甘蔗渣的不同预处理方法比较及其酶水解的类分形动力学

分别采用NaOH（0.45 mol/L aq.）、HCl（0.034 mol/L aq.）和高温液态水（LHW）三种方法对甘蔗渣进行预处理,并对其组分变化和酶解效果进行了比较。NaOH预处理方法获得最高的木质素去除率,达91.1%,糖损失率达23.5%;HCl和LHW预处理结果类似,木聚糖溶解率分别为85.2%和79.7%,糖损失率均约为15%,木质素去除率均小于16%。三种方法处理的甘蔗渣经纤维素酶水解后得到总单糖（葡萄糖 + 木糖）浓度分别为38.7 g/L（NaOH）、16.1 g/L（HCl）和15.6 g/L（LHW）。综合比较预处理和酶水解工艺,NaOH水溶液预处理法的糖回收率最高,其次为HCl水溶液预处理法,LHW预处理法的糖回收率最低。作为描述纤维素酶反应动力学的有力工具,类分形理论的研究表明,各种预处理后底物的不规则性依次为：HCl＞LHW＞NaOH,其与酶的有效吸附大小依次为：NaOH＞HCl＞LHW。     

渗滤与间歇高温液态水预处理甘蔗渣的对比研究

以甘蔗渣为原料,采用自主搭建的连续渗滤试验台,考察反应温度、反应时间、反应液流量对甘蔗渣水解情况的影响。实验发现水解液中木聚糖主要以低聚木糖的形式存在,其所占总木糖的比例主要与反应温度和反应液流量相关,高处理温度、低流量易于生成木糖并进而生成副产物,说明木聚糖在高温液态水中的水解路径为:木聚糖—低聚木糖—木糖—糠醛等。通过实验确定180℃是最合适的反应温度,反应液流量为30 m L/min,15 min获得具有4.17 g/L总木糖的水解液,此时总木糖收率可达93.95%。与间歇搅拌反应形式中的水解情况进行对比分析发现,在相同处理效果的前提下连续渗滤反应形式耗水量更大,但渗滤反应形式可获得更高的木糖收率,残渣酶解率、总糖收率均高于间歇法,副产物生成量也低于间歇反应形式。     

二倍体酿酒酵母表面展示半纤维素酶进行木质纤维素乙醇发酵的研究

拟构建一株可利用木糖产乙醇的二倍体酿酒酵母Y5,并通过a凝集素表面展示系统实现β-D-1,4内切木聚糖酶(XYNII)、β-D-1,4外切木糖苷酶(Xyl A)表面固定化。通过对菌株免疫荧光验证及酶催化活性的测定,证明融合蛋白以活性形式固定在细胞表面。以木聚糖为底物进行乙醇发酵实验,96 h时最大乙醇产量达0.62 g/L,乙醇产率为0.19 g/g,相当于理论值的37.3%。结果证明表面展示半纤维素酶的酿酒酵母可自主降解木聚糖生成木糖,并且利用木糖生产乙醇。     

诱变选育拜氏梭菌利用碱处理木糖渣水解液生产丁醇

试验以拜氏梭菌(Clostridium beijerinckii)NCIMB 8052为出发菌株,采用EMS诱变,并利用含淀粉、2-脱氧-D-葡萄糖的固体培养基对诱变菌株进行多次筛选,最终获得一株诱变菌株Y-3,其在葡萄糖培养基中总溶剂产量为15.8 g/L,丁醇产量为9.4 g/L,较出发菌株分别提高了30.6%和40.3%。利用以木糖渣为底物生产的纤维素酶对经2%氢氧化钠预处理后的木糖渣进行水解,所得的水解液作为Y-3丁醇发酵培养基中的碳源,总溶剂和丁醇产量分别达到16.3 g/L和8.8 g/L。结果表明,诱变菌株Y-3可利用碱处理木糖渣水解液作为发酵底物生产丁醇,且丁醇产量较高,适合于生物质发酵,大大降低了生产成本。     

高温液相水耦合湿磨预处理酶水解玉米秸秆的研究

以酶水解玉米秸秆获得高葡萄糖收率为目标,通过实验研究高温液相水耦合湿法打磨预处理技术对葡萄糖产率的影响。研究结果表明:将200℃,40 min条件下高温液相水预处理的玉米秸秆再湿磨预处理,经15 FPU/g的纤维素酶水解得到的葡萄糖收率高达83.1%。同时,以5 FPU/g的纤维素酶处理时湿磨效果更为明显,葡萄糖收率由湿磨前的58.4%上升至湿磨后的70.7%。     

纤维素温和条件下一步水热法制备甲烷的研究

生物质发酵法制备甲烷存在甲烷收率低、CO₂含量高等问题。本研究以纤维素为原料,在温和条件下采用水热催化转化的方法制备甲烷。对一系列催化剂进行了考察,发现Ru/C对该反应的催化活性最高。采用Ru/C催化剂进一步考察了一系列反应条件,结果表明,升高反应温度、延长反应时间、增加催化剂用量以及提高氢气初始压力对甲烷的生成具有促进作用。在1 MPa H₂、220℃、12 h反应条件下,甲烷碳摩尔收率最高,达88%,反应过程中无CO₂产生。采用TEM、BET、XRD和FT-IR等对催化剂进行了表征,结果表明,Ru/C催化剂的高催化活性可能与催化剂本身比表面积大、钌粒子颗粒小且分散均匀的特性有关。本研究采用的催化转化方法具有甲烷收率高、CO₂排放量小（<5%）、反应条件更为温和等特点。     

联合CO2捕集的生物质化学链气化制备合成气系统分析

本文提出以Fe₂O₃为载氧体、以CaO捕集CO₂的生物质化学链气化系统,利用Aspen Plus软件对该系统进行了模拟,以合成气组成（干基）、合成气氢碳比、含碳产物的碳摩尔分布、冷气效率及收率等为系统性能评价指标,重点分析了燃料反应器温度（T_FR）、载氧体Fe₂O₃与生物质碳摩尔比（Fe₂O₃/C）、水蒸气与生物质碳摩尔比（Steam/C）、CaO与生物质碳摩尔比（CaO/C）等系统参数对固体生物质化学链气化系统的影响。结果表明,在T_FR = 825℃、Fe₂O₃/C = 0.5、Steam/C = 0.71和CaO/C = 0.26条件下,合成气制备系统性能较优,合成气中H₂和CO₂含量分别为55.2%和15.4%,氢碳比为1.93,冷气效率为78.2%,被CaCO₃捕集的生物质碳为18.2%,收率（湿气基）为1.95 Nm³/kg_biomass,其中合成气中H₂和CO收率为1.24 Nm³/kg_biomass。     

酶酸联合水解玉米秸秆的实验研究

开发了一种通过酸酶联合水解处理玉米秸秆以得到可发酵单糖的工艺方法,进行了稀硫酸预处理玉米秸秆的研究。得到最佳的工艺条件,木糖收率达到84.90%。用纤维素酶水解酸处理过的玉米秸秆,考察了pH值、温度、时间对酶水解率的影响,结果表明:酶解温度为50℃,pH值为4.8,水解时间为60h时,酶水解率达到91.71%。该工艺达到了节能高效地转化玉米秸秆为可发酵单糖的目的。     

高温液态水处理杂交狼尾草提高总糖收率

以杂交狼尾草为原料,采用高温液态水预处理方法研究其对能源草酶解效果的影响。研究结果表明:180℃,40 min,固液比1∶20,饱和蒸汽压的水解条件为最优预处理条件,此时总木糖收率为98.09%。高温液态水可降解87.99%的半纤维素和41.28%的木质素;破坏原本平整的表面,使更多的纤维素裸露,使纤维素结晶指数增大,破坏纤维素、木质素中的化学基团(如分子内与分子间氢键等),这些变化均有利于后续酶解。当加酶量为40 FPU/g,杂交狼尾草72 h时的酶解率由62.50%提高到99.36%,高温液态水和酶解后总糖收率达到90.40%。     

碱改性HZSM-5热解生物质模型化合物影响研究

实验采用Py-GC/MS在500 ℃下对NaOH、Na₂CO₃和有机碱（CTAB/TPAOH）改性HZSM-5催化热解生物质模型化合物的产物分布影响机制进行探究。结果表明,利用0.1 mol/L NaOH/Na₂CO₃改性HZSM-5使热解油中小分子酮、酚和酯类物质的收率有所提高,有利于碳链长度≥5产物（C_≥5）的生成;0.2 mol/L NaOH/Na₂CO₃改性HZSM-5催化剂有助于脱羰和脱羟基反应的进行,促使环状化合物开裂转化为链状化合物。TPAOH的加入使NaOH改性HZSM-5催化热解产物中酮类产物收率降至18.56%、醛类产物收率增至3.01%,并促使C_≥9产物向C_≤4转化,链状产物增加;经CTAB改性后C_≥9产物向C_5-8转化,环状产物增加。     

Two-step steam pretreatment of softwood by dilute H2SO4 impregnation for ethanol production

Fuel ethanol can be produced from softwood through hydrolysis in an enzymatic process. Prior to enzymatic hydrolysis of the softwood, pretreatment is necessary. In this study two-step steam pretreatment by dilute H₂SO₄ impregnation to improve the overall sugar and ethanol yield has been investigated. The first pretreatment step was performed under conditions of low severity (180°C, 10 min, 0.5% H₂SO₄) to optimise the amount of hydrolysed hemicellulose. In the second step the washed solid material from the first pretreatment step was impregnated again with H₂SO₄ and pretreated under conditions of higher severity to hydrolyse a portion of the cellulose, and to make the cellulose more accessible to enzymatic attack. A wide range of conditions was used to determine the most favourable combination. The temperatures investigated were between 180°C and 220°C, the residence times were 2, 5 and 10 min and the concentrations of H₂SO₄ were 1% and 2%.

The effects of pretreatment were assessed by both enzymatic hydrolysis of the solids and with simultaneous saccharification and fermentation (SSF) of the whole slurry, after the second pretreatment step. For each set of pretreatment conditions the liquid fraction was fermented to determine any inhibiting effects. The ethanol yield using the SSF configuration reached 65% of the theoretical value while the sugar yield using the SHF configuration reached 77%. Maximum yields were obtained when the second pretreatment step was performed at 200°C for 2 min with 2% H₂SO₄. This form of two-step steam pretreatment is a promising method of increasing the overall yield in the wood-to-ethanol process.     

改进的柳枝稷预处理方法及乙醇发酵研究

为了提高柳枝稷中纤维素和半纤维素糖的转化率,降低水解液中抑制剂的浓度,首先,用稀酸在温和条件下对柳枝稷进行水解,然后用碱对酸水解后的固体物进行预处理,接着用纤维素酶酶解并分别对稀酸水解液和酶解液进行乙醇发酵.结果表明:纤维素转化率达到94.26%,半纤维素转化率为60.93%,稀酸水解液乙醇发酵的乙醇产率为0.441g乙醇/g糖,达到最高理论值的86.47%.酶解液乙醇发酵的乙醇产率为0.486g乙醇/g葡萄糖,达到最高理论值的95.29%.     

Construction and demolition lignocellulosic wastes to bioethanol

This work deals with conversion of four construction and demolition (C&D) lignocellulosic wastes including OSB, chipboard, plywood, and wallpaper to ethanol by separate enzymatic hydrolysis and fermentation (SHF). Similar to other lignocelluloses, the wastes were resistant to the enzymatic hydrolysis, in which only up to 7% of their cellulose was hydrolyzed. Therefore, the lignocellulosic wastes were treated with phosphoric acid, sodium hydroxide, or N-methylmorpholine-N-oxide (NMMO), which resulted in improving the subsequent enzymatic hydrolysis to 38.2–94.6% of the theoretical yield. The best performance was obtained after pretreatment by concentrated phosphoric acid, followed by NMMO. The pretreated and hydrolyzed C&D wastes were then successfully fermented by baker’s yeast to ethanol with 70.5–84.2% of the theoretical yields. The results indicate the possibility of producing 160 ml ethanol from each kg of the C&D wastes at the best conditions.     

Pretreatment of Miscanthus for hydrogen production by Thermotoga elfii

Pretreatment methods for the production of fermentable substrates from Miscanthus, a lignocellulosic biomass, were investigated. Results demonstrated an inverse relationship between lignin content and the efficiency of enzymatic hydrolysis of polysaccharides. High delignification values were obtained by the combination of mechanical, i.e. extrusion or milling, and chemical pretreatment (sodium hydroxide). An optimized process consisted of a one-step extrusion-NaOH pretreatment at moderate temperature (70°C). A mass balance of this process in combination with enzymatic hydrolysis showed the following: pretreatment resulted in 77% delignification, a cellulose yield of more than 95% and 44% hydrolysis of hemicellulose. After enzymatic hydrolysis 69% and 38% of the initial cellulose and hemicellulose fraction, respectively, was converted into glucose, xylose and arabinose. Of the initial biomass, 33% was converted into monosaccharides. Normal growth of Thermotoga elfii on hydrolysate was observed and high amounts of hydrogen were produced.     

Empty fruit bunches from oil palm as a potential raw material for fuel ethanol production

In this paper the fuel ethanol production from empty fruit bunches was experimentally evaluated using alkaline pretreatment and enzymatic hydrolysis for sugars release. Fermentation was accomplished using a native Saccharomyces cerevisiae strain. Ethanol concentration was carried on using a glass bench-scale distillation column. Experimental results were used for planning and designing the process scheme using Aspen Plus. Process simulation allowed calculating the mass and energy balances. It was found that coupling alkaline pretreatment with a later autoclaving improved the sugars yield in enzymatic hydrolysis. However, the use of the remaining soaking solution from pretreatment as hydrolysis medium had negative effects on sugars yield suggesting that there exist inhibit substance for the enzyme. Better results for enzymatic hydrolysis were obtained when sodium acetate buffer was used. Ethanol yield obtained from both experiments and simulation were very similar (66.50 and 65.84 dm³ of ethanol per each t of empty fruit bunches, respectively). These low ethanol yields were obtained because the native S. cerevisiae does not assimilate all reducing sugars, suggesting that those sugars were pentoses. Simulated alkaline and autoclaving pretreatment contributed only with 2% of the total energy consumption (198.4 GJ m⁻³ ethanol) while product recovery represented 57% of the total energy.     

Influence of impregnation with lactic acid on sugar yields from steam pretreatment of sugarcane bagasse and spruce, for bioethanol production

Lignocellulosic biomass can be utilized to produce ethanol, a promising alternative energy source produced through fermentation of sugars. However, in order to achieve high sugar and ethanol yields, the lignocellulosic material must be pretreated before the enzymatic hydrolysis and fermentation. Dilute acid pretreatment, using SO₂, is one of the most promising methods of pretreatment for softwood and agricultural residues. However, handling the high acidity of the slurry obtained from pretreatment and difficulty in recycling/degradation of the impregnating agent are some of the drawbacks of the dilute acid processes. In the present study the influence of utilization of a weak organic acid (lactic acid), as impregnating agent, on the sugar yield from pretreatment, with and without addition of SO₂, was investigated. The efficiency of pretreatment was assessed by enzymatic hydrolysis of the slurry obtained by pretreatment, using sugarcane bagasse and spruce, stored for one and two months in the presence of lactic acid separately, as feedstocks. Pretreatment of bagasse after storage with 0.5% lactic acid resulted in an overall glucose yield, i.e. after enzymatic hydrolysis, of 79% of theoretical based on the amount available in the raw material. This was as good as pretreatment using SO₂ as impregnating agent. However, storage of spruce with lactic acid before pretreatment, with and without addition of SO₂, was not efficient and resulted in lower sugar yields than pretreatment using SO₂ only.     

Study on saccharification techniques of seaweed wastes for the transformation of ethanol

Floating residue (FR), a surplus by-product from the alginate extraction process, contains large amount of cellulosic materials. The technical feasibility of FR utilization as a resource of renewable energy was investigated in this paper. The production of yeast-fermentable sugars (glucose) from FR was studied by dilute sulfuric acid pretreatment and further enzymatic hydrolysis. Dilute sulfuric acid pretreatment was conducted by using sulfuric acid at concentration of 0, 0.1, 0.2, 0.5 and 1.0%(w/v) for 0.5, 1.0 and 1.5 h respectively at 121 °C. The system of enzymatic hydrolysis consisted of cellulase and cellobiase. Results showed that FR might be a perfect bioenergy resource, containing high content of cellulose (30.0 ± 0.07%) and little hemicellulose (2.2 ± 0.86%). The acid pretreatment improved the hydrolysis efficiency of cellulase and cellobiase by increasing the reaction surface area of FR and enhanced the final yield of glucose for fermentation. The maximum yield of glucose reached 277.5 mg/g FR under the optimal condition of dilute sulfuric acid pretreatment (0.1% w/v, 121 °C, 1.0 h) followed by enzymatic hydrolysis (50 °C, pH 4.8, 48 h). After fermentation by Saccharomyces cerevisiae at 30 °C for 36 h, the ethanol conversion rate of the concentrated hydrolysates reached 41.2%, which corresponds to 80.8% of the theoretical yield. It indicates that cellulose in seaweed processing wastes including FR is easily hydrolyzed to produce glucose in comparison with that in terrestrial plants. FR shows excellent prospects as a potential feedstock for the production of bioethanol.     

Sugarcane hybrids with original low lignin contents and high field productivity are useful to reach high glucose yields from bagasse

Five sugarcane hybrids plus a reference material were evaluated according to the glucose yields obtained after alkaline-sulfite pretreatment and enzymatic hydrolysis. Sugarcane hybrids with varied original chemical compositions were used to assess how contrasting samples might influence the integrated pretreatment and hydrolysis process. The hydrolysis efficiency of six samples treated at three different chemical loads, suggested that lignin and hemicellulose removals during the pretreatment were not the single factor necessary to reach high cellulose conversion levels in the enzymatic hydrolysis step. Pretreated samples with the highest total acid contents (mainly sulfonic acids) were also the most digestible materials. The glucose yields were heavily dependent not only on the digestibility of the pretreated materials but also on the field productivity of the plants. One of the hybrids, presenting high glucan yields after pretreatment and high digestibility, produced low glucose yields because it presented very low biomass productivity. In contrast, one of the hybrids that presented low glucan yield after pretreatment, but was highly digestible and presented high biomass productivity, provided the highest glucose yields in the data set, producing 4192 and 5629 kg of glucose per hectare after enzymatic hydrolysis for 24 h and 72 h, respectively.     

Batch-based enzymatic saccharification of sweet sorghum bagasse

To improve enzymatic digestibility and sugar concentration, sweet sorghum bagasse was pretreated with alkali and liquid hot water, and then subjected to fed-batch enzymatic hydrolysis. Scanning electron microscopy assay suggested that different pretreatment methods resulted in different composition and structure of residues; these changes had a significant influence on cellulose hydrolysis. Fresh substrate was pretreated and then added at different amounts during the first 48 h to yield a final dry matter content of 30% (w/v). For liquid hot water pretreatment, a maximal glucose concentration of 95.71 g/L, corresponding to 52.85% xylan removal, was obtained with the sweet sorghum bagasse pretreated at 184°C for 18 min. NaOH soaking at ambient conditions removed lignin up to 60%, and the subsequent hydrolysis with cellulase loading of less than 10 FPU/g DM, and substrate supplementation every few hours yield the high glucose and xylose concentrations of 114.89L and 29.93 g/L, respectively after 144 h.     

Lipid production from sweet sorghum bagasse through yeast fermentation

Cryptococcus curvatus has great potential in fermenting unconditioned hydrolysates of sweet sorghum bagasse. With hydrolysates obtained by enzymatic hydrolysis of the solid pretreated by microwave with lime, the maximal yeast cell dry weight and lipid content were 10.83 g/l and 73.26%, respectively. For hydrolysates obtained in the same way but without lime, these two parameters were 15.50 g/l and 63.98%, respectively. During yeast fermentation, glucose and xylose were consumed simultaneously while cellobiose was released from the residual bagasse. The presence of lime, on one hand, made cellulose more accessible to enzymes as evidenced by higher total reducing sugar release compared to that without during enzymatic hydrolysis step; on the other hand, it caused the degradation of sugars to non-sugar chemicals during pretreatment step. As a result, higher lipid yield of 0.11 g/g bagasse or 0.65 ton/hectare of land was achieved from the pathway of microwave pretreatment and enzymatic hydrolysis while 0.09 g/g bagasse or 0.51 ton/hectare of land was attained from the process of lime-assisted microwave pretreatment followed by the same enzymatic saccharification.