乙醇预处理麦秆的酶解性能研究简

采用H2 SO4催化和自催化乙醇法对麦秆进行预处理,比较预处理后麦秆的主要化学组成、纤维素酶解性能和半同步糖化发酵生产乙醇特性,并进行物料衡算。结果表明：H2 SO4催化和自催化乙醇预处理过程中纤维素固体回收率大于90％。添加非离子表面活性剂吐温20和吐温80没有显著提高H2 SO4催化乙醇预处理后纤维素的酶解葡萄糖得率及半同步糖化发酵过程中乙醇的产量,而对自催化乙醇处理后麦秆的酶解和半同步糖化发酵过程有一定程度的促进作用,相应的酶解葡聚糖转化率由72.7％提高到85.0％,而半同步糖化发酵过程中乙醇质量浓度提高了11.4％。物料衡算结果表明：酸催化和自催化乙醇预处理后葡聚糖回收率分别为91.0％和95.4％;半同步糖化发酵生产乙醇的得率分别为10.4和11.6 g（按100 g原料计）。     

微波条件下LiCl/DMAc溶解稻秆纤维素制备还原糖

利用氯化锂/N,N-二甲基乙酰胺(LiCl/DMAc)溶剂体系在微波控制条件下对稻秆纤维素进行溶解预处理以提高纤维素酶解糖化效率。考察了微波时间和微波强度对产糖量及还原糖转化率的影响。通过扫描电镜(SEM)和热重分析仪(TG)对预处理前后稻秆纤维素的微观形貌及热稳定性进行表征,并利用高效液相色谱仪(HPLC)对酶解糖液进行糖成分鉴定和含量分析。结果表明,微波加热能够有效促进LiCl/DMAc对稻秆纤维素的溶解。与原生稻秆相比,经微波-LiCl/DMAc法溶解后再生纤维素出现明显解聚,热分解温度由290℃降至220℃。在微波功率为385 W、加热溶解时间为7 min时,所得稻秆纤维素还原糖转化率由30.90%上升至98.67%;HPLC谱图表明,糖液中主要成分为葡萄糖和木糖,分别占所得还原糖总量的43.74%和48.55%。     

乙醇预处理麦秆的酶解性能研究

采用H2SO4催化和自催化乙醇法对麦秆进行预处理,比较预处理后麦秆的主要化学组成、纤维素酶解性能和半同步糖化发酵生产乙醇特性,并进行物料衡算。结果表明:H2SO4催化和自催化乙醇预处理过程中纤维素固体回收率大于90%。添加非离子表面活性剂吐温20和吐温80没有显著提高H2SO4催化乙醇预处理后纤维素的酶解葡萄糖得率及半同步糖化发酵过程中乙醇的产量,而对自催化乙醇处理后麦秆的酶解和半同步糖化发酵过程有一定程度的促进作用,相应的酶解葡聚糖转化率由72.7%提高到85.0%,而半同步糖化发酵过程中乙醇质量浓度提高了11.4%。物料衡算结果表明:酸催化和自催化乙醇预处理后葡聚糖回收率分别为91.0%和95.4%;半同步糖化发酵生产乙醇的得率分别为10.4和11.6 g(按100 g原料计)。     

氨基磺酸应用于甘蔗渣预处理的研究

本研究尝试将氨基磺酸应用于甘蔗渣预处理,探究其作为酸预处理试剂对甘蔗渣成分和酶解的影响。氨基磺酸预处理最优条件为浓度3%,温度121℃,预处理1 h。在该条件下,甘蔗渣的固体回收率为64.45%,半纤维素和木质素去除率分别为70.81%和25.10%,纤维素损失率仅7.56%。与硫酸、盐酸预处理相比,氨基磺酸的半纤维素和木质素去除率不如硫酸、盐酸预处理,但固体回收率更高,纤维素损失率低,能保留更多纤维素有效成分。进一步酶解显示,氨基磺酸预处理的纤维素转化率高于硫酸、盐酸预处理。氨基磺酸作为一种新的酸预处理试剂,在木质纤维素降解上有良好应用前景。     

菊糖芽孢乳杆菌利用麸皮水解液发酵生产D-乳酸

考察菊糖芽孢乳杆菌YBS1-5利用麸皮的水解液发酵生产D-乳酸的性能。首先研究了不同蛋白酶对麸皮中蛋白组分的水解效率,优选酸性蛋白酶并对其进行水解工艺的优化,最终其水解液中的含氮量为4.6 g/L,水解效率为85.8%。对酸性蛋白酶的水解液残渣进行稀酸预处理后,利用纤维素酶对其进行酶解。通过批次补料酶解,水解液中的还原糖质量浓度达141.2 g/L,其中葡萄糖质量浓度为138.1 g/L、木糖质量浓度为1.4 g/L。利用麸皮的蛋白酶水解液和纤维素酶水解液替代葡萄糖和酵母粉发酵制备D-乳酸。在96 h内,D-乳酸产量达99.5 g/L,生产速率达1.04 g/(L·h),转化率89.1%。     

用普通纯水代替缓冲液时蔗渣酶法水解反应的考察

使用Onozuk RS型纤维素酶,以普通的城市自来水和纯水代替缓冲液,直接加热膨化甘蔗渣进行酶法水解实验研究,结果表明:所使用的各种水代替缓冲液时,蔗渣酶解还原糖得率基本不变,还原糖得率最高时的最适宜pH为4.2～4.9,当蔗渣浓度大于1％时,反应液的pH恰好处于最适宜pH范围内,还原糖得率最高的酶解反应温度为50℃。     

玉米秸秆分批补料获得高还原糖浓度酶解液的条件优化

木质纤维素高浓度还原糖水解液的获得是纤维乙醇产业化发展的方向。在发酵工业领域,分批补料法是实现这一目标的重要研究途径。本研究采用分批补料法对获得高浓度玉米秸秆酶解还原糖的条件进行了优化。以稀硫酸预处理的玉米秸秆为原料,考察了液固比、补加量与补加时间对分批补料糖化的影响。结果表明,秸秆高浓度酶解液条件的初始物料为20％ (重量/体积),木聚糖酶220 U/g (底物),纤维素酶6 FPU/g (底物),果胶酶50 U/g (底物),在24 h、48 h后分批补加8％预处理后的物料,同时添加与补料量相应的木聚糖酶20 U/g (底物),纤维素酶2 FPU/g (底物),72 h后,最终糖化结果与非补料法相比,还原糖浓度从48.5 g/L提高到138.5 g/L,原料的酶解率最终达到理论值的62.5％。试验结果表明补料法可以显著提高秸秆水解液还原糖浓度。     

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.     

纤维素酶水解稻草生产单细胞蛋白（SCP）：1.纤维素酶水解稻草的研究

对不同预处理方法与底物得糖率,纤维素转化率的关系进行了研究,以1％NaOH在100℃煮沸1h效果最好。研究了稻草酶解的适宜条件,在pH5、50℃、底物浓度7％、酶解48h的条件下,底物得糖率达53.8％,含糖量5.0％,全纤维素转化率80.2％。     

玉米秸秆酶水解过程中的酶复配条件优化

降低酶解成本是纤维素乙醇生产的关键。利用酶复配技术优化蒸汽爆破处理后玉米秸秆的酶水解工艺条件,以提高纤维素的转化率。通过单因素实验和正交实验,研究了纤维素酶、木聚糖酶和β-葡萄糖苷酶对酶解效率的影响规律。结果表明,汽爆玉米秸秆,纤维素含量达42.21%,半纤维素仅为3.65%。纤维素酶对酶解过程起决定性作用,添加40 FPU/g时,酶解率为75.45%;木聚糖酶可促使更多的纤维素暴露出来,添加1 500 IU/g时,酶解率最高为78.03%;β-葡萄糖苷酶有助于消除纤维二糖积累造成的反馈抑制,用量40 IU/g时,纤维二糖浓度为0.330 4 g/100 m L,酶解率达76.45%。正交实验确定最佳工艺为:纤维素酶用量30 FPU/g,木聚糖酶用量800 IU/g,β-葡萄糖苷酶用量40 IU/g;该条件下,进行底物质量浓度25%的验证实验,葡萄糖达9.3g/100 m L,若用单一天冠纤维素酶,葡萄糖仅5.9 g/100 m L,提高了57.63%。三种酶的影响顺序为:纤维素酶木聚糖酶β-葡萄糖苷酶。     

Effectiveness of microbial pretreatment by Ceriporiopsis subvermispora on different biomass feedstocks

Different types of feedstocks, including corn stover, wheat straw, soybean straw, switchgrass, and hardwood, were tested to evaluate the effectiveness of fungal pretreatment by Ceriporiopsis subvermispora. After 18-d pretreatment, corn stover, switchgrass, and hardwood were effectively delignified by the fungus through manganese peroxidase and laccase. Correspondingly, glucose yields during enzymatic hydrolysis reached 56.50%, 37.15%, and 24.21%, respectively, which were a 2 to 3-fold increase over those of the raw materials. A further 10-30% increase in glucose yields was observed when pretreatment time extended to 35 d. In contrast, cellulose digestibility of wheat straw and soybean straw was not significantly improved by fungal pretreatment. When external carbon sources and enzyme inducers were added during fungal pretreatment of wheat straw and soybean straw, only glucose and malt extract addition improved cellulose digestibility of wheat straw. The cellulose digestibility of soybean straw was not improved.     

Low temperature alkali pretreatment for improving enzymatic digestibility of sweet sorghum bagasse for ethanol production

A low temperature alkali pretreatment method was proposed for improving the enzymatic hydrolysis efficiency of lignocellulosic biomass for ethanol production. The effects of the pretreatment on the composition, structure and enzymatic digestibility of sweet sorghum bagasse were investigated. The mechanisms involved in the digestibility improvement were discussed with regard to the major factors contributing to the biomass recalcitrance. The pretreatment caused slight glucan loss but significantly reduced the lignin and xylan contents of the bagasse. Changes in cellulose crystal structure occurred under certain treatment conditions. The pretreated bagasse exhibited greatly improved enzymatic digestibility, with 24-h glucan saccharification yield reaching as high as 98% using commercially available cellulase and β-glucosidase. The digestibility improvement was largely attributed to the disruption of the lignin-carbohydrate matrix. The bagasse from a brown midrib (BMR) mutant was more susceptible to the pretreatment than a non-BMR variety tested, and consequently gave higher efficiency of enzymatic hydrolysis.     

Comparative studies on hydrothermal pretreatment and enzymatic saccharification of leaves and internodes of alamo switchgrass

Hydrothermal pretreatment was performed on the leaves and internodes portions of Alamo switchgrass, Panicum virgatum L., to enhance the digestibility of cellulose towards cellulase. It was observed that extractives free leaves portion provided 18.1% lower pretreatment gravimetrical yield and 33.8% greater cellulose-to-glucose yield than internodes portion. The degree of polymerization (DP) and ultrastructure of cellulose were determined by gel-permeation chromatography and solid-state cross polarization/magic angle spinning ¹³C NMR experiments. The results suggested that hydrothermal pretreatment hydrolyzed amorphous cellulose and yielded a product enriched in paracrystalline cellulose. Furthermore, the DP of cellulose was reduced to one third of the origin value after hydrothermal pretreatment. The resulting biomass after pretreatment for leaves and internodes has similar cellulose ultrastructure and chemical profiles. The results of the enzymatic hydrolysis studies of cellulose suggest that the reduced DP of cellulose of pretreated switchgrass was an important factor influencing the enhanced digestibility of pretreated switchgrass.     

Reconstitution of cellulose and lignin after [C2mim][OAc] pretreatment and its relation to enzymatic hydrolysis

Although the effects of cellulose crystallinity and lignin content as two major structural features on enzymatic hydrolysis have been extensively studied, debates regarding their effects still exist. In this study, reconstitution of cellulose and lignin after 1‐ethyl‐3‐methylimidazolium acetate ([C₂mim][OAc]) pretreatment was proposed as a new method to study their effects on enzymatic digestibility. Different mechanisms of lignin content for reduction of cellulose hydrolysis were found between the proposed method and the traditional method (mixing of cellulose and lignin). The results indicated that a slight change of the crystallinity of the reconstituted materials may play a minor role in the change of enzyme efficiency. In addition, the present study suggested that the lignin content does not significantly affect the digestibility of cellulose, whereas the conversion of cellulose fibers from the cellulose I to the cellulose II crystal phase plays an important role when an ionic liquid pretreatment of biomass was conducted. Biotechnol. Bioeng. 2013; 110: 729–736. © 2012 Wiley Periodicals, Inc.     

Increasing cellulose accessibility is more important than removing lignin: A comparison of cellulose solvent‐based lignocellulose fractionation and soaking in aqueous ammonia

While many pretreatments attempt to improve the enzymatic digestibility of biomass by removing lignin, this study shows that improving the surface area accessible to cellulase is a more important factor for achieving a high sugar yield. Here we compared the pretreatment of switchgrass by two methods, cellulose solvent‐ and organic solvent‐based lignocellulose fractionation (COSLIF) and soaking in aqueous ammonia (SAA). Following pretreatment, enzymatic hydrolysis was conducted at two cellulase loadings, 15 filter paper units (FPU)/g glucan and 3 FPU/g glucan, with and without BSA blocking of lignin absorption sites. The hydrolysis results showed that the lignin remaining after SAA had a significant negative effect on cellulase performance, despite the high level of delignification achieved with this pretreatment. No negative effect due to lignin was detected for COSLIF‐treated substrate. SEM micrographs, XRD crystallinity measurements, and cellulose accessibility to cellulase (CAC) determinations confirmed that COSLIF fully disrupted the cell wall structure, resulting in a 16‐fold increase in CAC, while SAA caused a 1.4‐fold CAC increase. A surface plot relating the lignin removal, CAC, and digestibility of numerous samples (both pure cellulosic substrates and lignocellulosic materials pretreated by several methods) was also developed to better understand the relative impacts of delignification and CAC on glucan digestibility. Biotechnol. Bioeng. 2011; 108:22–30. © 2010 Wiley Periodicals, Inc.     

Comparative study on chemical pretreatment methods for improving enzymatic digestibility of crofton weed stem

In order to utilize and control the invasive weed, crofton weed (Eupatorium adenophorum Spreng), a potential pathway was proposed by using it as a feedstock for production of fermentable sugars. Three chemical pretreatment methods were used for improving enzymatic saccharification of the weed stem. Mild H2SO4 pretreatment could obtain a relatively high yield of sugars in the pretreatment (32.89%, based on initial holocellulose), however, it led to only a slight enhancement of enzymatic digestibility. NaOH pretreatment could obtain a higher enzymatic conversion ratio of cellulose compared with H2SO4 pretreatment. Peracetic acid (PAA) pretreatment seemed to be the most effective for improving enzymatic saccharification of the weed stem in the three chemical pretreatment methods under the same conditions. The conversion ratio of cellulose in the sample pretreated by PAA under the "optimal" condition was increased to 50% by cellulase loading of 80 FPU/g cellulose for 72 h incubation. A number of empirical quadratic models were successfully developed according to the experimental data to predict the yield of sugar and degree of delignification.     

Structural changes of corn stover lignin during acid pretreatment

In this study, raw corn stover was subjected to dilute acid pretreatments over a range of severities under conditions similar to those identified by the National Renewable Energy Laboratory (NREL) in their techno-economic analysis of biochemical conversion of corn stover to ethanol. The pretreated corn stover then underwent enzymatic hydrolysis with yields above 70?% at moderate enzyme loading conditions. The enzyme exhausted lignin residues were characterized by (31)P NMR spectroscopy and functional moieties quantified and correlated to enzymatic hydrolysis yields. Results from this study indicated that both xylan solubilization and lignin degradation are important for improving the enzyme accessibility and digestibility of dilute acid pretreated corn stover. At lower pretreatment temperatures, there is a good correlation between xylan solubilization and cellulose accessibility. At higher pretreatment temperatures, lignin degradation correlated better with cellulose accessibility, represented by the increase in phenolic groups. During acid pretreatment, the ratio of syringyl/guaiacyl functional groups also gradually changed from less than 1 to greater than 1 with the increase in pretreatment temperature. This implies that more syringyl units are released from lignin depolymerization of aryl ether linkages than guaiacyl units. The condensed phenolic units are also correlated with the increase in pretreatment temperature up to 180?°C, beyond which point condensation reactions may overtake the hydrolysis of aryl ether linkages as the dominant reactions of lignin, thus leading to decreased cellulose accessibility.     

Effect of biological pretreatment with Trametes hirsuta yj9 on enzymatic hydrolysis of corn stover

In this study, a newly isolated Trametes hirsuta yj9 was used to pretreat corn stover in order to enhance enzymatic digestibility. T. hirsuta yj9 preferentially degraded lignin to be as high as 71.49% after 42-day pretreatment. Laccase and xylanase was the major ligninolytic and hydrolytic enzyme, respectively and filter paper activity (FPA) increased gradually with prolonged pretreatment time. Sugar yields increased significantly after pretreatment with T. hirsuta yj9, reaching an enzymatic digestibility of 73.99% after 42 days of pretreatment. Scanning electron microscopy (SEM) showed significant structural changes in pretreated corn stover, the surface of pretreated corn stover became increasingly coarse, the gaps between cellulose fibers were visible, and many pores were developed. Correlation analysis showed that sugar yields were inversely proportional to the lignin contents, less related to cellulose and hemicellulose contents.     

Features of promising technologies for pretreatment of lignocellulosic biomass

Cellulosic plant material represents an as-of-yet untapped source of fermentable sugars for significant industrial use. Many physio-chemical structural and compositional factors hinder the enzymatic digestibility of cellulose present in lignocellulosic biomass. The goal of any pretreatment technology is to alter or remove structural and compositional impediments to hydrolysis in order to improve the rate of enzyme hydrolysis and increase yields of fermentable sugars from cellulose or hemicellulose. These methods cause physical and/or chemical changes in the plant biomass in order to achieve this result. Experimental investigation of physical changes and chemical reactions that occur during pretreatment is required for the development of effective and mechanistic models that can be used for the rational design of pretreatment processes. Furthermore, pretreatment processing conditions must be tailored to the specific chemical and structural composition of the various, and variable, sources of lignocellulosic biomass. This paper reviews process parameters and their fundamental modes of action for promising pretreatment methods.     

Pretreatment of corn stover by aqueous ammonia

Corn stover was pretreated with aqueous ammonia in a flow-through column reactor, a process termed ammonia recycled percolation (ARP). This method was highly effective in delignifying of the biomass, reducing the lignin content by 70-85%. Most lignin removal occurred within the first 20 min of the process. Lignin removal by ARP was further confirmed by FTIR analysis and lignin staining. The ARP process solubilized 40-60% of the hemicellulose but left the cellulose intact. The solubilized carbohydrate existed in oligomeric form. Carbohydrate decomposition during the pretreatment was insignificant. Corn stover treated for 90 min exhibited enzymatic digestibility of 99% with 60 FPU/g of glucan enzyme loading, and 92.5% with 10 FPU/g of glucan. The digestibility of ARP treated corn stover was substantially higher than that of alpha-cellulose. The enzymatic digestibility was related with the removal of lignin and hemicellulose, perhaps due to increased surface area and porosity. The SEM pictures indicated that the biomass structure was deformed and its fibers exposed by the pretreatment. The crystallinity index increased with pretreatment reflecting removal of the amorphous portion of biomass. The crystalline structure of the cellulose in the biomass, however, was not changed by the ARP treatment.