Optimization of reverse-flow,two-temperature,dilute-acid pretreatment to enhance biomass conversion to ethanol

Applied Biochemistry and Biotechnology - A reverse-flow, two-temperature dilute-acid prehydrolysis process of commercial yellow poplar sawdust using two percolation reactors was designed to...     

Measurement of the inhibitory potential and detoxification of biomass pretreatment hydrolysate for ethanol production

Applied Biochemistry and Biotechnology - The Microtox assay represents a rapid, accurate, and reproducible method for determining general microbial toxicity. This assay was used to evaluate the...     

Inulin-containing biomass for ethanol production

The use of stalks instead of tubers as a source of carbohydrates for ethanol production has been investigated. The inulin present in the stalks of Jerusalem artichoke was extracted with water and the effect of solid-liquid ratio, temperature, and acid addition was studied and optimized in order to attain a high-fructose fermentable extract. The maximum extraction efficiency (corresponding to 35 g/L) of soluble sugars was obtained at 1/6 solidliquid ratio. Fermentations of hydrolyzed extracts by baker's yeast and direct fermentation by an inulinease activity yeast were also performed and the potential to use this feedstock for bioethanol production assessed. The results show that the carbohydrates derived from Jerusalem artichoke stalks can be converted efficiently to ethanol by acidic hydrolysis followed by fermentation with Saccharomyces cerevisiae or by direct fermentation of inulin using Kluyveromyces marxianus strains. In this last case about 30 h to complete fermentation was required in comparison with 8–9 h obtained in experiments with S. cerevisiae growth on acid extracted juices.     

SO2 prehydrolysis for high yield ethanol production from biomass

The advantages of the use of SO₂ in steam pretreatment are described. Two different large scale continuous reactors, the Stake and the Wenger, have been used for this purpose. Pine, aspen and corn stover were prehydrolysed by SO₂ in these reactors and hydrolysed by enzymes. The solution of hexoses and pentoses so obtained were fermented byPichia stipitis R, yielding 372, 346 and 388 L ethanol/tonne for the 3 feedstocks, respectively. When a mixed culture ofP. stipitis R, which is an excellent pentose fermenter, andBrettanomyces clausenii which is an excellent cellobiose fermenter, was used in a simultaneous saccharification-fermentation made with SO₂-prehydrolysed aspen, the yield rose to 384 L/tonne. These are higher yields than have been reported in the literature to date.     

Genetic improvement of poplar feedstock quality for ethanol production

Opportunities for matching chemical and physical properties of woody feedstocks to ethanol production process requirements via genetic improvement have long been recognized. Exploitation is now feasible owing to advances in trait measurement, breeding, and gene transfer technologies. Poplar genetic parameters are favorable largely for reducing lignin and increasing cellulose contents and specific gravity. Transgenic poplars with decreased lignin and increased cellulose contents, but otherwise normal growth and development, have been produced via genetic transformation. The long-standing debate on feasibility has thus become one of when, not if, designer varieties will become available.

     

Coal-induced enhancement of ethanol and biomass production

Applied Biochemistry and Biotechnology - Inclusion of coal inSaccharomyces cerevisiae cultures enhanced ethanol and biomass production (from dextrose) similarly to yeast extract. Coal-induced...     

Effect of organosolv ethanol pretreatment variables on physical characteristics of hybrid poplar substrates

Hybrid poplar (Populus nigra x P. maximowiczii) chips were pretreated using an organosolv ethanol process. The effect of pretreatment conditions (temperature, time, catalyst, and ethanol concentration) on the substrate characteristics, including fiber size, crystallinity, and degree of polymerization of cellulose, was investigated using an experimental matrix designed with response surface methodology. The conditions ranged 155-205 degrees C, 26-94 min, 0.83-1.67% catalyst (H(2)SO(4)) on oven-dry wood chip (w/w), and 25-75% ethanol concentration (v/v). The results indicated that the substrate characteristics are controllable and predictable. Desirable substrates can be prepared by fine-tuning the processing parameters. The regression models developed, allowed the quantitative prediction of the substrate characteristics from the pretreatment conditions used.     

Effect of surfactants and zeolites on simultaneous saccharification and fermentation of steam-exploded poplar biomass to ethanol

In this work, the effect of the addition of different concentrations of Tween-80 and three different zeolite-like products on enzymatic hydrolysis, ethanol fermentation, and simultaneous saccharification and fermentation (SSF) process has been investigated. The ability of these products to enhance the effectiveness of the SSF process to ethanol of steam-exploded poplar biomass using the thermotolerant strainKluyveromyces marxianus EMS-26 has been tested. Tween-80 (0.4 g/L) increased enzymatic hydrolysis yield by 20% when compared to results obtained in hydrolysis in absence of the additive. Zeolite-like products (ZESEP-56 and ZECER-56) (2.5 g/L) improved rates of conversion and ethanol yields in the fermentation of liquid fraction recovered from steam-exploded poplar. The periods required for the completion of fermentation were approx 10 h in the presence of zeolite-like products and 24 h in the absence of additives. The probable mode of action is through lowered levels of inhibitory substances because of adsorption by the additive.     

Conversion of biomass to ethanol

In theEscherichia coli cell-free system, the modification of cell extract can be achieved by preparation of the strains carrying additional property or those being induced with a certain gene expression prior to harvesting. In this study, we analyzed the cell-free system with S30 extract containing T7 RNA polymerases (S30 extract-T7pol) prepared from E.coli BL21(DE3) strain, which includes T7 RNA polymerase from extrinsic genes by IPTG induction, as a model for the improvement of the cell-free system. The fact that a significant degree of mRNA degradation was observed in the cell-free system with S30 extract-T7pol indicates the increase of ribonuclease activity was an unfavorable influence derived from the cell-extract modification process. We also showed that this influence was settled by the addition of an effective ribonuclease inhibitor, such as copper (II) ion, to the reaction mixture.     

Dilute sulfuric acid hydrolysis of biomass for ethanol production

A series of pilot plant experiments have been conducted to compare the performance of a system utilizing two percolation reactors in series to a single reactor system. Although theoretically capable of producing higher glucose yields or concentrations, the two-reactor system concentrations were approximately the same and the yields were considerably lower than those from the single reactor study. An associated kinetics study found the glucose degradation kinetics to be accelerated by chromium ions, but this effect was greatly reduced in the presence of wood. The presence of metal surfaces also increased the rate of degradation even without large ion concentrations. The poor performance of the reactor system is proposed to be caused by intraparticle glucose diffusion effects and the catalytic effect on glucose degradation reactions of chromium ions from the corrosion of stainless steel by the acid. Strategies for reducing the effects of these phenomena on PBR performance are presented.

     

Application of fentonś reaction to steam explosion prehydrolysates from poplar biomass

The application of Fenton’s reaction to enhance the fermentability of prehydrolysates obtained from steam explosion pretreatment of poplar biomass was studied. Reaction conditions of temperature and H₂O₂ and Fe(II) concentrations were studied. The fermentability of prehydrolysate treated by Fenton’s reaction was tested by using different inoculum sizes of thermotolerant strain Kluyveromyces marxianus CECT 10875. The highest percentages of toxic compound degradation (ranging from 71 to 93% removal) were obtained at the highest H₂O₂ concentration tested (50 mM). However, a negative effect on fermentability was observed at this H₂O₂ concentration at the lower inoculum loading. An increase in inoculum size to 0.6 g/L resulted in an enhanced ethanol fermentation yield of 95% relative to control.     

Alkaline and peracetic acid pretreatments of biomass for ethanol production

Prehydrolysis with dilute acid and steam explosion constitute the most promising methods for improving enzymatic digestibility of biomass for ethanol production. Despite world wide acceptance, these methods of pretreatment are quite expensive considering costs for the reactor, energy, and fractionation. Using peracetic acid is a lignin-oxidation pretreatment with low-energy input by which biomass can be treated in a silo-type system without need for expensive capitalization. Experimentally, ground hybrid poplar and sugar cane bagasse are placed in plastic bags and a peracetic acid solution is added to the biomass in different concentrations based on ovendried biomass. The ratio of solution to biomass is 6∶1 and a 7-d storage period at ambient temperature (20°C) has been used. As an auxiliary method, a series of pre-pretreatments using stoichiometri camounts of sodium hydroxide and ammonium hydroxide based on 4-methyl-glucuronic acid and acetyl content in the biomass are performed before addition of peracetic acid. The basic solutions are added to the biomass in a ratio of 14∶1 solution to biomass, and mixed for 24 h at the same ambient temperature. Biomass is filtered and washed to a neutral pH before peracetic acid addition. The aforementioned procedures give high xylan content substrates as a function of the selectivity of peracetic acid for lignin oxidation and the mild conditions of the process. Consequently, xylanase/β-glucosidase combinations were more effective than cellulase preparations in hydrolyzing these materials. The pretreatment efficiency was evaluated through enzymatic hydrolysis and simultaneous saccharification and cofermentation (SSCF) tests. Peracetic ac treatment improves enzymatic digestibility of hybrid poplar and sugar cane bagasse with no need of high temperatures. Alkaline treatments are helpful in reducing peracetic acid requirements in the pretreatment.     

Processing and economic impacts of biomass delignification for ethanol production

Applied Biochemistry and Biotechnology - The need for chemical pretreatment of biomass for the enzyme-catalyzed production of ethanol from lignocellulosic feedstocks has been established. Past...     

Hydrothermal pretreatment of rice straw at relatively lower temperature to improve biogas production via anaerobic digestion

The performances of rice straw (RS) degradation and biogas production were examined at different pretreatment temperatures from 90℃ to 130℃ to improve biogas fermentation efficiency and net energy production in whole slurry. Test at 100℃ pretreatment, which achieved 12.8% higher net energy production from RS than that observed in the control, could be considered as the optimal choice.     

Continuous fermentation of cellulosic biomass to ethanol

Experimental results are presented for continuous conversion of pretreated hardwood flour to ethanol. A simultaneous saccharification and fermentation (SSF) system comprised ofTrichoderma reesei cellulase supplemented with additional β-glucosidase and fermentation bySaccharomyces cerevisiae was used for most experiments, with data also presented for a direct microbial conversion (DMC) system comprised ofClostridium thermocellum. Using a batch SSF system, dilute acid pretreatment of mixed hardwood at short residence time(10 s, 220°C, 1% H₂SO₄) was compared to poplar wood pretreated at longer residence time (20 min, 160°C, 0.45% H₂SO₄). The short residence time pretreatment resulted in a somewhat (10–20%) more reactive substrate, with the reactivity difference particularly notable at low enzyme loadings and/or low agitation. Based on a preliminary screening, inhibition of SSF by byproducts of short residence time pretreatment was measurable, but minor. Both SSF and DMC were carried out successfully in well-mixed continuous systems, with steady-state data obtained at residence times of 0.58–3 d for SSF as well as 0.5 and 0.75 d for DMC. The SSF system achieved substrate conversions varying from 31% at a 0.58-d residence time to 86% at a 2-d residence time. At comparable substrate concentrations (4–5 g/l) and residence times (0.5–0.58 d), substrate conversion in the DMC system (77%) was significantly higher than that in the SSF system (31%). Our results suggest that the substrate conversion in SSF carried out in CSTR is relatively insensitive to enzyme loading in the range 7–25 U/g cellulose and to substrate concentration in the range of 5–60 g/L cellulose in the feed.     

Dilute sulfuric acid pretreatment of agricultural and agro-industrial residues for ethanol production

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

Steam pretreatment of acid-sprayed and acid-soaked barley straw for production of ethanol

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

Two-step steam pretreatment of softwood with SO2 impregnation for ethanol production

Two-step steam pretreatment of softwood was investigated with the aim of improving the enzymatic digestibility for ethanol production. In the first step, softwood was impregnated with SO₂ and steam pretreated at different severities. The first step was performed at low severity to hydrolyze the hemicellulose and release the sugars into the solution. The combination of time and temperature that yielded the highest amount of hemicellulosic sugars in the solution was determined. In the second step, the washed solid material from the optimized first step was impregnated once more with SO₂ and steam pretreated under more severe conditions to enhance the enzymatic digestibility. The investigated temperature range was between 180 and 220°C, and the residence times were 2, 5 and 10 min. The effectiveness of pretreatment was assessed by both enzymatic hydrolysis of the solids and simultaneous saccharification and fermentation (SSF) of the whole slurry after the second pretreatment step, in the presence of antibiotics. For each pretreatment combination, the liquid fraction was fermented to determine any inhibiting effects. At low severity in the second pretreatment step, a high conversion of cellulose was obtained in the enzymatic hydrolysis step, and at a high severity a high conversion of cellulose was obtained in the second pretreatment step. This resulted in an overall yield of sugars that was nearly constant over a wide range of severity. Compared with the one-step steam pretreatment, the two-step steam pretreatment resulted in a higher yield of sugar and in a slightly higher yield of ethanol. The overall sugar yield, when assessed by enzymatic hydrolysis, reached 80%. In the SSF configuration, an overall ethanol yield of 69% was attained.     

Electrochemical hydrogen production from biomass

Thermodynamic data indicate that the oxidation of oxygenated organic species originating from biomass instead of water at the anode of an electrolysis cell should allow decreasing the cell voltage below 1.23 V. Biosourced alcohols, polyols, sugars, lignocellulosic compounds, and their derivatives are then electroreformed to produce clean hydrogen at the cathode and compounds at the anode of electrolysis cells. The reported studies highlight the main challenges to make electroreforming a future industrial process: higher reaction kinetics and hydrogen evolution rate; better selectivity of anode catalysts toward the formation of CO₂ or added-value compounds; and utilization of nonstrategical metals. An attractive solution to decrease hydrogen production costs and to make bankable other economic activities consists in directly valuing wastes from agriculture/forestry (lignocellulosic raw materials) and/or wastes from biofuel industries.     

Optimization of SO2-catalyzed steam pretreatment of corn fiber for ethanol production

A batch reactor was employed to steam explode corn fiber at various degrees of severity to evaluate the potential of using this feedstock as part of an enzymatically mediated cellulose-to-ethanol process. Severity was controlled by altering temperature (150–230°C), residence time (1–9 min), and SO₂ concentration (0–6% [w/w] dry matter). The effects of varying the different parameters were assessed by response surface modeling. The results indicated that maximum sugar yields (hemicellulose-derived water soluble, and cellulose-derived following enzymatic hydrolysis) were recovered from corn fiber pretreated at 190°C for 5 minutes after exposure to 3% SO₂. Sequential SO₂-catalyzed steam explosion and enzymatic hydrolysis resulted in a conversion efficiency of 81% of the combined original hemicellulose and cellulose in the corn fiber to monomeric sugars. An additional posthydrolysis step performed on water soluble hemicellulose stream increased the concentration of sugars available for fermentation by 10%, resulting in the high conversion efficiency of 91%. Saccharomyces cerevisiae was able to ferment the resultant corn fiber hydrolysates, perhydrolysate, and liquid fraction from the posthydrolysis steps to 89, 94, and 85% of theoretical ethanol conversion, respectively. It was apparent that all of the parameters investigated during the steam explosion pretreatment had a significant effect on sugar recovery, inhibitory formation, enzymatic conversion efficiency, and fermentation capacity of the yeast.