Oscillating Cellulase Adsorption and Enhanced Lignocellulose Hydrolysis upon Ultrasound Treatment

We investigated the effects of ultrasound treatment on cellulase adsorption and lignocellulose hydrolysis. The activity of cellulase remained constant upon low-power ultrasound treatment (280 W). Oscillating cellulase adsorption occurred upon ultrasound treatment with any intensity. The maxima for desorption and adsorption were 41.9 and 83.1%, respectively, during 1 h of 90 W ultrasound treatment at 50 °C. A comparison between the short-time with long-time ultrasound experiments indicated that ultrasound treatment tended to desorb cellulase from substrate. However, ultrasound treatment also led to further surface erosion of biomass, which increased cellulase accessibility. These joint actions of ultrasound treatment induced the oscillating adsorption of cellulase. The increase in cellulase accessibility caused by ultrasound treatment led to a significant enhancement in lignocellulose hydrolysis. © 2016, Tianjin University and Springer-Verlag Berlin Heidelberg.

Production of Cellulase and Xylanase by Aspergillus terreus KJ829487 Using Cassava Peels as Subtrates

Cassava (Manihot esculenta, Crantz) is one of the most important food plants in West Africa. Its peels are made up of cellulose, hemicellulose and lignin. This lignocellulolytic biomass can be converted using microbial enzymes to fermentable sugars which have wide range of biotechnological relevance in many fermentation processes. The aim of this study is to screen filamentous fungi from decaying cassava peels that are good producers of xylanases and cellulases. Decaying parts of cassava peels were obtained and brought to the laboratory for further work. Fungi were isolated, identified and screened for cellulase and xylanase production. Isolate with highest frequency of occurrence and enzyme production was identified using phenotypic and molecular method. Optimisation of growth conditions for enzymes production was monitored using the DNSA method, also saccharification of cassava peel were carried out using the enzymes obtained from the isolate. Aspergillus terreus KJ829487 was the predominant fungus. It produces cellulases and xylanases optimally at 40°C, pH 6 and 8, utilising carboxymethylcellulose (CMC) or xylose and yeast extracts as its carbon and nitrogen sources respectively. Saccharification of the peels yielded 584 mg/L glucose, 78 mg/L xylose and 66 mg/L rhamnose. Aspergillus terreus KJ829487 obtained from cassava peels have the ability to produce high concentration cellulases and xylanases which effectively hydrolysed the lignocelluloses’ biomass to fermentable sugars.

Preliminary Characterization of a Cellulase Producing Bacterial Strain Isolated from a Romanian Hypersaline Lake

Cellulases are a group of enzymes that are used in many biotechnological processes. Since most of the enzymes synthesised by mesophilic microorganisms are unstable in industrial environments, it is necessary to direct research towards extremophile cellulolytic microorganisms because the enzymes synthesised by them are stable and active even in harsh physicochemical conditions. In the present investigation, our aim was to isolate and identify some microbial cellulolytic strains from a hypersaline lake located in Romania and to determine their optimal growth conditions. Of a total of 25 microbial strains isolated, only one extreme halotolerant bacterial strain was able to produce an endoglucanase. Based on molecular identification, we identified this cellulolytic strain as a species of Bacillus genus, most closely related to Bacillus zhangzhouensis. Optimal growth conditions were found to be at 15°C, pH 7.5 and 2 M NaCl. Endoglucanase activity of this bacterial strain is influenced by both salinity and temperature. The most significant endoglucanase activity was detected in the presence of 3 M NaCl, after 72 h of incubation at 15°C. In this situation, the amount of glucose released from a volume of 0.5 mL of 2% (w/v) carboxymethyl cellulose substrate is equivalent to 2.05 mg. In conclusion, this study represents the first preliminary characterization of a B. zhangzhouensis strain that has the ability to degrade cellulose and that demonstrates tolerance to high salt concentrations.

Lignin Interaction with Cellulase during Enzymatic Hydrolysis

The conversion of lignocellulosic biomass into biofuels or biochemicals typically involves a pretreatment process followed by the enzyme-catalyzed hydrolysis of cellulose and hemicellulose components to fermentable sugars.Many factors can contribute to the recalcitrance of biomass,e.g.,the lignin content and structure,crystallinity of cellulose,degree of fiber polymerization,and hemicellulose content,among others.However,nonproductive binding between cellulase and lignin is the factor with the greatest impact on enzymatic hydrolysis.To reduce the nonproductive adsorption of enzymes on lignin and improve the efficiency of enzymatic hydrolysis,this review comprehensively summarized the progress that has been made in understanding the interactions between lignin and enzymes.Firstly,the effects of pretreatment techniques on lignin content and enzymatic hydrolysis were reviewed.The effects of lignin content and functional groups on enzymatic hydrolysis were then summarized.Methods for the preparation and characterization of lignin films were assessed.Finally,the methods applied to characterize the interactions between lignin and cellulase were reviewed,and methods for decreasing the nonproductive binding of enzymes to lignin were discussed.This review provides an overview of the current understanding of how lignin hinders the enzymatic hydrolysis of lignocellulosic biomass,and provides a theoretical basis for the development of more economical and effective methods and additives to reduce the interaction of lignin and enzymes to improve the efficiency of enzymatic hydrolysis.

Application of cellulase in producing reducing sugar with waste lignocellulosic

The cellulase expands the use of waste lignocellulosic and improves the feasibility of ethanol production with waste lignocellulosic. In this paper the types, mechanisms of cellulase and its application in reducing sugar production were presented in detail. The strains that produce cellulase and methods for improving the cellulase activity in reducing sugar production with waste lignocellulosic were described.

Integrated engineering of enzymes and microorganisms for improving the efficiency of industrial lignocellulose deconstruction

Bioconversion of lignocellulosic biomass to fuels and chemicals represents a new manufacturing paradigm that can help address society’s energy,resource,and environmental problems.However,the low efficiency and high cost of lignocellulolytic enzymes currently used hinder their use in the industrial deconstruction of lignocellu-lose.To overcome these challenges,research efforts have focused on engineering the properties,synergy,and production of lignocellulolytic enzymes.First,lignocellulolytic enzymes’catalytic efficiency,stability,and toler-ance to inhibitory compounds have been improved through enzyme mining and engineering.Second,synergistic actions between different enzyme components have been strengthened to construct customized enzyme cocktails for the degradation of specific lignocellulosic substrates.Third,biological processes for protein synthesis and cell morphogenesis in microorganisms have been engineered to achieve a high level and low-cost production of lignocellulolytic enzymes.In this review,the relevant progresses and challenges in these fields are summa-rized.Integrated engineering is proposed to be essential to achieve cost-effective enzymatic deconstruction of lignocellulose in the future.

Cellulase production by Aspergillus unguis in solid state fermentation

Lignocellulosic substrates are a good carbon source and provide rich growth media for a variety of microorganisms which prodLuce industrially important enzymes. Cellulases are a group of hydrolytic enzymes such as filter paperase (FPase), carboxymethyl cellulase(CMCase) andβ-glucosidase-responsible for release of sugars in the bioconversion of the lignocellulosic biomass into a variety of value-added products. This study examined cellulase production by a newly isolated Aspergillus unguis on individual lignocellulosic substrates in solid state fermentation (SSF). The maximum peak production of enzymes varied from one substrate to another, however,based on the next best solid support and local availability of groundnut fodder supported maximum enzyme yields compared with other solid supports used in this study.Groundnut fodder supported significant production of FPase (5.9 FPU/g of substrate), CMCase (1.1 U/g of substrate) andβ-glucosidase activity (6.5 U/g of substrate) in SSF. Considerable secretion of protein (27.0 mg/g of substrate) on groundnut fodder was recorded. Constant increment of protein content in groundnut fodder due to cultivation of A. unguis is an interesting observation and it has implications for the improvement of nutritive value of groundnut fodder for cattle.

Cellulase Production from Species of Fungi and Bacteria from Agricultural Wastes and Its Utilization in Industry: A Review

In energy deficient world, cellulases play a major role for the production of alternative energy resources utilizing lignocellulosic waste materials for bioethanol and biogas production. This study highlights fungal and bacterial strains for the production of cellulases and its industrial applications. Solid State Fermentation (SSF) is more suitable process for cellulase production as compared to submerge fermentation techniques. Fungal cellulosomes system for the production of cellulases is more desirable and resistant to harsh environmental conditions. Trichoderma species are considered as most suitable candidate for cellulase production and utilization in industry as compared to Aspergillus and Humicola species. However, genetically modified strains of Aspergillus have capability to produce cellulase in relatively higher amount. Bacterial cellulase are more resistant to alkaline and thermophile conditions and good candidate in laundries. Cellulases are used in variety of industries such as textile, detergents and laundries, food industry, paper and pulp industry and biofuel production. Thermally stable modified strains of fungi and bacteria are good future prospect for cellulase production.

植物细胞壁抗降解屏障研究进展与展望

纤维素是地球上最丰富的可再生资源,经酶促水解反应生成葡萄糖是纤维素转化的理想方式,但转化效率低,探究相关的抑制因素及其特点成为纤维素高值化应用的首要基础科学问题。本研究首先从宏观形态、显微结构、超微结构、分子、基团、元素、化学键和基因水平,多角度、多层次总结制约纤维素酶促水解反应效率的关键抑制因素,阐述抗降解屏障的内涵;其次明确抗降解屏障表现出的在不同生物质的特有异质性、在植物发育不同阶段的高度动态性以及在预处理过程的复杂联动性;最后展望生物质抗降解屏障破解研究的新趋势和新策略:细胞壁界面抗降解屏障的新认识,细胞壁修饰与改造方法,细胞壁精准解构与组分分级利用的新技术和多酶协同酶解体系的建立等。

提高木质纤维生物炼制效率的纤维素酶生产策略

纤维素酶是木质纤维生物炼制过程中的关键要素,木质纤维的高效酶解为其生物炼制提供了巨大的机会。如何提高纤维素酶的效价、降低酶解成本,从而实现高效的木质纤维生物炼制是当前关注热点之一。本文综述了纤维素酶的种类及其作用机理,重点分析了纤维素酶的微生物来源,生产纤维素酶的发酵方式及诱导剂,产纤维素酶菌株的筛选与改造,并对利用纤维素酶的复配提升纤维素酶活力和利用纤维素酶的在线生产降低纤维素酶生产成本进行阐述,并提出了更好地实现纤维素酶生产成本控制的策略。

纤维素酶在木质纤维素生物质转化中的应用研究

选育得到纤维素酶高产菌株里氏木霉突变菌株 (Trichodermareesei) 81 3A ,优化了其发酵产酶条件。利用该菌株所产纤维素酶对天然木质纤维素的水解糖化过程进行研究 ,确定了实验条件下最优的糖化条件 (温度 5 0℃ ,pH 4 5 ,酶浓度 6～ 8FPU mL ,底物浓度 2 % )。以玉米叶和杨树叶为天然纤维素原料 ,水解糖化率分别达到 86 2 %和 5 6 0 %。通过酿酒酵母 (Saccharomycescerevisiae)将糖化液转化为酒精 ,产乙醇浓度达到 5 %～ 5 8% ,转化率为79 4 %～ 92 1 %。

直接生物转化纤维素类资源生产燃料乙醇的研究进展

随着全球性能源危机、粮食危机和环境危机的到来 ,人们对废弃物的有效利用更加关注 ,尤其是可再生的纤维素类资源的有效利用 ,已引起世界各国的高度重视。本文从纤维素类物质的化学结构出发 ,阐述了纤维素类物质的组成、预处理、纤维素酶及酶解机理和直接生物转化为酒精等方面的研究发展情况。

木质纤维生产燃料乙醇的生物转化技术

木质纤维生物转化乙醇可以避免以粮食为原料生产燃料乙醇所带来的“与农争地”和“与人争食”的弊端,兼得经济、生态、环保、社会多重效益。木质纤维原料的物理化学特性决定其具有不同于淀粉和糖类原料转化的难度和复杂性,这也是木质纤维生物转化乙醇成本居高难下、商业化裹足不前的主要技术原因。本文以国际(特别是美国)的研究和开发进展为背景,探讨木质纤维生物转化乙醇的几个富有挑战性的技术问题,并在同步糖化共发酵(SSCF)概念的基础上,提出优化的并行糖化共发酵生产模型。

生物转化木质纤维素原料生产乳酸的研究进展

以木质纤维素生物原料转化生产乳酸对节约粮食、保护环境和促进乳酸产业的发展意义重大。对纤维素生产乳酸的几个技术环节:原料预处理、酶促水解、乳酸发酵的研究进展作了概要介绍和评述。并强调提高酶解效率和降低生产成本仍是现阶段研究的主要目标,指出各种新技术的集成与优化是加快其产业化进程的必由之路。

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)。

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。

生物转化食用菌菌糠木质纤维素产燃料乙醇的研究进展

近年来,食用菌生产技术在世界各国得以广泛普及,全球食用菌菌糠(spent mushroom substrate,SMS)总产量也随之大幅增长。随着全球性能源危机的到来,利用可再生纤维素类物质生产燃料乙醇已引起世界各国的高度重视。食用菌菌糠是食用菌子实体采收后的固体废弃物,其含有纤维素、半纤维素、木质素、抗营养因子和胞外纤维素降解酶类等组分,具备了作为第二代生物乙醇转化基质的潜力,基于此,该文对当前利用食用菌菌糠生物转化生产乙醇的研究进展和应用前景进行了阐述。

用于生物质酶解过程的纤维素酶研究进展

纤维素酶解效率是木质纤维素经济、高效生化转化的限制瓶颈。该文讨论了影响纤维素酶酶解经济性与高效性的多个要素,如:高滤纸酶活菌株的选育、发酵、酶解机理、酶解影响因素及酶解混合体系的优化等。该文研究表明,纤维二糖水解酶可能是决定发酵体系中滤纸酶活高低的关键单酶组分,同时,该酶可能也是预处理后生物质酶解体系中决定滤纸酶活效率的关键单酶组分。酶解过程中关键限速反应的认识及关键限制因素形成机制的揭示将成为纤维素酶生化转化研究的重点,这些机理机制的建立可为构建高比活力纤维素酶提供理论依据。

粗糙脉孢菌木质纤维素降解利用研究进展

粗糙脉孢菌作为木质纤维素降解真菌,不仅具有完整的木质纤维素降解酶系,而且还拥有全基因组基因敲除突变体库,是研究丝状真菌纤维素酶表达分泌和木质纤维素降解机制的优秀体系。近年来,国内外利用粗糙脉孢菌系统,在木质纤维素降解机制方面取得了显著进展,包括纤维素酶信号传导、调控以及生物质降解后糖的转运利用等。笔者就相关方面的进展进行综述,并对利用粗糙脉孢菌研究木质纤维素降解利用进行展望,总结和分析木质纤维素降解机制研究的国际前沿动态,有助于加深本领域研究人员对真菌体系纤维素降解机制的理解。

假木质素沉积及对纤维素酶解的影响研究进展

木质纤维素类生物质是地球上最丰富的可再生资源。为提高木质纤维素类生物质的转化率,提升纤维素酶的水解效率和可发酵性糖产量,降低纤维素酶的使用量和生物质转化成本,对木质纤维素类生物质进行预处理十分必要;然而,木质素、纤维素和半纤维素之间的天然屏障限制了纤维素酶对纤维素组分的酶解。木质纤维素类生物质预处理主要有物理法、化学法、物理化学法和生物法,目前更多采用质量分数小于4%的稀酸法(如盐酸、硫酸和硝酸等,120~210℃)、高温热水法、蒸汽爆破法和液相水热法等,不同预处理方法对木质素或大部分半纤维素的溶解和去除有利于提高纤维素酶的可及性。木质素对纤维素酶解具有明显抑制作用,通过预处理降低木质素含量有利于提高纤维素酶解效率。木质纤维经稀酸或高温热水等预处理后,Klason木质素相对含量反而会增加。在木质纤维素类生物质预处理过程中,木质素液滴可能以假木质素形式沉积于纤维素表面,使其比天然木质素更加抑制纤维素酶解。本研究首先概述生物质预处理过程中木质素液滴和假木质素的形成过程,提出假木质素产生的可能机制,并对其组成和性质进行综述;然后阐述木质素液滴和假木质素对木质纤维酶解的影响;最后总结假木质素形成的调控策略。假木质素的形成过程属于非均相反应过程,受传质扩散(分子水平)和流动(宏观统计水平)的影响,可从介尺度行为研究假木质素的形成机制,同时建立相关模型和理论实现其科学的定量描述和定向调控,这不仅有利于木质纤维素类生物质炼制工艺的发展,也有利于促进跨学科科学规模的形成。