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At least 19 recordsLinked to original sources
PubMed2002

Combined sedimentation and filtration process for cellulase recovery during hydrolysis of lignocellulosic biomass.

A combined sedimentation and ultrafiltration process was investigated for recovering cellulase enzymes during the hydrolysis of lignocellulosic biomass. Lignocellulosic particles larger than approx 50 microm in length were first removed via sedimentation using an inclined settler. Ultrafiltration was then used to retain the remaining lignocellulosic particles and the cellulose enzymes, while transmitting fermentable sugars and other small molecules. The permeate flux from the ultrafiltration step for a feed consisting of 0.22 w/v% cellulase is 64+/-5 L/m2-h, while that for a feed consisting of the settler overflow from a mixture 0.22 w/v% cellulase and 10 wt% lignocellulose fed to the settler is 130+/-20 L/m2-h. The higher permeate flux in the latter case is presumably due to binding of a portion of the cellulase enzymes to the lignocellulosic particles during hydrolysis and filtration, preventing the enzymes from fouling the membrane. A filter paper activity assay shows little loss in enzymatic activity throughout the combined sedimentation/ultrafiltration separation process.

Biomass↗
PubMed2026

Multistrategy metabolic engineering of Talaromyces pinophilus for α-amylase production from lignocellulosic biomass.

Filamentous fungi are important hosts for industrial enzyme production. Growing demand for α-amylase has increased reliance on food-derived carbon substrates, necessitating fungal strains that efficiently utilize nongrain biomass. In this study, Talaromyces pinophilus Y117 was metabolically engineered to produce α-amylase from lignocellulosic biomass. A strong cellobiohydrolase I gene (cbh1) promoter (Pcbh1Tru) was identified to drive expression. Multiple rounds of multilocus integration of the α-amylase gene were performed using homologous multicopy genomic sequences as recombination arms with a Cre/loxP-based recyclable selection system, yielding the multicopy strain Tp4, which achieved 4124.5 U/mL α-amylase activity in shake-flask fermentation with corncob powder as the sole carbon source. To minimize enzyme degradation, the protease gene 8538 was deleted using the Cre/lox2272 system, generating Tp4Δp. This strain showed a 50% increase in shake-flask α-amylase activity (6208.4 U/mL). In 3-L bioreactor cultivation, Tp4Δp exhibited excellent production performance, achieving 26 712.2 U/mL α-amylase activity. When corncob powder was used as the sole substrate, the cellulose and hemicellulose degradation rates reached 90.00% and 70.01%, respectively, and the enzyme yield reached 213 697.5 U per gram of corncob powder. This engineered strain demonstrates strong potential for industrial applications. The synthesis-degradation synergistic optimization strategy provides a practical approach for engineering filamentous fungal cell factories to produce enzymes directly from lignocellulosic biomass. One sentence summary Metabolic engineering of Talaromyces pinophilus through promoter optimization, multicopy integration, and protease deletion enables efficient α-amylase production from lignocellulosic biomass, achieving 26 712 U/mL in bioreactor fermentation.

Talaromyces↗
PubMed2001

Submerged cultivation of scytalidium thermophilum on complex lignocellulosic biomass for endoglucanase production.

Scytalidium thermophilum endoglucanase production was analyzed on lignocellulosic biomass in submerged cultures at 45 degrees C and 155 rpm for 8 days. Endoglucanase, adsorbability of endoglucanase onto avicel, as well as exoglucanase, and filter paper activities were determined and compared with those on microcrystalline cellulose (avicel) as the main source of carbon. Lentil bran and sunflower seed bagasse yielded c. 1.5 fold more endoglucanase and avicel-adsorbable endoglucanase activity than avicel, and activities on grass clippings were similar. Grass clippings yielded the highest percentage of avicel-adsorbable endoglucanase among all lignocellulosic substrates tested. By the time when endoglucanase activities reached maximal levels, exoglucanase activities on lentil bran, sunflower seed bagasse and grass clippings were c. 1.5-3 fold lower than those on avicel, although a significant difference in filter paper activities was not observed. On lignocellulosic biomass, maximum levels of endoglucanase activity were reached within 3-4 days, and within 6-7 days on avicel.

Journal Article↗
PubMed2001

Pyrolysis-GC-MS analysis of the formation and degradation stages of charred residues from lignocellulosic biomass.

The structural transformations undergone by lignocellulosic biomass (freeze-dried rye grass, Lolium rigidum) subjected to progressive isothermal heating (burning at 350 degrees C under oxidizing conditions for 30, 45, 60, 75, and 90 s) have been monitored by Curie-point pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). The pyrograms suggest that even charred residues after severe heating (carbon loss ca. 50%) still contain substantial concentrations of some resistant plant structural components. Several trends were observed when monitoring the relative concentrations of the different groups of pyrolysis compounds released during successive charring stages: (i) the tetrapyrrole moiety of chlorophylls is rapidly destroyed as indicated by the decreasing yields of pyrroles and pyrrolines, whereas the phytol backbone is comparatively more resistant, leading to phytadienes after dehydration and reduction; (ii) the increasing yields of imidazoles from progressively heated samples (maximum at 45 s stage) suggest accumulation of newly formed nitrogen-containing compounds that may survive natural fires; (iii) the lignin backbone shows a relative resistance, the yields of aromatic products pointing to progressive demethoxylation; and, (iv) a selective accumulation of recalcitrant alkyl material occurred, which is interpreted as the result of thermal condensation of hydrocarbons and fatty acids into macromolecular materials in the charred residue. In terms of the intensity of the isothermal heating, the yields of the different classes of alkyl compounds follow the order phytadienes < fatty acids < alkanes < wax esters < sterols.

Biomass↗
PubMed2005

Features of promising technologies for pretreatment of lignocellulosic biomass.

Cellulosic plant material represents an as-of-yet untapped source of fermentable sugars for significant industrial use. Many physio-chemical structural and compositional factors hinder the enzymatic digestibility of cellulose present in lignocellulosic biomass. The goal of any pretreatment technology is to alter or remove structural and compositional impediments to hydrolysis in order to improve the rate of enzyme hydrolysis and increase yields of fermentable sugars from cellulose or hemicellulose. These methods cause physical and/or chemical changes in the plant biomass in order to achieve this result. Experimental investigation of physical changes and chemical reactions that occur during pretreatment is required for the development of effective and mechanistic models that can be used for the rational design of pretreatment processes. Furthermore, pretreatment processing conditions must be tailored to the specific chemical and structural composition of the various, and variable, sources of lignocellulosic biomass. This paper reviews process parameters and their fundamental modes of action for promising pretreatment methods.

Biomass↗
PubMed1997

Biological conversion of lignocellulosic biomass to ethanol.

The important key technologies required for the successful biological conversion of lignocellulosic biomass to ethanol have been extensively reviewed. The biological process of ethanol fuel production utilizing lignocellulose as substrate requires: (1) delignification to liberate cellulose and hemicellulose from their complex with lignin, (2) depolymerization of the carbohydrate polymers (cellulose and hemicellulose) to produce free sugars, and (3) fermentation of mixed hexose and pentose sugars to produce ethanol. The development of the feasible biological delignification process should be possible if lignin-degrading microorganisms, their echophysiological requirements, and optimal bioreactor design are effectively coordinated. Some thermophilic anaerobes and recently-developed recombinant bacteria have advantageous features for direct microbial conversion of cellulose to ethanol, i.e. the simultaneous depolymerization of cellulosic carbohydrate polymers with ethanol production. The new fermentation technology converting xylose to ethanol needs also to be developed to make the overall conversion process more cost-effective. The bioconversion process of lignocellulosics to ethanol could be successfully developed and optimized by aggressively applying the related novel science and technologies to solve the known key problems of conversion process.

Biomass↗
PubMed1997

Evaluation of Candida acidothermophilum in ethanol production from lignocellulosic biomass.

A Saccharomyces-cerevisiae-based simultaneous saccharification and fermentation (SSF) of lignocellulosic biomass is limited to an operating temperature of about 37 degrees C, and even a small increase in temperature can have a deleterious effect. This points to a need for a more thermotolerant yeast. To this end, S. cerevisiae D5A and a thermotolerant yeast, Candida acidothermophilum, were tested at 37 degrees C, 40 degrees C, and 42 degrees C using dilute-acid-pretreated poplar as substrate. At 40 degrees C, C. acidothermophilum produced 80% of the theoretical ethanol yield, which was higher than the yield from S. cerevisiae D5A at either 37 degrees C or 40 degrees C. At 42 degrees C, C. acidothermophilum showed a slight drop in performance. On the basis of preliminary estimates, SSF with C. acidothermophilum at 40 degrees C can reduce cellulase costs by about 16%. Proportionately greater savings can be realized at higher temperatures if such a high-temperature SSF is feasible. This demonstrates the advantage of using thermophilic or thermotolerant yeasts.

Biomass↗
PubMedDate not supplied

Development and validation of a kinetic model for enzymatic saccharification of lignocellulosic biomass.

A multireaction kinetic model was developed for closed-system enzymatic hydrolysis of lignocellulosic biomass such as corn stover. Three hydrolysis reactions were modeled, two heterogeneous reactions for cellulose breakdown to cellobiose and glucose and one homogeneous reaction for hydrolyzing cellobiose to glucose. Cellulase adsorption onto pretreated lignocellulose was modeled via a Langmuir-type isotherm. The sugar products of cellulose hydrolysis, cellobiose and glucose, as well as xylose, the dominant sugar prevalent in most hemicellulose hydrolyzates, were assumed to competitively inhibit the enzymatic hydrolysis reactions. Model parameters were estimated from experimental data generated using dilute acid pretreated corn stover as the substrate. The model performed well in predicting cellulose hydrolysis trends at experimental conditions both inside and outside the design space used for parameter estimation and can be used for in silico process optimization.

Carbohydrates↗
PubMed1998

Nonisothermal simultaneous saccharification and fermentation for direct conversion of lignocellulosic biomass to ethanol.

The enzymatic reaction in the simultaneous saccharification and fermentation (SSF) is operated at a temperature much lower than its optimum level. This forces the enzyme activity to be far below its potential, consequently raising the enzyme requirement. To alleviate this problem, a nonisothermal simultaneous saccharification and fermentation process (NSSF) was investigated. The NSSF is devised so that saccharification and fermentation occur simultaneously, yet in two separate reactors that are maintained at different temperatures. Lignocellulosic biomass is retained inside a column reactor and hydrolyzed at the optimum temperature for the enzymatic reaction (50 degrees C). The effluent from the column reactor is recirculated through a fermenter, which runs at its optimum temperature (20-30 degrees C). The cellulase enzyme activity is increased by a factor of 2-3 when the hydrolysis temperature is raised from 30 to 50 degrees C. The NSSF process has improved the enzymatic reaction in the SSF to the extent that it reduces the overall enzyme requirement by 30-40%. The effect of temperature on beta-glucosidase activity was the most significant among the individual cellulase compounds. Both ethanol yield and productivity in the NSSF are substantially higher than those in the SSF at the enzyme loading of 5 IFPU/g glucan. With 10 IFPU/g glucan, improvement in productivity was more discernible for the NSSF. The terminal yield attainable in 4 d with the SSF was reachable in 40 h with the NSSF.

Biomass↗
PubMed2004

Kinetics of glucose decomposition during dilute-acid hydrolysis of lignocellulosic biomass.

Recent research work in-house both at Auburn University and National Renewable Energy Laboratory has demonstrated that extremely low concentrations of acid (e.g., 0.05-0.2 wt% sulfuric acid) and high temperatures (e.g., 200-230 degrees C) are reaction conditions that can be effectively applied for hydrolysis of the cellulosic component of biomass. These conditions are far from those of the conventional dilute-acid hydrolysis processes, and the kinetic data for glucose decomposition are not currently available. We investigated the kinetics of glucose decomposition covering pH values of 1.5-2.2 and temperatures of 180-230 degrees C using glass ampoule reactors. The primary factors controlling glucose decomposition are the reaction medium, acid concentration, and temperature. Based on the experimental data, a kinetic model was developed and the best-fit kinetic parameters were determined. However, a consistent discrepancy in the rate of glucose disappearance was found between that of the model based on pure glucose data and that observed during the actual process of lignocellulosic biomass hydrolysis. This was taken as an indication that glucose recombines with acid-soluble lignin during the hydrolysis process, and this conclusion was incorporated accordingly into the overall model of glucose decomposition.

Acids↗
PubMed1991

Anaerobic digestion of lignocellulosic biomass and wastes. Cellulases and related enzymes.

Anaerobic digestion represents one of several commercially viable processes to convert woody biomass, agricultural wastes, and municipal solid wastes to methane gas, a useful energy source. This process occurs in the absence of oxygen, and is substantially less energy intensive than aerobic biological processes designed for disposal purposes. The anaerobic conversion process is a result of the synergistic effects of various microorganisms, which serve as a consortium. The rate-limiting step of this conversion process has been identified as the hydrolysis of cellulose, the major polymeric component of most biomass and waste feedstocks. Improvements in process economics therefore rely on improving the kinetic and physicochemical characteristics of cellulose degrading enzymes. The most thoroughly studied cellulase enzymes are produced by aerobic fungi, namely Trichoderma reesei. However, the pH and temperature optima of fungal cellulases make them incompatible for use in anaerobic digestion systems, and the major populations of microorganisms involved in cellulase enzyme production under anaerobic digestion conditions are various bacterial producers. The current state of understanding of the major groups of bacterial cellulase producers is reviewed in this paper. Also addressed in this review are recently developed methods for the assessment of actual cellulase activity levels, reflective of the digester "hydrolytic potential," using a series of detergent extractive procedures.

Anaerobiosis↗
PubMed2002

Diffusion of sulfuric acid within lignocellulosic biomass particles and its impact on dilute-acid pretreatment.

Intra-particle diffusion of sulfuric acid into sugarcane bagasse, corn stover, rice straw and yellow poplar was investigated to determine the effective diffusivity of sulfuric acid within the porous biomass structure. Diffusion experiments were conducted over 25-75 degrees C for two different biomass sizes using dynamic diffusion test cells. Diffusivities of sulfuric acid in agricultural residues were significantly higher than those of hard wood. Diffusivity data for each biomass were fitted into the Arrhenius equation for extrapolation to higher temperatures. The diffusivity data were subsequently incorporated into a theoretical model to determine acid profile within the biomass matrix. The modeling results indicate that intra-particle diffusion of acid influences the rate of dilute-acid pretreatment if unground biomass feedstock is used under normal pretreatment conditions. A criterion was set up to determine the critical biomass size at which the intra-particle acid diffusion becomes a rate-influencing factor for a given pretreatment condition.

Acids↗
PubMed2004

Production of cellulases and hemicellulases by Aspergillus niger KK2 from lignocellulosic biomass.

To investigate the production of cellulases and hemicellulases from Aspergillus niger KK2, solid state fermentation (SSF) was performed by using different ratios of rice straw and wheat bran. When A. niger KK2 was grown on rice straw alone as a solid support in SSF, the maximum FPase activity was 19.5 IU g(-1) in 4 days. Also, CMCase (129 IU g(-1)), beta-glucosidase (100 IU g(-1)), xylanase (5070 IU g(-1)) and beta-xylosidase (193 IU g(-1)) activities were concurrently obtained after 5-6 days of fermentation. The higher enzyme activities produced by A. niger KK2 is a significant advantage from the viewpoint of practical saccharification reaction. Cellulases and hemicellulases produced by A. niger KK2 might be applied to pulp and paper industry, feed industry and chemical industry.

Aspergillus niger↗
PubMedDate not supplied

Effects of pressing lignocellulosic biomass on sugar yield in two-stage dilute-acid hydrolysis process.

Dilute sulfuric acid catalyzed hydrolysis of biomass such as wood chips often involves pressing the wood particles in a dewatering step (e.g., after acid impregnation) or in compression screw feeders commonly used in continuous hydrolysis reactors. This study addresses the effects of pressing biomass feedstocks using a hydraulic press on soluble sugar yield obtained from two-stage dilute-acid hydrolysis of softwood. The pressed acid-impregnated feedstock gave significantly lower soluble sugar yields than the never-pressed (i.e., partially air-dried or filtered) feedstock. Pressing acid-impregnated feedstocks before pretreatment resulted in a soluble hemicellulosic sugar yield of 76.9% from first-stage hydrolysis and a soluble glucose yield of 33.7% from second-stage hydrolysis. The dilute-acid hydrolysis of partially air-dried feedstocks having total solids and acid concentrations similar to those of pressed feedstocks gave yields of 87.0% hemicellulosic sugar and 46.9% glucose in the first and second stages, respectively. Microscopic examination of wood structures showed that pressing acid-impregnated wood chips from 34 to 54% total solids (TS) did not cause the wood structure to collapse. However, pressing first-stage pretreated wood chips (i.e., feedstock for second-stage hydrolysis) from approximately 30 to 43% TS caused the porous wood matrix to almost completely collapse. It is hypothesized that pressing alters the wood structure and distribution of acid within the cell cavities, leading to uneven heat and mass transfer during pretreatment using direct steam injection. Consequently, lower hydrolysis yield of soluble sugars results. Dewatering of corn stover by pressing did not impact negatively on the sugar yield from single-stage dilute-acid pretreatment.

Acids↗
PubMed2000

Optimization of simultaneous saccharification and fermentation for the production of ethanol from lignocellulosic biomass.

Simultaneous saccharification and fermentation (SSF) of alkaline hydrogen peroxide pretreated Antigonum leptopus (Linn) leaves to ethanol was optimized using cellulase from Trichoderma reesei QM-9414 (Celluclast from Novo) and Saccharomyces cerevisiae NRRL-Y-132 cells. Response surface methodology (RSM) and a three-level four-variable design were employed to evaluate the effects of SSF process variables such as cellulase concentration (20-100 FPU/g of substrate), substrate concentration (5-15% w/v), incubation time (24-72 h), and temperature (35-45 degrees C) on ethanol production efficiency. Cellulase and substrate concentrations were found to be the most significant variables. The optimum conditions arrived at are as follows: cellulase = 100 FPU/g of substrate, substrate = 15% (w/v), incubation time = 57.2 h, and temperature = 38.5 degrees C. At these conditions, the predicted ethanol yield was 3.02% (w/v) and the actual experimental value was 3.0% (w/v).

Biomass↗
PubMed2004

Cellulase retention and sugar removal by membrane ultrafiltration during lignocellulosic biomass hydrolysis.

Technologies suitable for the separation and reuse of cellulase enzymes during the enzymatic saccharification of pretreated corn stover are investigated to examine the economic and technical viability of processes that promote cellulase reuse while removing inhibitory reaction products such as glucose and cellobiose. The simplest and most suitable separation is a filter with relatively large pores on the order of 20-25 mm that retains residual corn stover solids while passing reaction products such as glucose and cellobiose to form a sugar stream for a variety of end uses. Such a simple separation is effective because cellulase remains bound to the residual solids. Ultrafiltration using 50-kDa polyethersulfone membranes to recover cellulase enzymes in solution was shown not to enhance further the saccharification rate or overall conversion. Instead, it appears that the necessary cellulase enzymes, including beta-glucosidase, are tightly bound to the substrate; when fresh corn stover is contacted with highly washed residual solids, without the addition of fresh enzymes, glucose is generated at a high rate. When filtration was applied multiple times, the concentration of inhibitory reaction products such as glucose and cellobiose was reduced from 70 to 10 g/L. However, an enhanced saccharification performance was not observed, most likely because the concentration of the inhibitory products remained too high. Further reduction in the product concentration was not investigated, because it would make the reaction unnecessarily complex and result in a product stream that is much too dilute to be useful. Finally, an economic analysis shows that reuse of cellulase can reduce glucose production costs, especially when the enzyme price is high. The most economic performance is shown to occur when the cellulase enzyme is reused and a small amount of fresh enzyme is added after each separation step to replace lost or deactivated enzyme.

Biomass↗
PubMed2000

Development of new ethanologenic Escherichia coli strains for fermentation of lignocellulosic biomass.

Two new ethanologenic strains (FBR4 and FBR5) of Escherichia coli were constructed and used to ferment corn fiber hydrolysate. The strains carry the plasmid pLOI297, which contains the genes from Zymomonas mobilis necessary for efficiently converting pyruvate into ethanol. Both strains selectively maintained the plasmid when grown anaerobically. Each culture was serially transferred 10 times in anaerobic culture with sugar-limited medium containing xylose, but no selective antibiotic. An average of 93 and 95% of the FBR4 and FBR5 cells, respectively, maintained pLOI297 in anaerobic culture. The fermentation performances of the repeatedly transferred cultures were compared with those of cultures freshly revived from stock in pH-controlled batch fermentations with 10% (w/v) xylose. Fermentation results were similar for all the cultures. Fermentations were completed within 60 h and ethanol yields were 86-92% of theoretical. Maximal ethanol concentrations were 3.9-4.2% (w/v). The strains were also tested for their ability to ferment corn fiber hydrolysate, which contained 8.5% (w/v) total sugars (2.0% arabinose, 2.8% glucose, and 3.7% xylose). E. coli FBR5 produced more ethanol than FBR4 from the corn fiber hydrolysate. E. coli FBR5 fermented all but 0.4% (w/v) of the available sugar, whereas strain FBR4 left 1.6% unconsumed. The fermentation with FBR5 was completed within 55 h and yielded 0.46 g of ethanol/g of available sugar, 90% of the maximum obtainable.

Biomass↗