Onsite bio-detoxification of steam-exploded corn stover for cellulosic ethanol production

In the process of ethanol production from steam-exploded corn stover (SECS), a cellulose-degradation strain of Aspergillus nidulans (FLZ10) was investigated whether it could remove the inhibitors released from steam exploded pretreatment , and thereby be used for biological detoxification on Saccharomycescerevisiae. The results showed that FLZ10 removed 75.2% formic acid, 53.6% acetic acid, and 100% hydroxymethyl furfural (5-HMF) and furfural from the hydrolysate washed from SECS after 72 h cultivation. A cellulase activity of 0.49 IU/ml was simultaneously produced while the biological detoxification occurred. An ethanol yield of 0.45 g/g on glucose was obtained in the hydrolysate biodetoxified by FLZ10. The glucose consumption rate of FLZ10 was much lower than that of S. cerevisiae, thereby it had little competition with S. cerevisiae on glucose consumption. Based on SECS to ethanol mass balance analysis, with the onsite bio-detoxification, fermentation using S. cerevisiae effectively converted monomeric glucose with 94.4% ethanol yield.     

秸秆汽爆炼制木质素制备酚醛泡沫材料

以木质纤维素为原料的生物炼制不仅需要考虑到纤维素和半纤维素,同时也需要考虑到木质素的利用,以提高木质纤维素炼制的整体经济效益。利用汽爆处理的玉米秸秆为原料,通过优化碱提取的温度以及碱浓度,获得木质素得率较高,糖含量较少的提取液,同时该提取液经过浓缩后直接部分替代苯酚与甲醛反应制备酚醛泡沫。结果表明:当提取温度120℃,碱浓度1%,固液比1∶10,提取时间2 h,木质素的提取率达到79.67%。由该提取液替代苯酚制备的酚醛泡沫随着替代率的增加,其泡沫的密度逐渐增加,其压缩强度相对于纯酚醛泡沫也得到了提高。而木质素的加入并没有显著影响其热导率以及阻燃性能,同时由于其利用较为低廉的可再生资源木质素替代不可再生的苯酚原料,成本低,环保性好,具有更好的市场应用前景。采用木质素提液直接制备酚醛泡沫材料,工艺流程简单;增加了副产物木质素的经济价值,提高了整个木质纤维素炼制的经济性。     

Laccase-initiated cross-linking of lignocellulose fibres using a ultra-filtered lignin isolated from kraft black liquor

In this work, the effect of Trametes pubescens laccase (TpL) used in combination with a low-molecular-weight ultra-filtered lignin (UFL) to improve mechanical properties of kraft liner pulp and chemi-thermo-mechanical pulp was studied. UFL was isolated by ultra-filtration from the kraft cooking black liquor obtained from softwood pulping. This by-product from the pulp industry contains an oligomeric lignin with almost twice the amount of free phenolic moieties than residual kraft pulp lignin. The reactivity of TpL on UFL and kraft pulp was studied by nuclear magnetic resonance spectroscopy and size exclusion chromatography. Laccase was shown to polymerise UFL and residual kraft pulp lignin in the fibres, seen by the increase in their average molecular weight and in the case of UFL as a decrease in the amount of phenolic hydroxyls. The laccase initiated cross-linking of lignin, mediated by UFL, which gives rise to more than a twofold increase in wet strength of kraft liner pulp handsheets without loosing other critical mechanical properties. Hence, this could be an interesting path to decrease mechano-sorptive creep that has been reported to lessen in extent as wet strength is given to papers. The laccase/2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) mediator system showed a greater increase in wet tensile strength of the resulting pulp sheets than the laccase/UFL system. However, other mechanical properties such as dry tensile strength, compression strength and Scott Bond internal strength were negatively affected by the laccase/ABTS system.     

The influence of lignin migration and relocation during steam pretreatment on the enzymatic hydrolysis of softwood and corn stover biomass substrates

To be effective, steam pretreatment is typically carried out at temperatures/pressures above the glass transition point (Tg) of biomass lignin so that it can partly fluidize and relocate. The relocation of Douglas-fir and corn stover derived lignin was compared with the expectation that, with the corn stover lignin's lower hydrophobicity and molecular weight, it would be more readily fluidized. It was apparent that the Tg of lignin decreased as the moisture increased, with the easier access of steam to the corn stover lignin promoting its plasticization. Although the softwood lignin was more recalcitrant, when it was incorporated onto filter paper, it too could be plasticized, with its relocation enhancing enzymatic hydrolysis. When lignin recondensation was minimized, the increased hydrophobicity suppressed lignin relocation. It was apparent that differences in the accessibility of the lignin present in Douglas-fir and corn stover to steam significantly impacted lignin fluidization, relocation, and subsequent cellulose hydrolysis.     

High solid simultaneous saccharification and fermentation of wet oxidized corn stover to ethanol

In this study ethanol was produced from corn stover pretreated by alkaline and acidic wet oxidation (WO) (195 degrees C, 15 min, 12 bar oxygen) followed by nonisothermal simultaneous saccharification and fermentation (SSF). In the first step of the SSF, small amounts of cellulases were added at 50 degrees C, the optimal temperature of enzymes, in order to obtain better mixing condition due to some liquefaction. In the second step more cellulases were added in combination with dried baker's yeast (Saccharomyces cerevisiae) at 30 degrees C. The phenols (0.4-0.5 g/L) and carboxylic acids (4.6-5.9 g/L) were present in the hemicellulose rich hydrolyzate at subinhibitory levels, thus no detoxification was needed prior to SSF of the whole slurry. Based on the cellulose available in the WO corn stover 83% of the theoretical ethanol yield was obtained under optimized SSF conditions. This was achieved with a substrate concentration of 12% dry matter (DM) acidic WO corn stover at 30 FPU/g DM (43.5 FPU/g cellulose) enzyme loading. Even with 20 and 15 FPU/g DM (corresponding to 29 and 22 FPU/g cellulose) enzyme loading, ethanol yields of 76 and 73%, respectively, were obtained. After 120 h of SSF the highest ethanol concentration of 52 g/L (6 vol.%) was achieved, which exceeds the technical and economical limit of the industrial-scale alcohol distillation. The SSF results showed that the cellulose in pretreated corn stover can be efficiently fermented to ethanol with up to 15% DM concentration. A further increase of substrate concentration reduced the ethanol yield significant as a result of insufficient mass transfer. It was also shown that the fermentation could be followed with an easy monitoring system based on the weight loss of the produced CO2.     

Simultaneous saccharification and ethanol fermentation at high corn stover solids loading in a helical stirring bioreactor

The higher ethanol titer inevitably requires higher solids loading during the simultaneous enzymatic saccharification and fermentation (SSF) using lignocellulose as the feedstock. The mixing between the solid lignocellulose and the liquid enzyme is crucially important. In this study, a bioreactor with a novel helical impeller was designed and applied to the SSF operation of the steam explosion pretreated corn stover under different solids loadings and different enzyme dosages. The performances using the helical impeller and the common Rushton impeller were compared and analyzed by measuring rheological properties and the mixing energy consumption. The results showed that the new designed stirring system had better performances in the saccharification yield, ethanol titer, and energy cost than those of the Rushton impeller stirring. The mixing energy consumption under different solids loadings and enzyme dosages during SSF operation were analyzed and compared to the thermal energy in the ethanol produced. A balance for achieving the optimal energy cost between the increased mixing energy cost and the reduced distillation energy cost at the high solids loading should be made. The potentials of the new bioreactor were tested under various SSF conditions for obtaining optimal ethanol yield and titer. Biotechnol. Bioeng. 2010. 105: 718–728. © 2009 Wiley Periodicals, Inc.     

Effect of xylan and lignin removal by batch and flowthrough pretreatment on the enzymatic digestibility of corn stover cellulose

Compared with batch systems, flowthrough and countercurrent reactors have important potential advantages for pretreating cellulosic biomass, including higher hemicellulose sugar yields, enhanced cellulose digestibility, and reduced chemical additions. Unfortunately, they suffer from high water and energy use. To better understand these trade-offs, comparative data are reported on xylan and lignin removal and enzymatic digestibility of cellulose for corn stover pretreated in batch and flowthrough reactors over a range of flow rates between 160 degrees and 220 degrees C, with water only and also with 0.1 wt% sulfuric acid. Increasing flow with just water enhanced the xylan dissolution rate, more than doubled total lignin removal, and increased cellulose digestibility. Furthermore, adding dilute sulfuric acid increased the rate of xylan removal for both batch and flowthrough systems. Interestingly, adding acid also increased the lignin removal rate with flow, but less lignin was left in solution when acid was added in batch. Although the enzymatic hydrolysis of pretreated cellulose was related to xylan removal, as others have shown, the digestibility was much better for flowthrough compared with batch systems, for the same degree of xylan removal. Cellulose digestibility for flowthrough reactors was related to lignin removal as well. These results suggest that altering lignin also affects the enzymatic digestibility of corn stover.     

An economic comparison of different fermentation configurations to convert corn stover to ethanol using Z. mobilis and Saccharomyces

Numerous routes are being explored to lower the cost of cellulosic ethanol production and enable large‐scale production. One critical area is the development of robust cofermentative organisms to convert the multiple, mixed sugars found in biomass feedstocks to ethanol at high yields and titers without the need for processing to remove inhibitors. Until such microorganisms are commercialized, the challenge is to design processes that exploit the current microorganisms' strengths. This study explored various process configurations tailored to take advantage of the specific capabilities of three microorganisms, Z. mobilis 8b, S. cerevisiae, and S. pastorianus. A technoeconomic study, based on bench‐scale experimental data generated by integrated process testing, was completed to understand the resulting costs of the different process configurations. The configurations included whole slurry fermentation with a coculture, and separate cellulose simultaneous saccharification and fermentation (SSF) and xylose fermentations with none, some or all of the water to the SSF replaced with the fermented liquor from the xylose fermentation. The difference between the highest and lowest ethanol cost for the different experimental process configurations studied was $0.27 per gallon ethanol. Separate fermentation of solid and liquor streams with recycle of fermented liquor to dilute the solids gave the lowest ethanol cost, primarily because this option achieved the highest concentrations of ethanol after fermentation. Further studies, using methods similar to ones employed here, can help understand and improve the performance and hence the economics of integrated processes involving enzymes and fermentative microorganisms. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Production of bioethanol, methane and heat from sugarcane bagasse in a biorefinery concept

The potential of biogas production from the residues of second generation bioethanol production was investigated taking into consideration two types of pretreatment: lime or alkaline hydrogen peroxide. Bagasse was pretreated, enzymatically hydrolyzed and the wastes from pretreatment and hydrolysis were used to produce biogas. Results have shown that if pretreatment is carried out at a bagasse concentration of 4% DM, the highest global methane production is obtained with the peroxide pretreatment: 72.1 L methane/kg bagasse. The recovery of lignin from the peroxide pretreatment liquor was also the highest, 112.7 ± 0.01 g/kg of bagasse. Evaluation of four different biofuel production scenarios has shown that 63-65% of the energy that would be produced by bagasse incineration can be recovered by combining ethanol production with the combustion of lignin and hydrolysis residues, along with the anaerobic digestion of pretreatment liquors, while only 32-33% of the energy is recovered by bioethanol production alone.     

玉米秸秆基纤维素乙醇生命周期能耗与温室气体排放分析

生命周期评价是目前分析产品或工艺的环境负荷唯一标准化工具,利用其生命周期分析方法可以有效地研究纤维素乙醇生命周期能耗与温室气体排放问题。为了定量解释以玉米秸秆为原料的纤维素乙醇的节能和温室气体减排潜力,利用生命周期分析方法对以稀酸预处理、酶水解法生产的玉米秸秆基乙醇进行了生命周期能耗与温室气体排放分析,以汽车行驶1 km为功能单位。结果表明：与汽油相比,纤维素乙醇E100 (100％乙醇) 和E10 (乙醇和汽油体积比=1∶9) 生命周期化石能耗分别减少79.63％和6.25％,温室气体排放分别减少53.98％和6.69％;生物质阶段化石能耗占到总化石能耗68.3％,其中氮肥和柴油的生命周期能耗贡献最大,分别占到生物质阶段的45.78％和33.26％;工厂电力生产过程的生命周期温室气体排放最多,占净温室气体排放量的42.06％,提升技术减少排放是降低净排放的有效措施。     

Ethanolic fermentation of hydrolysates from ammonia fiber expansion (AFEX) treated corn stover and distillers grain without detoxification and external nutrient supplementation

External nutrient supplementation and detoxification of hydrolysate significantly increase the production cost of cellulosic ethanol. In this study, we investigated the feasibility of fermenting cellulosic hydrolysates without washing, detoxification or external nutrient supplementation using ethanologens Escherichia coli KO11 and the adapted strain ML01 at low initial cell density (16 mg dry weight/L). The cellulosic hydrolysates were derived from enzymatically digested ammonia fiber expansion (AFEX)-treated corn stover and dry distiller's grain and solubles (DDGS) at high solids loading (18% by weight). The adaptation was achieved through selective evolution of KO11 on hydrolysate from AFEX-treated corn stover. All cellulosic hydrolysates tested (36-52 g/L glucose) were fermentable. Regardless of strains, metabolic ethanol yields were near the theoretical limit (0.51 g ethanol/g consumed sugar). Volumetric ethanol productivity of 1.2 g/h/L was achieved in fermentation on DDGS hydrolysate and DDGS improved the fermentability of hydrolysate from corn stover. However, enzymatic hydrolysis and xylose utilization during fermentation were the bottlenecks for ethanol production from corn stover at these experimental conditions. In conclusion, fermentation under the baseline conditions was feasible. Utilization of nutrient-rich feedstocks such as DDGS in fermentation can replace expensive media supplementation.     

Xanthan gum and ethanol production by Xanthomonas campestris and Zymomonas mobilis from peach pulp

The wild type of Xanthomonas campestris and a mutant strain of Zymomonas mobilis CP4, tolerant to sucrose up to 40% (w/v), were used to produce either xanthan gum or ethanol, respectively, from peach pulp supplemented with different salts. Both bacteria grew well (2.7 mg/ml for X. campestris and 1.45 mg/ml for Z. mobilis) in fine peach pulp and the production of xanthan gum or ethanol was 0.1–0.2 g/l or 110 g/l, respectively.     

响应面法优化玉米秸秆同步酶解发酵产乙醇条件

对汽爆玉米秸秆同步酶解发酵生产乙醇的条件进行优化。首先利用Fractional Factorial设计法对影响乙醇产量的7个因素进行评价,筛选出具有显著效应的3个因素,即反应温度、酶添加量、总反应时间,再以Box—Behnken设计法及响应面分析法确定主要因素的最佳水平,即反应温度37℃,每g纤维素添加纤维素酶32u,反应时间87h,此时乙醇体积分数达到3．69％。新工艺条件实验结果表明,乙醇体积分数在87h可达到3．76％,和原工艺相比,反应时间缩短了9h,乙醇体积分数提高了13％。     

Effect of crude protein concentration and sugar-beet pulp on nutrient digestibility, nitrogen excretion, intestinal fermentation and manure ammonia and odour emissions from finisher pigs

A 2 × 2 factorial experiment was conducted to investigate the interaction between high and low dietary crude protein (CP) (200 v. 150 g/kg) and sugar-beet pulp (SBP) (200 v. 0 g/kg) on nutrient digestibility, nitrogen (N) excretion, intestinal fermentation and manure ammonia and odour emissions from 24 boars (n = 6, 74.0 kg live weight). The diets were formulated to contain similar concentrations of digestible energy (13.6 MJ/kg) and lysine (10.0 g/kg). Pigs offered SBP-containing diets had a reduced (P < 0.05) digestibility of dry matter, ash, N, gross energy and an increased (P < 0.001) digestibility of neutral-detergent fibre compared with pigs offered diets containing no SBP. There was an interaction between CP and SBP on urinary N excretion and the urine : faeces N ratio. Pigs offered the 200 g/kg CP SBP-based diet had reduced urine : faeces N ratio (P < 0.05) and urinary N excretion (P < 0.05) compared with those offered the 200 g/kg CP diet without SBP. However, there was no effect of SBP in pigs offered 150 g/kg CP diets. Manure ammonia emissions were reduced by 33% from 0 to 240 h (P < 0.01); however, odour emissions were increased by 41% (P < 0.05) when pigs were offered SBP diets. Decreasing dietary CP to 150 g/kg reduced total N excretion (P < 0.001) and ammonia emissions from 0 to 240 h (P < 0.05). There was an interaction between dietary CP and SBP on branched-chain fatty acids (P < 0.001) in caecal digesta. Pigs offered the 200 g/kg CP SBP-containing diet reduced branched-chain fatty acids in the caecum compared with pigs offered the 200 g/kg CP diet containing no SBP. However, there was no effect of SBP in the 150 g/kg CP diet. In conclusion, pigs offered SBP-containing diets had a reduced manure ammonia emissions and increased odour emissions compared with diets containing no SBP. Pigs offered the 200 g/kg CP SBP-containing diet had a reduced urine : faeces N ratio and urinary N excretion compared with those offered the 200 g/kg CP diet containing no SBP.     

Acetic acid removal from corn stover hydrolysate using ethyl acetate and the impact on Saccharomyces cerevisiae bioethanol fermentation

Acetic acid is introduced into cellulose conversion processes as a consequence of composition of lignocellulose feedstocks, causing significant inhibition of adapted, genetically modified and wild‐type S. cerevisiae in bioethanol fermentation. While adaptation or modification of yeast may reduce inhibition, the most effective approach is to remove the acetic acid prior to fermentation. This work addresses liquid–liquid extraction of acetic acid from biomass hydrolysate through a pathway that mitigates acetic acid inhibition while avoiding the negative effects of the extractant, which itself may exhibit inhibition. Candidate solvents were selected using simulation results from Aspen Plus?, based on their ability to extract acetic acid which was confirmed by experimentation. All solvents showed varying degrees of toxicity toward yeast, but the relative volatility of ethyl acetate enabled its use as simple vacuum evaporation could reduce small concentrations of aqueous ethyl acetate to minimally inhibitory levels. The toxicity threshold of ethyl acetate, in the presence of acetic acid, was found to be 10 g L^?1. The fermentation was enhanced by extracting 90% of the acetic acid using ethyl acetate, followed by vacuum evaporation to remove 88% removal of residual ethyl acetate along with 10% of the broth. NRRL Y‐1546 yeast was used to demonstrate a 13% increase in concentration, 14% in ethanol specific production rate, and 11% ethanol yield. This study demonstrated that extraction of acetic acid with ethyl acetate followed by evaporative removal of ethyl acetate from the raffinate phase has potential to significantly enhance ethanol fermentation in a corn stover bioethanol facility. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:929–937, 2016     

Characterization of sugars from model and enzyme-mediated pulp hydrolyzates using high-performance liquid chromatography coupled to evaporative light scattering detection

High-performance liquid chromatography (HPLC) coupled to an evaporative light scattering detector was used to quantitatively determine glucose and cellobiose in hydrolyzates from the production of cellulose nanofillers from modified lignocellulosic materials. Prevail Carbohydrate ES 5 μ column proved more suitable for achieving the chromatographic separation of the model pulp hydrolyzate into its constituent sugars than the YMC-Pack Polyamine column. Linear calibration curves for the various sugars in the mixtures were developed. Glucose and cellobiose were clearly detectable in pulp hydrolyzates obtained from enzyme-mediated hydrolysis of recycled pulp, pine and hardwood dissolving pulps. Finally, the amount of glucose in the pulp hydrolyzates was generally higher than cellobiose.     

Remote engineering for a cheese whey biorefinery: an Internet-based application for process design,economic analysis,monitoring, and control of multiple plant sites

The proteolysis of cheese whey with the aid of immobilized enzymes is an attractive alternative for this by-product of the dairy industry. Among some possible applications for whey protein hydrolysates, one may cite their use as protein source for individuals with reduced capacity of digestion, or with genetic metabolic disorders (phenylketonuria patients, for instance). The multipurpose plant that processes whey is named here as a cheese whey biorefinery. This work presents the remote control and monitoring of the whey biorefineries using the Internet. In an integrated environment, the web application also enables simulation and economic analyses of the process. This technology might allow small companies to access a remote “engineering centre”, with know-how on plant design and advanced control techniques. The idea can also be extended to large dairy companies, providing the remote control of geographically spread sites of production.     

从源头防治制浆造纸废水污染的新策略

制浆造纸工业废水对环境造成了严重污染。本文概述了微生物生物技术在防治造纸污染中的应用,同时着重介绍了木质素生物合成途径与转基因植物降低造纸原料木质素含量的研究进展,对我国发展从源头防治制浆造纸污染的策略提出了建议     

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