纤维素酶水解农田纤维素废弃物生产还原糖的研究

以不同农田纤维素废弃物为原料,利用纤维素酶水解生产还原糖。考察了反应时间、底物浓度、酶用量、反应温度及反应pH值等因素对纤维素酶水解农田废弃物生产还原糖的影响,确立了各自的最适反应条件。     

粗纤维素酶液对微晶纤维素酶解的影响因素

栗生灰黑孔菌多被用于产漆酶的研究,很少有利用其纤维素酶的报道。为了降低在纤维素水解中纤维素酶的使用成本,利用栗生灰黑孔菌发酵制备的粗纤维素酶液,以微晶纤维素为底物模型,研究粗纤维素酶液水解微晶纤维素的最佳pH、温度和最佳表面活性剂助剂种类及浓度,并对不同表面活性剂存在条件下的纤维素酶解动力学、紫外和荧光光谱进行了研究。结果表明,粗纤维素酶水解微晶纤维素的最佳条件为pH 4.8,温度50℃,最佳表面活性剂助剂为吐温80,添加剂量为1.12mg/g底物;吐温80的添加可提高粗纤维素酶解的最大反应速度常数V_max,降低米氏常数K_m;表面活性剂改变了纤维素酶的紫外和荧光最大吸收峰,酰胺Ⅰ带和酰胺Ⅲ带的谱峰,可能通过与纤维素酶中的氨基酸残基发生反应影响了纤维素酶的结构,进而影响了微晶纤维素的水解反应。该研究为进一步降低纤维素水解成本提供了理论指导。     

植物纤维原料纤维素酶水解的研究

以麦草为原料,探讨纤维素酶水解植物纤维的适宜条件。麦草含大量的纤维素和聚戊糖,其中的纤维素在纤维素酶的作用下分解生成葡萄糖和纤维二糖。对温度,pH值,酶解时间,酶用量分别进行单因素实验,通过测定葡萄糖含量和总还原糖含量,找出酶水解麦草的适宜条件为:pH4.6,温度47℃,酶解时间48h,酶用量7.5IU(每克绝干原料)。对不同底物浓度的实验表明,在尽可能高的底物浓度下连续水解,产物浓度高,得率也高。     

酸碱复合处理和酶浓度对药渣纤维素水解效率的影响

为了研究酸碱复合处理对药渣木质纤维素水解效率的作用效果,探究药渣纤维素转化酶解转化乙醇的可行性,以水提药渣为原料,用NaOH-PAA进行预处理,在不同反应体系下用纤维素酶进行水解。实验结果表明,预处理后药渣酶解的葡萄糖浓度比预处理前提高3～4倍;在酶总量不变的情况下,增加底物数量和酶浓度能显著提高反应体系中的葡萄糖浓度。83～116 mg/mL的底物浓度和5.8 U/mL的酶浓度可使反应体系中的葡萄糖浓度达到12 mg/mL以上。在83 mg/mL的底物浓度和5.8 U/mL的酶浓度下,对预处理后的丹参药渣、甘草药渣及混合药渣进行纤维素酶解,60 h时,其糖产率分别为29.07 g/kg、49.31 g/kg、52.83 g/kg。结论:预处理能显著提高纤维素的酶解效率,药渣的葡萄糖产率主要取决于其纤维素含量,与药渣类型没有密切关系。     

中性纤维素酶糖化中性汽爆玉米秸秆工艺优化

提出一种在非缓冲系统中水解中性汽爆秸秆的工艺。首先选取具有较好协同降解木质纤维素能力的特异腐质霉(Humicola insolens)所产中性纤维素酶进行工艺优化,确定其水解工艺可以在非缓冲体系中进行。在此基础上,通过添加β-葡萄糖苷酶、木聚糖酶、漆酶和表面活性剂与中性纤维素酶制剂中进行复配后在非缓冲体系中(自然pH值)水解中性汽爆秸秆,并用分批加酶水解提高了复合酶的酶解效率。结果表明,每克中性汽爆秸秆底物中加入10 FPU中性纤维素酶,75 IU β-葡萄糖苷酶,3 000 IU木聚糖酶和体积分数为0.5 % Triton-100,以100 g/L底物浓度水解120 h后,综合水解率为48.4 %。每克底物中复合酶以15+5 FPU,分批加酶水解120 h后综合水解率、纤维素水解率和半纤维素水解率分别为56.0 %、64.9 %和42.5 %。这有助于拓宽木质纤维素糖化工艺研究的思路,为木质纤维素材料高效糖化及后续乙醇发酵提供参考。     

蔗渣酶法水解在非缓冲溶剂体系中的反应机理探讨

通过测定蔗渣酶法水解在水系介质中的反应过程的pH随各种反应条件的变化,探讨了酶解液的各种特性,并测定了甘蔗渣、纤维素及纤维素酶相互作用的红外图谱。通过分析认为,纤维素在酶水解过程中,纤维素和纤维素酶相互作用形成了中间产物,使纤维素酶的立体构象发生变化而放出H^ ,使酶解液的pH发生变化,并形成了一种微弱的缓冲体系,致使甘蔗渣酶解反应在采用普通纯水代替缓冲液时,酶解反应的还原糖得率不变。     

木薯酒精渣的预处理及补料同步糖化发酵制取乙醇

木薯酒精渣的处置是制约木薯燃料乙醇大规模产业化的问题之一。本文立足于探索木薯酒精渣利用途径,分析了木薯酒精渣的主要成分,对比了氨水、氢氧化钠、氨水组合稀硫酸3种预处理方式对于木薯酒精渣纤维素和木素含量及纤维素酶水解效率的影响,分析了处理前后木薯酒精渣的表面结构及纤维素结晶度,并以氨水处理后的木薯酒精渣为底物,进行了同步糖化发酵。结果表明,3种预处理方法中组合预处理能更好地增加纤维素含量和提高纤维素酶水解效率,与未处理原料相比,组合预处理后纤维素含量增加了111.26%,木素下降了35.05%,酶水解72h纤维素转化率从42.10%增加到61.71%。氨水预处理后,原料的木素含量降低,处理后木薯酒精渣的表面变得更加粗糙,纤维素结晶度有所增加,以氨水处理后的木薯酒精渣为底物进行分批补料同步糖化发酵,当初始底物浓度为100.0g/L,分别在20h、40h、60h进行补料至最终底物浓度为400.0g/L时,发酵120h乙醇浓度达到51.0g/L。     

碱性过氧化氢预处理对稻草秸秆酶解的影响

木质纤维素生物转化的关键是预处理和酶解,为了提高木质纤维素的生物转化效率,在温和条件下对稻草秸秆进行弱碱性过氧化氢处理。研究了稻草秸秆处理固液比、过氧化氢和纤维素酶添加量对酶解糖化的影响,结果表明:稻草秸秆固液比为800 g/L,过氧化氢添加量为0.125 g/g,酶解时纤维素酶浓度25 FPU/g底物为较好,红外分析表明稻草秸秆处理后,秸秆纤维素的结晶度和木质素含量都有所下降,促进了生物转化后续过程中的酶解糖化。     

非离子表面活性剂对木质纤维素酶解的促进作用

纤维素酶单位酶活力较低、酶用量较高及酶自身易失活等因素依然是木质纤维素工业生产能源和生物基产品的瓶颈性问题。本文尝试在木质纤维素基质水解时添加一些非离子型表面活性剂以减少纤维素酶用量,并对这些非离子型表面活性剂促进酶解效率提高的原因进行了初步探讨。研究发现,添加非离子性表面活性剂能提高木质纤维素的酶解,添加浓度为0.05 g/g底物,常压甘油自催化预处理麦草经过添加两种非离子表面活性剂Tween-80和PEG 6000后葡萄糖产量分别可提高20%左右;非离子表面活性剂对不含木质素的原料酶解产糖也有较大的提高,以滤纸为底物时葡萄糖产量提高近90%,以微晶纤维素为底物时分别提高70%以上;添加非离子表面活性剂使得酶解体系中扩散系数k升高,异相反应效率提高,酶促反应动力学Km值明显减小,显著提高底物对纤维素酶的亲和力。     

半纤维素酶添加对碱处理甘蔗渣结构及酶解的影响

半纤维素作为木质纤维素的重要组分之一,通过氢键与纤维素的微纤丝结合,严重阻碍了纤维素表面与纤维素酶的接触,降低了酶解的效率。该试验以碱处理甘蔗渣作为底物,通过添加不同量的半纤维素酶去除不同比例的半纤维素。通过成分分析、X射线衍射(XRD)和扫描电镜(SEM)等手段分析添加半纤维素酶前后残渣的结构和酶解特性变化,发现随着半纤维素酶添加量的增大,残渣中木质素所占的比例逐渐增大,结晶指数逐渐增大,电镜表面沟壑逐渐加深,纤维束之间结构变得疏松。以半纤维素酶处理过的甘蔗渣作为底物,按照5FPU/g底物加入纤维素酶水解72h,与不添加半纤维素酶对照组相比,添加1600U/g半纤维素酶处理的试验组木聚糖的转化率提高了74.24%,葡聚糖转化率提高了35.30%。通过半纤维素酶添加可以有效促进纤维素酶解过程的进行,节约反应时间提高酶解转化率。     

Cellulolytic properties of Chaetomium crispatum

The cellulolytic properties of a Chaetomium crispatum strain were investigated. The cellulolytic enzyme complex i.e.: exo-1,4-β-glucosidase (EC 3.2.1.74); endo-1,4-β-glucanase (EC 3.2.1.4.) and β-glucosidase or cellobiase (EC 3.2.1.21) displayed optimal activity at pH 5.0 and 25°C. Although carboxymethyl-celluloses are the usual pseudo-substrates for this enzyme complex, those with a high degree of substitution gave rise to poor growth and low cellulase activity. Insoluble crude cellulosics such as newsprint, recycled paper, rice and flax straw were substantially solubilised at 28°C within 3–5 days of fermentation. A study of the cellulase-complex formation during the growth cycle revealed that β-glucosidase was produced mainly intracellularly in the early exponential phase, while the overall exo-1,4-β-glucosidase and endo-1,4-β-glucanase formation gradually increased during the total fermentation cycle. The mycelial protein of Chaetomium crispatum grown on crude cellulosics displayed a favourable amino acid pattern, indicating its potential value as a source of single cell protein (SCP).     

Laboratory- and commercial-scale investigations into the action of cellulase enzymes on Tencel

The surface appearance and feel of fabrics produced from the new cellulosic fibre Tencel can be enhanced by cellulase treatment. Laboratory-scale studies have been carried out using purified Trichoderma reesei cellulase components to determine which enzymes provide a benefit and which cause detrimental effects on Tencel fibre. Two enzymes were identified as beneficial and this information was used to produce new commercial cellulase products, using genetic engineering to remove detrimental components and increase amounts of the beneficial enzyme components. The effects of the new cellulases on Tencel fibre (both 100% pure and blends) were then evaluated. The observed advantages included improved hand-feel and reduced garment damage, especially in blends with other cellulosics.     

Evaluation of acids and cellulase enzyme for the effective hydrolysis of agricultural lignocellulosic residues

This paper reports the results of experimentation carried out to compare the ability of mineral acids (HCl and H₂SO₄) and cellulase enzyme (from Trichoderma reesei QM 9414) in the saccharification of corn-cob, groundnut shell, sugarcane bagasse and wheat straw. With the exception of corn-cobs, acids proved to be better saccharifying agents than the cellulase complex, but the former gave a poor substrate for alcoholic fermentation since the saccharified mashes contained large amounts of pentoses which are not metabolized by most strains of yeast. In addition, both acids and enzymes have been found to be substrate specific. Maximal saccharification of groundnut shell, sugarcane bagasse and wheat straw were obtained with sulphuric acid at 15.0, 5.0 and 5.0% (v/v) under 15, 15 and 15 psi pressure for 15, 30 and 30 min, respectively; whereas hydrochloric acid at 7.5% (v/v) with. autoclaving for 30 min at 10 psi resulted in maximum saccharification of corn-cob. However, the order of susceptibility of substrates to enzymatic attack was corn-cob > wheat straw > sugarcane bagasse > groundnut shell. Increase in enzyme concentration (1–4 IU ml^?1) and treatment duration (12–72 h) improved saccharification, but increases in substrate concentration (>5.0%, w/v) had an inhibitory effect on the hydrolytic ability of the cellulase enzyme complex. Of the various substrate-acid ratios tested, a ratio of 1:8 was found to be optimal for the eflcient hydrolysis of the substrates under study.     

Antimicrobial and Antioxidant Activity of Chitosan/Hydroxypropyl Methylcellulose Film-Forming Hydrosols Hydrolyzed by Cellulase

The aim of this study was to evaluate the impact of cellulase (C) on the biological activity of chitosan/hydroxypropyl methylcellulose (CH/HPMC) film-forming hydrosols. The hydrolytic activity of cellulase in two concentrations (0.05% and 0.1%) was verified by determination of the progress of polysaccharide hydrolysis, based on viscosity measurement and reducing sugar-ends assay. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging effect, the ferric reducing antioxidant power (FRAP), and microbial reduction of Pseudomonas fluorescens, Yersinia enterocolitica, Bacillus cereus, and Staphylococcus aureus were studied. During the first 3 h of reaction, relative reducing sugar concentration increased progressively, and viscosity decreased rapidly. With increasing amount of enzyme from 0.05% to 0.1%, the reducing sugar concentration increased, and the viscosity decreased significantly. The scavenging effect of film-forming solutions was improved from 7.6% at time 0 and without enzyme to 52.1% for 0.1% cellulase after 20 h of reaction. A significant effect of cellulase addition and reaction time on antioxidant power of the tested film-forming solutions was also reported. Film-forming hydrosols with cellulase exhibited a bacteriostatic effect on all tested bacteria, causing a total reduction.     

里氏木霉Rut-C30产纤维素酶培养基优化及其酶解特性

以廉价的工业纤维素诱导里氏木霉Rut-C30产纤维素酶,并对液体深层发酵培养基进行优化,采用响应面中心组合设计,以滤纸酶活为响应值,考察工业纤维素、麦麸、大豆粉浓度对纤维素酶活的影响. 结果表明,优化后的培养基组成为：工业纤维素35.62 g/L、麦麸19.37 g/L、大豆粉38.49 g/L,该条件下滤纸酶活达9.13 IU/mL,比优化前提高了72.26%,葡萄糖苷酶酶活提高了80.39%. 在121℃下用2% NaOH对玉米秸秆预处理45 min,物料中纤维素含量达64.94%,用该粗酶液酶解后酶解得率为94.68%.     

木质纤维原料生物转化燃料乙醇的研究进展

木质纤维生物量能够用来生产一种可替代有限的石油产品的能源——乙醇。木质纤维的转化主要分两个步骤木质纤维生物量中纤维素水解生成还原糖;糖发酵成乙醇。基于目前的技术,木质纤维原料生产乙醇的主要问题是得率低、水解成本高。促进木质纤维水解的方法包括木质纤维原料预处理脱除木素和半纤维素;纤维素酶的优化;同步糖化发酵法(SSF)。     

木质素结构以及表面活性剂对木质素吸附纤维素酶的影响

引言随着世界经济的快速发展和人口的增加,石油以及其他不可再生资源已经濒临枯竭。出于经济发展、国家安全以及环境保护等各个方面的需要,各个国家都在寻找石油的代替能源。生物乙醇这一新     

外切-β-葡聚糖酶基因重组里氏木霉的筛选及其产酶性能

外切-b-葡聚糖酶是纤维素酶的重要组分之一,提高该组分的活力是增强纤维素酶协同降解性能、降低纤维素水解成本的关键。分别采用微晶纤维素琼脂平板法和滤纸崩解法,对已有的基因重组转化子进行筛选试验,获得了6个优良转化子,其滤纸崩解速率和微晶纤维素琼脂平板上的生长速率都较大。进一步在摇瓶条件下进行复筛试验,获得了外切-β-葡聚糖酶（C₁）高产转化子Trichoderma reesei ZU-101,液体培养48 h,其C₁酶活力可达18.24 U·ml^-1,是出发菌株的2.16倍;分析结果表明：重组转化子的纤维素酶体系中内切-b-葡聚糖酶和纤维二糖酶的活力与出发菌株相比变化不大,但由于外切-b-葡聚糖酶活力得到了大幅度提高,纤维素酶的总活力（滤纸酶活力FPA）也提高了61.9%。采用纤维素酶对碱预处理玉米秸秆进行酶解试验,当酶用量为20 FPIU·（g底物）^-1,水解48 h,重组转化子T.reesei ZU-101纤维素酶的酶解得率高达94.4%。本文的研究结果在可再生纤维素资源的生物转化与利用方面具有广阔的应用前景。     

微孔淀粉的工艺研究

微孔淀粉是一种新型的变性淀粉。介绍了酸法和高温α-淀粉酶与糖化酶联用的方法水解玉米淀粉制备微孔淀粉,通过比较发现酶法优于酸法。并且当高温α-淀粉酶与糖化酶联用重量比为1∶3、pH=4.5、50℃、水解18 h时,可获得吸油率较高的微孔淀粉。