Effect of initial moisture content and chip size on the bioconversion efficiency of softwood lignocellulosics

Previous optimization strategies for the bioconversion of lignocellulosics by steam explosion technologies have focused on the effects of temperature, pH, and treatment time, but have not accounted for changes in severity brought about by properties inherent in the starting feedstock. Consequently, this study evaluated the effects of chip properties, feedstock size (40-mesh, 1.5 x 1.5 cm, 5 x 5 cm), and moisture content (12% and 30%) on the overall bioconversion process, and more specifically on the efficacy of removal of recalcitrant lignin from the lignocellulosic substrates following steam explosion. Increasing chip size resulted in an improvement in the solids recovery, with concurrent increases in the water soluble, hemicellulose-derived sugar recovery (7.5%). This increased recovery is a result of a decrease in the "relative severity" of the pretreatment as chip size increases. Additionally, the decreased relative severity minimized the condensation of the recalcitrant residual lignin and therefore increased the efficacy of peroxide fractionation, where a 60% improvement in lignin removal was possible with chips of larger initial size. Similarly, increased initial moisture content reduced the relative severity of the pretreatment, generating improved solids and hemicellulose-derived carbohydrate recovery. Both increased chip size and higher initial moisture content results in a substrate that performs better during peroxide delignification, and consequently enzymatic hydrolysis. Furthermore, a post steam-explosion refining step increased hemicellulose-derived sugar recovery and was most effectively delignified (to as low as 6.5%). The refined substrate could be enzymatically hydrolyzed to very high levels (98%) and relatively fast rates (1.23 g/L/h).     

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.     

木质素复合水凝胶性能及应用的研究进展北大核心CSCD

木质素是自然界中储量仅次于纤维素的木质纤维素资源,也是唯一的天然芳香族聚合物,其衍生的高值化产品可以应用于多个领域。木质素的高效高值高质生产是木质纤维素生物炼制的关键所在,但木质素大分子结构复杂多变、反应的活性差、官能团冗杂,制备出性能稳定的高分子材料有一定的难度。随着木质素改性的研究越来越深入,木质素复合水凝胶的应用也受到了极大的关注。本文从木质素的基本结构组成与反应特性出发,简要概括了木质素复合水凝胶的制备方法;具体介绍了木质素复合水凝胶的应用现状,包括生物传感器、控制释放材料、环境响应材料、吸附材料、电极材料以及其他材料的应用;综述了木质素复合水凝胶的最新研究与应用进展,并对木质素制备复合水凝胶的发展前景进行了评述。     

Effect of phenolic acids on manganese peroxidase-dependent peroxidation of linoleic acid and degradation of a non-phenolic lignin model compound

The influence of aromatic phenolic and non-phenolic acids on manganese peroxidase (MnP)-dependent peroxidation of linoleic acid, and oxidation of a non-phenolic lignin model compound (LMC) was studied. Phenolic compounds inhibited both the MnP-dependent lipid peroxidation (LPO) and non-phenolic LMC degradation in the system. The antioxidant activity of the aromatic compounds in the enzymatic system with MnP-dependent LPO depends on the presence of the phenolic hydroxyl groups attached to the aromatic ring structure, the methoxylation of the hydroxyl group in the ortho position in diphenolics, and number of carbon atoms in the side chain. Natural phenolic compounds inhibit the oxidation of non-phenolic lignin in the system based on MnP-mediated LPO, but do not prevent it. This result indicates that MnP-mediated LPO may play an important role in lignin degradation even in the presence of the phenolic antioxidant compounds, and supports the possibility of the involvement of LPO in the degradation of lignin in wood.     

Density functional theory for the selective adsorption of small molecules on a surface modified with polymer brushes

A density functional method based on weighted density approximation is extended to study the selective adsorption of small molecules on a surface modified with end-grafted square-well chains. The excess part of the Helmholtz free energy functional is divided into two components: the hard sphere repulsion and the square-well attraction. The equation of state for hard sphere chain fluids developed by Liu et al. is used to calculate the repulsive part of the excess Helmholtz free energy functional, and the equation of state for square-well chain fluid with variable range developed by Li et al. is employed to calculate the attractive part. With this theoretical model, we examine the physical properties of the grafted polymer and the selective adsorption of small molecules on the modified surface.     

Electrogenerated chemiluminescence of ruthenium complex immobilized in a highly cross‐linked polymer and its analytical applications

Electrogenerated chemiluminescence (ECL) of a ruthenium complex polymer modified carbon paste electrode and its analytical applications were investigated. The ruthenium complex polymer was prepared using bis(2,2‐bipyridine) (4,4‐dicarboxy‐2,2‐bipyridine) ruthenium(II). The ECL behaviours of ruthenium complex polymer modified carbon paste electrode were investigated in the absence and presence of tripropylamine (TPA). The modified carbon paste electrode exhibited long‐term stability and fine reproducibility. The ECL intensity of the modified carbon paste electrode was linear with the concentration of TPA in the range 2.0 × 10^–6–3.8 × 10^–3 mol/L, with a detection limit (S:N = 3) of 6 × 10^–7 mol/L. It was also found that raceanisodamine could enhance the ECL intensity of the modified electrode. The ECL intensity of the modified carbon paste electrode was linear with the concentration of raceanisodamine in the range 1.1 × 10^–5–6.0 × 10^–4 mol/L, with a detection limit (S:N = 3) of 6 × 10^–6 mol/L. This work demonstrates that the entrapment of ruthenium complex in a highly cross‐linked polymer is a promising approach to construct an ECL modified electrode with long‐term stability and fine reproducibility. The modified electrode designed has a potential application in the ECL detector. Copyright © 2008 John Wiley & Sons, Ltd.     

Renewable bio ionic liquids‐water mixtures‐mediated selective removal of lignin from rice straw: Visualization of changes in composition and cell wall structure

Pretreatment of rice straw by using renewable cholinium amino acids ionic liquids ([Ch][AA] ILs)‐water mixtures and the subsequent enzymatic hydrolysis of the residues were conducted in the present work. Of the eight mixtures composed of ILs and water, most were found to be effective for rice straw pretreatment. After pretreatment with 50% ILs‐water mixtures, the enzymatic digestion of the lignocellulosic biomass was enhanced significantly, thus leading to satisfactory sugar yields of >80% for glucose and approximately 50% for xylose. To better understand the ILs pretreatment mechanism, confocal laser scanning microscopy combined with immunolabeling and transmission electron microscopy were used to visualize changes in the contents and distribution of two major components—lignin and xylan. The results coupled with changes in chemical structures (infrared spectra) of the substrates indicated occurrence of extensive delignification, especially in cell corner and compound middle lumen of cell walls, which made polysaccharides more accessible to enzymes. This pretreatment process is promising for large‐scale application because of the high sugar yields, easy handling, being environmentally benign and highly tolerant to moisture, and significantly reduced cost and energy consumption. Biotechnol. Bioeng. 2013; 110: 1895–1902. © 2013 Wiley Periodicals, Inc.     

Enzyme-mediated coupling of a bi-functional phenolic compound onto wool to enhance its physical,mechanical and functional properties

Wool fibres have been modified with nordihydroguaiaretic acid (NDGA) to improve their performance at use. This water insoluble bi-functional phenolic compound has been grafted on wool through a laccase enzyme catalyzed reaction in an aqueous–ethanol mixture. The capacity of laccase to oxidise NDGA in this aqueous–organic medium has been studied electrochemically. The increase of CH₂, CH₃ and aromatic groups signal in the DRIFT spectra, together with SEM images of the enzymatically modified fabrics confirmed the covalent grafting of NDGA on wool. This one step enzymatic process for grafting of NDGA improved the physical and mechanical properties of wool fabrics such as shrink resistance, crease recovery and tensile strength. Furthermore, the NDGA imparted to the textile material strong antioxidant activity and UV protection.     

吉林蛟河针阔混交林功能性状分布格局及其对地形因素的响应

森林群落的构建过程及其内在机制是生态学研究的热点问题。植物功能性状是指能够代表植物的生活史策略,反映植物对环境变化响应的一系列植物属性。通过植物功能性状的分布格局及其对环境因素的响应有助于推测群落的构建过程及其内在作用机制。以吉林蛟河21.12hm²温带针阔混交林样地为研究对象,采集并测量了样地内34种木本植物的6种不同的功能性状。以20m×20m的样方为研究单元,通过计算平均成对性状距离指数（mean pairwise trait distance;PW）和平均最近邻体性状距离指数（mean nearest neighbor trait distance;NN）来探讨群落中单个性状和综合性状的分布格局。同时结合地形因子采用回归分析探讨功能性状的分布格局对局域生境变化的响应。基于PW的结果显示：单个性状中除叶面积外,其余性状的分布格局均为聚集分布多于离散分布;基于NN的结果显示：除叶面积和最大树高外,其余性状的分布格局为聚集分布多于离散分布。此外,由6种单个性状组成的综合性状的分布格局同样为聚集分布多于离散分布。基于回归分析的结果显示：森林群落中功能性状的分布格局受到海拔、坡度和坡向等因素的显著影响,而凹凸度的影响则不显著。研究结果表明包括环境过滤和生物相互作用的非随机过程能够影响温带针阔混交林的群落构建过程,中性过程对该区域群落构建过程的影响不显著。     

Production of lignin peroxidase by Phanerochaete chrysosporium immobilized on porous poly(styrene-divinylbenzene) carrier and its application to the degrading of 2-chlorophenol

Porous poly(styrene-divinylbenzene) carriers, for the immobilization of white rot fungus Phanerochaete chrysosporium have been prepared by the concentrated emulsion polymerization method. The concentrated emulsion consists of a mixture of styrene and divinylbenzene containing a suitable surfactant and an initiator as the continuous phase, and water as the dispersed phase. The polymerization of the monomers of the continuous phase generated the polymer carrier with a porcus structure. The white rot fungus Phanerochaete chrysosporium has been immobilized on porous poly(styrene-divinylbenzene) carriers and used for the batch production and the repeated batch production of lignin peroxidase in shake cultures based on a carbon-limited medium containing veratryl alcohol. The best results were achieved when a spore inoculum was used for immobilization instead of 1-day-old mycelial pellets, for both the batch production and the repeated batch production. The porous poly(styrene-divinylbenzene) immobilized Phanerochaete chrysosporium and freely suspended mycelial pellets were used as biocatalysts for the degradation of 2-chilorophenol in a 2-L bioreactor. The porous poly(styrene-divinylbenzene) particle (diameter congruent with 0.2 cm) immobilized spores exhibited a much higher activity in the degradation of 2-chlorophenol than the freely suspended mycelial pellets. (c) 1994 John Wiley & Sons, Inc.     

亚热带次生常绿阔叶林植物功能性状及多样性对人工抚育的响应

亚热带次生常绿阔叶林普遍存在生长滞缓、固碳功能和经济价值低下等问题,迫切需要通过人工抚育提高其生态系统功能。对植物功能性状及其多样性的深入研究,能够找寻植物共存与群落稳定发展的内在机制,为提升森林生态系统功能的实践提供基础。为了探究人工抚育对亚热带次生常绿阔叶林质量提升的影响,以杭州午潮山国家森林公园内的次生常绿阔叶林为研究对象,实施了不同强度的抚育措施,通过群落植被复查,分析群落植物功能性状以及功能多样性对不同强度人工抚育的响应。结果发现:抚育2年后,群落最大潜在树高与叶片氮、磷、钾含量显著上升,而叶片干物质含量和叶片叶绿素含量均有显著下降,比叶面积有下降趋势,两种强度的抚育之间差异不显著;叶片干物质含量、叶绿素含量与叶片氮含量间的相关性由不显著变为了显著负相关,而叶片干物质含量与叶片叶绿素含量,叶片磷、钾含量与叶片氮含量,叶片磷含量与叶片钾含量间的相关性由不显著变为显著正相关;在20%强度抚育下,功能丰富度指数均值从对照的0.52显著下降为0.16,功能均匀度指数与功能分离度指数的均值则分别上升到0.61和0.67,显著高于对照组的0.51和0.56;在10%强度抚育下,功能丰富度有所下降,功能均匀度与功能分离度均有所上升,但差异不显著。研究表明,人工抚育在一定程度上改变了群落水平的植物功能性状与功能多样性,提高了群落植物利用资源的能力和效率;而适度的抚育强度更能解放资源空间,对群落产生的影响更加明显。该结果为亚热带次生常绿阔叶林的经营与管理提供依据与参考。     

Degradation of softwood [14C lignin] lignocelluloses and its relation to the formation of humic substances in river and pond environments

The fate of lignin in water and sediment of the Garonne river (France) and of a pond in its floodplain was examined using specifically labeled [¹⁴C-lignin] lignocelluloses. No significant differences appeared in the mineralization rate of alder, poplar or willow [¹⁴C-lignin] in running water samples. Conversion of total radioactivity to ¹⁴CO₂ ranged between 18.7% and 24.4% after 120 days of incubation. Degree of ¹⁴C-labeled lignin mineralization in standing water and sediments was clearly lower, especially in submerged sediments, and was correlated with oxygen supply. After 60 days of incubation 3.3% to 7.9% of the ¹⁴C-labeled lignin was recovered in water samples as dissolved organic carbon originating from microbial metabolism. In water extracts from sediment the percentage of dissolved organic ¹⁴C was only 0.4% to 1.3% of the applied activity. In the humic fraction extracted from sediments it did not exceed 4.4% which was much lower than in soils. No significant difference appeared between river and pond conditions for humic substances formation.     

Conversion of a primary amine to a labeled secondary amine by the addition of phenolic group and radioiodination

To preserve the nucleophilicity of amino compounds during conjugative radioiodination, a new method for converting primary amines to phenolic secondary amines was developed. Amino acids were used as model compounds for establishing optimal conditions for the reductive amination. In the first step of the reaction, the aldehyde group of 4-hydroxybenzaldehyde (formylphenol) was reacted reversibly with an amino group to form an imine. The irreversible attachment of formylphenol to the amino group was accomplished by reduction of the imine with sodium cyanoborohydride. The pH optimum for the reaction was 5.0. Higher temperature has favorable effects on the rate and extent of the conjugation. Phenolic derivatives of amino compounds suitable for radioiodination are produced by the reactions described.     

Functional and structural insight into the flexibility of cytochrome P450 reductases from Sorghum bicolor and its implications for lignin composition

Plant NADPH-dependent cytochrome P450 reductase (CPR) is a multidomain enzyme that donates electrons for hydroxylation reactions catalyzed by class II cytochrome P450 monooxygenases involved in the synthesis of many primary and secondary metabolites. These P450 enzymes include trans-cinnamate-4-hydroxylase, p-coumarate-3′-hydroxylase, and ferulate-5-hydroxylase involved in monolignol biosynthesis. Because of its role in monolignol biosynthesis, alterations in CPR activity could change the composition and overall output of lignin. Therefore, to understand the structure and function of three CPR subunits from sorghum, recombinant subunits SbCPR2a, SbCPR2b, and SbCPR2c were subjected to X-ray crystallography and kinetic assays. Steady-state kinetic analyses demonstrated that all three CPR subunits supported the oxidation reactions catalyzed by SbC4H1 (CYP73A33) and SbC3′H (CYP98A1). Furthermore, comparing the SbCPR2b structure with the well-investigated CPRs from mammals enabled us to identify critical residues of functional importance and suggested that the plant flavin mononucleotide–binding domain might be more flexible than mammalian homologs. In addition, the elucidated structure of SbCPR2b included the first observation of NADP⁺ in a native CPR. Overall, we conclude that the connecting domain of SbCPR2, especially its hinge region, could serve as a target to alter biomass composition in bioenergy and forage sorghums through protein engineering.     

Structural and functional characterization of solute binding proteins for aromatic compounds derived from lignin: p‐Coumaric acid and related aromatic acids

Lignin comprises 15–25% of plant biomass and represents a major environmental carbon source for utilization by soil microorganisms. Access to this energy resource requires the action of fungal and bacterial enzymes to break down the lignin polymer into a complex assortment of aromatic compounds that can be transported into the cells. To improve our understanding of the utilization of lignin by microorganisms, we characterized the molecular properties of solute binding proteins of ATP‐binding cassette transporter proteins that interact with these compounds. A combination of functional screens and structural studies characterized the binding specificity of the solute binding proteins for aromatic compounds derived from lignin such as p‐coumarate, 3‐phenylpropionic acid and compounds with more complex ring substitutions. A ligand screen based on thermal stabilization identified several binding protein clusters that exhibit preferences based on the size or number of aromatic ring substituents. Multiple X‐ray crystal structures of protein–ligand complexes for these clusters identified the molecular basis of the binding specificity for the lignin‐derived aromatic compounds. The screens and structural data provide new functional assignments for these solute‐binding proteins which can be used to infer their transport specificity. This knowledge of the functional roles and molecular binding specificity of these proteins will support the identification of the specific enzymes and regulatory proteins of peripheral pathways that funnel these compounds to central metabolic pathways and will improve the predictive power of sequence‐based functional annotation methods for this family of proteins.Proteins 2013; 81:1709–1726. © 2013 Wiley Periodicals, Inc.     

Synthesis and characterization of a novel three-dimensional photoluminescent coordination polymer generated from unsymmetrically bridging ligand

A novel three-dimensional metal-organic coordination polymer, [Zn₂(HBTC)₂(H₂O)₃]_n (1) (BTC=1,2,4-benzenetricarboxylate), has been prepared by aqueous solution reaction of Zn(NO₃)₂ · 6H₂O and BTC at moderate temperature and characterized by IR, TGA and single-crystal X-ray diffraction analysis. The three-dimensional architecture of 1 not only possesses rectangular cavities but also has ordered one-dimensional straight channels. Furthermore, compound 1 shows intense photoluminescent property at room temperature.     

Cloning of the promoter region of plasma membrane aquaporin BnPIP1 from Brassica napus and its functional analysis

Aquaporins make water-selective channels in plants, facilitating the permeation of water through membranes and adjusting water fast transport during seed germination, cell elongation, stoma movement, fertilization and responses to environmental stresses. They belong to the MIP (major intrinsic protein) family with molecular weight of 2629 kD and are characterized by six membrane-spanning a-helixes connected by five loops and short N-terminal and C-terminal domains in the cytoplasm[13]. The p…     

Differences in functional traits between invasive and native Amaranthus species under simulated acid deposition with a gradient of pH levels

Co-occurring invasive plant species (invaders hereafter) and natives receive similar or even the same environmental selection pressures. Thus, the differences in functional traits between natives and invaders have become widely recognized as a major driving force of the success of plant invasion. Meanwhile, increasing amounts of acid are deposited into ecosystems. Thus, it is important to elucidate the potential effects of acid deposition on the functional traits of invaders in order to better understand the potential mechanisms for the successful invasion. This study aims to address the differences in functional traits between native red amaranth (Amaranthus tricolor L.; amaranth hereafter) and invasive redroot pigweed (A. retroflexus L.; pigweed hereafter) under simulated acid deposition with a gradient of pH levels. Pigweed was significantly taller than amaranth under most treatments. The greater height of pigweed can lead to greater competitive ability for resource acquisition, particularly for sunlight. Leaf shape index of pigweed was also significantly greater than that of amaranth under all treatments. The greater leaf shape index of pigweed can enhance the efficiency of resource capture (especially sunlight capture) via adjustments to leaf shape and size. Thus, the greater height and leaf shape index of pigweed can significantly enhance its competitive ability, especially under acid deposition. Acid deposition of pH 5.6 significantly increased amaranth leaf width in the co-cultivation due to added nutrients. The pH 4.5 acid deposition treatment significantly increased the specific leaf area of amaranth in the monoculture compared with the pH 5.6 acid deposition treatment and the control. The main mechanism explaining this pattern may be due to acid deposition mediating a hormesis effect on plants, promoting plant growth. The values of the relative competition intensity between amaranth and pigweed for most functional traits were lower than zero under most treatments. Thus, competitive performance arose in most treatments when the two species were grown together. This may be due to the enhanced competitive intensity under interspecific coexistence. However, the values of the relative competition intensity of the leaf functional traits between amaranth and pigweed were all higher than zero under the pH 5.6 simulated acid deposition treatment. Thus, interspecific facilitation occurs when the two species are co-cultivated under the pH 5.6 simulated acid deposition treatment. This may be due the positive nutritional effects induced in the pH 5.6 simulated acid deposition treatment.