzyme hydrolysis in a separate experiment. The sulfuric acid concentration and the substrate loading were maintained at 1.0 vol% and 15.0 wt.%, respectively, and the temperature was maintained at 120 degrees C in all the experiments. Experiments were also performed to study the effect of removing oil from the samples prior to the pretreatment. The highest concentration of monomeric sugars (MS) was observed when three stages of pretreatment were followed by the enzyme reaction. The enzyme hydrolysis of the three-stage pretreated dried distillers' grains and corn fiber yielded 122.6 +/- 5.8 and 184.5 +/- 4.1 mg/mL of MS, respectively. The formation of inhibitory products was also monitored.

Topics: beta-Glucosidase; Cellulase; Distillation; Edible Grain; Ethanol; Hydrolysis; Lignin; Sulfuric Acids; Zea mays

In general, pretreatments are designed to enhance the accessibility of cellulose to enzymes, allowing for more efficient conversion. In this study, we have detected the penetration of major cellulases present in a commercial enzyme preparation (Spezyme CP) into corn stem cell walls following mild-, moderate- and high-severity dilute sulfuric acid pretreatments. The Trichoderma reesei enzymes, Cel7A (CBH I) and Cel7B (EG I), as well as the cell wall matrix components xylan and lignin were visualized within digested corn stover cell walls by immuno transmission electron microscopy (TEM) using enzyme- and polymer-specific antibodies. Low severity dilute-acid pretreatment (20 min at 100 degrees C) enabled <1% of the thickness of secondary cell walls to be penetrated by enzyme, moderate severity pretreatment at (20 min at 120 degrees C) allowed the enzymes to penetrate approximately 20% of the cell wall, and the high severity (20 min pretreatment at 150 degrees C) allowed 100% penetration of even the thickest cell walls. These data allow direct visualization of the dramatic effect dilute-acid pretreatment has on altering the condensed ultrastructure of biomass cell walls. Loosening of plant cell wall structure due to pretreatment and the subsequently improved access by cellulases has been hypothesized by the biomass conversion community for over two decades, and for the first time, this study provides direct visual evidence to verify this hypothesis. Further, the high-resolution enzyme penetration studies presented here provide insight into the mechanisms of cell wall deconstruction by cellulolytic enzymes.

Topics: Caustics; Cell Wall; Cellulase; Lignin; Microscopy, Immunoelectron; Sulfuric Acids; Xylans; Zea mays

Due to concerns with biomass collection systems and soil sustainability there are opportunities to investigate the optimal plant fractions to collect for conversion. An ideal feedstock would require a low severity pretreatment to release a maximum amount of sugar during enzymatic hydrolysis. Corn stover fractions were separated manually and analyzed for glucan, xylan, acid soluble lignin, acid insoluble lignin, and ash composition. The stover fractions were also pretreated with either 0%, 0.4%, or 0.8% NaOH for 2 h at room temperature, washed, autoclaved and saccharified. In addition, dilute sulfuric acid pretreated samples underwent simultaneous saccharification and fermentation (SSF) to ethanol. In general, the two pretreatments produced similar trends with cobs, husks, and leaves responding best to the pretreatments, the tops of stalks responding slightly less, and the bottom of the stalks responding the least. For example, corn husks pretreated with 0.8% NaOH released over 90% (standard error of 3.8%) of the available glucan, while only 45% (standard error of 1.1%) of the glucan was produced from identically treated stalk bottoms. Estimates of the theoretical ethanol yield using acid pretreatment followed by SSF were 65% (standard error of 15.9%) for husks and 29% (standard error of 1.8%) for stalk bottoms. This suggests that integration of biomass collection systems to remove sustainable feedstocks could be integrated with the processes within a biorefinery to minimize overall ethanol production costs.

Topics: Biomass; Carbohydrate Metabolism; Cellulase; Chemical Fractionation; Chromatography, High Pressure Liquid; Elements; Fermentation; Glucans; Hydrolysis; Lignin; Sodium Hydroxide; Solubility; Sulfuric Acids; Water; Xylans; Zea mays

A combined sulfuric acid-free ethanol cooking and pulverization process was developed in order to achieve the complete saccharification of the cellulosic component of woody biomass, thereby avoiding the problems associated with the use of strong acid catalysts. Eucalyptus wood chips were used as a raw material and exposed to an ethanol/water/acetic acid mixed solvent in an autoclave. This process can cause the fibrillation of wood chips. During the process, the production of furfural due to an excessive degradation of polysaccharide components was extremely low and delignification was insignificant. Therefore, the cooking process is regarded not as a delignification but as an activation of the original wood. Subsequently, the activated solid products were pulverized by ball-milling in order to improve their enzymatic digestibility. Enzymatic hydrolysis experiments demonstrated that the conversion of the cellulosic components into glucose attained 100% under optimal conditions. Wide-angle X-ray diffractometry and particle size distribution analysis revealed that the scale affecting the improvement of enzymatic digestibility ranged from 10 nm to 1 microm. Field emission scanning electron microscopy depicted that the sulfuric acid-free ethanol cooking induced a pore formation by the removal of part of the lignin and hemicellulose fractions in the size range from a few of tens nanometers to several hundred nanometers.

Topics: Cellulase; Enzyme Activation; Ethanol; Eucalyptus; Hot Temperature; Polysaccharides; Sulfuric Acids; Wood

In order to utilize and control the invasive weed, crofton weed (Eupatorium adenophorum Spreng), a potential pathway was proposed by using it as a feedstock for production of fermentable sugars. Three chemical pretreatment methods were used for improving enzymatic saccharification of the weed stem. Mild H2SO4 pretreatment could obtain a relatively high yield of sugars in the pretreatment (32.89%, based on initial holocellulose), however, it led to only a slight enhancement of enzymatic digestibility. NaOH pretreatment could obtain a higher enzymatic conversion ratio of cellulose compared with H2SO4 pretreatment. Peracetic acid (PAA) pretreatment seemed to be the most effective for improving enzymatic saccharification of the weed stem in the three chemical pretreatment methods under the same conditions. The conversion ratio of cellulose in the sample pretreated by PAA under the "optimal" condition was increased to 50% by cellulase loading of 80 FPU/g cellulose for 72 h incubation. A number of empirical quadratic models were successfully developed according to the experimental data to predict the yield of sugar and degree of delignification.

Topics: Ageratina; Agriculture; Biotechnology; Cellulase; Cellulose; Peracetic Acid; Plant Stems; Sodium Hydroxide; Sulfuric Acids; Temperature; Time Factors

Our previous research has shown that saline Creeping Wild Ryegrass (CWR), Leymus triticoides, has a great potential to be used for bioethanol production because of its high fermentable sugar yield, up to 85% cellulose conversion of pretreated CWR. However, the high cost of enzyme is still one of the obstacles making large-scale lignocellulosic bioethanol production economically difficult. It is desirable to use reduced enzyme loading to produce fermentable sugars with high yield and low cost. To reduce the enzyme loading, the effect of addition of non-ionic surfactants and non-catalytic protein on the enzymatic hydrolysis of pretreated CWR was investigated in this study. Tween 20, Tween 80, and bovine serum albumin (BSA) were used as additives to improve the enzymatic hydrolysis of dilute sulfuric-acid-pretreated CWR. Under the loading of 0.1 g additives/g dry solid, Tween 20 was the most effective additive, followed by Tween 80 and BSA. With the addition of Tween 20 mixed with cellulase loading of 15 FPU/g cellulose, the cellulose conversion increased 14% (from 75 to 89%), which was similar to that with cellulase loading of 30 FPU/g cellulose and without additive addition. The results of cellulase and BSA adsorption on the Avicel PH101, pretreated CWR, and lignaceous residue of pretreated CWR support the theory that the primary mechanism behind the additives is prevention of non-productive adsorption of enzymes on lignaceous material of pretreated CWR. The addition of additives could be a promising technology to improve the enzymatic hydrolysis by reducing the enzyme activity loss caused by non-productive adsorption.

Topics: Cellulase; Cellulose; Hydrolysis; Ions; Lignin; Poaceae; Serum Albumin, Bovine; Sulfuric Acids; Surface-Active Agents

Utilization of ethanol produced from biomass has the potential to offset the use of gasoline and reduce CO(2) emissions. This could reduce the effects of global warming, one of which is the current outbreak of epidemic proportions of the mountain pine beetle (MPB) in British Columbia (BC), Canada. The result of this is increasing volumes of dead lodgepole pine with increasingly limited commercial uses. Bioconversion of lodgepole pine to ethanol using SO(2)-catalyzed steam explosion was investigated. The optimum pretreatment condition for this feedstock was determined to be 200 degrees C, 5 min, and 4% SO(2) (w/w). Simultaneous saccharification and fermentation (SSF) of this material provided an overall ethanol yield of 77% of the theoretical yield from raw material based on starting glucan, mannan, and galactan, which corresponds to 244 g ethanol/kg raw material within 30 h. Three conditions representing low (L), medium (M), and high (H) severity were also applied to healthy lodgepole pine. Although the M severity conditions of 200 degrees C, 5 min, and 4% SO(2) were sufficiently robust to pretreat healthy wood, the substrate produced from beetle-killed (BK) wood provided consistently higher ethanol yields after SSF than the other substrates tested. BK lodgepole pine appears to be an excellent candidate for efficient and productive bioconversion to ethanol.

Topics: Cellulase; Ethanol; Fermentation; Hydrolysis; Pinus; Saccharomyces cerevisiae; Steam; Sulfuric Acids

The potential of dilute-acid prehydrolysis as a pretreatment method for sugarcane bagasse, rice hulls, peanut shells, and cassava stalks was investigated. The prehydrolysis was performed at 122 degrees C during 20, 40, or 60 min using 2% H(2)SO(4) at a solid-to-liquid ratio of 1:10. Sugar formation increased with increasing reaction time. Xylose, glucose, arabinose, and galactose were detected in all of the prehydrolysates, whereas mannose was found only in the prehydrolysates of peanut shells and cassava stalks. The hemicelluloses of bagasse were hydrolyzed to a high-extent yielding concentrations of xylose and arabinose of 19.1 and 2.2 g/L, respectively, and a xylan conversion of more than 80%. High-glucose concentrations (26-33.5 g/L) were found in the prehydrolysates of rice hulls, probably because of hydrolysis of starch of grain remains in the hulls. Peanut shells and cassava stalks rendered low amounts of sugars on prehydrolysis, indicating that the conditions were not severe enough to hydrolyze the hemicelluloses in these materials quantitatively. All prehydrolysates were readily fermentable by Saccharomyces cerevisiae. The dilute-acid prehydrolysis resulted in a 2.7- to 3.7-fold increase of the enzymatic convertibility of bagasse, but was not efficient for improving the enzymatic hydrolysis of peanut shells, cassava stalks, or rice hulls.

Topics: Agriculture; Cellulase; Cellulose; Ethanol; Industrial Waste; Saccharomyces cerevisiae; Sulfuric Acids

Barley is an abundant crop in Europe, which makes its straw residues an interesting cellulose source for ethanol production. Steam pretreatment of the straw followed by enzymatic hydrolysis converts the cellulose to fermentable sugars. Prior to pretreatment the material is impregnated with a catalyst, for example, H2SO4, to enhance enzymatic digestibility of the pretreated straw. Different impregnation techniques can be applied. In this study, soaking and spraying were investigated and compared at the same pretreatment condition in terms of overall yield of glucose and xylose. The overall yield includes the soluble sugars in the liquid from pretreatment, including soluble oligomers, and monomer sugars obtained in the enzymatic hydrolysis. The yields obtained differed for the impregnation techniques. Acid-soaked barley straw gave the highest overall yield of glucose, regardless of impregnation time (10 or 30 min) or acid concentration (0.2 or 1.0 wt%). For xylose, soaking gave the highest overall yield at 0.2 wt% H2SO4. An increase in acid concentration resulted in a decrease in xylose yield for both acid-soaked and acid-sprayed barley straw. Optimization of the pretreatment conditions for acid-sprayed barley straw was performed to obtain yields using spraying that were as high as those with soaking. For acid-sprayed barley straw the optimum pretreatment condition for glucose, 1.0 wt% H2SO4 and 220 degrees C for 5 min, gave an overall glucose yield of 92% of theoretical based on the composition of the raw material. Pretreatment with 0.2 wt% H2SO4 at 190 degrees C for 5 min resulted in the highest overall xylose yield, 67% of theoretical based on the composition of the raw material.

Topics: beta-Glucosidase; Cellulase; Chemical Fractionation; Ethanol; Hordeum; Plant Components, Aerial; Steam; Sulfuric Acids; Temperature

The cellulose reactivity of two lignocellulosic feedstocks, switchgrass and poplar, was evaluated under straight saccharification (SS) and simultaneous saccharification and fermentation (SSF) conditions following dilute sulfuric acid pretreatments designed for optimum xylose yields. The optimum pretreatment conditions, within the constraints of the experimental system (Parr batch reactor), were 1.2% acid, 180 degrees C, and 0.5 min for switchgrass and 1% acid, 180 degrees C, and 0.56 min for poplar. The cellulase enzyme preparation was from Trichoderma reesei and fermentations were done with Saccharomyces cerevisiae. Time courses for SS were monitored as the sum of glucose and cellobiose; those for SSF as the sum of glucose, cellobiose, and ethanol. Percentage conversions under SS conditions were 79.1% and 91.4% for the pretreated poplar and switchgrass feedstocks, respectively. Analogous values under SSF conditions were 73.0% and 90.3% for pretreated poplar and switchgrass, respectively.

Topics: Cell Culture Techniques; Cell Proliferation; Cell Survival; Cellulase; Cellulose; Ethanol; Fermentation; Poaceae; Populus; Saccharomyces cerevisiae; Sulfuric Acids; Trichoderma; Xylose

Animal manure is an underutilized biomass resource containing a large amount of organic carbon that is often wasted with the existing manure disposal practices. A research project funded by the US Department of Energy explored the feasibility of using manure via the sugar platform in a biorefinery, converting the carbon from fiber to biochemicals. The results showed that (1) fiber was the major component of manure dry material making up approx 50%, 40%, and 36% of the dry dairy, swine, and poultry manure material, respectively; within dairy manure, more than 56% of the dry matter was in particles larger than 1.680 mm; (2) in addition to being a carbon source, manure could provide a variety of nutrient for fungi T. reesei and A. phoenicis to produce cellulase; (3) the hemicellulose component in the manure fiber could be readily converted to sugar through acid hydrolysis; while concentrated acid decrystallization treatment was most effective in manure cellulose hydrolysis; (4) purification and separation was necessary for further chemical conversion of the manure hydrolysate to polyols through hydrogenation; and (5) the manure utilization strategy studied in this work is currently not profitable.

Topics: Agriculture; Animals; Carbohydrates; Cellulase; Chemical Industry; Conservation of Natural Resources; Feasibility Studies; Hydrolysis; Industrial Waste; Manure; Refuse Disposal; Species Specificity; Sulfuric Acids; Trichoderma

We have previously demonstrated that pretreatment of corn stover with dilute sulfuric acid can achieve high digestibility and efficient recovery of hemicellulose sugars with high yield and concentration. Further improvement of this process was sought in this work. A modification was made in the operation of the percolation reactor that the reactor is preheated under atmospheric pressure to remove moisture that causes autohydrolysis. This eliminated sugar decomposition during the preheating stage and led to a considerable improvement in overall sugar yield. In addition, liquid throughput was minimized to the extent that only one reactor void volume of liquid was collected. This was done to attain a high xylose concentration in the hydrolyzate. The optimum reaction and operating conditions were identified wherein near quantitative enzymatic digestibilities are obtained with enzyme loading of 15 FPU/g glucan. With a reduced enzyme loading of 5 FPU/g glucan, the enzymatic digestibility was decreased, but still reached a level of 92%. Decomposition of carbohydrates was extremely low as indicated by the measured glucan and xylan mass closures (recovered sugar plus unreacted) which were 98% and 94%, respectively. The data obtained in this work indicate that the digestibility is related to the extent of xylan removal.

Topics: Bioreactors; Cellulase; Chemical Fractionation; Culture Media; Glucans; Hydrolysis; Plant Components, Aerial; Solutions; Sulfuric Acids; Xylans; Xylose; Zea mays

For the first time, a single source of cellulosic biomass was pretreated by leading technologies using identical analytical methods to provide comparative performance data. In particular, ammonia explosion, aqueous ammonia recycle, controlled pH, dilute acid, flowthrough, and lime approaches were applied to prepare corn stover for subsequent biological conversion to sugars through a Biomass Refining Consortium for Applied Fundamentals and Innovation (CAFI) among Auburn University, Dartmouth College, Michigan State University, the National Renewable Energy Laboratory, Purdue University, and Texas A&M University. An Agricultural and Industrial Advisory Board provided guidance to the project. Pretreatment conditions were selected based on the extensive experience of the team with each of the technologies, and the resulting fluid and solid streams were characterized using standard methods. The data were used to close material balances, and energy balances were estimated for all processes. The digestibilities of the solids by a controlled supply of cellulase enzyme and the fermentability of the liquids were also assessed and used to guide selection of optimum pretreatment conditions. Economic assessments were applied based on the performance data to estimate each pretreatment cost on a consistent basis. Through this approach, comparative data were developed on sugar recovery from hemicellulose and cellulose by the combined pretreatment and enzymatic hydrolysis operations when applied to corn stover. This paper introduces the project and summarizes the shared methods for papers reporting results of this research in this special edition of Bioresource Technology.

Topics: beta-Glucosidase; Biomass; Calcium Compounds; Cellulase; Cellulose; Conservation of Energy Resources; Fermentation; Hydrolysis; Oxides; Saccharomyces cerevisiae; Sulfuric Acids; Water; Zea mays

A number of previous studies determined dilute acid pretreatment conditions that maximize xylose yields from pretreatment or glucose yields from subsequent digestion of the pretreated cellulose, but our emphasis was on identifying conditions to realize the highest yields of both sugars from both stages. Thus, individual xylose and glucose yields are reported as a percentage of the total potential yield of both sugars over a range of sulfuric acid concentrations of 0.22%, 0.49% and 0.98% w/w at 140, 160, 180 and 200 degrees C. Up to 15% of the total potential sugar in the substrate could be released as glucose during pretreatment and between 15% and 90+% of the xylose remaining in the solid residue could be recovered in subsequent enzymatic hydrolysis, depending on the enzyme loading. Glucose yields increased from as high as 56% of total maximum potential glucose plus xylose for just enzymatic digestion to 60% when glucose released in pretreatment was included. Xylose yields similarly increased from as high as 34% of total potential sugars for pretreatment alone to between 35% and 37% when credit was taken for xylose released in digestion. Yields were shown to be much lower if no acid was used. Conditions that maximized individual sugar yields were often not the same as those that maximized total sugar yields, demonstrating the importance of clearly defining pretreatment goals when optimizing the process. Overall, up to about 92.5% of the total sugars originally available in the corn stover used could be recovered for coupled dilute acid pretreatment and enzymatic hydrolysis. These results also suggest that enhanced hemicellulase activity could further improve xylose yields, particularly for low cellulase loadings.

Topics: Catalysis; Cellulase; Cellulose; Glucose; Hydrogen-Ion Concentration; Hydrolysis; Sulfuric Acids; Xylose; Zea mays

Corn stover, a well-known example of lignocellulosic biomass, is a potential renewable feed for bioethanol production. Dilute sulfuric acid pretreatment removes hemicellulose and makes the cellulose more susceptible to bacterial digestion. The rheologic properties of corn stover pretreated in such a manner were studied. The Power Law parameters were sensitive to corn stover suspension concentration becoming more non-Newtonian with slope n, ranging from 0.92 to 0.05 between 5 and 30% solids. The Casson and the Power Law models described the experimental data with correlation coefficients ranging from 0.90 to 0.99 and 0.85 to 0.99, respectively. The yield stress predicted by direct data extrapolation and by the Herschel-Bulkley model was similar for each concentration of corn stover tested.

Topics: Biomass; Biotechnology; Calibration; Cellulase; Cellulose; Ethanol; Kinetics; Lignin; Models, Statistical; Polysaccharides; Rheology; Stress, Mechanical; Sulfuric Acids; Time Factors; Zea mays

Among the available agricultural byproducts, corn stover, with its yearly production of 10 million t (dry basis), is the most abundant promising raw material for fuel ethanol production in Hungary. In the United States, more than 216 million t of corn stover is produced annually, of which a portion also could possibly be collected for conversion to ethanol. However, a network of lignin and hemicellulose protects cellulose, which is the major source of fermentable sugars in corn stover (approx 40% of the dry matter [DM]). Steam pretreatment removes the major part of the hemicellulose from the solid material and makes the cellulose more susceptible to enzymatic digestion. We studied 12 different combinations of reaction temperature, time, and pH during steam pretreatment. The best conditions (200 degrees C, 5 min, 2% H2SO4) increased the enzymatic conversion (from cellulose to glucose) of corn stover more then four times, compared to untreated material. However, steam pretreatment at 190 degrees C for 5 min with 2% sulfuric acid resulted in the highest overall yield of sugars, 56.1 g from 100 g of untreated material (DM), corresponding to 73% of the theoretical. The liquor following steam explosion was fermented using Saccharomyces cerevisiae to investigate the inhibitory effect of the pretreatment. The achieved ethanol yield was slightly higher than that obtained with a reference sugar solution. This demonstrates that baker's yeast could adapt to the pretreated liquor and ferment the glucose to ethanol efficiently.

Topics: Biotechnology; Carbohydrates; Cellulase; Cellulose; Chromatography, High Pressure Liquid; Enzymes; Ethanol; Fermentation; Glucose; Hydrogen-Ion Concentration; Hydrolysis; Polysaccharides; Saccharomyces cerevisiae; Sulfuric Acids; Temperature; Time Factors; Zea mays

Corn stover is emerging as a viable feedstock for producing bioethanol from renewable resources. Dilute-acid pretreatment of corn stover can solubilize a significant portion of the hemicellulosic component and enhance the enzymatic digestibility of the remaining cellulose for fermentation into ethanol. In this study, dilute H2SO4 pretreatment of corn stover was performed in a steam explosion reactor at 160 degrees C, 180 degrees C, and 190 degrees C, approx 1 wt % H2SO4, and 70-s to 840-s residence times. The combined severity (Log10 [Ro] - pH), an expression relating pH, temperature, and residence time of pretreatment, ranged from 1.8 to 2.4. Soluble xylose yields varied from 63 to 77% of theoretical from pretreatments of corn stover at 160 and 180 degrees C. However, yields >90% of theoretical were found with dilute-acid pretreatments at 190 degrees C. A narrower range of higher combined severities was required for pretreatment to obtain high soluble xylose yields when the moisture content of the acidimpregnated feedstock was increased from 55 to 63 wt%. Simultaneous saccharification and fermentation (SSF) of washed solids from corn stover pretreated at 190 degrees C, using an enzyme loading of 15 filter paper units (FPU)/ g of cellulose, gave ethanol yields in excess of 85%. Similar SSF ethanol yields were found using washed solid residues from 160 and 180 degrees C pretreatments at similar combined severities but required a higher enzyme loading of approx 25 FPU/g of cellulose.

Topics: Animal Feed; Bioreactors; Biotechnology; Cellulase; Humidity; Hydrolysis; Kinetics; Sulfuric Acids; Temperature; Thermodynamics; Xylose; Zea mays

Sugarcane bagasse is a potential lignocellulosic feedstock for ethanol production, since it is cheap, readily available, and has a high carbohydrate content. In this work, bagasse was subjected to steam explosion pretreatment with different impregnation conditions. Three parallel pretreatments were carried out, one without any impregnation, a second with sulfur dioxide, and a third with sulfuric acid as the impregnating agent. The pretreatments were performed at 205 degrees C for 10 min. The pretreated material was then hydrolyzed using cellulolytic enzymes. The chemical composition of the hydrolyzates was analyzed. The highest yields of xylose (16.2 g/100 g dry bagasse), arabinose (1.5 g/100 g), and total sugar (52.9 g/100 g) were obtained in the hydrolysis of the SO2 -impregnated bagasse. The H2SO4 -impregnated bagasse gave the highest glucose yield (35.9 g/100 g) but the lowest total sugar yield (42.3 g/100 g) among the three methods. The low total sugar yield from the H2SO4-impregnated bagasse was largely due to by-product formation, as the dehydration of xylose to furfural. Sulfuric acid impregnation led to a three-fold increase in the concentration of the fermentation inhibitors furfural and 5-hydroxymethylfurfural (HMF) and a two-fold increase in the concentration of inhibitory aliphatic acids (formic, acetic, and levulinic acids) compared to the other two pretreatment methods. The total content of phenolic compounds was not strongly affected by the different pretreatment methods, but the quantities of separate phenolic compounds were widely different in the hydrolyzate from the H2SO4-impregnated bagasse compared with the other two hydrolyzates. No major differences in the content of inhibitors were observed in the hydrolyzates obtained from SO2-impregnated and non-impregnated bagasse. The fermentability of all three hydrolyzates was tested with a xylose-utilizing Saccharomyces cerevisiae strain with and without nutrient supplementation. The hydrolyzates of SO2-impregnated and nonimpregnated bagasse showed similar fermentability, whereas the hydrolyzate of H2SO4-impregnated bagasse fermented considerably poorer.

Topics: Cellulase; Cellulose; Ethanol; Fermentation; Furaldehyde; Furans; Glycoside Hydrolases; Saccharum; Sulfur Dioxide; Sulfuric Acids

Whole tree chips obtained from softwood forest thinnings were pretreated via single- and two-stage dilute-sulfuric acid pretreatment. Whole-tree chips were impregnated with dilute sulfuric acid and steam treated in a 4-L steam explosion reactor. In single-stage pretreatment, wood chips were treated using a wide range of severity. In two-stage pretreatment, the first stage was carried out at low severity to maximize hemicellulose recovery. Solubilized sugars were recovered from the first-stage prehydrolysate by washing with water. In the second stage, water-insoluble solids from first-stage prehydrolysate were impregnated with dilute sulfuric acid, then steam treated at more severe conditions to hydrolyze a portion of the remaining cellulose to glucose and to improve the enzyme digestibility. The total sugar yields obtained after enzymatic hydrolysis of two-stage dilute acid-pretreated samples were compared with sugar yields from single-stage pretreatment. The overall sugar yield from two-stage dilute-acid pretreatment was approx 10% higher, and the net enzyme requirement was reduced by about 50%. Simultaneous saccharification and fermentation using an adapted Saccharomyces cerevisiae yeast strain further improved cellulose conversion yield and lowered the enzyme requirement.

Topics: Cellulase; Cellulose; Cycadopsida; Fermentation; Hydrolysis; Polysaccharides; Saccharomyces cerevisiae; Steam; Sulfuric Acids; Trees; Wood

Cassava wastes--the peel and the root fibre were taken through various pretreatment procedures before being subjected to solid state fermentation with Trichoderma harzianum. Most of the pretreatment processes increased the cellulose and hemicellulose content of the cassava peel and fibre by as high as 155% while sulfuric acid treatment resulted in 25.3% loss in the peel hemicellulose. The best pretreatment found to be 1% NaOH at 120 degrees C gave the highest production of the Cx, the Cl and xylanase enzymes with the cassava root fibre. Xylanase and cellulase production with the exception of the Cx was found to be affected by age while an improved cassava variety TMS(2) 1425 peel and fibre rated highest in terms of production of the enzymes. Percentage hydrolysis within range of 56.52-67.64% were recorded for the enzymes on sorghum grains.

Topics: Age Factors; Cellulase; Edible Grain; Fermentation; Manihot; Sodium Hydroxide; Sulfuric Acids; Trichoderma; Xylan Endo-1,3-beta-Xylosidase; Xylosidases