New lignocellulose pretreatments using cellulose solvents: a review

Non‐food lignocellulosic biomass is the most abundant renewable bioresource as a collectable, transportable, and storable chemical energy that is far from fully utilized. The goal of biomass pretreatment is to improve the enzymatic digestibility of pretreated lignocellulosic biomass. Many substrate factors, such as substrate accessibility, lignin content, particle size and so on, contribute to its recalcitrance. Cellulose accessibility to hydrolytic enzymes is believed to be the most important substrate characteristic limiting enzymatic hydrolysis. Cellulose solvents effectively break linkages among cellulose, hemicellulose and lignin, and also dissolve highly‐ordered hydrogen bonds in cellulose fibers accompanied with great increases in substrate accessibility. Here the history and recent advances in cellulose solvent‐based biomass pretreatment are reviewed and perspectives provided for addressing remaining challenges. The use of cellulose solvents, new and existing, provides opportunities for emerging biorefineries to produce a few precursors (e.g. monosaccharides, oligosaccharides, and lignin) for the production of low‐value biofuels and value‐added biochemicals. © 2012 Society of Chemical Industry     

木质纤维素预处理技术研究进展

木质纤维素转化燃料乙醇一般需要经过原料预处理、酶水解和发酵过程。由于木质纤维原料化学结构复杂、直接酶解效率非常低,一般在酶水解之前需要进行适当的预处理以打破其致密结构,增加纤维表面积,提高后续纤维素酶的可及性。预处理程度直接影响纤维底物后续酶水解的效果。本文在木质纤维素常用预处理技术分析的基础上,重点讨论了3种相对高效的预处理技术：微波辅助离子液体预处理、两阶段深度共熔溶剂（DES）预处理和氯化铁预处理技术,分析了它们的优势、不足及发展现状。文中指出微波辅助离子液体预处理可有效解构木质素和半纤维素,破坏纤维素结晶区域,利于后续酶解,但微波加热过程会使离子液体分解和部分底物碳化。两阶段DES预处理可有效提高酶水解效率,但是预处理后原料中残留的DES可能会对后续反应中纤维素酶和微生物产生抑制作用。氯化铁预处理可有效破坏木质素与碳水化合物间的结合键,脱除底物中的半纤维素,而对木质素和纤维素降解较少,具有很好的发展前景。由于单一预处理技术的局限性,寻求低成本高效的联合预处理技术将是未来重点发展的方向。     

微波烘焙预处理降解玉米秸秆

用微波可高效对生物质烘焙预处理,考察了不同微波烘焙过程对玉米秸秆主要组分的降解作用及酸、碱、甘油催化剂对纤维素转化效率的影响,并对预处理的玉米秸秆进行酶解实验。结果表明,单纯的微波预处理对玉米秸秆中主要组分纤维素、半纤维素和木质素均有强烈的转化作用。无催化剂微波烘焙后,样品中纤维素含量降低了30%。在微波烘焙中添加酸、碱、甘油催化剂,可选择性降解玉米秸秆中的半纤维素或木质素,有效提高预处理后玉米秸秆中的纤维素含量,添加NaOH后纤维素含量增加最明显,由33%增至42%,纤维素最高转化率达65%。     

木质纤维素转化为燃料乙醇的研究进展

以木质纤维素为原料生产燃料乙醇的生物转化方法包括预处理、酶水解和发酵过程,对这些过程中的技术进展以及解决现存问题的方法进行了评述。氨法爆破技术是较好的预处理方法,超声波、微波处理等新技术有助于改善酶水解。阐述了酶水解机理、纤维素酶的生产以及酶水解过程的优化方法。指出固定化酶糖化发酵技术在生物转化木质纤维原料技术中的前景广阔;选择合适的发酵方法,优化发酵过程,以及解决抑制问题对于提高乙醇产率尤为重要;利用基因重组技术构建旨在发酵混合糖的重组菌对于生产生物乙醇具有里程碑意义。     

甘蔗渣的酸催化常压甘油有机溶剂预处理及其酶解

预处理可以打破木质纤维素原料纤维素、半纤维素和木质素三大组分间的顽抗结构,从而提升纤维素基质可酶解性。本文针对目前常压甘油有机溶剂预处理花费时间过长的问题,尝试开展酸催化的常压甘油有机溶剂预处理研究以缩短预处理时间。实验通过单因素选择和响应面Box-Behnken设计优化,获得酸催化常压甘油有机溶剂预处理的最佳条件为：预处理温度245℃,预处理时间38min,硫酸添加质量0.1%。在此条件下获得基质48h酶解率的响应面预测值为94.0%,实际值为91.4%。结果表明响应面优化方案和回归模型适用于本实验,预处理显著提高了基质可酶解性。高浓度基质（15%～20%）酶解进一步证明了预处理后基质具有突出的可酶解性,20%浓度基质在酶载量5FPU/g干基质条件下批次酶解72h,酶解率达60%,葡萄糖浓度达83.4g/L。酸催化常压甘油有机溶剂酸预处理在明显缩短预处理时间的同时,能显著提高木质纤维素基质可酶解性,使后续工业化意义的浓醪酶解糖化成为可能。     

木质纤维素燃料乙醇生物转化预处理技术

由丰富的木质纤维素资源制备乙醇有利于缓解能源紧缺、减少环境污染、实现可持续发展.然而某些物理、化学因素阻碍了木质纤维素中纤维素和半纤维素的转化和利用.预处理引起物理和/或化学上的变化,主要目的是改变或去除各种结构和(或)化学障碍,增加纤维素酶解率和转化效果,是一系列纤维素乙醇转化技术中的关键和核心.本文就纤维素乙醇生物...     

Effects of ball milling on structural changes and hydrolysis of lignocellulosic biomass in liquid hot-water compressed carbon dioxide

Mechanical activation is an effective method for destroying the crystalline structure. Biomass, especially its hemicellulose, can be degraded in the green solvent of liquid hot-water compressed carbon dioxide. To improve the degradation of crystalline cellulose in liquid hot-water compressed carbon dioxide, pretreatment of camphorwood sawdust by mechanical activation with a stirring ball mill was studied. Ball milling parameters and their effects on structure were determined by SEM, XRD and FT-IR. The influence of milling parameters on cellulose conversion can be ranked as follows: ball milling speed>activation time>the mass ratio of ball to biomass. The optimum milling condition was obtained at ball milling speed of 450 rpm and mass ratio of 30: 1 of ball to biomass for 2 h. In this condition, cellulose crystallinity of sawdust decreased from 60.93% to 21.40%. The cellulose conversion was 37.8%, which was nearly four times of raw material (10.2%). The glucose yield in the hydrolysate was 1.49 g·L^?1, which was nearly three times of that of raw material. It showed mechanical activation can destroy the crystalline structure of cellulose to promote degradation and the damage of lignocellulosic internal structure caused by ball milling is irreversible.     

Utilization of waste cellulose. IV. Comparative study of the reactivity of different substrates in the enzymatic hydrolysis with trichoderma viride

Different cellulosic substrates are compared. Cellulose II (grinded Cellophane®) was demonstrated to be a very reactive substrate without any other pretreatment. Indeed, Cellophane®, even coated one side with PVDC or nitrocellulose, can be hydrolyzed to a very high yield of glucose in a reasonable time. Suspensions with an initial high content of substrate can be used. Different pretreatment (ball milling, γ-irradiation, and Fe²⁺/H₂O₂) were found to modify the reactivity of cellulose I. The pretreatments which result in the chemical transformation of the glucose units of the initial cellulosic substrate are shown to inhibit the enzymatic hydrolysis.     

废弃棉织物浓磷酸预处理条件与高固酶水解基质浓度的关系

以浓磷酸预处理废弃棉织物,获得再生纤维素,对其进行酶水解,研究了预处理温度、时间和磷酸浓度与高固酶水解基质浓度的关系. 结果表明,温度和时间与高固酶水解潜力基质浓度均呈线性正相关性,温度从30℃提高至60℃,潜力基质浓度由4.6%提高至18.0%;时间由3.0 h延长至12.0 h,潜力基质浓度由10.8 %提高25.1%,但预处理时间过长不利于提高酶水解基质浓度. 磷酸浓度为85.0%时,潜力基质浓度达17.6%,改变浓度对潜力基质浓度提高影响较小. 废弃棉织物在60℃下用85.0%磷酸处理12.0 h后,高固酶水解的潜力基质浓度可达25.1%.     

Enzymatic hydrolysis from carbohydrates of barley straw pretreated by ionic liquids

BACKGROUND: Lignocellulosic biomass offers many potential advantages in comparison with the traditionally used sugars or starchy biomass since it is very widely available and does not compete with food and feed production. The abundance and high carbohydrates content of barley straw make it a good candidate for bioethanol production in Europe. Since biomass must be pretreated before enzymatic hydrolysis to improve the digestibility of both the cellulose and the hemicellulose biomass, the use of ionic liquids (IL) has been proposed as an environment‐friendly pretreatment of biomass. RESULTS: Different pretreatment conditions were investigated to determine the effects of the experimental conditions (temperature and time) on the enzymatic digestibility of pretreated material. The pretreatment of barley straw with 1‐ethyl‐3‐methyl imidazolium acetate treatment resulted in up to a 9‐fold increase in the cellulose conversion and a 13‐fold increase in the xylan conversion when compared with the untreated barley straw. CONCLUSION: Ionic liquid pretreatment of barley straw at 110°C for 30 min, followed by enzymatic hydrolysis, leads to a sugar yield of 53.5 g per 100 g raw material. It is then ready available for conversion into ethanol and is equivalent to more than 86% from potential sugars. The increase in saccharification was possible due to rupture of the lignin–hemicellulose linkages by treatment with 1‐ethyl‐3‐methyl imidazolium acetate. © 2012 Society of Chemical Industry     

水热预处理竹子促进酶解的效果及其影响因素

采用间歇式水热预处理装置,研究了水热预处理用于竹子的酶解规律,探讨了不同温度、处理时间、纤维素酶添加量及原料种类对促进竹子酶解的效果及其影响。结果表明水热预处理能显著提升竹子的酶解率,在优化条件190℃、10 min水热预处理,添加15 FPU·(g葡聚糖)^-1纤维素酶,72 h葡聚糖与木聚糖酶解率分别为74.3%、54.0%,提高到原来的3.5倍和4.7倍。过高的预处理温度与过长的预处理时间都将导致木糖大量降解和部分葡萄糖降解,使单糖总量下降。纤维素酶的添加量从15 FPU·(g 葡聚糖)^-1提高到60 FPU·(g 葡聚糖)^-1,可使未作预处理和水热预处理竹子的总糖回收率分别提高21.5%和9.9%,其促进酶解的作用远低于预处理的效果,通过预处理增大酶的可及性是提高酶解率的关键。水热预处理对于生物质原料具有选择性,不同的竹子原料具有显著不同的效果。     

Alkaline sulfite/anthraquinone pretreatment followed by disk refining of Pinus radiata and Pinus caribaea wood chips for biochemical ethanol production

BACKGROUND: Alkaline sulfite/anthraquinone (ASA) cooking of Pinus radiata and Pinus caribaea wood chips followed by disk refining was used as a pretreatment for the production of low lignified and high fibrillated pulps. The pulps produced with different delignification degrees and refined at different energy inputs (250, 750 and 1600 Wh) were saccharified with cellulases and fermented to ethanol with Saccharomyces cerevisiae using separated hydrolysis and fermentation (SHF) or semi‐simultaneous saccharification and fermentation (SSSF) processes. RESULTS: Delignification of ASA pulps was between 25% and 50%, with low glucans losses. Pulp yield was from 70 to 78% for pulps of P. radiata and 60% for the pulp of P. caribaea. Pulps obtained after refining were evaluated in assays of enzymatic hydrolysis. Glucans‐to‐glucose conversion varied from 20 to 70%, depending on the degree of delignification and fibrillation of the pulps. The best ASA pulp of P. radiata was used in SHF and SSSF experiments of ethanol production. Such experiments produced maximum ethanol concentration of 20 g L^?1, which represented roughly 90% of glucose conversion and an estimated amount of 260 L ethanol ton^?1 wood. P. caribaea pulp also presented good performance in the enzymatic hydrolysis and fermentation but, due to the low amount of cellulose present, only 140 L ethanol would be obtained from each ton of wood. CONCLUSION: ASA cooking followed by disk refining was shown to be an efficient pretreatment process, which generated a low lignified and high‐fibrillated substrate that allowed the production of ethanol from the softwoods with high conversion yields. Copyright © 2012 Society of Chemical Industry     

低共熔溶剂预处理提高甘草渣酶解效果优化

近年来，低共熔溶剂（deep eutectic solvent，DES）以易制备、成本低、易回收等优势，在生物质预处理方面受到广泛关注。本研究以氯化胆碱为氢键受体，乙醇胺为氢键供体，合成DES，研究了不同温度、时间和固液比预处理条件对中药渣组分和酶解效果的影响。结果表明：固液比1∶20、120℃、预处理4h后原料中木质素去除率达到78.42%，纤维素回收率为83.89%。随后对不同条件下所得底物进行酶水解，反应96h后发现，较优条件下所得底物酶解效率为78.57%，较未处理中药渣（30.40%）提高了1.58倍。类分形动力学分析表明，预处理温度对酶解效果影响最大。SEM、XRD和FTIR检测发现，预处理后底物形貌、结晶指数和官能团变化有利于酶解效果的提高。     

Catalytic organosolv fractionation of willow wood and wheat straw as pretreatment for enzymatic cellulose hydrolysis

BACKGROUND: Ethanol‐based organosolv fractionation of lignocellulosic biomass is an effective pretreatment technology for enzymatic cellulose hydrolysis to produce sugars and lignin within a biorefinery. This study focuses on the catalytic effect of H₂SO₄, HCl, and MgCl₂ on organosolv pretreatment of willow wood and wheat straw. RESULTS: The use of catalysts improved fractionation of both feedstocks. The maximum enzymatic cellulose digestibility obtained was 87% for willow wood (using 0.01 mol L^?1 H₂SO₄ as catalyst) and 99% for wheat straw (0.02 mol L^?1 HCl). Non‐catalytic organosolv fractionation at identical conditions resulted in 74% (willow wood) and 44% (wheat straw) glucose yield by enzymatic hydrolysis. Application of catalysts in organosolv pretreatment was particularly effective for wheat straw. The influence of the acid catalysts was found to be primarily due to their effect on the pH of the organosolv liquor. Acid catalysts particularly promoted xylan hydrolysis. MgCl₂ was less effective than the acid catalysts, but it seemed to more selectively improve delignification of willow wood. CONCLUSION: Application of catalysts in organosolv pretreatment of willow wood and wheat straw was found to substantially improve fractionation and enzymatic digestibility. The use of catalysts can contribute to achieving maximum utilization of lignocellulosic biomass in organosolv‐based biorefineries. Copyright © 2011 Society of Chemical Industry     

Application of a microassay method to study enzymatic hydrolysis of pretreated wheat straw

BACKGROUND: The conversion of lignocellulosic biomass to ethanol includes a disruptive pretreatment process followed by enzyme‐catalyzed hydrolysis of the cellulose and hemicellulose components to fermentable sugars. As the cost and hydrolytic efficiency of enzymes are major factors that restrict the commercialization of biomass conversion processes, significant efforts are made nowadays to improve the enzymatic mixtures and make the process cost‐effective. RESULTS: In this work, enzymatic microassays have been developed and validated to test new different enzymatic formulations on real lignocellulosic substrates. Homogeneous handsheets from steam pretreated wheat straw were elaborated to be used as substrate. The microassay was adapted to test both water‐insoluble solids and the whole slurry as substrates. Results in hydrolysis microassays were comparable with those obtained in standard flask assays using pretreated wheat straw. Moreover, using the enzymatic microassays, two novel preparations have been evaluated, demonstrating the ability of microassays to discriminate between different enzymatic mixtures. CONCLUSIONS: This enzymatic microassay represents a rapid method to test the performance of new selected cellulase enzymes on real pretreated lignocellulosic substrates. This microassay will enable evaluation of enzyme components separately, or optimized mixtures. Copyright © 2010 Society of Chemical Industry     

NaOH/AQ预处理对玉米秸秆脱木质素及酶解性能的影响

对玉米秸秆进行氢氧化钠/蒽醌（NaOH/AQ）去木质化预处理,考察了预处理温度、时间和NaOH用量对玉米秸秆脱木质素程度的影响,并探讨了脱木质素程度对提高预处理后物料酶解性能的影响。L₉（3⁴）正交试验得出较适宜预处理工艺条件为：温度160℃,时间60 min,NaOH用量（以绝干原料质量计）2.8%;其他条件为AQ用量0.05%,固液比1：5（g：mL）,此时木质素脱除率为75%,酶解后聚糖转化率达到73.79%。随着物料脱木质素程度的提高,其酶解效率相应增加;当木质素脱除率达到一定程度后,预处理后的聚糖转化率达到最大值,继续提高木质素脱除率,聚糖转化率反而降低。响应面优化的酶水解工艺条件为纤维素酶用量30 FPU/g,β-葡萄糖苷酶10 IU/g,反应时间72 h,温度50℃,底物质量分数2.5%,此时还原糖得率为85.62%。对酶解液进行HPLC分析,酶解液中的葡萄糖质量浓度为14.83 g/L,木糖质量浓度为4.83 g/L。XRD分析显示,预处理前后纤维素的晶型没有变化,而结晶度由31.40%提高至46.91%,表明物料中木质素和半纤维素发生了不同程度的溶出。     

A study on enzymatic hydrolysis of cellulose in an attrition bioreactor

In the biological conversion of cellulose, a physical or chemical pretrealment proceeds the hydrolysis by an enzyme. The hydrolysis rate however, is slowed down as the active sites in the pretreated substrates are reduced. In this study, attempts were made to use an attrition bioreactor in which the pretreatment and the hydrolysis were carried out at the same time, where higher hydrolysis rales were achieved. Glass beads of 0.3-cm-diameter and sand were used as the milling media in a batch reactor where pure cellulose, Solka Floc BW200 was hydrolyzed by cellulase secreted from a fungus mutant,Trichoderma reesei. The higher rates observed are believed due to the synergistic effects of the size reduction and the conversion of the crystalline to the amorphous form of cellulose which was observed by comparing the X-ray diffractorgrams of the cellulose hydrolyzed in the reactors with and without milling medium. A simple kinetic model was found satisfactory in depicting the hydrolysis mechanism, and the kinetic parameters were estimated. Higher power consumption as compared to a reqular stirred reactor was observed and a quantitative expression was derived for its estimation.     

慈竹机械浆（RMP）氨法预处理工艺研究

研究了在氨水中添加少量KOH或K3PO4预处理慈竹机械浆(RMP)的工艺。考察了预处理条件对酶解还原糖产率的影响。单因素实验得到氨水加KOH最佳预处理条件为:预处理温度120℃、固液比〔即1 g绝干浆加入液体的体积(mL),下同〕1∶6、时间3.5 h、氨水用量70%(即氨水质量占绝干浆质量的百分数,下同)、KOH用量5%(即KOH质量占绝干浆质量的百分数,下同)。在此最佳条件下,慈竹RMP的纤维素保留率为90.66%,半纤维素保留率为92.90%,木质素脱除率为41.05%;在pH=4.8、加酶量20 FPU/g预处理后底物、反应温度50℃的条件下酶解24 h,还原糖产率为23.95%,纤维素转化率为44.61%。虽然氨水加K3 PO4预处理酶解纤维素转化率可达56.95%,但是纤维素保留率仅为74.59%,与氨水加KOH相比,纤维素损失较大。     

Comparison of atmospheric aqueous glycerol and steam explosion pretreatments of wheat straw for enhanced enzymatic hydrolysis

BACKGROUND: The oversupply of cheap glycerol by the oleochemicals industry together with problems occurring in low‐boiling‐point organosolv pretreatments, has generated an interest in the use of glycerol in the organosolv pretreatment of lignocellulosic biomass. Atmospheric aqueous glycerol autocatalytic organosolv pretreatment (AAGAOP) is a promising strategy that can effectively enhance enzymatic hydrolysis of lignocellulosic biomass. As a cost‐effective technique, steam explosion pretreatment (SEP) is being adopted in industrial applications. Accordingly, work has been carried out to investigate how AAGAOP enhanced enzymatic hydrolysis of lignocellulosic biomass compares with the SEP method. RESULTS: Under controlled laboratory conditions, based on ≥ 90% cellulose recovery, AAGAOP removed ≥ 60% hemicellulose and ≥ 60% lignin from wheat straw while SEP led to ～80% hemicellulose and 10% lignin removal. Enzymatic hydrolysis yields of AAGAOP and SEP reached ～90% and ～70%, respectively. Physical‐chemical structural characterization by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT‐IR), helped explain the above results. The two methods gave priority to dissociating the guaiacyl lignin and had a relatively small effect on syringyl units. However, AAGAOP exhibited a superior performance. CONCLUSION: The two methods enhanced the enzymatic hydrolysis of lignocellulosic biomass by removing and/or altering physical‐chemical structural impediments. The AAGAOP technique, with some special advantages, was more effective than SEP in enhancing the recovery and enzymatic digestibility of cellulose. Copyright © 2008 Society of Chemical Industry     

Effect of LHW,HCl, and NaOH pretreatment on enzymatic hydrolysis of sugarcane bagasse: sugar recovery and fractal-like kinetics

Conversion of lignocellulose to sugars involves two main processes, namely pretreatment and enzymatic hydrolysis. Lignocellulose pretreatment leads to the degradation of enzymatic recalcitrance of substrate for achieving efficient saccharification. In this study, liquid hot water (LHW), hydrochloric acid (HCl), and sodium hydroxide (NaOH) were as reagents used to pretreat sugarcane bagasse (SB). Results showed that LHW, HCl, and NaOH pretreatment could solubilize 95.3%, 94.7% xylan and 88.7% lignin, respectively. Enzymatic hydrolysis of pretreated SB showed that the maximum glucose (26.0?g/L) and xylose (12.7?g/L) concentration were produced by NaOH pretreatment, and slightly more glucose and less xylose were produced after HCl pretreatment compared to LHW pretreatment. Addition of Tween 80 or xylanase could significantly improve both glucose and xylose production. At 48?h, the glucose increase for LHW, HC1 and NaOH pretreatment was 38.3%, 26.4% and 8.0%, respectively, and the xylose increase for them was 35.0%, 24.9% and 1.7%, respectively. Fractal-like kinetics showed that the value of rate constant increased after the addition of Tween 80 or xylanase, and the efficiency of enzymatic hydrolysis mainly depended on rate constant other than fractal dimension of substrate. Totally, substrate accessibility was dominated for efficient of lignocellulose to sugar compared to enzyme loading. The application of fractal-like theory on the heterogeneous enzymatic hydrolysis of lignocellulose was quite successful.