Steam-pretreated Wheat Straw Research Articles

Co-Digestion and Mono-Digestion of Sewage Sludge and Steam-Pretreated Winter Wheat Straw in Continuous Stirred-Tank Reactors—Nutrient Composition and Process Performance

Wheat straw (WS) constitutes a considerable biomass resource and can be used to produce the energy carrier methane through anaerobic digestion. Due to the low contents of several nutrient elements and water in harvested WS, the use of sewage sludge (SS), consisting of primary sludge and waste-activated sludge, as a nutrient source in co-digestion with steam-pretreated wheat straw (PWS) was investigated theoretically and practically. WS was steam-pretreated, with acetic acid as the catalyst, at 190 °C for 10 min, ending with a rapid reduction in pressure. Process stability and specific methane production were studied for the mono-digestion and co-digestion of PWS and SS in continuous stirred-tank reactors for 208 days. The HRT was 22 days and the OLR 2.1 gVS L−1 d−1. In co-digestion, the OLR was increased to 2.8 gVS L−1 d−1 for one week. Nutrient elements were added to PWS mono-digestion at two different concentration levels. Co-digestion was stable, with a total concentration of short-chain fatty acids (SCFAs) at a safe level below 0.35 g L−1 at both OLRs. The higher OLR during co-digestion would require an increase in reactor volume of 14%, compared to the mono-digestion of SS, but would increase the annual production of methane by 26%. The specific methane production levels for PWS mono-digestion, SS mono-digestion, and co-digestion were 170, 320, and 260 mL g−1VS, respectively. Co-digestion did not result in a synergistic increase in the methane yield. SCFAs accumulated in the mono-digestion of PWS when using lower levels of nutrient supplements, and the concentrations fluctuated at higher nutrient levels. The main conclusion is that PWS and SS can be co-digested with long-term process stability, without the addition of chemicals other than water and acetic acid. The specific methane production for mono-digestion of PWS was relatively low. The effect of using higher concentrations of micronutrients in PWS mono-digestion should be evaluated in future studies.

Counter-Current Suspension Extraction Process of Lignocellulose in Biorefineries to Reach Low Water Consumption, High Extraction Yields, and Extract Concentrations

The processing of large quantities of water in biorefining processes can lead to immense costs for heating, evaporation, and wastewater disposal. These costs may prohibit the exploitation of alternative products, e.g., xylooligosaccharides from straw, which are regarded as too costly. A new counter-current extractions method is proposed that aims at low solvent (water) consumption, as well as high yields and extract concentrations. This process was evaluated with suspension extraction experiments with steam pretreated wheat straw and the process window analysis based on a mass balance for a washing and a leaching scenario. The latter was conducted with two other suspension extraction processes as a comparison. The equilibration time was found to be well below 10 min. While the suspension extraction with and without recycling need to be designed to achieve a high yield or a high concentration and low solvent consumption, the proposed extraction method can reach all three simultaneously. Thus, this new process is evaluated as a potential method to spare water and downstream costs and allow new processing pathways in second-generation biorefineries.

Storage and handling of pretreated lignocellulose affects the redox chemistry during subsequent enzymatic saccharification

The decomposition of lignocellulose in nature, as well as when used as feedstock in industrial settings, takes place in a dynamic system of biotic and abiotic reactions. In the present study, the impact of abiotic reactions during the storage of pretreated lignocellulose on the efficiency of subsequent saccharification was investigated. Abiotic decarboxylation was higher in steam-pretreated wheat straw (SWS, up till 1.5% CO2) than in dilute-acid-catalysed steam-pretreated forestry residue (SFR, up till 3.2% CO2) which could be due to higher iron content in SFR and there was no significant CO2 production in warm-water-washed slurries. Unwashed slurries rapidly consumed O2 during incubation at 50 °C; the behaviour was more dependent on storage conditions in case of SWS than SFR slurries. There was a pH drop in the slurries which did not correlate with acetic acid release. Storage of SWS under aerobic conditions led to oxidation of the substrate and reduced the extent of enzymatic saccharification by Cellic® CTec3. Catalase had no effect on the fractional conversion of the aerobically stored substrate, suggesting that the lower fractional conversion was due to reduced activity of the lytic polysaccharide monooxygenase component during saccharification. The fractional conversion of SFR was low in all cases, and cellulose hydrolysis ceased before the first sampling point. This was possibly due to excessive pretreatment of the forest residues. The conditions at which pretreated lignocellulose are stored after pretreatment significantly influenced the extent and kind of abiotic reactions that take place during storage. This in turn influenced the efficiency of subsequent saccharification. Pretreated substrates for laboratory testing must, therefore, be stored in a manner that minimizes abiotic oxidation to ensure that the properties of the substrate resemble those in an industrial setting, where pretreated lignocellulose is fed almost directly into the saccharification vessel.

Diversity and Abundance of Microbial Communities in UASB Reactors during Methane Production from Hydrolyzed Wheat Straw and Lucerne.

The use of straw for biofuel production is encouraged by the European Union. A previous study showed the feasibility of producing biomethane in upflow anaerobic sludge blanket (UASB) reactors using hydrolyzed, steam-pretreated wheat straw, before and after dark fermentation with Caldicellulosiruptor saccharolyticus, and lucerne. This study provides information on overall microbial community development in those UASB processes and changes related to acidification. The bacterial and archaeal community in granular samples was analyzed using high-throughput amplicon sequencing. Anaerobic digestion model no. 1 (ADM1) was used to predict the abundance of microbial functional groups. The sequencing results showed decreased richness and diversity in the microbial community, and decreased relative abundance of bacteria in relation to archaea, after process acidification. Canonical correspondence analysis showed significant negative correlations between the concentration of organic acids and three phyla, and positive correlations with seven phyla. Organic loading rate and total COD fed also showed significant correlations with microbial community structure, which changed over time. ADM1 predicted a decrease in acetate degraders after a decrease to pH ≤ 6.5. Acidification had a sustained effect on the microbial community and process performance.

Wheat straw pre-treatments using eco-friendly strategies for enhancing the tensile properties of bio-based polylactic acid composites

Individual and hybrid pre-treatments, including hot water, steam and microwave were used on wheat straw to mitigate the surface quality deficiencies for an intimate interfacial bond between substrate and Polylactic Acid matrix. Microstructural analysis showed the expansion of cells due to the pre-treatments which in turn promotes mechanical entanglement between wheat straw and PLA, thus, improving the tensile properties of bio-based composites. The tensile strength, elongation at break, elastic modulus, and toughness increased by 166 %, 18 %, 68 %, and 285 %, respectively, for hot water followed by steam pre-treated wheat straw bio-based composites compared to untreated.

The Potential of Using Immobilized Xylanases to Enhance the Hydrolysis of Soluble, Biomass Derived Xylooligomers.

Earlier work had indicated that enzyme-mediated hydrolysis of xylooligomer-rich water-soluble streams (derived from steam pre-treated wheat straw) resulted in the effective production of xylose which was subsequently used to produce bio-glycol. In the work reported here, both the thermostability and recyclability of xylanases were significantly improved by covalent immobilizing the enzymes onto alginate beads. The immobilized xylanases showed a lower hydrolytic potential (~55% xylooligomer conversion) compared to the commercial xylanase cocktail HTec3 (~90% xylooligomer conversion) when used at the same protein loading concentration. This was likely due to the less efficient immobilization of key higher molecular weight enzymes (>75 kDa), such as β-xylosidases. However, enzyme immobilization could be improved by lowering the glutaraldehyde loading used to activate the alginate beads, resulting in improved hydrolysis efficacy (~65% xylooligomer conversion). Enzyme immobilization improved enzyme thermostability (endoxylanase and β-xylosidase activities were improved by 80% and 40%, respectively, after 24 h hydrolysis) and this allowed the immobilized enzymes to be reused/recycled for multiple rounds of hydrolysis (up to five times) without any significant reduction in their hydrolytic potential.

Redox processes acidify and decarboxylate steam-pretreated lignocellulosic biomass and are modulated by LPMO and catalase

BackgroundThe bioconversion of lignocellulosic feedstocks to ethanol is being commercialised, but further process development is required to improve their economic feasibility. Efficient saccharification of lignocellulose to fermentable sugars requires oxidative cleavage of glycosidic linkages by lytic polysaccharide monooxygenases (LPMOs). However, a proper understanding of the catalytic mechanism of this enzyme class and the interaction with other redox processes associated with the saccharification of lignocellulose is still lacking. The in-use stability of LPMO-containing enzyme cocktails is increased by the addition of catalase implying that hydrogen peroxide (H2O2) is generated in the slurry during incubation. Therefore, we sought to characterize the effects of enzymatic and abiotic sources of H2O2 on lignocellulose hydrolysis to identify parameters that could improve this process. Moreover, we studied the abiotic redox reactions of steam-pretreated wheat straw as a function of temperature and dry-matter (DM) content.ResultsAbiotic reactions in pretreated wheat straw consume oxygen, release carbon dioxide (CO2) to the slurry, and decrease the pH. The magnitude of these reactions increased with temperature and with DM content. The presence of LPMO during saccharification reduced the amount of CO2 liberated, while the effect on pH was insignificant. Catalase led to increased decarboxylation through an unknown mechanism. Both in situ-generated and added H2O2 caused a decrease in pH.ConclusionsAbiotic redox processes similar to those that occur in natural water-logged environments also affect the saccharification of pretreated lignocellulose. Heating of the lignocellulosic material and adjustment of pH trigger rapid oxygen consumption and acidification of the slurry. In industrial settings, it will be of utmost importance to control these processes. LPMOs interact with the surrounding redox compounds and redirect abiotic electron flow from decarboxylating reactions to fuel the oxidative cleavage of glycosidic bonds in cellulose.

Sequential Targeting of Xylose and Glucose Conversion in Fed-Batch Simultaneous Saccharification and Co-fermentation of Steam-Pretreated Wheat Straw for Improved Xylose Conversion to Ethanol

Efficient conversion of both glucose and xylose in lignocellulosic biomass is necessary to make second-generation bioethanol from agricultural residues competitive with first-generation bioethanol and gasoline. Simultaneous saccharification and co-fermentation (SSCF) is a promising strategy for obtaining high ethanol yields. However, with this method, the xylose-fermenting capacity and viability of yeast tend to decline over time and restrict the xylose utilization. In this study, we examined the ethanol production from steam-pretreated wheat straw using an established SSCF strategy with substrate and enzyme feeding that was previously applied to steam-pretreated corn cobs. Based on our findings, we propose an alternative SSCF strategy to sustain the xylose-fermenting capacity and improve the ethanol yield. The xylose-rich hydrolyzate liquor was separated from the glucose-rich solids, and phases were co-fermented sequentially. By prefermentation of the hydrolyzate liquor followed fed-batch SSCF, xylose, and glucose conversion could be targeted in succession. Because the xylose-fermenting capacity declines over time, while glucose is still converted, it was advantageous to target xylose conversion upfront. With our strategy, an overall ethanol yield of 84% of the theoretical maximum based on both xylose and glucose was reached for a slurry with higher inhibitor concentrations, versus 92% for a slurry with lower inhibitor concentrations. Xylose utilization exceeded 90% after SSCF for both slurries. Sequential targeting of xylose and glucose conversion sustained xylose fermentation and improved xylose utilization and ethanol yield compared with fed-batch SSCF of whole slurry.

Hybrid SSF/SHF Processing of SO2 Pretreated Wheat Straw\u2014Tuning Co-fermentation by Yeast Inoculum Size and Hydrolysis Time

Wheat straw is one of the main agricultural residues of interest for bioethanol production. This work examines conversion of steam-pretreated wheat straw (using SO2 as a catalyst) in a hybrid process consisting of a short enzymatic prehydrolysis step and a subsequent simultaneous saccharification and fermentation (SSF) step with a xylose-fermenting strain of Saccharomyces cerevisiae. A successful process requires a balanced design of reaction time and temperature in the prehydrolysis step and yeast inoculum size and temperature in the SSF step. The pretreated material obtained after steam pretreatment at 210 °C for 5 min using 2.5 % SO2 (based on moisture content) showed a very good enzymatic digestibility at 45 °C but clearly lower at 30 °C. Furthermore, the pretreatment liquid was found to be rather inhibitory to the yeast, partly due to a furfural content of more than 3 g/L. The effect of varying the yeast inoculum size in this medium was assessed, and at a yeast inoculum size of 4 g/L, a complete conversion of glucose and a 90 % conversion of xylose were obtained within 50 h. An ethanol yield (based on the glucan and xylan in the pretreated material) of 0.39 g/g was achieved for a process with this yeast inoculum size in a hybrid process (10 % water-insoluble solid (WIS)) with 4 h prehydrolysis time and a total process time of 96 h. The obtained xylose conversion was 95 %. A longer prehydrolysis time or a lower yeast inoculum size resulted in incomplete xylose conversion.

Functional characterization of the native swollenin from Trichoderma reesei: study of its possible role as C1 factor of enzymatic lignocellulose conversion.

BackgroundThrough binding to cellulose, expansin-like proteins are thought to loosen the structural order of crystalline surface material, thus making it more accessible for degradation by hydrolytic enzymes. Swollenin SWO1 is the major expansin-like protein from the fungus Trichoderma reesei. Here, we have performed a detailed characterization of a recombinant native form of SWO1 with respect to its possible auxiliary role in the enzymatic saccharification of lignocellulosic substrates.ResultsThe swo1 gene was overexpressed in T. reesei QM9414 Δxyr1 mutant, featuring downregulated cellulase production, and the protein was purified from culture supernatant. SWO1 was N-glycosylated and its circular dichroism spectrum suggested a folded protein. Adsorption isotherms (25 °C, pH 5.0, 1.0 mg substrate/mL) revealed SWO1 to be 120- and 20-fold more specific for binding to birchwood xylan and kraft lignin, respectively, than for binding to Avicel PH-101. The SWO1 binding capacity on lignin (25 µmol/g) exceeded 12-fold that on Avicel PH-101 (2.1 µmol/g). On xylan, not only the binding capacity (22 µmol/g) but also the affinity of SWO1 (Kd = 0.08 µM) was enhanced compared to Avicel PH-101 (Kd = 0.89 µM). SWO1 caused rapid release of a tiny amount of reducing sugars (<1 % of total) from different substrates (Avicel PH-101, nanocrystalline cellulose, steam-pretreated wheat straw, barley β-glucan, cellotetraose) but did not promote continued saccharification. Atomic force microscopy revealed that amorphous cellulose films were not affected by SWO1. Also with AFM, binding of SWO1 to cellulose nanocrystallites was demonstrated at the single-molecule level, but adsorption did not affect this cellulose. SWO1 exhibited no synergy with T. reesei cellulases in the hydrolysis of the different celluloses. However, SWO1 boosted slightly (1.5-fold) the reducing sugar release from a native grass substrate.ConclusionsSWO1 is a strongly glycosylated protein, which has implications for producing it in heterologous hosts. Although SWO1 binds to crystalline cellulose, its adsorption to xylan is much stronger. SWO1 is not an auxiliary factor of the enzymatic degradation of a variety of cellulosic substrates. Effect of SWO1 on sugar release from intact plant cell walls might be exploitable with certain (e.g., mildly pretreated) lignocellulosic feedstocks.

Combined production of biogas and ethanol at high solids loading from wheat straw impregnated with acetic acid: experimental study and techno-economic evaluation

Production of ethanol and biogas from acetic acid-impregnated steam-pretreated wheat straw was investigated. The solid fraction after pretreatment was used at high solids concentrations to generate ethanol by simultaneous saccharification and fermentation (SSF). The residual streams were evaluated with regard to biogas production. The experimental results were used to perform a techno-economic evaluation of a biorefinery concerning ethanol, raw biogas, pellet, and electricity production at various solid contents and residence times in the fermentation step. The configurations were also altered to include biogas upgrading to vehicle fuel quality or fermentation of xylose to ethanol. At a water-insoluble solids (WIS) content in the SSF of between 10 and 20 %, the ethanol yields exceeded 80 %, the highest being 86 % at 12.5 % WIS, expressed as % of the theoretical maximum, based on the glucan content in SSF. Anaerobic digestion of wheat straw hydrolysate and stillage yielded 4.8 and 1.0–1.2 g methane/100 g dry straw, respectively, in 7-day experiments. Maximum recovery of overall product was achieved when SSF was run at between 10 and 15 % initial WIS content, for which the product yields from 100 g dry wheat straw were 16.1–16.3 g ethanol, 5.8–6.0 g methane, and 25 g lignin-rich solid residue. The net present value (NPV) was negative at discount rate of 11 % but positive at 5 % discount rate for all configurations. The 20 % WIS configuration with a residence time of 96 h in the fermentation stage attained the highest NPV. The minimum ethanol selling price varied between 0.72 and 0.87 EUR/L ethanol when the biogas was unchanged and it decreased to between 0.46 and 0.60 EUR/L ethanol when the biogas was upgraded to vehicle fuel quality or when xylose was converted to ethanol. According to the techno-economic assessment, a process that is based on the fermentation of only hexoses to ethanol, and production of raw biogas from xylose is not profitable under the economic assumptions including 11 % discount rate in the evaluation. However, the profitability of a plant can be improved by biogas upgrading to vehicle fuel quality or fermentation of xylose to ethanol.

Prefermentation improves ethanol yield in separate hydrolysis and cofermentation of steam-pretreated wheat straw

Agricultural residues, such as wheat straw, are feasible substrates for ethanol fermentation provided that pentoses and hexoses can be converted efficiently. Separate hydrolysis and cofermentation (SHCF) constitute a framework for improvement of conversion efficiency, because it permits independent optimization of the enzymatic hydrolysis and cofermentation steps. A drawback is that the high glucose concentrations present in SHCF repress xylose utilization and constrain ethanol yields. To improve xylose utilization the xylose-rich hydrolyzate liquor was separated from glucose-rich solids and the phases were cofermented sequentially. Prefermentation of the xylose-rich hydrolyzate liquor followed by fed-batch cofermentation of glucose-rich prehydrolyzed solids enabled sequential targeting of xylose and glucose conversion. The aim was to improve the xylose conversion by lowering the glucose repression of the xylose uptake. Various prefermentation configurations and feed patterns for prehydrolyzed solids were examined. Prefermentation increased ethanol yields overall, and fed-batch prefermentation reduced xylitol production. The best results were obtained by balancing promotion of efficient xylose conversion with maintained yeast viability. Fed-batch prefermentation and a single addition of prehydrolyzed solids, elicited an ethanol yield of 0.423 g·g−1 and a xylitol yield of 0.036 g·g−1.

Model-based optimization and scale-up of multi-feed simultaneous saccharification and co-fermentation of steam pre-treated lignocellulose enables high gravity ethanol production.

BackgroundHigh content of water-insoluble solids (WIS) is required for simultaneous saccharification and co-fermentation (SSCF) operations to reach the high ethanol concentrations that meet the techno-economic requirements of industrial-scale production. The fundamental challenges of such processes are related to the high viscosity and inhibitor contents of the medium. Poor mass transfer and inhibition of the yeast lead to decreased ethanol yield, titre and productivity. In the present work, high-solid SSCF of pre-treated wheat straw was carried out by multi-feed SSCF which is a fed-batch process with additions of substrate, enzymes and cells, integrated with yeast propagation and adaptation on the pre-treatment liquor. The combined feeding strategies were systematically compared and optimized using experiments and simulations.ResultsFor high-solid SSCF process of SO2-catalyzed steam pre-treated wheat straw, the boosted solubilisation of WIS achieved by having all enzyme loaded at the beginning of the process is crucial for increased rates of both enzymatic hydrolysis and SSCF. A kinetic model was adapted to simulate the release of sugars during separate hydrolysis as well as during SSCF. Feeding of solid substrate to reach the instantaneous WIS content of 13 % (w/w) was carried out when 60 % of the cellulose was hydrolysed, according to simulation results. With this approach, accumulated WIS additions reached more than 20 % (w/w) without encountering mixing problems in a standard bioreactor. Feeding fresh cells to the SSCF reactor maintained the fermentation activity, which otherwise ceased when the ethanol concentration reached 40–45 g L−1. In lab scale, the optimized multi-feed SSCF produced 57 g L−1 ethanol in 72 h. The process was reproducible and resulted in 52 g L−1 ethanol in 10 m3 scale at the SP Biorefinery Demo Plant.ConclusionsSSCF of WIS content up to 22 % (w/w) is reproducible and scalable with the multi-feed SSCF configuration and model-aided process design. For simultaneous saccharification and fermentation, the overall efficiency relies on balanced rates of substrate feeding and conversion. Multi-feed SSCF provides the possibilities to balance interdependent rates by systematic optimization of the feeding strategies. The optimization routine presented in this work can easily be adapted for optimization of other lignocellulose-based fermentation systems.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0500-7) contains supplementary material, which is available to authorized users.

Comparative net energy ratio analysis of pellet produced from steam pretreated biomass from agricultural residues and energy crops

A process model was developed to determine the net energy ratio (NER) for the production of pellets from steam pretreated agricultural residue (wheat straw) and energy crops (i.e., switchgrass in this case). The NER is a ratio of the net energy output to the total net energy input from non-renewable energy sources into a system. Scenarios were developed to measure the effects of temperature and level of steam pretreatment on the NER of steam pretreated wheat straw and switchgrass pellets. The NERs for the base case at 6 kg h−1 are 1.76 and 1.37 for steam-pretreated wheat straw and switchgrass-based pellets, respectively. The reason behind the difference is that more energy is required to dry switchgrass pellets than wheat straw pellets. The sensitivity analysis for the model shows that the optimum temperature for steam pretreatment is 160 °C with 50% pretreatment (i.e. 50 % steam treated material is blended with the raw biomass and then pelletised). The uncertainty results for NER for steam pretreated wheat straw and switch grass pellets are 1.62 ± 0.10 and 1.42 ± 0.11, respectively.

Use of Wheat Straw for Extracellular Cellulase Production from Aspergillus niger F8Isolated from Compost

An extracellular cellulase producing celluloytic Aspergillus niger F8 was isolated from compost sample expressing maximum cellulase activity of 6.75 IU/ml on basal salt medium, pH 4.5, temperature 30°C and inoculum size 10% with 1% substrate concentration on 7 days of incubation. 358.70% increase in enzyme activity was observed after optimisation of parameters under submerged fermentation. Further, enhancement in enzyme activity was done under solid-state fermentation using alkali+steam pretreated wheat straw. A. niger F8 exhibited enzyme activity of 71.22 U/g with pretreated (alkali+steam) biomass. It is a cost-effective enzyme production approach of utilising cheap and abundant lignocellulosic agricultural waste as carbon source.

Phenols and lignin: Key players in reducing enzymatic hydrolysis yields of steam-pretreated biomass in presence of laccase

Phenols are known as inhibitors for cellulases and fermentative microorganisms in bioethanol production processes. The addition of laccases removes the phenolic compounds and subsequently reduces the lag phase of the fermentative microorganism. However, the application of laccases diminishes glucose release during the enzymatic hydrolysis. In this study a model cellulosic substrate (Sigmacell) together with lignin extract, whole steam-pretreated wheat straw (slurry) and its water insoluble solid fraction (WIS) were subjected to enzymatic hydrolysis to evaluate the effects of laccase treatment in presence of lignin and phenols. The presence of laccase in enzymatic hydrolysis of Sigmacell with lignin extract reduced glucose yield by 37% compared with assays without laccase. Furthermore, this reduction was even more marked in presence of phenols (55% reduction). Interestingly, when hydrolyzing WIS, the addition of phenols coupled with laccase treatment did not show a reduction when compared with only laccase addition. This fact suggests the key role of lignin in the hydrolysis inhibition since in WIS the ratio cellulase per gram of lignin was much lower than in Sigmacell experiments. Finally, the lower cellobiose and xylose recoveries point out that phenolic oligomers formed by laccase oxidation play important roles in the inhibition of endoglucanases, cellobiohydrolases and xylanases. To conclude, the proportion of lignin and the composition of phenols are key players in the inhibition of cellulases when the enzymatic hydrolysis is combined with laccases detoxification.

Catalase improves saccharification of lignocellulose by reducing lytic polysaccharide monooxygenase-associated enzyme inactivation.

ObjectivesEfficient enzymatic saccharification of plant cell wall material is key to industrial processing of agricultural and forestry waste such as straw and wood chips into fuels and chemicals.ResultsSaccharification assays were performed on steam-pretreated wheat straw under ambient and O2-deprived environments and in the absence and presence of a lytic polysaccharide monooxygenase (LPMO) and catalase. A kinetic model was used to calculate catalytic rate and first-order inactivation rate constants of the cellulases from reaction progress curves. The addition of a LPMO significantly (P < 0.01, Student’s T test) enhanced the rate of glucose release from 2.8 to 6.9 h−1 under ambient O2 conditions. However, this also significantly (P < 0.01, Student’s T test) increased the rate of inactivation of the enzyme mixture, thereby reducing the performance half-life from 65 to 35 h. Decreasing O2 levels or, strikingly, the addition of catalase significantly reduced (P < 0.01, Student’s T test) enzyme inactivation and, as a consequence, higher efficiency of the cellulolytic enzyme cocktail was achieved.ConclusionOxidative inactivation of commercial cellulase mixtures is a significant factor influencing the overall saccharification efficiency and the addition of catalase can be used to protect these mixtures from inactivation.Electronic supplementary materialThe online version of this article (doi:10.1007/s10529-015-1989-8) contains supplementary material, which is available to authorized users.

Simultaneous saccharification and co-fermentation of whole wheat in integrated ethanol production

Two of the most important ways of reducing the production cost of lignocellulosic ethanol are to increase the ethanol yield and the concentration in the fermentation broth. This can be facilitated by co-fermentation of glucose and xylose from agricultural residues such as wheat straw, due to the high amount of xylose in the hemicelluloses in these materials.Simultaneous saccharification and co-fermentation (SSCF) of steam-pretreated wheat straw (SPWS) with and without the addition of liquefied wheat meal (LWM) was performed using the pentose-fermenting yeast, TMB3400. The highest overall ethanol yield in batch operation, of around 70%, equivalent to an ethanol concentration of 43.7 g L−1, was achieved using SPWS with 7.5% water-insoluble solids (WIS) and addition of LWM with 1% WIS. Using SPWS with a higher WIS (10%) resulted in a decreased yield, 60%, although the concentration of ethanol increased to 53.0 g L−1. SSCF of 7.5% straw was also performed with a single (after 20 h) or fed-batch addition of 1% WIS LWM (after 20, 24 and 28 h) resulting in an increase in both ethanol yield and concentration compared to the reference, without wheat meal addition, but no significant difference compared to the batch experiments.The addition of wheat meal to SSCF did not improve xylose utilization significantly, probably due to the instant release of glucose from the liquefied meal, which hampers the uptake of xylose. The instant release of glucose was shown to be caused by the high amylase activity of the β-glucosidase enzyme preparation.

SSF of steam-pretreated wheat straw with the addition of saccharified or fermented wheat meal in integrated bioethanol production

BackgroundIntegration of second-generation (2G) bioethanol production with existing first-generation (1G) production may facilitate commercial production of ethanol from cellulosic material. Since 2G hydrolysates have a low sugar concentration and 1G streams often have to be diluted prior to fermentation, mixing of streams is beneficial. Improved ethanol concentrations in the 2G production process lowers energy demand in distillation, improves overall energy efficiency and thus lower production cost. There is also a potential to reach higher ethanol yields, which is required in economically feasible ethanol production. Integrated process scenarios with addition of saccharified wheat meal (SWM) or fermented wheat meal (FWM) were investigated in simultaneous saccharification and (co-)fermentation (SSF or SSCF) of steam-pretreated wheat straw, while the possibility of recovering the valuable protein-rich fibre residue from the wheat was also studied.ResultsThe addition of SWM to SSF of steam-pretreated wheat straw, using commercially used dried baker’s yeast, S. cerevisiae, resulted in ethanol concentrations of about 60 g/L, equivalent to ethanol yields of about 90% of the theoretical. The addition of FWM in batch mode SSF was toxic to baker’s yeast, due to the ethanol content of FWM, resulting in a very low yield and high accumulation of glucose. The addition of FWM in fed-batch mode still caused a slight accumulation of glucose, but the ethanol concentration was fairly high, 51.2 g/L, corresponding to an ethanol yield of 90%, based on the amount of glucose added.In batch mode of SSCF using the xylose-fermenting, genetically modified S. cerevisiae strain KE6-12, no improvement was observed in ethanol yield or concentration, compared with baker’s yeast, despite the increased xylose utilization, probably due to the considerable increase in glycerol production. A slight increase in xylose consumption was seen when glucose from SWM was fed at a low feed rate, after 48 hours, compared with batch SSCF. However, the ethanol yield and concentration remained in the same range as in batch mode.ConclusionEthanol concentrations of about 6% (w/v) were obtained, which will result in a significant reduction in the cost of downstream processing, compared with SSF of the lignocellulosic substrate alone. As an additional benefit, it is also possible to recover the protein-rich residue from the SWM in the process configurations presented, providing a valuable co-product.

Glucose and xylose co-fermentation of pretreated wheat straw using mutants of S. cerevisiae TMB3400

Wheat straw was pretreated and fermented to ethanol. Two strains, which had been mutated from the genetically modified Saccharomyces cerevisiae TMB3400, KE6-12 and KE6-13i, have been used in this study and the results of performance were compared to that of the original strain. The glucose and xylose co-fermentation ability was investigated in batch fermentation of steam-pretreated wheat straw (SPWS) liquid (undiluted, and diluted 1.5 and 2 times). Both strains showed improved xylose uptake in diluted SPWS liquid, and increased ethanol yields compared with the original TMB3400 strain, although xylitol formation also increased slightly. In undiluted SPWS liquid, however, only KE6-13i performed better than the original strain regarding xylose utilization.Fed-batch fermentation of 1.5 and 2 times diluted liquid was performed by adding the glucose-rich hydrolysates from enzymatic hydrolysis of the solid fraction of SPWS at a constant feed rate after 5h of fermentation, when the glucose had been depleted. The modified strains showed improved xylose conversion; however, the ethanol yield was not significantly improved due to increased glycerol production. Fed-batch fermentation resulted in faster xylose utilization than in the batch cases.