Second-generation ethanol production from steam exploded barley straw by Kluyveromyces marxianus CECT 10875

Barley straw is nowadays being considered a potential lignocellulosic raw material for fuel-ethanol production as an alternative to starch- or sugar-containing feedstock. In this work, several configuration strategies for ethanol production from steam-exploded barley straw by Kluyveromyces marxianus CECT 10875 have been studied with the aim of obtaining higher ethanol concentrations.Different substrate loading (2-15%, w/v) were studied during enzymatic hydrolysis. The xylanase contribution on glucose production and glucan conversion at different substrate loading was also investigated. In addition, three different process configurations, separate hydrolysis and fermentation, simultaneous saccharification and fermentation and presaccharification and simultaneous saccharification, were compared at different water insoluble solids concentration (5%, 10% and 15%). The influence of xylanase addition on the ethanol yield was studied as well.Results show that endo-xylanases improved glucan conversion and ethanol yield compared with a standard enzymatic mixture, markedly at low substrate concentration. The positive effect of added xylanase was most evident at early stages of enzymatic hydrolysis. Regarding process configurations for the period of 72 h, SSF with endo-xylanases provided the best ethanol yield, nearly 70%, for 10% WIS. Nonetheless, the higher ethanol concentration, 29.4 g/l, was obtained at 15% WIS.     

Bioethanol production from corn meal by simultaneous enzymatic saccharification and fermentation with immobilized cells of Saccharomyces cerevisiae var. ellipsoideus

The simultaneous enzymatic saccharification and fermentation (SSF) of corn meal using immobilized cells of Saccharomycescerevisiae var. ellipsoideus yeast in a batch system was studied. The yeast cells were immobilized in Ca-alginate by electrostatic droplet generation method. The process kinetics was assessed and determined and the effect of addition of various yeast activators (mineral salts: ZnSO₄ · 7H₂O and MgSO₄ · 7H₂O, and vitamins: Ca-pantothenate, biotin and myo-inositol) separately or mixed, was investigated. Taking into account high values of process parameters (such as ethanol concentration, ethanol yield, percentage of the theoretical ethanol yield, volumetric productivity and utilized glucose) and significant energy savings the SSF process was found to be superior compared to the SHF process. Further improvement in ethanol production was accomplished with the addition of mineral salts as yeast activators which contributed to the highest increase in ethanol production. In this case, the ethanol concentration of 10.23% (w/w), percentage of the theoretical ethanol yield of 98.08%, the ethanol yield of 0.55 g/g and the volumetric productivity of 2.13 g/l·h were obtained.     

A comprehensive ligninolytic pre-treatment approach from lignocellulose green biotechnology to produce bio-ethanol

In the present study, we developed a novel ligninolytic enzymes based pre-treatment method for lignocellulosic wheat straw to depolymerize lignin and expose the cellulose polymers to produce bio-ethanol. Wheat straw was pre-treated with ligninolytic enzymes extract produced from Ganoderma lucidum under optimum solid state fermentation conditions. The pre-treated biomass was further subjected to the enzymatic hydrolysis by the crude unprocessed cellulases (β-1,4 endoglucanase, 53.5 ± 1.24 U/mL; β-1,4 exoglucanase, 41.3 ± 1.31 U/mL; β-1,4 glucosidase, 46.8 ± 1.43 U/mL; and xylanase 39 ± 2.2 U/mL) produced by Trichoderma harzaianum. Under optimal conditions for enzymatic saccharification, 10% (w/v) of pre-treated biomass was hydrolyzed completely and converted to 72.5 and 2.4 g/L of glucose and xylose, respectively. Initial time screening Sequential Saccharification and Fermentation (SSF) of the concentrated enzymatic hydrolyzate (10%, w/v) by Saccharomyces cerevisiae produced 22.6 g/L ethanol in a fermented medium after 72 h of temperature controlled incubation at 37 °C. For maximum ethanol production, different physical and nutritional parameters like pH, temperature, substrate level and inoculum sizes were optimized. Under optimal conditions ethanol production of 33.5 g/L was obtained from ligninolytic treated residual (wheat straw) biomass.     

Determination of optimal pre-treatment conditions for ethanol production from olive-pruning debris by simultaneous saccharification and fermentation

Ethanol generation from lignocellulose materials provides an alternative energy-production system. This study investigates the effect of pre-treatment conditions: maximum temperature (range 423.15-483.15 K) and sulfuric-acid concentration (interval 0.002-0.059 kmol/m³) on fuel-ethanol production from simultaneous saccharification and fermentation (SSF) of olive-pruning debris by Saccharomyces cerevisiae IR2-9a (a thermal acclimatized microorganism, 313.15 K). The influence of these two variables was determined by using a response-surface methodology. Cellulose percentage in pre-treated solids reached a maximum of 71.6% of the content in raw material at 483.15 K and 0.010 kmol/m³ of acid concentration. The conversion of hemicellulose into monosaccharides and oligosaccharides also was analyzed. After the wash and filtration of solids, a significant quantity of d-glucose was obtained in the liquid fraction. For ethanol generation, the bio-fuel yield (maximum of 9.6 kg from 100 kg olive-pruning debris), and volumetric ethanol productivity (maximum of 0.27 kg/(m³ h)), strongly depended on pre-treatments conditions. According to statistical optimization, the highest ethanol yield (9.9 kg ethanol from 100 kg olive-pruning debris) is achieved at 480.15 K using a catalyst concentration of 0.016 kmol/m³. A maximum overall process yield of 15.3 kg ethanol/100 kg olive-pruning debris may result when taking into account ethanol from SSF and d-glucose present in the pre-hydrolysate, assuming its theoretical conversion (22.8 kg ethanol/100 kg raw material, also considering the total conversion of d-xylose in the filtrate).     

Production of fuel ethanol from softwood by simultaneous saccharification and fermentation at high dry matter content

BACKGROUND: The production of bio‐ethanol from softwood is considered a promising alternative to fossil fuels in Sweden. In order to make fuel ethanol economically competitive with fossil fuels, it is important to reduce the production cost, which can be done by increasing the dry matter content of the fermentation medium, thus reducing the energy demand in the final distillation of the fermentation broth. Running simultaneous saccharification and fermentation (SSF) at higher dry matter content has, however, been found to decrease the ethanol yield. RESULTS: The use of different stirrer types and stirring speeds in the present study has shown to have an influence on the final ethanol yield in SSF with 10% water‐insoluble solids (WIS). Also, higher concentration of pretreatment hydrolysate, i.e., with increased inhibitor concentration, at the same WIS resulted in a decreased ethanol yield. However, despite stirring problems and high inhibitor concentration, ethanol was produced at 12% WIS with an ethanol yield in the SSF step of 81% of the theoretical based on the content of fermentable sugars in the fermentor. CONCLUSION: The decrease in ethanol yield in SSF at high dry matter content has been shown to be a combined effect of increased mass transfer resistance and increased inhibitor concentration in the fermentation broth. Copyright © 2008 Society of Chemical Industry     

Comparison of bioethanol production of simultaneous saccharification & fermentation and separation hydrolysis & fermentation from cellulose-rich barley straw

Cellulose rich barley straw, which has a glucan content of 62.5%, followed by dilute acid pretreatment, was converted to bioethanol by simultaneous saccharification and fermentation (SSF). The optimum fractionation conditions for barley straw were an acid concentration of 1% (w/v), a reaction temperature of 158 °C and a reaction time of 15 min. The maximum saccharification of glucan in the fractionated barley straw was 70.8% in 72 h at 60 FPU/gglucan, while the maximum digestibility of the untreated straw was only 18.9%. With 6% content WIS (water insoluble solid) for the fractionated barley straw, 70.5 and 83.2% of the saccharification yield were in SHF and SSF (representing with glucose equivalent), respectively, and a final ethanol concentration of 18.46 g/L was obtained under the optimized SSF conditions: 34 °C with 15 FPU/g-glucan enzyme loading and 1 g dry yeast cells/L. The results demonstrate that the SSF process is more effective than SHF for bioethanol production by around 18%.     

Extraction of phenolic fraction from guava seeds (Psidium guajava L.) using supercritical carbon dioxide and co-solvents

In this work the supercritical fluid extraction (SFE) with carbon dioxide (CO₂) and with ethyl acetate (EtAc) and ethanol (EtOH) as co-solvents was applied to obtain the phenolic fraction from guava seeds (Psidium guajava L.). The extraction was explored at various operating conditions, using 10, 20 and 30 MPa and 40, 50 and 60 °C. The use of EtAc and EtOH as co-solvents in SFE was also studied. The supercritical process was compared with traditional techniques such as Soxhlet extraction using EtAc and EtOH as solvents. The quality of the different extracts, obtained using SFE and Soxhlet methods and different solvents, was evaluated through the antioxidant activity, obtained by the collection methods of scavenging DPPH and bleaching of β-carotene, and also through the total phenolic content (TPC) of the samples, by the Folin-Ciocalteu method. The antioxidant potential indicates the use of ethanol as co-solvent as the best modifier in SFE, used in concentration of 10% (w/w) at 50 °C and 30 MPa. The quality of the extracts obtained by SFE with EtOH varied with the operating conditions of temperature and pressure, with higher values obtained at 10 and 20 MPa for TPC results and also antioxidant methods. The process yield of the phenolic fraction was also evaluated for all the extraction procedures studied (SFE and Soxhlet), with results varying from 0.380 to 1.738% (w/w).     

FED-BATCH SIMULTANEOUS SACCHARIFICATION AND FERMENTATION OF MICROWAVE/ACID/ALKALI/H2O2 PRETREATED RICE STRAW FOR PRODUCTION OF ETHANOL

The batch simultaneous saccharification and fermentation (SSF) of microwave/acid/alkali/H₂O₂ pretreated rice straw to ethanol was optimized using cellulase from Trichoderma reesei and Saccharomyces cerevisiae YC-097 cells prior to the fed-batch SSF studies. The batch SSF optima were 10% w/v substrate, 40°C, 15 mg cellulase/g substrate, initial pH 5.3, and 72 hours. Under the optimum conditions the ethanol concentration and its yield were 29.1 g/L and 61.3% respectively. Based on the optimal batch SSF, the fed-batch SSF was investigated and its operation parameters were optimized. Under its optimal conditions the ethanol concentration reached 57.3 g/L, while its productivity and yield were only slightly less than those in the batch SSF. This suggests that fed-batch SSF is a potential operation mode for effective ethanol production from microwave/acid/alkali/H₂O₂ pretreated rice straw.     

Extraction of biologically active compounds from Sideritis ssp. L.

In this study extraction of polyphenols and flavonoids from cultivated hybrid Sideritis scardica × Sideritis syriaca, known for its rich content of phenolics and flavonoids with antioxidant activity, was investigated. Extractions have been done by ethanol and water-ethanol, respectively. High equilibrium values of the extracted species were obtained—17.55 mg/(g solid) total phenolics and 5.7 mg/(g solid) total flavonoids with ethanol as solvent. The influence of the solvent on the total yield and the content of biologically active compounds were studied. Maximum polyphenolics and flavonoids extraction was observed for water-ethanol solvent ratio 20/80. Increase of the content of ethanol in the solvents led to lower total yield of extracts but higher percentage of polyphenolics. The extraction kinetics showed that 90% of the phenolic compounds were extracted during the first 2.5 h. The experimental kinetics was described by a constant effective diffusion coefficient D_e = 1.5 × 10⁻¹² m²/s in the solid, accounting for the actual particle size distribution.     

Aluminium (III)-selective PVC membrane sensor based on a Schiff base complex of N,N′-bis (salicylidene)-1, 2-cyclohexanediamine

A potentiometric aluminium sensor, based on the use N,N′-bis(salicylidene)-1,2-cyclohexanediamine (NBSC) as a neutral carrier, in poly(vinyl chloride) (PVC) matrix, is reported. Effect of various plasticizers; 2-nitrophenyloctylether (o-NPOE), tri-n-butyl phosphates (TBP), dioctylpththalate (DOP) & chloronapthalen (CN), and anion excluder, sodium tetraphenylborate (NaTPB) was studied. The best performance was obtained with a membrane composition of PVC: o-NPOE: NBSC: NaTPB ratio (w/w; mg) of 150:150:5:5. The sensor exhibits significantly enhanced selectivity toward Al³⁺ ions over the concentration range 1.0 × 10⁻⁸-1.0 × 10⁻¹ M with a lower detection limit of 5.0 × 10⁻⁹ M and a Nernstian slope of 20.3 ± 0.1 mV decade⁻¹ of activity. Influence of the membrane composition and possible interfering ions was investigated on the response properties of the electrode. Fast and stable response, good reproducibility and long-term stability are demonstrated. The sensor shows response time of <5 s and can be used for about 3 months without any considerable divergence in their potential response. Selectivity coefficients determined by matched potential method (MPM) indicate high selectivity for aluminium (III) ion. The proposed electrode shows fairly good discrimination of aluminium (III) from many metal ions. It was successfully applied for direct determination of aluminium (III) in biological, industrial and environmental samples. The electrode can be used in the pH range of 2.0-9.0 and mixtures containing up to 20% (v/v) non-aqueous content. It was used as an indicator electrode in potentiometric titration of aluminium ion vs. EDTA.     

Sago starch saccharification and simultaneous ethanol fermentation by amyloglucosidase and Zymomonas mobilis

Simultaneous saccharification and ethanol fermentation (SSF) of sago starch was studied using amyloglucosidase (AMG) and Zymomonas mobilis. The optimal concentration of AMG and operating temperature for the SSF process were found to be 0.5% (v/w) and 35°C, respectively. Under these conditions with 150 g dm^?3 sago starch as a substrate, the final ethanol concentration obtained was 69.2 g dm^?3 and ethanol yield, Y_P/S, 0.50 g g^?1 (97% of theoretical yield). Sago starch in the concentration range of 100–200 g dm^?3 was efficiently converted into ethanol. When compared to a two-step process involving separate saccharification and fermentation stages, the SSF reduced the total process time by half.     

Supercritical fluid extraction of flavonoids from Maydis stigma and its nitrite-scavenging ability

Supercritical fluid carbon dioxide (SF-CO₂) extraction (SFE) of flavonoids from Maydis stigma and its nitrite-scavenging ability were investigated. The effects of extraction time, particle size and co-solvent composition in terms of water content in ethanol were first optimized. Then, a Box-Behnken design combined with response surface methodology (RSM) was employed to study the effects of three independent variables (temperature, pressure and co-solvent amount) on the extraction yield of flavonoids. A maximal extraction yield of flavonoids of approximately 4.24 mg/g of M. stigma by SFE was obtained under optimal conditions (a temperature of 50.88 °C, a pressure of 41.80 MPa, a co-solvent amount of 2.488 mL/g and an extraction time of 120 min with 0.4-mm particle sizes and 20% aqueous ethanol as the co-solvent). Furthermore, the nitrite-scavenging ability of the flavonoid-enriched SFE extracts was assessed using the Griess reagent. The flavonoid-enriched SFE extracts exhibited the highest scavenging ability on nitrite (88.1 ± 3.04%) at the concentration of 500 μg/mL and at pH 3.0. The nitrite-scavenging ability of the extracts appeared to be concentration dependent but negatively correlated with the pH.     

Brazilian Ginseng extraction via LPSE and SFE: Global yields, extraction kinetics, chemical composition and antioxidant activity

Brazilian Ginseng extracts of two species, Pfaffia paniculata and Pfaffia glomerata, were obtained by supercritical fluid extraction (SFE) with CO₂ and by low-pressure solvent extraction (LPSE) with methanol, hexane and ethanol. The SFE assays were conducted at pressures of 100, 200 and 300 bar, and temperatures of 30 and 50 °C. The qualitative chemical compositions of the extracts were determined by thin layer chromatography (TLC). One of the active principles of interest from P. glomerata extract, β-ecdysone, was identified and quantified by HPLC. The antioxidant activities of Brazilian Ginseng extracts were determined by the coupled reaction of linolenic acid and β-carotene. For P. paniculata, the highest SFE yield was obtained at 200 bar/50 °C (0.22%, dry basis—d.b.), while the best extraction condition for P. glomerata was obtained at 200 bar/30 °C (0.18%, d.b.). The higher extract yields obtained by LPSE were 2.0% and 5.8% (w/w, d.b.) for P. paniculata and P. glomerata, respectively, both obtained with methanol as extraction solvent. From the overall extraction curve of P. glomerata, it was possible to obtain the kinetic parameters of extraction; the duration of the CER (constant extraction rate) period was determined as 134 min. The TLC plates showed the possible presence of flavonoids in the ethanolic extract for both Pfaffia species. The antioxidant activity analysis detected that LPSE extracts had higher activity than SFE extracts.     

Friedel–Crafts green alkylation of xylenes with tert-butanol over mesoporous superacid UDCaT-5

Friedel–Crafts green alkylation of xylenes with tert-butanol was investigated in the presence of mesoporous superacidic catalysts named as UDCaT-4, UDCaT-5 and UDCaT-6. The catalysts are modified versions of zirconia showing high catalytic activity, stability and reusability. The catalytic activity is in the order: UDCaT-5 (most active) > UDCaT-6 > UDCaT-4 > sulfated zirconia (least active). Synergistic effect of very high sulfur content present (9% (w/w) S) and preservation of tetragonal phase in UDCaT-5, in comparison with sulfated zirconia (4% (w/w) S), were responsible for higher catalytic activity. The performance of UDCaT-5 in alkylation of xylenes was studied with tert-butanol with reference to selectivity and stability. Alkylation of m-xylene over UDCaT-5 gives 96% conversion of tert-butanol with 82% selectivity towards 5-tert-butyl-m-xylene (5-TBMX) under optimum reaction conditions. The formation of products is correlated with the acidity of the catalyst. The reactions were conducted in liquid phase at relatively low reaction temperatures (130–160 °C). A systematic investigation of the effects of various operating parameters was done to describe the reaction pathway. The reaction was carried out without any solvent in order to make the process cleaner and greener. An overall second order kinetic equation was used to fit the experimental data, under the assumption that both xylene and tert-butanol are weakly adsorbed. An independent study of dehydration of tert-butanol (TBA) was also done. Alkylation of o-xylene and p-xylene with tert-butanol was also studied. The overall process is green and clean.     

FED-BATCH SIMULTANEOUS SACCHARIFICATION AND FERMENTATION OF MICROWAVE/ACID/ALKALI/H2O2 PRETREATED RICE STRAW FOR PRODUCTION OF ETHANOL

The batch simultaneous saccharification and fermentation (SSF) of microwave/acid/alkali/H₂O₂ pretreated rice straw to ethanol was optimized using cellulase from Trichoderma reesei and Saccharomyces cerevisiae YC-097 cells prior to the fed-batch SSF studies. The batch SSF optima were 10% w/v substrate, 40°C, 15 mg cellulase/g substrate, initial pH 5.3, and 72 hours. Under the optimum conditions the ethanol concentration and its yield were 29.1 g/L and 61.3% respectively. Based on the optimal batch SSF, the fed-batch SSF was investigated and its operation parameters were optimized. Under its optimal conditions the ethanol concentration reached 57.3 g/L, while its productivity and yield were only slightly less than those in the batch SSF. This suggests that fed-batch SSF is a potential operation mode for effective ethanol production from microwave/acid/alkali/H₂O₂ pretreated rice straw.     

Gel polymer electrolyte based on poly(acrylonitrile-co-styrene) and a novel organic iodide salt for quasi-solid state dye-sensitized solar cell

A gel polymer electrolyte based on poly(acrylonitrile-co-styrene) as polymer matrix and N-methyl pyridine iodide salt as I⁻ source was prepared. Controlling the concentration of polymer matrix of poly(acrylonitrile-co-styrene) at 17.5 wt.%, mixing the binary organic solvents mixture ethylene carbonate and propylene carbonate with 6:4 (w/w), and the concentration of N-methyl pyridine iodide and iodine with 0.5 and 0.05 M, respectively, the gel polymer electrolyte attains the maximum ionic conductivity (at 30 °C) of 4.63 mS cm⁻¹. Based on the gel polymer electrolyte, a quasi-solid state dye-sensitized solar cell was fabricated and its overall energy conversion efficiency of light-to-electricity of 3.10% was achieved under irradiation of 100 mW cm⁻².     

Production of biodiesel by ultrasound assisted esterification of Oreochromis niloticus oil

This paper evaluates the production of methyl esters from Oreochromis niloticus (Nile tilapia) oil and methanol. The reaction was carried out applying low-frequency high-intensity ultrasound (40 kHz) under atmospheric pressure and ambient temperature. Response surface methodology (RSM) was used to evaluate the influence of alcohol to oil molar ratio, catalyst concentration (sulfuric acid) and temperature on the yield of O. niloticus oil into methyl esters. Analysis of the operating conditions by RSM showed that the most important operating condition affecting the reaction was the alcohol to FFA molar ratio. The highest yield observed was of 98.2% after 90 min of reaction. The optimal operating condition was obtained applying an alcohol to oil molar ratio of 9.0 and a catalyst concentration of 2.0% w/w and temperature of 30 °C.     

Study of the influence of processing parameters on the production of carboxymethylchitin

Carboxymethylchitin was prepared at different reaction temperatures and from alkali chitin with different concentrations of alkali. Properties of the product were studied. Alkali chitin were prepared using freshly prepared sodium hydroxide of 45, 50, 55, 60 and 65% (w/w) concentration and carboxymethylated using monochloroacetic acid at controlled (35-40 °C) and uncontrolled (30-80 °C) temperature conditions. Molecular weight, viscosity, degree of deacetylation, etc. of the resultant product, i.e. carboxymethylchitin were determined. It was found that the reaction temperature has a profound influence on the property of the product than alkali concentration. A hygroscopic and completely water-soluble product was formed. Optimum conditions for the production of carboxymethylchitin were found to be 60% NaOH concentration and at 35-40 °C reaction temperature. At these conditions, it was obtained with a molecular weight of 4.11×10⁶ Da, viscosity 1926 cP and degree of deacetylation 45.02%.     

Guava (Psidium guajava L.) seed oil obtained with a homemade supercritical fluid extraction system using supercritical CO2 and co-solvent

In this work we designed and built a homemade supercritical fluid extraction (HM-SFE) system, in which pure CO₂ and CO₂ with co-solvents were used. The HM-SFE was made by means of thermal dilatation-contraction (TDC). This HM-SFE system was used for obtaining guava (Psidium guajava L.) seed oil, using supercritical CO₂ adding ethanol as co-solvent (CO₂ SC/EtOH), extractions were performed at 313 K and different pressures (10, 20 and 30 MPa), each one in four stages of 30 min, the extract with higher yield was subjected to transesterification and high-resolution gas chromatography (HRGC) analysis. The highest extraction yield was obtained at 30 MPa (17.30% w/w), this yield was higher than one observed in a previous work using SC-CO₂, and near to the one obtained by Soxhlet extraction (20.2% w/w). HRGC enabled the identification of components of the derivatized extract as methyl esters of palmitic, oleic, linoleic, and stearic fatty acids. The results obtained with HM-SFE system was compared with a commercial SFE system, obtained very similar results. In this work was possible to construct a low cost and simple manner HM-SFE system which was employed for obtaining guava seed oil, using CO₂ SC/EtOH.     

Kinetics of ultrasound assisted extraction of anthocyanins from Aronia melanocarpa (black chokeberry) wastes

Black chokeberry fruits are very rich in antioxidant phenolics. After juice extraction, the wastes of pressed berries (juice production by-products) still contain important amount of phenolic species and namely anthocyanins. Ultrasound assisted extraction (UAE) from such wastes was studied. The influence of extraction time (0–240 min), temperature (20–70 °C), solvent composition (0–50% ethanol in water) and ultrasound power (0–100 W) on anthocyanins and polyphenols yields and on antioxidant capacity of the extracts was studied. High temperature and ethanol content in the solvent improved greatly the extraction yields. Ultrasounds improved mainly the extraction kinetics. Ultrasound effect was higher in the beginning of extraction process and at low temperatures. Suitability of UAE for preparation of antioxidant-rich plant extracts at reduced time and energy was confirmed obtaining high extraction efficiencies and high antioxidant capacities of Aronia extracts. To optimize extraction conditions a kinetic mathematical model, based on Peleg's equation and considering also the rate of anthocyanins thermal degradation, was proposed. The influences of extraction time, temperature, solvent composition and ultrasound power on polyphenols and anthocyanins yields and on antioxidant capacity were considered. The optimal conditions for anthocyanins extraction predicted by this model were experimentally validated.