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Background Bamboo, a lignocellulosic feedstock, is considered as a potentially excellent raw material and evaluated for lignocellulose degradation and bioethanol production, with a focus on using physical and chemical pre-treatment. However, studies reporting the biodegradation of bamboo lignocellulose using microbes such as bacteria and fungi are scarce. Results In the present study, Bacillus velezensis LC1 was isolated from Cyrtotrachelus buqueti, in which the symbiotic bacteria exhibited lignocellulose degradation ability and cellulase activities. We performed genome sequencing of B. velezensis LC1, which has a 3929,782-bp ring chromosome and 46.5% GC content. The total gene length was 3,502,596 bp using gene prediction, and the GC contents were 47.29% and 40.04% in the gene and intergene regions, respectively. The genome contains 4018 coding DNA sequences, and all have been assigned predicted functions. Carbohydrate-active enzyme annotation identified 136 genes annotated to CAZy families, including GH, GTs, CEs, PLs, AAs and CBMs. Genes involved in lignocellulose degradation were identified. After a 6-day treatment, the bamboo shoot cellulose degradation efficiency reached 39.32%, and the hydrolysate was subjected to ethanol fermentation with Saccharomyces cerevisiae and Escherichia coli KO11, yielding 7.2 g/L of ethanol at 96 h. Conclusions These findings provide an insight for B. velezensis strains in converting lignocellulose into ethanol. B. velezensis LC1, a symbiotic bacteria, can potentially degrade bamboo lignocellulose components and further transformation to ethanol, and expand the bamboo lignocellulosic bioethanol production.

Phenolic aldehydes from lignocellulose pretreatment harshly inhibit the viability and metabolism of ethanol fermenting strains. Direct conversion of phenolic aldehydes is usually incomplete due to their low water solubility and recalcitrance to bioconversion. Here we consolidated phenolic aldehydes bioconversion and ethanol fermentation in a typical ethanologenic bacterium Zymomonas mobilis by constructing an intracellular oxidative pathway. The gene PP_2680 encoding NAD+-dependent aldehyde dehydrogenase from Pseudomonas putida KT2440 was expressed in Z. mobilis ZM4. The expression significantly improved both aldehyde inhibitor conversion and ethanol fermentability in corn stover hydrolysate. The purified PP_2680 aldehyde dehydrogenase showed strong in vitro oxidative capacity on phenolic aldehydes and its in vivo expression significantly up-regulated the key genes in the ED pathway and the oxidative phosphorylation. This study provided an important concept of simultaneous biodetoxification and fermentation in ethanologenic strains for the improvement of ethanol fermentability.

Thermophilic methane fermentation was a valid approach for handling the stillage eluted from ethanol fermentation of waste paper and kitchen waste. The wide organic loading rate (OLR) range (2-14 g VTS/(L d)) for stable performance and relatively high energy recovery efficiency (79.0%) were achieved, and OLR of 8 g VTS/(L d) was optimum for achievement of highest biogas evolution and VTS removal efficiency. Microbial community analysis revealed that hydrolysis of cellulose was the critical step for methane production from the stillage.

Lignocellulosic biomass is a sustainable feedstock for fuel ethanol production, but it is characterized by low mass and energy densities, and distributed production with relatively small scales is more suitable for cellulosic ethanol, which can better balance cost for the feedstock logistics. Lignocellulosic biomass is recalcitrant to degradation, and pretreatment is needed, but more efficient pretreatment technologies should be developed based on an in-depth understanding of its biosynthesis and regulation for engineering plant cell walls with less recalcitrance. Simultaneous saccharification and co-fermentation has been developed for cellulosic ethanol production, but the concept has been mistakenly defined, since the saccharification and co-fermentation are by no means simultaneous. Lignin is unreactive, which not only occupies reactor spaces during the enzymatic hydrolysis of the cellulose component and ethanol fermentation thereafter, but also requires extra mixing, making high solid loading difficult for lignocellulosic biomass and ethanol titers substantially compromised, which consequently increases energy consumption for ethanol distillation and stillage discharge, presenting another challenge for cellulosic ethanol production. Pentose sugars released from the hydrolysis of hemicelluloses are not fermentable with Saccharomyces cerevisiae used for ethanol production from sugar- and starch-based feedstocks, and engineering the brewing yeast and other ethanologenic species such as Zymomonas mobilis with pentose metabolism has been performed within the past decades. However strategies for the simultaneous co-fermentation of pentose and hexose sugars that have been pursued overwhelmingly for strain development might be modified for robust ethanol production. Finally, unit integration and system optimization are needed to maximize economic and environmental benefits for cellulosic ethanol production. In this article, we critically reviewed updated progress, and highlighted challenges and strategies for solutions.

The aim of this study was to develop an effective thermochemical method for treatment of industrial hemp, in order to increase its bioconversion to biofuels and bio-products. Industrial hemp was subjected to various thermochemical pretreatments using: alkaline (3 % NaOH), oxidative (3 % H2O2 at pH 11.5) and glycerol-based methods (70-90 % of glycerol, 1-3 % NaOH), prior to enzymatic hydrolysis with Cellic® CTec2/Cellic® HTec2 (15 FPU∙g−1 glucan). Innovative pretreatment with glycerol fraction (80 % glycerol content, 2 % NaOH, 12.5 % biomass loading) showed to be superior over commonly used alkaline and oxidative methods with respect to by-products generation and sugar losses. Integrated process of ethanol production from enriched cellulose fraction (172 kg EtOH∙Mg−1 of dry hemp) and succinic production from xylose-rich residue after ethanol fermentation (59 kg∙Mg−1 of dry hemp) allowed to convert about 97 % of sugars released (glucose and xylose) during enzymatic hydrolysis of pre-treated biomass. The present study showed that it is possible to replace 50 % of the costly yeast extract, used during succinic fermentation as nitrogen source, by alternative nitrogen source (rapeseed cakes) without significant deterioration of succinic yield. Pretreatment liquor after lignin precipitation (52 kg∙Mg−1 of biomass treated) exhibited a high biodegradability (92 %) and allowed to produce 420 m3 CH4/Mg VS). Results obtained in this study clearly document the possibility of biofuels (bioethanol, biogas) and bio-chemicals production from industrial hemp, in a biorefinery approach.

This study investigates the feasibility of producing ethanol from date palm seeds. The chemical compositions of three varieties of date seeds were first studied, showing mainly the presence of cellulose and hemicellulose. Ethanol was produced after a pre-treatment of date seeds using acid hydrolysis to extract the cellulosic fraction and to remove the lignin. Producing ethanol by fermentation was performed using the yeast Saccharomyces cerevisiae for 24 h, during which ethanol yield, biomass concentration, and total reducing sugars were recorded. The results obtained showed that the sugar content decreased over time, while ethanol production increased. Indeed, date seeds gave the highest ethanol concentration of 21.57 g/L after 6 h of alcoholic fermentation. These findings proved the feasibility of producing ethanol from date seeds.

Made available in DSpace on 2020-12-12T02:16:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-08-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Cellulases have been investigated for their potential application in lignocellulosic residues, used to produce second-generation ethanol (2 GE), as these enzymes can hydrolyze cellulose and release glucose for alcoholic fermentation by yeasts. The aim of this project was to characterize the endoglucanases produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid-state cultivation. Tests were carried out with the crude enzyme extract (CEE) to assess the effects of pH, temperature, and various chemicals including cations and phenolic compounds on endoglucanolytic activity. The molecular weight estimation was performed by polyacrylamide gel electrophoresis (SDS-PAGE), and it was possible to observe seven endoglucanases detected by zymography, with estimated Mw in the range between 26 and 82 kDa. The optimum pH for the CEE was 5.5 and the optimum incubation temperature was 70 °C. Concerning endoglucanase stability, the highest activities were achieved in the pH range of 6.0–6.5, and between temperatures of 30 and 40 °C up to 120 min. Among the tested reagents, Triton, dithiotreitol (DTT), and Isopropanol increased the enzymatic activity on the crude extract by about 10%. Among cations, only Sr2+ significantly increased the endoglucanases’ activity. Of the phenolic compounds, although all induced a decrease of the endoglucanolytic activity, none could nullify it after a pre-incubation for 24 h. The apparent kinetic parameters were determined using carboxymethylcellulose (CMC), being Vmax = 51.7 ± 2.7 μmol min−1 and Km = 0.99 ± 0.15 g% CMC (9.9 mg mL−1). Graduate Program in Microbiology Department of Biology Department of Chemistry and Environmental Sciences Biosciences Languages and Exact Sciences Institute São Paulo State University (IBILCE-UNESP) Department of Chemistry and Environmental Sciences Biosciences Languages and Exact Sciences Institute São Paulo State University (IBILCE-UNESP)

The objective of this study was to obtain two types of fuels, i.e., bioethanol and biogas, in a sequential combination of biochemical processes from lignocellulosic biomass (corn straw). Waste from the agricultural sector containing lignocellulose structures was used to obtain bioethanol, while the post-fermentation (cellulose stillage) residue obtained from ethanol fermentation was a raw material for the production of high-power biogas in the methane fermentation process. The studies on obtaining ethanol from lignocellulosic substrate were based on the simultaneous saccharification and fermentation (SSF) method, which is a simultaneous hydrolysis of enzymatic cellulose and fermentation of the obtained sugars. Saccharomyces cerevisiae (D-2) in the form of yeast cream was used for bioethanol production. The yeast strain D-2 originated from the collection of the Institute of Agricultural and Food Biotechnology. Volatile compounds identified in the distillates were measured using gas chromatography with flame ionization detector (GC-FID). CH4 and CO2 contained in the biogas were analyzed using a gas chromatograph in isothermal conditions, equipped with thermal conductivity detector (katharometer) with incandescent fiber. Our results show that simultaneous saccharification and fermentation enables production of bioethanol from agricultural residues with management of cellulose stillage in the methane fermentation process.

Mixed biomass has the characteristics of stable supply and low cost of collection, and its bio-refinery could provide an effective contribution to the sustainable development of fuel ethanol production. Compared to traditional single energy crops, the key challenge of mixed biomass conversion lies in overcoming the non-uniformity of blended materials. In this study, the influence of blending on hydrolysis and ethanol fermentation of garden wastes was investigated. There was an unexpected result in that the total xylose yield during a liquid hot water process was enhanced when different garden biomasses were mixed. The peak of total xylose recovery appeared at the value of ∼0.5 for the ratio of xylan to lignin, which was further confirmed by blending multiple biomasses. Moreover, the non-uniformity of physio-chemical characteristics in mixed garden wastes restricted the co-hydrolysis of cellulose, particularly in the mixed-five-biomass. In spite of this disadvantage, blended feedstocks showed a better flow behavior, which could make them more suitable for enzymatic hydrolysis and ethanol fermentation with high substrate loading. The 72 h enzymatic digestibility and glucose concentration of treated mixed-three-biomass reached 100 ± 5% and 52.3 ± 2.6 g/l under the conditions of 15% (w/v) substrate loading, with the addition of 1% (v/v) Tween 20, which gave a maximum ethanol concentration of 16.9 ± 0.8 g/l. Overall, these results have shown that the complementary or synergistic effects of different individual biomasses make it feasible to produce ethanol from mixed biomass.

The exploitation of starchy wastes for the production of value-added products and the consolidated bioprocessing (CBP) have a positive environmental and cost-effective impact. For this reason, a cell factory (CF) was employed to perform three bioprocesses in one step (CBP) for alcoholic fermentation of starch without genetic modification. CF is a bilayer biocatalyst consisting of an inner layer of immobilized Saccharomyces cerevisiae on tubular cellulose (TC) and an external layer of immobilized Aspergillus awamori on alginates (ALG). In first, the suggested CF was proved to be more effective compared with co-immobilized S. cerevisiae and A. awamori on ALG beads for starch fermentation. Subsequently, the effect of the (i) S. cerevisiae concentration (g S. cerevisiae/g TC) during its immobilization, (ii) CaCl 2 concentration (% w/v), (iii) form of bilayer CF (wet or freeze-dried) and (iv) freeze-dried CF concentration on 5% (w/v) starch fermentation was examined. It was found that the higher ethanol production (32.17 mL/L), productivity (4.60 mL/L/d) and yield (0.51 g ethanol/g starch) reaching the theoretical one were obtained when the CF in freeze dried form fermented 100 mL of 5% (w/v) starch solution. Finally, two repeated fermentation batches were performed using the best CF. The ethanol yield was decreased during the repeated fermentation batches, but remained in acceptable levels. The successful preparation of CF was verified with FTIR spectroscopy, SEM and TEM analysis. The results indicated that the objective of designing CF was successfully achieved, and the results are promising to be the base for the exploitation of starchy wastes and the development of an innovation in brewing industry by eliminating the malting stage. This CF can be applied as model for different bioprocesses of White Biotechnology, e.g. substituting S. cerevisiae with the appropriate microorganism to produce other metabolites of added value.

A promising approach to help offset production costs for the cellulosic ethanol industry is to improve ethanol productivity while simultaneously generating value-added by-products. This study reports integration of an advanced fermentation approach (self-cycling fermentation) with the production of cellulose nanocrystals. Specifically, wood pulp was enzymatically hydrolyzed to yield dissolved sugars, which were fed to a self-cycling fermentation system for ethanol production, and residual solids were used for cellulose nanocrystals production via acid hydrolysis. Self-cycling fermentation achieved stable ethanol production for 10 cycles with significantly greater productivity than batch operation: ethanol volumetric productivity increased by 63–95% and annual ethanol productivity by 96 ± 5%. Additionally, the enzyme hydrolysis approach employed did not impede ethanol fermentation, and the cellulose nanocrystals generated displayed properties consistent with previous studies. Taken together, these results highlight the potential of this co-production strategy to produce both cellulosic ethanol and cellulose nanocrystals from a single feedstock.

Sugarcane bagasse (SCB) is an abundant, renewable and inexpensive agricultural byproduct for the production of biofuel and other biobased products. To effectively saccharify SCB with cellulases, combination with dilute alkali salts Na2SO3/Na3PO4 (0.4% Na3PO4, 0.03% Na2SO3) at 7.5% sulfidity and hot water (DASHW) in "one-pot" pretreatment media by autoclaving at 110°C for 40min was attempted to pretreat SCB in this study. Furthermore, FT-IR, XRD and SEM were employed to characterize the changes in the cellulose structural characteristics (porosity, morphology, and crystallinity) of the pretreated Na2SO3/Na3PO4-SCB solid residue, which indicated that combination pretreatment could effectively remove lignin and hemicellulose for enhancing enzymatic saccharification. After 72h, the reducing sugars and glucose from the enzymatic in situ hydrolysis of 50g/L Na2SO3/Na3PO4-SCB in dilute Na2SO3/Na3PO4 (0.27% Na3PO4, 0.02% Na2SO3) media were obtained at 33.8 and 21.8g/L, respectively. Finally, the SCB-hydrolysates containing 20g/L glucose were used for ethanol fermentation in the presence of dilute alkali salts. After 48h, the ethanol yield was 0.42g ethanol/g glucose, which represents 82.1% of the theoretical yield. In conclusion, this study provided an effective pretreatment strategy for enhancing SCB's saccharification, which has potential application of other lignocellulosic materials.

Numerical analyses of energy production processes are important for practical applications. Here, we established a model of enzymatic hydrolysis and ethanol fermentation. The kinetic model of these reactions were represented by Langmuir adsorption, Michaelis-Menten, and Shuler models. In our model, ethanol fermentation and enzymatic hydrolysis models were fit to experimental data separately, and their model equations were then combined for fitting of experimental data, including the size of cellulose, amount of adsorbed protein, and decreases in glucose as the theoretical production values for SSF. From these kinetic models, the theoretical values for saccharification and fermentation were calculated, and optimal parameters were determined. Using these parameters, theoretical curves for simultaneous saccharification and fermentation were predicted. Additionally, we identified changes in the radius of cellulose and the concentrations of cellulose, cellobiose, glucose, and ethanol during saccharification and fermentation.

Background: Common buckwheat (Fagopyrum esculentum Moench) is an annual spring-emerging crop that is classified among the dicotyledons, due to the manner of its cultivation, use, and chemical composition of seeds. The use of buckwheat straw for energy purposes—for example, for the production of second generation bioethanol—might enable its wider application and increase the cost-effectiveness of tillage. Methods: In this study, we examined the usability of buckwheat straw for the production of bioethanol. We pretreated the raw material with ionic liquids and subsequently performed enzymatic hydrolysis and alcoholic fermentation. The obtained chemometric data were analyzed using the Partial Least Squares (PLS) regression model. PLS regression in combination with spectral analysis within the near-infrared (NIR) spectrum allowed for the rapid determination of the amount of cellulose in the raw material and also provided information on the changes taking place in its structure. Results: We obtained good results for the combination of 1-ethyl-3-methylimidazolium acetate as the ionic liquid and Cellic CTec2 as the enzymatic preparation for the pretreatment of buckwheat straw. The highest concentration of glucose following 72 h of enzymatic hydrolysis was found to be around 5.5 g/dm3. The highest concentration of ethanol (3.31 g/dm3) was obtained with the combination of 1-butyl-3-methylimidazolium acetate for the pretreatment and cellulase from Trichoderma reesei for enzymatic hydrolysis. Conclusions: In summary, the efficiency of the fermentation process is strictly associated with the pool of available fermenting sugars, and it depends on the type of ionic liquid used during the pretreatment and on the enzymatic preparation. It is possible to obtain bioethanol from buckwheat straw using ionic liquid for pretreatment of the raw material prior to the enzymatic hydrolysis and alcoholic fermentation of the material.

FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais This study evaluated the production of lignocellulose-degrading enzymes by solid-state fermentation (SSF) using a microbial consortium of Aspergillus fumigatus SCBM6 and A. niger SCBM1 (AFN extract). The fungal strains were cultivated in sugarcane bagasse (SCB) and wheat bran (WB) as lignocellulosic substrates for 7 days at 30 °C. After SSF, the highest peaks of enzyme production were 150 and 80 U g−1 for β-xylosidase and β-glucosidase at 48 h, 375 U g−1 for xylanase at 96 h, and 80 U g−1 for endoglucanase and 4 U g−1 for cellulase activity on filter paper (FPase) at 144 h. The efficiency of the produced AFN extract was investigated in the enzymatic hydrolysis of crude biomass sorghum (BS) and after the removal of extractives (ES). After saccharification, the glucose and xylose concentrations were 10× superior in ES than in BS hydrolysate (2.5 g L−1 after 12 h). The presence of inhibitors of alcoholic fermentation, such as formic acid, was also reduced in ES hydrolysates, indicating that the removal of extractives positively contributed to the effectiveness of enzymatic hydrolysis of biomass sorghum using AFN extract

Rice husk (RH) is the main biomass byproduct of the rice industry, so much that it corresponds to 20% of the total cereal production and is generally considered a waste. Due to its content of cellulose and hemicellulose, the RH has a great potential for its alternative use in the production of bioethanol, since it has a great advantage with respect to the conventional raw materials used (sugar cane, corn, beet and sweet sorghum, among others), given that this lignocellulosic material is abundant and does not compete with the food industry. The objective of this work was to obtain bioethanol from rice husk at a pilot scale, taking advantage of its cellulose and hemicellulose content. For this, a sample of 450g of RH was taken, which was pretreated with a 2.00% w/v NaOH solution and subsequently hydrolyzed with acid cellulase (CFB3S). The alcoholic fermentation of total reducing sugars (TRS) obtained in the enzymatic hydrolysis was carried out with Saccharomyces cerevisiae, finally, the fractional distillation of the must was carried out, by means of which four bioethanol samples were obtained with concentrations of 71.3% v/v, 30.0% v/v, 10.3% v/v, and 5.3% v/v of alcohol, which when mixed produced a sample of 95mL of solution with a concentration of 27% v/v alcohol (4.70g/100g of RH).

The aim of this study was to remove 5-hydroxymethyl furfural (5-HMF) and furfural, known as fermentation inhibitors, in acid pretreated hydrolysates (APH) obtained from Scenedesmus obliquus using activated carbon. Microwave-assisted pretreatment was used to produce APH containing glucose, xylose, and fermentation inhibitors (5-HMF, furfural). The response surface methodology was applied to optimize key detoxification variables such as temperature (16.5–58.5 °C), time (0.5–5.5 h), and solid–liquid (S-L) ratio of activated carbon (0.6–7.4 w/v%). Three variables showed significant effects on the removal of fermentation inhibitors. The optimum detoxification conditions with the maximum removal of fermentation inhibitors and the minimum loss of sugars (glucose and xylose) were as follows: temperature of 36.6 °C, extraction time of 3.86 h, and S-L ratio of 3.3 w/v%. Under these conditions, removal of 5-HMF, furfural, and sugars were 71.6, 83.1, and 2.44%, respectively, which agreed closely with the predicted values. When the APH and detoxified APH were used for ethanol fermentation by S. cerevisiae, the ethanol produced was 38.5% and 84.5% of the theoretical yields, respectively, which confirmed that detoxification using activated carbon was effective in removing fermentation inhibitors and increasing fermentation yield without significant removal of fermentable sugars.

The aim of this investigation was to study the use of nano/micro-porous cellulosic materials (tubular cellulose, TC) produced by delignification of mango (Magnifera indica L.), sal (Shorea robusta G.) sawdust and rice husk (Oryza sativa L.) in various food bioprocesses. Specifically, the porous structure of TC renders it suitable for use as filter material in continuous cold pasteurization processes as well as biocatalyst (carrier for yeast immobilization) for alcoholic fermentation. Water contaminated with Saccharomyces cerevisiae or Lactobacillus bulgaricus cells was continuously pumped through TC filters. The effect of TC origin, type of contamination and microbial load on the operational stability of the system was assessed. The microbial load removal ranged between 99.9-88.2% and 92–77% for yeast and bacteria, respectively. The system operated efficiently for up to 55 days. Filter regeneration was done with hot water. Scanning electron micrographs showed the entrapment of cells in the TC filters. The TCs were further used as S. cerevisiae immobilization carriers for alcoholic fermentation of grape must and glucose media at 15 °C. Satisfactory fermentation rates, high ethanol content, ethanol productivity, and volatile by-products formation were observed. The TCs showed great potential for water pretreatment and food bioprocessing with simultaneous valorization of lignocellulosic biomass.

The stacking fermentation (SF) process is a bridge between Daqu making and alcoholic fermentation in the pits. Revealing the dynamic diversification of the microbial functional groups in this process is of vital importance to unfold the mechanism of Baiyunbian liquor flavor, and improve the quantity and production of the traditional Chinese liquor fermentation process. A total of 142 strains of bacteria were isolated from the Baiyunbian liquor SF process, which were divided into 17 species and two subspecies. These isolated bacteria could hydrolyze the main carbon resources such as starch, protein, cellulose, lignin, and fat as well as produce acetic acid, lactic acid, or butyric acid. Our results indicated that the dynamic variations of the bacterial functional groups revealed their specific functions in the Baiyunbian SF process. Interestingly, although the bacterial bio-diversity decreased during the SF process, the total bacterial biomass increased at the end, in which the biomass of the starch degrading bacteria was dominate throughout the whole SF process. Among the identified bacteria, B.licheniformis and B.subtilis contributed the most biomass and functionalities during the Baiyunbian SF process, playing important roles in the formation of the Maotai-flavor. Our study revealed the bio-diversity of the microbial functional groups in the Baiyunbian liquor SF process and the mechanism of the Baiyunbian liquor flavor formation, providing valuable guidance to improve the traditional Chinese liquor fermentation process. It could also act as an attempt to turn the present manufacturing process to standardized procedures.

Peanut shells (i.e., an abundant industrial by-product) were subjected to an innovative hydrothermal pretreatment approach using high-pressure CO2 to enhance their enzymatic hydrolysis conversion into glucose. This pretreatment led to a reduction in hemicellulose content in the pretreated peanut shells from 12.4% to as low as 1.8%, which facilitated subsequent conversion into glucose by enzymatic hydrolysis. This pretreatment approach was assessed within a 170–200 °C temperature range and a 20–60 bar CO2 pressure range, after which the results of these conditions were compared to those of conventional hot water pretreatment. Treatment at 190 °C and a 60-bar CO2 pressure was determined to be optimal, resulting in the highest glucose yield (80.7%) from subsequent enzymatic hydrolysis. Acidic conditions resulting from CO2-derived carbonic acid significantly reduced the hemicellulose content of the peanut shells and weakened the interaction between cellulose, hemicellulose, and lignin, improving enzyme accessibility to the cellulose. Furthermore, high-pressure CO2 increased the pore size and porosity of the resulting pretreated peanut shells, improving their enzyme adsorption capacities, as confirmed by cellulase adsorption and mercury intrusion porosimetry tests. The dual effect from high-pressure CO2 led to significant hemicellulose reduction and improved adsorption of enzymes on the cellulose, which in turn increased glucose yield from the subsequent enzymatic hydrolysis of pretreated peanut shells. Alcoholic fermentation of the hydrolyzed glucose resulted in a 12.4% increase in bio-ethanol production compared to a glucose control, thus highlighting the potential of pre-treated peanut shells as a glucose precursor used in biofuel industry.

Corn fiber is a co-product of commercial ethanol dry-grind plants, which is processed into distillers dried grains with solubles (DDGS) and used as animal feed, yet it holds high potential to be used as feedstock for additional ethanol production. Due to the tight structural make-up of corn fiber, a pretreatment step is necessary to make the cellulose and hemicellulose polymers in the solid fibrous matrix more accessible to the hydrolytic enzymes. A pretreatment process was developed in which whole corn kernels were soaked in aqueous solutions of 2.5, 5.0, 7.5, and 10.0 wt% ammonia at 105 °C for 24 h. The pretreated corn then was subjected to a conventional mashing procedure and subsequent ethanol fermentation using a commercial strain of natural Saccharomyces cerevisiae with addition of a commercial cellulase. Pretreatment of the corn with 7.5 wt% ammonia solution plus cellulase addition gave the highest ethanol production, which improved the yield in fermentation using 25 wt% solid from 334 g ethanol/kg corn obtained in the control (no pretreatment and no cellulase addition) to 379 g ethanol/kg corn (a 14% increase). The process developed can potentially be implemented in existing dry-grind ethanol facilities as a “bolt-on” process for additional ethanol production from corn fiber, and this additional ethanol can then qualify as “cellulosic ethanol” by the Environmental Protection Agency’s (EPA’s) Renewable Fuels Standard and thereby receive RINs (Renewable Identification Numbers).

Background Pretreatment of biomass to maximize the recovery of fermentable sugars as well as to minimize the amount of enzyme inhibitors formed during the pretreatment is a challenge in biofuel process. We develop a modified Fenton pretreatment in a mixed solvent (water/DMSO) to combine the advantages of organosolv and Fenton pretreatments. The hemicellulose and cellulose in corncob were effectively degraded into xylose, glucose, and soluble glucose oligomers in a few hours. This saccharide solution, separated from the solid lignin simply by filtration, can be directly applied to the subsequent enzymatic hydrolysis and ethanol fermentation. Results After the pretreatment, 94% carbohydrates were recovered as soluble monosaccharide (xylose and glucose) and glucose oligomers in the filtrates, and 87% of solid lignin was recovered as the filter residue. The filtrates were directly applied to enzymatic hydrolysis, and 92% of raw corncob glucose was recovered. The hydrolysates containing the glucose and xylose from the enzymatic hydrolysis were directly applied to ethanol fermentation with ethanol yield equals 79% of theoretical yield. The pretreatment conditions (130 °C, 1.5 bar; 30 min to 4 h) are mild, and the pretreatment reagents (H2O2, FeCl3, and solvent) had low impact to environment. Using ferrimagnetic Fe3O4 resulted in similar pretreatment efficiency and Fe3O4 could be removed by filtration. Conclusions A modified Fenton pretreatment of corncob in DMSO/water was developed. Up to 94% of the carbohydrate content of corncob was recovered as a saccharide solution simply by filtration. Such filtrate was directly applied to the subsequent enzymatic hydrolysis and where 92% of the corncob glucose content was obtained. The hydrolysate so obtained was directly applied to ethanol fermentation with good fermentability. The pretreatment method is simple, and the additives and solvents used have a low impact to the environment. This method provides the opportunity to substantially maximize the carbohydrate and solid lignin recovery of biomass with a comparatively green process, such that the efficiency of biorefinery as well as the bioethanol production process can be improved. The pretreatment is still relatively energy intensive and expensive, and further optimization of the process is required in large-scale operation. Electronic supplementary material The online version of this article (10.1186/s13068-018-1288-4) contains supplementary material, which is available to authorized users.

Soybean straw is a renewable resource in agricultural residues that can be used for lignocellulosic bioethanol production. To enhance enzymatic digestibility and fermentability, the biomass was prepared with an alkali-thermal pretreatment (sodium hydroxide, 121°C, 60 min). The delignification yield was 34.1~53%, in proportion to the amount of sodium hydroxide, from 0.5 to 3.0 M. The lignin and hemicellulose contents of the pretreated biomass were reduced by the pretreatment process, whereas the proportion of cellulose was increased. Under optimal condition, the pretreated biomass consisted of 74.0±0.1% cellulose, 10.3±0.1% hemicellulose, and 10.1±0.6% lignin. During enzymatic saccharification using Cellic® CTec2 cellulase, 10% (w/v) of pretreated soybean straw was hydrolyzed completely and converted to 67.3±2.1 g/L glucose and 9.4±0.5 g/L xylose with a 90.9% yield efficiency. Simultaneous saccharification and fermentation of the pretreated biomass by Saccharomyces cerevisiae W303-1A produced 30.5±1.2 g/L ethanol in 0.5 L fermented medium containing 10% (w/v) pretreated biomass after 72 h. The ethanol productivity was 0.305 g ethanol/g dry biomass and 0.45 g ethanol/g glucose after fermentation, with a low concentration of organic acid metabolites. Also, 82% of fermentable sugar was used by the yeast for ethanol fermentation. These results show that the combination of alkaline pretreatment and biomass hydrolysate is useful for enhancing bioethanol productivity using delignified soybean straw.

Background: Future biorefineries will focus on converting low value waste streams to chemical products that are derived from petroleum or refined sugars. Feedstock pretreatment in a simple, cost effective, agnostic manner is a major challenge. Methods: In this work, beechwood sawdust was delignified via an organosolv process, assisted by homogeneous inorganic acid catalysis. Mixtures of water and several organic solvents were evaluated for their performance. Specifically, ethanol (EtOH), acetone (AC), and methyl- isobutyl- ketone (MIBK) were tested with or without the use of homogeneous acid catalysis employing sulfuric, phosphoric, and oxalic acids under relatively mild temperature of 175 &deg, C for one hour. Results: Delignification degrees (DD) higher than 90% were achieved, where both AC and EtOH proved to be suitable solvents for this process. Both oxalic and especially phosphoric acid proved to be good alternative catalysts for replacing sulfuric acid. High gravity simultaneous saccharification and fermentation with an enzyme loading of 8.4 mg/gsolids at 20 wt.% initial solids content reached an ethanol yield of 8.0 w/v%. Conclusions: Efficient delignification combining common volatile solvents and mild acid catalysis allowed for the production of ethanol at high concentration in an efficient manner.

Об’єктом розробки є технологія виробництва біоетанолу з целюлозовмісної сировини. Мета: розробка модернізованої технології виробництва біоетанолу з целюлозовмісної сировини на базі відомої гідролізної технології. Розроблено проект технології виробництва біоетанолу з целюлозовмісної сировини з метою отримання біопалива, що відповідатиме нормам євростандарту і задовольнить потребу країні у біоетанолі. Технологічна схема базується на зброджуванні гідролізату целюлозовмісної сировини за використання дріжджів Saccharomyces cerevisiae, з подальшим виділення біоетанолу з бражки. Новизною проекту є технологічна стадія попередньої обробки сировини реактивом Фентона в водно-органічній суміші розчинників. Матеріали даної магістерської дисертації можна використовувати при проектуванні підприємств по виробництву спирту, які в якості сировини використовують целюлозовмісні відходи. Актуальність проекту полягає в вирішенні перш за все енергетичної проблеми – нестачі моторних палив в країні, а також частково економічної і екологічно. Master’s dissertation: pages,figures, tables, sources. The object of development is technology of bioethanol production from cellulose - containing materials. Objective: development an improved technology of bioethanol production from cellulose - containing materials based on known hydrolysis technology. A project of bioethanol production from cellulose - containing materials technology to obtain biofuel that meet the standards of standarts and meet the needs of the country in bioethanol was designed. Technological scheme is based on the fermentation of hydrolyzate from cellulose by Saccharomyces cerevisiae, with further receiving of bioethanol by rectification. The novelty of the project is using a new technological stage for cellulose organosolv pretreatment through Fenton reaction. Materials for this master’s dissertation can be used in the projection of ethanol-producing factories, that use cellulose as raw material. The topicality of the project is primarily a solving of energy problem – lacking of motor fuels in the country, and partly solving economic and environmental problem.

Choline acetate (ChOAc), a cholinium ionic liquid (IL), was compared with 1-ethyl-3-methylimidazolium acetate (EmimOAc) with regard to biomass pretreatment, inhibition on cellulase and yeast, residuals in pretreated biomass, and saccharification and fermentation of pretreated biomass. Irrespective of ChOAc and EmimOAc, cellulose and hemicellulose saccharification of the IL-pretreated bagasse were over 90% and 60%, respectively. Median effective concentrations (EC50) based on cellulase activity were 32 wt% and 16 wt% for ChOAc and EmimOAc, respectively. The EC50 based on yeast growth were 3.1 wt% and 0.3 wt% for ChOAc and EmimOAc respectively. The residuals in IL-pretreated bagasse were 10% and 23% for ChOAc and EmimOAc, respectively, when washed 2 times after pretreatment. Ethanol yield on a bagasse basis were 60% and 24% for ChOAc and EmimOAc, respectively, in the saccharification and fermentation of IL-pretreated bagasse when washed 2 times. ChOAc-pretreated bagasse could be saccharified and fermented with fewer wash times than EmimOAc-pretreated bagasse.

Jerusalem artichoke is a potential energy crop. While its tubers are being used to extract inulin, its stalks could be used for biofuels production. In this article, fed-batch saccharification and fermentation of Jerusalem artichoke stalks (JAS) was studied. Pretreatment with 2.0% (w/v) NaOH not only retained most inulin, the unique component in JAS, but also increased cellulose content from 42.3% to 58.2% due to the removal of lignin. Batch-feeding of both the pretreated JAS and cellulases was an effective strategy for saccharification, through which 115.8 g L−1 total sugars including glucose, xylose, fructose and inulin were released from 20% (w/v) solids uploading under the supplementation of cellulases at 20 FPU (filter paper unit) g−1 dry biomass. An inulinase-producing yeast Saccharomyces cerevisiae MK01 was developed by the cell surface display of inulinase for ethanol fermentation from the pretreated JAS under the fed-batch conditions, and 38.3 g L−1 ethanol was produced at 96 h, with an ethanol yield of 0.361 g g−1 total sugars consumed, about 71% of the theoretical yield of 0.511, indicating that JAS would be a promising feedstock for ethanol production.

Several yeast strains have been engineered to express different cellulases to achieve simultaneous saccharification and fermentation of lignocellulosic materials. However, successes in these endeavors were modest, as demonstrated by the relatively low ethanol titers and the limited ability of the engineered yeast strains to grow using cellulosic materials as the sole carbon source. Recently, substantial enhancements to the breakdown of cellulosic substrates have been observed when lytic polysaccharide monooxygenases (LPMOs) were added to traditional cellulase cocktails. LPMOs are reported to cleave cellulose oxidatively in the presence of enzymatic electron donors such as cellobiose dehydrogenases. In this study, we coexpressed LPMOs and cellobiose dehydrogenases with cellobiohydrolases, endoglucanases, and β-glucosidases in Saccharomyces cerevisiae . These enzymes were secreted and docked onto surface-displayed miniscaffoldins through cohesin-dockerin interaction to generate pentafunctional minicellulosomes. The enzymes on the miniscaffoldins acted synergistically to boost the degradation of phosphoric acid swollen cellulose and increased the ethanol titers from our previously achieved levels of 1.8 to 2.7 g/liter. In addition, the newly developed recombinant yeast strain was also able to grow using phosphoric acid swollen cellulose as the sole carbon source. The results demonstrate the promise of the pentafunctional minicellulosomes for consolidated bioprocessing by yeast.

Optimum utilization of fermentable sugars from lignocellulosic biomass to deliver multiple products under biorefinery concept has been reported in this work. Alcohol fermentation has been carried out with multiple cell recycling of Kluyveromyces marxianus IIPE453. The yeast utilized xylose-rich fraction from acid and steam treated biomass for cell generation and xylitol production with an average yield of 0.315±0.01g/g while the entire glucose rich saccharified fraction had been fermented to ethanol with high productivity of 0.9±0.08g/L/h. A detailed insight into its genome illustrated the strain's complete set of genes associated with sugar transport and metabolism for high-temperature fermentation. A set flocculation proteins were identified that aided in high cell recovery in successive fermentation cycles to achieve alcohols with high productivity. We have brought biomass derived sugars, yeast cell biomass generation, and ethanol and xylitol fermentation in one platform and validated the overall material balance. 2kg sugarcane bagasse yielded 193.4g yeast cell, and with multiple times cell recycling generated 125.56g xylitol and 289.2g ethanol (366mL).

Several papers have reported various technical aspects of lignocellulosic bioethanol production. Recalcitrance to saccharification is a major limitation for conversion of lignocellulosic biomass to ethanol. The biological process for converting lignocellulose to fuel ethanol includes delignification in order to liberate cellulose and hemicelluloses, depolymerization of carbohydrate polymers to produce free sugars, and sugar fermentation to produce ethanol. Access of plant cell wall polysaccharides to chemical, enzymatic and microbial digestion is limited by many factors, including the presence of lignin and hemicellulose that cover cellulose microfibrils. An effort to support the fuel ethanol fermentation industry using the Indonesian woody plant species Shorea uliginosa Foxw., was undertaken with regard to the established efficient bioethanol production process. This paper relates to a method for producing bioethanol from the lignocellulose of S. uliginosa Foxw. by saccharification and fermentation of xylem. A literature study of previous research on cellulose hydrolysis as a method for producing bioethanol was necessary. The objective of this study was to gain a deeper understanding of the degradation mechanisms of cellulose by enzymes through a study of previous research, which were then compared to the new method.

The purpose of this paper was to analyze the utilization of pulp and residues from pineapple processing as substrates for alcoholic fermentation. The fruits were cleaned, ground, and hydrolyzed with pectinase, cellulase, hemicellulase, and amylase. Pineapple pulp and whole fruit were analyzed to determine the amounts of amide, hemicellulose, total reducing sugars, °Brix, pectin, cellulose, and lignin present. Fermentation experiments were conducted with 0, 100, 200, and 300 g/kg of peel. Wines were analyzed for total or volatile acidity, total reducing sugars, and ethanol. The assays with the highest fermentation kinetic parameters were analyzed to determine the secondary compounds in the wine. The results showed that the enzymatic hydrolysis decreased the polysaccharide amount and increased reducing sugars and soluble solids, thereby improving the fermentation process, mainly in integral samples. In pineapple fermentation, 100 g/kg of peel addition did not influence the kinetic parameters and secondary c...

Direct ethanol fermentation from amorphous cellulose was achieved using an engineered industrial Saccharomyces cerevisiae strain. Two cellulase genes endoglucanase (eg3) and β-glucosidase (bgl1) were obtained from Trichoderma viride and integrated into the genome of S. cerevisiae. These two cellulases could be constitutively coexpressed and secreted by the recombinant strain S. cerevisiae-eb. The enzyme activities were analyzed in the culture supernatants, with the highest endoglucanase activity of 2.34 units/ml and β-glucosidase activity of 0.95 units/ml. The effects of pH, temperature and metal ions on enzyme activities were analyzed. The coexpression strain S. cerevisiae-eb could grow in carboxymethyl cellulose (CMC) and utilize it as the single carbon source. The 20 g/L CMC as a model substrate of amorphous cellulose was used in fermentation. The ethanol production reached 4.63 g/L in 24 h, with the conversion ratio of 64.2% compared with the theoretical concentration. This study demonstrated that the engineered industrial strain S. cerevisiae-eb could convert amorphous cellulose to ethanol simultaneously and achieve consolidated bioprocessing (CBP) directly.

Background Lignocellulosic biomass is an abundant and inexpensive resource for biofuel production. Alongside its biotechnological conversion, pretreatment is essential to enable efficient enzymatic hydrolysis by making cellulose susceptible to cellulases. Wet oxidation of biomass, such as acetone/water oxidation, that employs hot acetone, water, and oxygen, has been found to be an attractive pretreatment method for removing lignin while producing less degradation products. The remaining enriched cellulose fraction has the potential to be utilized under high gravity enzymatic saccharification and fermentation processes for the cost-competing production of bioethanol. Results Beech wood residual biomass was pretreated following an acetone/water oxidation process aiming at the production of high concentration of cellulosic ethanol. The effect of pressure, reaction time, temperature, and acetone-to-water ratio on the final composition of the pretreated samples was studied for the efficient utilization of the lignocellulosic feedstock. The optimal conditions were acetone/water ratio 1:1, 40 atm initial pressure of 40 vol% O2 gas, and 64 atm at reaction temperature of 175 °C for 2 h incubation. The pretreated beech wood underwent an optimization step studying the effect of enzyme loading and solids content on the enzymatic liquefaction/saccharification prior to fermentation. In a custom designed free-fall mixer at 50 °C for either 6 or 12 h of prehydrolysis using an enzyme loading of 9 mg/g dry matter at 20 wt% initial solids content, high ethanol concentration of 75.9 g/L was obtained. Conclusion The optimization of the pretreatment process allowed the efficient utilization of beech wood residual biomass for the production of high concentrations of cellulosic ethanol, while obtaining lignin that can be upgraded towards high-added-value chemicals. The threshold of 4 wt% ethanol concentration that is required for the sustainable bioethanol production was surpassed almost twofold, underpinning the efficient conversion of biomass to ethanol and bio-based chemicals on behalf of the biorefinery concept. Electronic supplementary material The online version of this article (doi:10.1186/s13068-017-0737-9) contains supplementary material, which is available to authorized users.

The yeast Kluyveromyces marxianus is considered as a potential alternative to Saccharomyces cerevisiae in producing ethanol as a biofuel. In this study, we investigated the ethanol fermentation properties of novel K. marxianus strain DMB1, isolated from bagasse hydrolysates. This strain utilized sorbitol as well as various pentoses and hexoses as single carbon sources under aerobic conditions and produced ethanol from glucose in hydrolysates of the Japanese cedar at 42 °C. Reference strains K. marxianus NBRC1777 and S. cerevisiae BY4743 did not assimilate sorbitol or ferment lignocellulosic hydrolysates to ethanol at this temperature. Thus strain DMB1 appears to be optimal for producing bioethanol at high temperatures, and might provide a valuable means of increasing the efficiency of ethanol fermentation.

The present study aimed to characterize and evaluate the hydrolysis of sisal fibers for ethanol production. Acid hydrolysis of the fiber was performed and main products determined were xylose and glucose. However, xylose and arabinose (pentoses) are not fermented by the conventional yeast Saccharomyces cerevisiae . Prehydrolysis at 105 °C was efficient, removing most of the pentoses. The main product formed in acid hydrolysis at 160 °C was glucose, which can be metabolized by the yeast S. cerevisiae through alcoholic fermentation. In these conditions ethanol was produced with 92% conversion of initial total reducing sugars (0.47 g/g of ethanol yield). Sisal has potential as a raw material for ethanol production since it is not a food source, has high cellulose content and low biomass cost.

The focus of this work was to develop a combined acid and alkaline hydrothermal pretreatment of lignocellulose that ensures high recovery of both hexose and pentose. Dilute sulfuric acid and lime pretreatments were employed sequentially. Process performance was optimized in terms of catalyst concentration, retention time, and temperature using response surface methodology. Medium operational conditions in the acid stage and harsh conditions in the alkaline stage were desirable with optimal performance at 0.73 wt% H(2)SO(4), 150 °C, 6.1 min in the first stage, and 0.024 g lime/g biomass, 202 °C, 30 min in the second stage. In comparison to single-stage pretreatments with high recovery of either glucose or xylose, two-stage process showed great promises with80 % glucose and70 % xylose recovery. In addition, the method greatly improved ethanol fermentation with yields up to 0.145 g/g Miscanthus, due to significantly reduced formation of inhibitory by-products such as weak acids, furans, and phenols. Supplementing biomimetic acids would further increase glucose yield by up to 15 % and xylose yield by 25 %.

This study aimed to produce a cellulase blend and to evaluate its application in a simultaneous saccharification and fermentation (SSF) process for second generation ethanol production from sugar cane bagasse. The sugar cane bagasse was subjected to pretreatments (diluted acid and alkaline), as for disorganizing the ligocellulosic complex, and making the cellulose component more amenable to enzymatic hydrolysis. The residual solid fraction was named sugar cane bagasse partially delignified cellulignin (PDC), and was used for enzyme production and ethanol fermentation. The enzyme production was performed in a bioreactor with two inoculum concentrations (5 and 10% v/v). The fermentation inoculated with higher inoculum size reduced the time for maximum enzyme production (from 72 to 48). The enzyme extract was concentrated using tangential ultrafiltration in hollow fiber membranes, and the produced cellulase blend was evaluated for its stability at 37°C, operation temperature of the simultaneous SSF process, and at 50°C, optimum temperature of cellulase blend activity. The cellulolytic preparation was stable for at least 300h at both 37°C and 50°C. The ethanol production was carried out by PDC fed-batch SSF process, using the onsite cellulase blend. The feeding strategy circumvented the classic problems of diffusion limitations by diminishing the presence of a high solid:liquid ratio at any time, resulting in high ethanol concentration at the end of the process (100g/L), which corresponded to a fermentation efficiency of 78% of the maximum obtainable theoretically. The experimental results led to the ratio of 380L of ethanol per ton of sugar cane bagasse PDC.

Among four isolated cellulolytic microorganisms, Streptomyces sp. MDS can utilize a broad range of cellulosic substrates including: carboxymethyl cellulose, avicel, xylan, cellobiose, filter paper, wood straw and rice straw by producing a large amount of endoglucanase, exoglucanase, cellobiase, xylanase and glucoamylase. Effects of different physicochemical parameters to achieve maximum production of cellulolytic enzymes were systematically investigated. It was found that the cellulolytic enzyme activities were mainly located at extracellular location. It was observed that supplementation of CaCl2 (5 mM) as a metal additive significantly induced the whole enzyme system. These cellulases also have high thermal stability as evidenced by retaining 60–70% activity at 90 °C and pH 5 and alkalotolerance as more than 55–60% of the activity at pH 10 and 40 °C and 60 °C (for endoglucanase) after 1 h of incubation. The enzymes also retained up to 50–70% of its initial activity upon incubation in the presence of commercials detergents for 1 h. Finally, the efficiency of fermentative conversion of the CMC and rice straw hydrolyzate by Saccharomyces cerevisiae (KCTC 7296) resulted in 55.8 and 57.2% of maximum theoretical ethanol yield based on sugar (glucose and xylose) fermentation, respectively.

Background Previous work has demonstrated that glycerol organosolv pretreatment can effectively improve the hydrolyzability of various lignocellulosic substrates. This pretreatment process strategy is ideal to integrate a commercially successful lignocellulosic and vegetable oil biorefinery industry. However, industrially relevant high-solid-loading hydrolyzability and fermentability of the pretreated substrates have yet to be considered for enzyme-based lignocellulosic biorefineries. Results In this study, an AGO pretreatment of sugarcane bagasse was evaluated with regard to the component selectivity, structural modification, hydrolyzability, and fermentation of pretreated substrates. The results showed that the AGO pretreatment presented good component selectivity, removing approximately 70 % lignin and hemicellulose, respectively, from sugarcane bagasse with a near-intact preservation (94 %) of the overall cellulose. The pretreatment deconstructed the recalcitrant architecture of natural lignocellulosic biomass, thereby modifying the structure at the macro-/micrometer level (fiber size, surface area, average size, roughness) and supermolecular level (key chemical bond dissociation) of lignocellulosic substrates towards good hydrolyzability. Notably, extraordinarily few fermentation inhibitors (

The effect of time and pressure on the selective extraction of sugar beet pectin using steam pre-treatment on unprocessed Sugar Beet Pulp was evaluated using a design of experiments approach. This process gave the highest solubilisation of pectin oligomers at a relatively low pressure and longer time (5 Bar, 24 min), whilst leaving the majority of the cellulose fraction intact. This method of steam pre-treatment fits into the concept of a sugar beet biorefinery as it valorises an existing waste stream without requiring any further physical processing such as milling or dilution with water. The residual cellulose fraction was enriched in cellulose and could be effectively fermented into ethanol by yeast after enzymatic digestion, producing 0.48 g ethanol per gram of glucose.

Accurate mass balance and conversion data from integrated operation is needed to fully elucidate the economics of biofuel production processes. This study explored integrated conversion of corn stover to ethanol and highlights techniques for accurate yield calculations. Acid pretreated corn stover (PCS) produced in a pilot-scale reactor was enzymatically hydrolyzed and the resulting sugars were fermented to ethanol by the glucose–xylose fermenting bacteria, Zymomonas mobilis 8b. The calculations presented here account for high solids operation and oligomeric sugars produced during pretreatment, enzymatic hydrolysis, and fermentation, which, if not accounted for, leads to overestimating ethanol yields. The calculations are illustrated for enzymatic hydrolysis and fermentation of PCS at 17.5% and 20.0% total solids achieving 80.1% and 77.9% conversion of cellulose and xylan to ethanol and ethanol titers of 63 g/L and 69 g/L, respectively. These procedures will be employed in the future and the resulting information used for techno-economic analysis.

The potential of cashew apple bagasse as a source of sugars for ethanol production by Kluyveromyces marxianus CE025 was evaluated in this work. This strain was preliminarily cultivated in a synthetic medium containing glucose and xylose and was able to produce ethanol and xylitol at pH 4.5. Next, cashew apple bagasse hydrolysate (CABH) was prepared by a diluted sulfuric acid pretreatment and used as fermentation media. This hydrolysate is rich in glucose, xylose, and arabinose and contains traces of formic acid and acetic acid. In batch fermentations of CABH at pH 4.5, the strain produced only ethanol. The effects of temperature on the kinetic parameters of ethanol fermentation by K. marxianus CE025 using CABH were also evaluated. Maximum specific growth rate (μmax), overall yields of ethanol based on glucose consumption \(Y_{{P \mathord{\left/ {\vphantom {P {S_1 }}} \right. \kern-\nulldelimiterspace} {S_1 }}}^{\text{G}}\) and based on glucose + xylose consumption (YP/S), overall yield of ethanol based on biomass (YP/X), and ethanol productivity (PE) were determined as a function of temperature. Best results of ethanol production were achieved at 30°C, which is also quite close to the optimum temperature for the formation of biomass. The process yielded 12.36 ± 0.06 g l−1 of ethanol with a volumetric production rate of 0.257 ± 0.002 g l−1 h−1 and an ethanol yield of 0.417 ± 0.003 g g−1 glucose.

Sugarcane bagasse is one of the most promising agricultural by-products for conversion to biofuels. Here, ethanol fermentation from bagasse has been achieved using an integrated process combining mechanical pretreatment by ball milling, with enzymatic hydrolysis and fermentation. Ball milling for 2 h was sufficient for nearly complete cellulose structural transformation to an accessible amorphous form. The pretreated cellulosic residues were hydrolyzed by a crude enzyme preparation from Penicillium chrysogenum BCC4504 containing cellulase activity combined with Aspergillus flavus BCC7179 preparation containing complementary beta-glucosidase activity. Saccharification yields of 84.0% and 70.4% for glucose and xylose, respectively, were obtained after hydrolysis at 45 degrees C, pH 5 for 72 h, which were slightly higher than those obtained with a commercial enzyme mixture containing Acremonium cellulase and Optimash BG. A high conversion yield of undetoxified pretreated bagasse (5%, w/v) hydrolysate to ethanol was attained by separate hydrolysis and fermentation processes using Pichia stipitis BCC15191, at pH 5.5, 30 degrees C for 24 h resulting in an ethanol concentration of 8.4 g/l, corresponding to a conversion yield of 0.29 g ethanol/g available fermentable sugars. Comparable ethanol conversion efficiency was obtained by a simultaneous saccharification and fermentation process which led to production of 8.0 g/l ethanol after 72 h fermentation under the same conditions. This study thus demonstrated the potential use of a simple integrated process with minimal environmental impact with the use of promising alternative on-site enzymes and yeast for the production of ethanol from this potent lignocellulosic biomass.

Banana is a major cash crop of many regions generating good amount of waste after harvest. This agro waste which is left for natural degradation is used as substrate for single step ethanol fermentation by thermophilic, cellulolytic, ethanologenic Clostridium thermocellum CT2, a new culture isolated from elephant droppings. Scanning electron microscopic pictures clearly indicate cellulolysis and close interaction of selected isolate CT2 with cellulose. The optimum conditions for cellulose fermentation were 60°C, pH 7.5, inoculums size 5% and incubation time 5 days. Ethanol produced and reducing sugars were estimated by gas chromatography. Clostridium thermosaccharolyticum HG8 and Thermoanaerobacter ethanolicus ATCC 31937 were used in coculture fermentation with CT2. Coculture fermentation of CT2 with HG8 was more efficient in terms of ethanol production, cellulose degradation and reducing sugars utilization. A maximum ethanol yield of 0.41g/g substrate used was obtained oncoculturing CT2 with HG8 on alkali treated banana waste. Coculture was active even at substrate concentrations up to 100 g/l, a maximum ethanol of 22 g/l was obtained at 100 g/l substrate concentration on coculturing CT2 with HG8. This is the first report on anaerobic single step conversionof banana waste to ethanol by C. thermocellum.

Renewable energy sources have received increased interest from the international community with biomass being one of the oldest and the most promising ones. In the concept of exploitation of agro-industrial residues, the present study investigates the pre-treatment and ethanol fermentation potential of the olive pulp, which is the semi solid residue generated from the two-phase processing of the olives for olive oil production. Wet oxidation and enzymatic hydrolysis have been applied aiming at the enhancement of carbohydrates' bioavailability. Different concentrations of enzymes and enzymatic durations have been tested. Both wet oxidation and enzymic treatment were evaluated based on the ethanol obtained in a subsequent fermentation step by Saccharomyces cerevisiae and Thermoanaerobacter mathranii. It was found that a four-day hydrolysis time was adequate for a satisfactory release of glucose and xylose. The combination of wet oxidation and enzymatic hydrolysis resulted in the glucose and xylose concentration increase of 138 and 444%, respectively, compared to 33 and 15% with only enzymes added. However, the highest ethanol production was obtained when only enzymic pre-treatment was applied, implying that wet oxidation is not a recommended pre-treatment process for olive pulp at the conditions tested. It was also showed that increased dry matter concentration did not have a negative effect on the release of sugars, indicating that the cellulose and xylan content of the olive pulp is relatively easily available. The results of the experiments in batch processes clearly emphasize that the simultaneous saccharification and fermentation (SSF) mode is advantageous in comparison with the separate hydrolysis and fermentation (SHF) mode concerning process contamination.

We examined the saccharification and fermentation of meals from Acacia mangium wood, Paraserianthes falcataria wood, and Elaeis guineensis trunk. The levels of enzymatic hydrolysis of cellulose and ethanol production were highest for P. falcataria wood and lowest for A. mangium wood. Ultrasonication pretreatment of meal further increased the rates of hydrolysis and ethanol production in meal from P. falcataria wood. Through this pretreatment, hemicelluloses (xylan and xyloglucan) and cellulose were released in the meal from P. falcataria wood. Loosening of hemicellulose associations can be expected to make P. falcataria wood more useful for bioethanol production.