Scale-up and integration of alkaline hydrogen peroxide pretreatment, enzymatic hydrolysis, and ethanolic fermentation

Alkaline hydrogen peroxide (AHP) has several attractive features as a pretreatment in the lignocellulosic biomass‐to‐ethanol pipeline. Here, the feasibility of scaling‐up the AHP process and integrating it with enzymatic hydrolysis and fermentation was studied. Corn stover (1 kg) was subjected to AHP pretreatment, hydrolyzed enzymatically, and the resulting sugars fermented to ethanol. The AHP pretreatment was performed at 0.125 g H₂O₂/g biomass, 22°C, and atmospheric pressure for 48 h with periodic pH readjustment. The enzymatic hydrolysis was performed in the same reactor following pH neutralization of the biomass slurry and without washing. After 48 h, glucose and xylose yields were 75% and 71% of the theoretical maximum. Sterility was maintained during pretreatment and enzymatic hydrolysis without the use of antibiotics. During fermentation using a glucose‐ and xylose‐utilizing strain of Saccharomyces cerevisiae, all of the Glc and 67% of the Xyl were consumed in 120 h. The final ethanol titer was 13.7 g/L. Treatment of the enzymatic hydrolysate with activated carbon prior to fermentation had little effect on Glc fermentation but markedly improved utilization of Xyl, presumably due to the removal of soluble aromatic inhibitors. The results indicate that AHP is readily scalable and can be integrated with enzyme hydrolysis and fermentation. Compared to other leading pretreatments for lignocellulosic biomass, AHP has potential advantages with regard to capital costs, process simplicity, feedstock handling, and compatibility with enzymatic deconstruction and fermentation. Biotechnol. Bioeng. 2012; 109:922–931. © 2011 Wiley Periodicals, Inc.     

Bacillus coagulans tolerance to 1‐ethyl‐3‐methylimidazolium‐based ionic liquids in aqueous and solid‐state thermophilic culture

The use of ionic liquids (ILs) to disrupt the recalcitrant structure of lignocellulose and make polysaccharides accessible to hydrolytic enzymes is an emerging technology for biomass pretreatment in lignocellulosic biofuel production. Despite efforts to reclaim and recycle IL from pretreated biomass, residual IL can be inhibitory to microorganisms used for downstream fermentation. As a result, pathways for IL tolerance are needed to improve the activity of fermentative organisms in the presence of IL. In this study, microbial communities from compost were cultured under high‐solids and thermophilic conditions in the presence of 1‐ethyl‐3‐methylimidazolium‐based ILs to enrich for IL‐tolerant microorganisms. A strain of Bacillus coagulans isolated from an IL‐tolerant community was grown in liquid and solid‐state culture in the presence of the ILs 1‐ethyl‐3‐methylimidazolium acetate ([C2mim][OAc]) or 1‐ethyl‐3‐methylimidazolium chloride ([C2mim][Cl]) to gauge IL tolerance. Viability and respiration varied with the concentration of IL applied and the type of IL used. B. coagulans maintained growth and respiration in the presence of 4 wt% IL, a concentration similar to that present on IL‐pretreated biomass. In the presence of both [C2mim][OAc] and [C2mim][Cl] in liquid culture, B. coagulans grew at a rate approximately half that observed in the absence of IL. However, in solid‐state culture, the bacteria were significantly more tolerant to [C2mim][Cl] compared with [C2mim][OAc]. B. coagulans tolerance to IL under industrially relevant conditions makes it a promising bacterium for understanding mechanisms of IL tolerance and discovering IL tolerance pathways for use in other microorganisms, particularly those used in bioconversion of IL‐pretreated plant biomass. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:311–316, 2014     

Renewable bio ionic liquids‐water mixtures‐mediated selective removal of lignin from rice straw: Visualization of changes in composition and cell wall structure

Pretreatment of rice straw by using renewable cholinium amino acids ionic liquids ([Ch][AA] ILs)‐water mixtures and the subsequent enzymatic hydrolysis of the residues were conducted in the present work. Of the eight mixtures composed of ILs and water, most were found to be effective for rice straw pretreatment. After pretreatment with 50% ILs‐water mixtures, the enzymatic digestion of the lignocellulosic biomass was enhanced significantly, thus leading to satisfactory sugar yields of >80% for glucose and approximately 50% for xylose. To better understand the ILs pretreatment mechanism, confocal laser scanning microscopy combined with immunolabeling and transmission electron microscopy were used to visualize changes in the contents and distribution of two major components—lignin and xylan. The results coupled with changes in chemical structures (infrared spectra) of the substrates indicated occurrence of extensive delignification, especially in cell corner and compound middle lumen of cell walls, which made polysaccharides more accessible to enzymes. This pretreatment process is promising for large‐scale application because of the high sugar yields, easy handling, being environmentally benign and highly tolerant to moisture, and significantly reduced cost and energy consumption. Biotechnol. Bioeng. 2013; 110: 1895–1902. © 2013 Wiley Periodicals, Inc.     

Co‐hydrolysis of lignocellulosic biomass for microbial lipid accumulation

The herbaceous perennial energy crops miscanthus, giant reed, and switchgrass, along with the annual crop residue corn stover, were evaluated for their bioconversion potential. A co‐hydrolysis process, which applied dilute acid pretreatment, directly followed by enzymatic saccharification without detoxification and liquid–solid separation between these two steps was implemented to convert lignocellulose into monomeric sugars (glucose and xylose). A factorial experiment in a randomized block design was employed to optimize the co‐hydrolysis process. Under the optimal reaction conditions, corn stover exhibited the greatest total sugar yield (glucose + xylose) at 0.545 g g^?1 dry biomass at 83.3% of the theoretical yield, followed by switch grass (0.44 g g^?1 dry biomass, 65.8% of theoretical yield), giant reed (0.355 g g^?1 dry biomass, 64.7% of theoretical yield), and miscanthus (0.349 g g^?1 dry biomass, 58.1% of theoretical yield). The influence of combined severity factor on the susceptibility of pretreated substrates to enzymatic hydrolysis was clearly discernible, showing that co‐hydrolysis is a technically feasible approach to release sugars from lignocellulosic biomass. The oleaginous fungus Mortierella isabellina was selected and applied to the co‐hydrolysate mediums to accumulate fungal lipids due to its capability of utilizing both C5 and C6 sugars. Fungal cultivations grown on the co‐hydrolysates exhibited comparable cell mass and lipid production to the synthetic medium with pure glucose and xylose. These results elucidated that combining fungal fermentation and co‐hydrolysis to accumulate lipids could have the potential to enhance the utilization efficiency of lignocellulosic biomass for advanced biofuels production. Biotechnol. Bioeng. 2013; 110: 1039–1049. © 2012 Wiley Periodicals, Inc.     

Ionic liquid pretreatment of cellulosic biomass: enzymatic hydrolysis and ionic liquid recycle

Ionic liquids (ILs) are promising solvents for the pretreatment of biomass as certain ILs are able to completely solubilize lignocellulose. The cellulose can readily be precipitated with an anti-solvent for further hydrolysis to glucose, but the anti-solvent must be removed for the IL to be recovered and recycled. We describe the use of aqueous kosmotropic salt solutions to form a three-phase system that precipitates the biomass, forming IL-rich and salt-rich phases. The phase behavior of [Emim][Ac] and aqueous phosphate salt systems is presented, together with a process for recycling the [Emim][Ac] and enzymatically hydrolyzing the cellulose. This process reduces the amount of water to be evaporated from recycled IL, permitting efficient recycle of the IL. Material balances on the process, with multiple recycles of the [Emim][Ac], quantify the major components from a Miscanthus feedstock through the pretreatment, separation, and enzymatic hydrolysis steps. A more rapid and higher yielding conversion of cellulose to glucose is obtained by use of the three-phase system as compared to the cellulose obtained from biomass pretreated with IL and precipitated with water. The addition of a kosmotropic salt during the precipitation results in partial delignification of the biomass, which makes the substrate more accessible, enhancing the enzymatic hydrolysis.     

Novel renewable ionic liquids as highly effective solvents for pretreatment of rice straw biomass by selective removal of lignin

Cholinium amino acids ionic liquids ([Ch][AA] ILs), a novel type of bio‐ILs that can easily be prepared from renewable biomaterials, were investigated for pretreatment of rice straw by selective extraction of lignin from this abundant lignocellulosic biomass material. Of the eight ILs examined, most were demonstrated to be excellent pretreatment solvents. Upon pretreatment using these ILs, the initial saccharification rates of rice straw residues were substantially improved as well as the extent to which polysaccharides could be digested (>90% for cellulose and >60% for xylan). Enzymatic hydrolysis of pretreated rice straw by Trichoderma reesei cellulase/xylanase furnished glucose and xylose with the yields in excess of 80% and 30%, respectively. Detailed spectroscopic characterization showed that the enhancement of polysaccharides degestibility derived mainly from delignification rather than changes in cellulose crystallinity. The yields of fermentable reducing sugars were significantly improved after individual optimization of pretreatment temperature and duration. With [Ch][Lys] as the solvent, the sugar yields of 84.0% for glucose and 42.1% for xylose were achieved after pretreatment at 90°C for 5 h. The IL [Ch][Lys] showed excellent reusability across five successive batches in pretreatment of rice straw. These bio‐ILs performed as well as or better than previously investigated non‐renewable ILs, and thus present a new and environmentally friendly way to pretreat lignocellulose for production of fermentable sugars and total utilization of the biomass. Biotechnol. Bioeng. 2012; 109: 2484–2493. © 2012 Wiley Periodicals, Inc.     

Facile pretreatment of lignocellulosic biomass at high loadings in room temperature ionic liquids

Ionic liquids (ILs) have emerged as attractive solvents for lignocellulosic biomass pretreatment in the production of biofuels and chemical feedstocks. However, the high cost of ILs is a key deterrent to their practical application. Here, we show that acetate based ILs are effective in dramatically reducing the recalcitrance of corn stover toward enzymatic polysaccharide hydrolysis even at loadings of biomass as high as 50% by weight. Under these conditions, the IL serves more as a pretreatment additive rather than a true solvent. Pretreatment of corn stover with 1‐ethyl‐3‐methylimidizolium acetate ([Emim] [OAc]) at 125 ± 5°C for 1 h resulted in a dramatic reduction of cellulose crystallinity (up to 52%) and extraction of lignin (up to 44%). Enzymatic hydrolysis of the IL‐treated biomass was performed with a common commercial cellulase/xylanase from Trichoderma reesei and a commercial β‐glucosidase, and resulted in fermentable sugar yields of ～80% for glucose and ～50% for xylose at corn stover loadings up to 33% (w/w) and 55% and 34% for glucose and xylose, respectively, at 50% (w/w) biomass loading. Similar results were observed for the IL‐facilitated pretreatment of switchgrass, poplar, and the highly recalcitrant hardwood, maple. At 4.8% (w/w) corn stover, [Emim][OAc] can be readily reused up to 10 times without removal of extracted components, such as lignin, with no effect on subsequent fermentable sugar yields. A significant reduction in the amount of IL combined with facile recycling has the potential to enable ILs to be used in large‐scale biomass pretreatment. Biotechnol. Bioeng. 2011;108: 2865–2875. © 2011 Wiley Periodicals, Inc.     

Enrichment,isolation and characterization of fungi tolerant to 1‐ethyl‐3‐methylimidazolium acetate

Aims: This work aimed to characterize microbial tolerance to 1‐ethyl‐3‐methylimidazolium acetate ([C2mim][OAc]), an ionic liquid that has emerged as a novel biomass pretreatment for lignocellulosic biomass. Methods and Results: Enrichment experiments performed using inocula treated with [C2mim][OAc] under solid and liquid cultivation yielded fungal populations dominated by Aspergilli. Ionic liquid‐tolerant Aspergillus isolates from these enrichments were capable of growing in a radial plate growth assay in the presence of 10% [C2mim][OAc]. When a [C2mim][OAc]‐tolerant Aspergillus fumigatus strain was grown in the presence of switchgrass, endoglucanases and xylanases were secreted that retained residual enzymatic activity in the presence of 20% [C2mim][OAc]. Conclusions: The results of the study suggest that tolerance to ionic liquids is a general property of the Aspergilli. Significance and Impact of the Study: Tolerance to an industrially important ionic liquid was discovered in a fungal genera that is widely used in biotechnology, including biomass deconstruction.     

Compatible ionic liquid-cellulases system for hydrolysis of lignocellulosic biomass

Ionic liquids (ILs) have been increasingly recognized as novel solvents for dissolution and pretreatment of cellulose. However, cellulases are inactivated in the presence of ILs, even when present at low concentrations. To more fully exploit the benefits of ILs it is critical to develop a compatible IL‐cellulases system in which the IL is able to effectively solubilize and activate the lignocellulosic biomass, and the cellulases possess high stability and activity. In this study, we investigated the stability and activity of a commercially available cellulases mixture in the presence of different concentrations of 1‐ethyl‐3‐methylimidazolium acetate ([Emim][OAc]). A mixture of cellulases and β‐glucosidase (Celluclast1.5L, from Trichoderma reesei, and Novozyme188, from Aspergillus niger, respectively) retained 77% and 65% of its original activity after being pre‐incubated in 15% and 20% (w/v) IL solutions, respectively, at 50°C for 3 h. The cellulases mixture also retained high activity in 15% [Emim][OAc] to hydrolyze Avicel, a model substrate for cellulose analysis, with conversion efficiency of approximately 91%. Notably, the presence of different amounts of yellow poplar lignin did not interfere significantly with the enzymatic hydrolysis of Avicel. Using this IL‐cellulase system (15% [Emim][OAc]), the saccharification of yellow poplar biomass was also significantly improved (33%) compared to the untreated control (3%) during the first hour of enzymatic hydrolysis. Together, these findings provide compelling evidence that [Emim][OAc] was compatible with the cellulase mixture, and this compatible IL‐cellulases system is promising for efficient activation and hydrolysis of native biomass to produce biofuels and co‐products from the individual biomass components. Bioeng. 2011; 108:1042–1048. © 2010 Wiley Periodicals, Inc.     

Inhibition of cellulose enzymatic hydrolysis by laccase‐derived compounds from phenols

The presence of inhibitors compounds after pretreatment of lignocellulosic materials affects the saccharification and fermentation steps in bioethanol production processes. Even though, external addition of laccases selectively removes the phenolic compounds from lignocellulosic prehydrolysates, when it is coupled to saccharification step, lower hydrolysis yields are attained. Vanillin, syringaldehyde and ferulic acid are phenolic compounds commonly found in wheat‐straw prehydrolysate after steam‐explosion pretreatment. These three phenolic compounds were used in this study to elucidate the inhibitory mechanisms of laccase‐derived compounds after laccase treatment. Reaction products derived from laccase oxidation of vanillin and syringaldehyde showed to be the strongest inhibitors. The presence of these products causes a decrement on enzymatic hydrolysis yield of a model cellulosic substrate (Sigmacell) of 46.6 and 32.6%, respectively at 24 h. Moreover, a decrease in more than 50% of cellulase and β‐glucosidase activities was observed in presence of laccase and vanillin. This effect was attributed to coupling reactions between phenoxyl radicals and enzymes. On the other hand, when the hydrolysis of Sigmacell was performed in presence of prehydrolysate from steam‐exploded wheat straw a significant inhibition on enzymatic hydrolysis was observed independently of laccase treatment. This result pointed out that the other components of wheat‐straw prehydrolysate are affecting the enzymatic hydrolysis to a higher extent than the possible laccase‐derived products. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:700–706, 2015     

Alkali‐based AFEX pretreatment for the conversion of sugarcane bagasse and cane leaf residues to ethanol

Sugarcane is one of the major agricultural crops cultivated in tropical climate regions of the world. Each tonne of raw cane production is associated with the generation of 130 kg dry weight of bagasse after juice extraction and 250 kg dry weight of cane leaf residue postharvest. The annual world production of sugarcane is ～1.6 billion tones, generating 279 MMT tones of biomass residues (bagasse and cane leaf matter) that would be available for cellulosic ethanol production. Here, we investigated the production of cellulosic ethanol from sugar cane bagasse and sugar cane leaf residue using an alkaline pretreatment: ammonia fiber expansion (AFEX). The AFEX pretreatment improved the accessibility of cellulose and hemicelluloses to enzymes during hydrolysis by breaking down the ester linkages and other lignin carbohydrate complex (LCC) bonds and the sugar produced by this process is found to be highly fermentable. The maximum glucan conversion of AFEX pretreated bagasse and cane leaf residue by cellulases was ～85%. Supplementation with hemicellulases during enzymatic hydrolysis improved the xylan conversion up to 95–98%. Xylanase supplementation also contributed to a marginal improvement in the glucan conversion. AFEX‐treated cane leaf residue was found to have a greater enzymatic digestibility compared to AFEX‐treated bagasse. Co‐fermentation of glucose and xylose, produced from high solid loading (6% glucan) hydrolysis of AFEX‐treated bagasse and cane leaf residue, using the recombinant Saccharomyces cerevisiae (424A LNH‐ST) produced 34–36 g/L of ethanol with 92% theoretical yield. These results demonstrate that AFEX pretreatment is a viable process for conversion of bagasse and cane leaf residue into cellulosic ethanol. Biotechnol. Bioeng. 2010;107: 441–450. © 2010 Wiley Periodicals, Inc.     

斑茅酶解转化可发酵单糖的液氨预处理及参数优化

斑茅(Saccharum arundinaceum Retz.)的生物产量高,对土壤条件要求低,可作为纤维素乙醇生产的原料作物在我国南方地区广泛种植.实验以斑茅为原料,采用液氨预处理法克服其水解顽抗性,并添加纤维素酶进行酶解,运用高效液相色谱(HPLC)测定了酶解液中的单糖含量.实验结果表明在纤维素酶添加量为15FPU/(g当量葡聚糖)、预处理原料含水率为80％、预处理温度为130℃、预处理驻留时间为10 min、液氨与生物质的质量比例为2∶1时,葡聚糖和木聚糖的总转化率分别为69.34％和82.60％,相比于未作预处理的原料分别提高了573％和1 056％,单糖产量提高8倍.实验结果表明液氨预处理对斑茅是一种有效的预处理方式,并优于稀酸或湿爆法预处理,与酸预处理和氨爆法(AFEX)处理效果接近.     

Biological pretreatment of corn stover with Phlebia brevispora NRRL‐13108 for enhanced enzymatic hydrolysis and efficient ethanol production

Biological pretreatment of lignocellulosic biomass by white‐rot fungus can represent a low‐cost and eco‐friendly alternative to harsh physical, chemical, or physico‐chemical pretreatment methods to facilitate enzymatic hydrolysis. In this work, solid‐state cultivation of corn stover with Phlebia brevispora NRRL‐13018 was optimized with respect to duration, moisture content and inoculum size. Changes in composition of pretreated corn stover and its susceptibility to enzymatic hydrolysis were analyzed. About 84% moisture and 42 days incubation at 28°C were found to be optimal for pretreatment with respect to enzymatic saccharification. Inoculum size had little effect compared to moisture level. Ergosterol data shows continued growth of the fungus studied up to 57 days. No furfural and hydroxymethyl furfural were produced. The total sugar yield was 442 ± 5 mg/g of pretreated corn stover. About 36 ± 0.6 g ethanol was produced from 150 g pretreated stover per L by fed‐batch simultaneous saccharification and fermentation (SSF) using mixed sugar utilizing ethanologenic recombinant Eschericia coli FBR5 strain. The ethanol yields were 32.0 ± 0.2 and 38.0 ± 0.2 g from 200 g pretreated corn stover per L by fed‐batch SSF using Saccharomyces cerevisiae D5A and xylose utilizing recombinant S. cerevisiae YRH400 strain, respectively. This research demonstrates that P. brevispora NRRL‐13018 has potential to be used for biological pretreatment of lignocellulosic biomass. This is the first report on the production of ethanol from P. brevispora pretreated corn stover. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:365–374, 2017     

Eastern gamagrass as an alternative cellulosic feedstock for bioethanol production

Eastern gamagrass (Trypsacum dactyloides) is a C₄ perennial grass, native to the USA with desirable characteristics that warrants further investigation as a new lignocellulosic crop for bioethanol production. Chemical composition assays showed that eastern gamagrass had comparable cellulose, hemicellulose and lignin compositions to those of switchgrass (Panicum virgatum). With the cellulose solvent-based lignocellulose fractionation (CSLF) pretreatment and subsequent enzymatic saccharification, 80.5–99.8% of cellulosic glucose was released from the gamagrass biomass, which was 10–17% greater than the glucose release efficiency from switchgrass (73.5–87.1%). Furthermore, the hydrolysate of gamagrass supported greater ethanol fermentation yield (up to 0.496 g/g glucose) than the hydrolysates of switchgrass. As such, in the whole process of biomass-to-ethanol conversion, gamagrass could yield 13–35% more ethanol per gram of biomass than switchgrass, indicating that gamagrass has high potential as an alternative energy feedstock for lignocellulosic ethanol production.     

High consistency enzymatic hydrolysis of hardwood substrates

The feasibility of using a laboratory peg mixer to carry out high consistency enzymatic hydrolysis of lignocellulosic substrates was investigated. Two hardwood substrates, unbleached hardwood pulp (UBHW) and organosolv pretreated poplar (OPP), were used in this study. Hydrolysis of UBHW and OPP at 20% substrate consistency led to a high glucose concentration in the final hydrolysate. For example, a 48 h enzymatic hydrolysis of OPP resulted in a hydrolysate with 158 g/L of glucose. This is the highest glucose concentration ever obtained from enzymatic hydrolysis of lignocellulosic substrates. Fermentation of UBHW and OPP hydrolysates with high glucose content led to high ethanol concentrations, 50.4 and 63.1 g/L, respectively after fermentation. Our results demonstrate that using common pulping equipment to carry out high consistency hydrolysis can overcome the rheological problems and greatly increase the sugar and ethanol concentrations after the hydrolysis and fermentation.     

Microwave‐based alkali pretreatment of switchgrass and coastal bermudagrass for bioethanol production

Switchgrass and coastal bermudagrass are promising lignocellulosic feedstocks for bioethanol production. However, pretreatment of lignocelluloses is required to improve production of fermentable sugars from enzymatic hydrolysis. Microwave‐based alkali pretreatment of switchgrass and coastal bermudagrass was investigated in this study. Pretreatments were carried out by immersing the biomass in dilute alkali reagents and exposing the slurry to microwave radiation at 250 W for residence times ranging from 5 to 20 min. Simons' stain method was used to quantify changes in biomass porosity as a result of the pretreatment. Pretreatments were evaluated based on yields of total reducing sugars, glucose, and xylose. An evaluation of different alkalis identified sodium hydroxide as the most effective alkali reagent for microwave‐based pretreatment of switchgrass and coastal bermudagrass. 82% glucose and 63% xylose yields were achieved for switchgrass and 87% glucose and 59% xylose yields were achieved for coastal bermudagrass following enzymatic hydrolysis of biomass pretreated under optimal conditions. Dielectric properties for dilute sodium hydroxide solutions were measured and compared with solid losses, lignin reduction, and reducing sugar levels in hydrolyzates. Results indicate that dielectric loss tangent of alkali solutions is a potential indicator of the severity of microwave‐based pretreatments. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Bioconversion of Lignocellulose into Bioethanol: Process Intensification and Mechanism Research

Biofuels produced from lignocellulosic biomass can significantly reduce the energy dependency on fossil fuels and the resulting effects on environment. In this respect, cellulosic ethanol as an alternative fuel has the potential to become a viable energy source in the near future. Over the past few decades, tremendous effort has been undertaken to make cellulosic ethanol cost competitive with conventional fossil fuels. The pretreatment step is always necessary to deconstruct the recalcitrant structures and to make cellulose more accessible to enzymes. A large number of pretreatment technologies involving physical, chemical, biological, and combined approaches have been developed and tested at the pilot scale. Furthermore, various strategies and methods, including multi-enzyme complex, non-catalytic additives, enzyme recycling, high solids operation, design of novel bioreactors, and strain improvement have also been implemented to improve the efficiency of subsequent enzymatic hydrolysis and fermentation. These technologies provide significant opportunities for lower total cost, thus making large-scale production of cellulosic ethanol possible. Meanwhile, many researchers have focused on the key factors that limit cellulose hydrolysis, and analyzing the reaction mechanisms of cellulase. This review describes the most recent advances on process intensification and mechanism research of pretreatment, enzymatic hydrolysis, and fermentation during the production of cellulosic ethanol.     

Conversion of rice straw to bio-based chemicals: an integrated process using Lactobacillus brevis

Commercialization of lignocellulosic biomass as a feedstock for bio-based chemical production is problematic due to the high processing costs of pretreatment and saccharifying enzymes combined with low product yields. Such low product yield can be attributed, in large part, to the incomplete utilization of the various carbohydrate sugars found in the lignocellulosic biomass. In this study, we demonstrate that Lactobacillus brevis is able to simultaneously metabolize all fermentable carbohydrates in acid pre-processed rice straw hydrolysate, thereby allowing complete utilization of all released sugars. Inhibitors present in rice straw hydrolysate did not affect lactic acid production. Moreover, the activity of exogenously added cellulases was not reduced in the presence of growing cultures of L. brevis. These factors enabled the use of L. brevis in a process termed simultaneous saccharification and mixed sugar fermentation (SSMSF). In SSMSF with L. brevis, sugars present in rice straw hydrolysate were completely utilized while the cellulase maintained its maximum activity due to the lack of feedback inhibition from glucose and/or cellobiose. By comparison to a sequential hydrolysis and fermentation process, SSMSF reduced operation time and the amount of cellulase enzyme necessary to produce the same amount of lactic acid.     

Evaluation of fermentative potential of <Emphasis Type="Italic">Kluyveromyces marxianus</Emphasis> ATCC 36907 in cellulosic and hemicellulosic sugarcane bagasse hydrolysates on xylitol and ethanol production

This paper evaluates the fermentative potential of Kluyveromyces marxianus grown in sugarcane bagasse cellulosic and hemicellulosic hydrolysates obtained by acid hydrolysis. Ethanol was obtained from a single glucose fermentation product, whereas xylose assimilation resulted in xylitol as the main product and ethanol as a by-product derived from the metabolism of this pentose. Fermentation performed in a simulated hydrolysate medium with a glucose concentration similar to that of the hydrolysate resulted in ethanol productivity (Qp?=?0.86 g L^?1 h^?1) that was tenfold higher than the one observed in the cellulosic hydrolysate. However, the use of hemicellulosic hydrolysate favored xylose assimilation in comparison with simulated medium with xylose and glucose concentrations similar to those found in this hydrolysate, without toxic compounds such as acetic acid and phenols. Under this condition, xylitol yield was 53.8 % higher in relation to simulated medium. Thus, the total removal of toxic compounds from the hydrolysate is not necessary to obtain bioproducts from lignocellulosic hydrolysates.     

Improved one‐step steam pretreatment of SO2‐Impregnated softwood with time‐dependent temperature profile for ethanol production

In the production of ethanol from lignocellulosic material, pretreatment of the raw material before enzymatic hydrolysis and fermentation is essential to obtain high overall yields of sugar and ethanol. Two‐step steam pretreatment results in higher ethanol yields from softwood than the standard one‐step pretreatment process. However, the difficulty with separation and washing of the material at high pressure between the two pretreatment steps is a major drawback. In this study, a new one‐step pretreatment procedure was investigated, in which the time‐temperature profile was varied during pretreatment. The efficiency of pretreatment was assessed by performing simultaneous saccharification and fermentation on the pretreated slurries. Pretreatment of SO₂‐impregnated softwood performed by varying the temperature (190–226°C), the residence time (5–10 min), and the mode of temperature increase (linear or stepwise), resulted in recovery of about 90% of the mannose and glucose present in the raw material. The highest ethanol yield, 75% of theoretical based on the glucan and mannan content of the raw material, was obtained at pretreatment conditions of 190°C for 12 min. Similar ethanol yields were achieved when running the pretreatment as one‐step (190–200°C), two levels of temperature, at shorter residence time (7 min), which results in lower capital costs for the process. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010