组合白腐真菌预处理木质纤维素原料的研究

木质纤维素原料种类多、分布广、数量巨大，通过燃料乙醇生产技术、厌氧沼气发酵技术将其转化成乙醇、沼气等二次能源，一定程度上可以缓解化石能源的不断消耗所带来的能源危机，也解决了农林废弃物引起的环境污染问题。其中以木质纤维素原料生产燃料乙醇，还可以避免以淀粉类和糖类原料生产燃料乙醇时带来的“与人争粮”等一系列问题。因此具有重要的经济效益、环境效益和社会效益。 然而，木质纤维素原料结构致密，木质素包裹在纤维素、半纤维素外围，导致其很难被降解利用，必须进行适当的预处理，去除木质素，打破原有的致密结构，利于原料的后续利用。因此，预处理成为木质纤维素原料能源化利用的关键。而目前预处理环节的费用过于昂贵，于是寻找一种高效、低成本的预处理方法是当今研究的热点。 本论文采用组合白腐真菌对木质纤维素原料进行生物预处理研究，与其他物理化学法相比，该法有着专一性较强、反应温和、不造成环境污染、成本低等优势。白腐真菌主要通过分泌木质素降解酶对木质素进行降解，从而破坏原料的致密结构，提高后续利用效率。所以木质素降解酶酶活的高低是影响原料预处理效果的一个关键因素。于是本论文首先通过将白腐真菌进行组合的方式提高木质素降解酶（漆酶，Lac）酶活；接着对组合菌的菌株相互作用机理进行研究，阐明组合菌Lac 酶活提高的原因，为菌株组合提高Lac 酶活这种方法的应用提供理论依据，同时也为后续组合白腐真菌预处理木质纤维素原料提供指导；进一步采用固态发酵和木质素降解酶两种方式对木质纤维素原料进行预处理研究，最大化去除木质素成分，破坏原料的致密结构；最终对预处理后原料的酶解糖化进行初步研究，为原料后续的能源化应用奠定基础。具体研究结果如下： （1） 以实验室保存的三株主要分泌Lac 的白腐真菌为出发菌株，筛选得到一组Lac 酶活明显提高的组合菌55+m-6，其中菌株55 为Trametes trogii sp.，m-6 为Trametes versicolor sp.，组合后Lac 酶活较单菌株分别提高24.13倍和4.07 倍。组合菌的最适产酶条件为pH 6.5、C/N 16:1、Tween 80 添加量为0.01%，在该条件下组合菌的Lac 酶活峰值比未优化时提高4.11倍。 （2） 对组合菌55+m-6 菌株间相互作用机理进行研究，发现菌株之间不存在抑制作用；平板培养时，菌丝交界处Lac 酶活最高并分泌棕色色素；液体培养时，菌株m-6 对组合后Lac 酶活的提高起着更为重要的作用：菌株m-6的菌块、过滤灭菌胞外物以及高温灭菌胞外物均能明显刺激菌株55 的Lac产生；菌株55、m-6 进行组合后，同工酶种类未发生增减，但有三种Lac同工酶浓度有所提高；对菌株胞外物进行薄层层析和质谱分析，结果表明组合前后菌株胞外物中各物质在浓度上存在较大的变化。推测组合菌Lac酶活的明显提高，主要是由于菌株m-6 胞外物中的一些物质能刺激菌株55 分泌大量Lac 进行代谢，且这些刺激物质并非菌株m-6 特有，菌株55自身也可以代谢生成，但是适当的浓度才能刺激Lac 的大量分泌。 （3） 将组合菌55+m-6 用于固态发酵预处理木质纤维素原料，发现其对玉米秆的降解程度最大，在粉碎度40 目、含水率65%的最优处理条件下，处理至第15d，秸秆失重率为41.24%，其中木质素、纤维素、半纤维素均有降解，且Lac 和纤维素酶（CMC）酶活以及还原糖量均达到峰值。 （4） 对玉米秆进行木质素降解酶预处理，发现Lac/1-羟基苯并三唑（HBT）系统对玉米秆木质素的降解效果最好，在最优处理条件时，即HBT 用量0.2%、处理时间1d、Lac 用量50U/g，木质素降解率可达12.60%。预处理后玉米秆的致密结构被破坏，比表面积增大，利于后续酶与纤维素、半纤维素成分的结合。 （5） 对预处理后的玉米秆进行酶解糖化，其中组合菌固态发酵预处理后玉米秆的糖化率比对照高4.33 倍；Lac/HBT 系统预处理后玉米秆的糖化率比对照高2.99％，糖化液中主要含有木糖、葡萄糖两种单糖。 There are many kinds and large quantities of lignocellulosic biomass widely distributed on the earth. They can be converted into secondary energy such as fuel ethanol, biogas, et al., which can relieve the energy crisis caused by consumption of fossil energy resources and solve the problem of environmental pollution caused by agriculture and forestry waste. Meanwhile, the production of fuel ethanol from lignocellulosic biomass can ensure food supply to human kind instead of starch- and sugar-containing raw materials. So the energy conversion of lignocellulosic biomass contributes considerable economic, environment and social benefits. However, lignocellulosic biomass has the compact structure, in which lignin surrounds cellulose and hemicellulose, so it must be pretreated before energy usage and pretreatment is one of the most critical steps in the energy conversion of lignocellulosic biomass. At present, the cost of pretreatment is too expensive, so looking for an efficient and low-cost pre-treatment method is one of recent research hot spots. In this research, combined white rot fungi pretreatment method was used, which had some advantages in low cost, high specificity, mild reacting conditions and friendly environmental effects compared with the other physical and chemical methods. White rot fungi secrete lignin degrading enzymes to degrade the content of lignin and damage the contact structure of lignocellulosic biomass, so the activity of the lignin degrading enzymes is the key factor to the degradation effect of raw materials. Firstly, the combined fungi with high laccase activity were screened; secondly, the interaction mechanism between strains was studied, and the cause of higher laccase activity after strains combination was also preliminary clarified; under the guidance of the mechanism, lignocellulosic biomass was pretreated by the combined fungi; lastly, the enzymatic hydrolysis of pretreated lignocellulosic biomass was also preliminary studied; all of the researches could lay the foundation for the energy application of lignocellulosic biomass. The specific research results were as follows: (1) The combined fungi 55+m-6 with significant higher laccase activity were screened from the three white rot fungi stored in our lab which mainly secreted laccase. Strain 55 and strain m-6 were Trametes trogii sp. and Trametes versicolor sp., respectively. The laccase activity of combined fungi was 24.13 and 4.07-fold than strain 55 and strain m-6, respectively. The optimized condition for laccase production of the combined fungi in liquid medium was pH 6.5, C/N 16:1 and Tween 80 0.01%. In this optimized condition, the laccase activity of combined fungi was 4.11-fold higher comparing with which in non-optimized medium. (2) The interaction mechanism between strain 55 and strain m-6 was further studied, and no inhibition effect was observed. Brown pigment was secreted on the junction of the two strains on the plate, where the highest laccase activity was detected. Strain m-6 was much important to boost laccase activity of combined fungi in liquid medium, and strain 55 was stimulated by fungal plug, filter sterilized extracellular substances and high temperature sterilized extracellular substances of strain m-6 to produce laccase. The types of laccase isozymes did not change after combining strain 55 and strain m-6, but the concentrations of three types increased. Mass Spectrometry and TLC analysis of extracellular substances of each strain showed that concentration of some substances considerably changed after strains were combined. It was supposed that the cause of higher laccase activity of combined fungi was mainly due to some extracellular substances of strain m-6 with the appropriate concentration which stimulated laccase secretion of strain 55 and generated not only by strain m-6 but also by strain 55. (3) Combined fungi 55+m-6 were used to lignocellulosic biomass pretreatment with the type of solid-state fermentation. The highest degree of degradation of corn straw was obtained, including the rate of weight loss was 41.24% and the lignin, cellulose and hemicellulose were degraded partially under the optimized condition of 40 mesh, 65% water content on 15th day. Laccase, CMCase activities and content of reducing sugar reached the maximum value on that day. (4) Lignin degrading enzymes from combined fungi 55+m-6 were used for corn straw pretreatment. The most remarkable degradation of lignin in corn straw with Lac/1-hydroxybenzotriazole (HBT) system was observed, and the 12.60% lignin degradation was obtained under the optimized condition of 0.2% HBT, 50 U/g laccase for 1 d. After pretreated by Lac/HBT, the tight structure of corn straw was demolished and specific surface area increased, which had advantages for accessible of enzyme to cellulose and hemicellulose. (5) The corn straws pretreated by combined fungi 55+m-6 with the type of solid-state fermentation and Lac/HBT were used for enzymatic hydrolysis, and the saccharification rates of each pretreatment type were 4.33 times and 2.99% higher than CK, respectively. The enzymatic hydrolysis liquid of corn straw pretreated by Lac/HBT mainly contained xylose and glucose.

Template-assisted syntheses of two novel porous zirconium methylphosphonates

Two porous zirconium methylphosphonates (designated as ZMPmi and ZMPme respectively) were synthesized by using dibutyl methylphosphonate (DBMP) as a template. Two efficient post-synthetic treatments were developed to remove the incorporated template without destroying the hybrid structures. The materials were characterized by SEM, EPMA, TG, DTA, FTIR, and NMR. Specific surface area and porosity were evaluated by BET, alpha(s)-plots and DFT methods based on N-2 adsorption-desorption isotherms. The specific surface areas of ZMPmi and ZMPme are determined to be 279 and 403 m(2) g(-1) and the maxima of pore size distributions are at 0.7 and 1.3 nm respectively. (c) 2005 Elsevier Inc. All rights reserved.

Effect of drying methods on the structure and catalytic combustion activity of Mn-substituted hexaaluminate catalysts

Conventional oven drying (COD) and supercritical drying (SCD) methods were applied to the preparation of Mn-substituted hexaaluminate (BaMnA(11)O(19-alpha)) catalysts. The effect of drying methods on phase composition, specific surface area, pore structure and combustion activity of the samples was investigated. The samples obtained by SCD have higher surface area, narrower pore size distribution, and higher combustion activity than those obtained by COD.

Fischer-Tropsch reaction over cobalt catalysts supported on zirconia-modified activated carbon

An attractive Fischer-Tropsch catalyst was prepared using an activated carbon as carrier to support cobalt based catalysts. Zr promoted Co/AC catalysts remarkably enhanced the activity and the selectivity toward diesel distillates and lower the methane selectivity. This modification may be attributed to specific behavior of activated carbon with high surface area and the weak interaction between metallic cobalt active sites and activated carbon. It was emphasized that the pore size of activated carbon played a very important role in restricting the growth of carbon chain to wax.

Preparation of Mn substituted La-hexaaluminate catalysts by using supercritical drying

LaMnxAl12-xO19 catalysts were prepared from NH4OH and metal nitrates solutions. Supercritical drying (SCD) and conventional oven drying (CD) methods were used to extract the water in the hydrogel. The effects of drying methods on properties of the catalysts were investigated by means of TEM, N-2-adsorption, thermogravimetry (TG)-differential thermal analysis (DTA) and X-ray diffraction. SCD method is beneficial to maintain high surface area and improving catalytic activity for methane combustion of the catalyst. The specific surface area and pore volume of LaMn1Al11O19 catalyst prepared by SCD method are 28 m(2)/g and 0.23 cm(3)/g, respectively, and the ignition of methane could be carried out at 450degreesC. However, those of the CD catalyst prepared from the same precursor are 15 m(2)/g, 0.11 cm(3)/g and 530 degreesC, respectively. Suitable Mn content (0 less than or equal to x less than or equal to 2) could promote the formation of LaMnAl11O19 hexaaluminate, while further addition of Mn (2 less than or equal to x less than or equal to 6) cause the formation of LaMnO3. (C) 2003 Elsevier B.V All rights reserved.

Formation of a novel type of reverse microemulsion system and its application in synthesis of the nanostructured La0.95Ba0.05MnAl11O19 catalyst

In the study, a novel microemulsion system, consisting of water, iso-propanol and n-butanol, was developed to synthesize the nanostructured La0.95Ba0.05MnAl11O19 catalyst with high surface area and catalytic activity for methane combustion.

Difference of ZSM-5 zeolites synthesized with various templates

ZSM-5 zeolites with similar SUM ratio were synthesized successfully using various templates (n-butylamine (BTA), ethylamine (ETA), isopropylamine (IPA), ethylenediamine (EDA), ethanol (ETL), ethanol-ammonium (ETL-AM) and no template (NT)) under hydrothermal conditions. The samples were characterized by XRD, SEM, XRF, NH3-TPD and BET surface area measurements in order to understand the template effects and the differences of the ZSM-5 samples. The synthesis of ZSM-5 with organic templates was relatively easier than those with inorganic templates and without template. SEM results revealed that ZSM-5 synthesized with different templates had different morphology and particle size. The Si/Al ratio and BET specific surface area of the sample with ethanol as template was the lowest. NH3-TPD results showed that the sample synthesized without template had fewer strong acid sites than others. n-Hexane cracking reaction was carried out over the samples to evaluate the catalytic properties. All ZSM-5 zeolites were effective in n-hexane cracking reaction, especially for the sample synthesized without template. (C) 2004 Elsevier B.V. All rights reserved.

Activated carbon supported bimetallic CoMo carbides synthesized by carbothermal hydrogen reduction

A carbothermal hydrogen reduction method was employed for the preparation of activated carbon supported bimetallic carbide. The resultant samples were characterized by BET surface area measurement, X-ray diffraction, and temperature-programmed reduction-mass spectroscopy. The results showed that nanostructured beta-Mo2C can be formed on the activated carbon by carbothermal hydrogen reduction above 700 degreesC. The particle sizes of beta-Mo2C increase with increasing reaction temperatures and Mo loading. The bimetallic CoMo carbide can be synthesized by the carbothermal hydrogen reduction even around 600 degreesC. The bimetallic CoMo carbide is from carbothermal hydrogen reduction of CoMoO4 precursor and is easily formed when the Co/Mo molar ratio is 1.0. Separation of the bimetallic CoMo carbide phase into Mo carbide and Co metal occurs when the temperature of the reduction is above 700 degreesC. The addition of a second metal such as Co and Ni, decreases the formation temperature of carbide because the second metal promotes formation of CHx species from reactive carbon atoms or groups on carbon material and hydrogen, which further carburizes oxide precursors. (C) 2003 Elsevier Science Ltd. All rights reserved.

Synthesis and spectroscopic study of mesoporous aluminum methylphosphonate foam templated by dibutyl methylphosphonate

New types of templates and novel interactive mechanisms between template and framework are very important for creating porous materials. In this work, by using neutral dibutyl methylphosphonate as a template, an inorganic-organic hybrid mesoporous material, aluminum methylphosphonate, was prepared. The as-synthesized material was studied by P-31 magnetic angle spinning nuclear magnetic resonance (MAS NMR), Al-27 MAS NMR, C-13 CP/MAS, FT-IR spectroscopy, thermogravimetry (TG), differential thermal analysis (DTA), and transmission electron microscopy. After thermal treatment at 673 K and 10 mmHg for 2 h, hybrid mesoporous foam was obtained. The transformation process was investigated by FT-IR. TG-DTA results indicate that the methyl group bonded to the framework keeps intact up to 792 K under air and 823 K under nitrogen. The characterization results from nitrogen gas adsorption-desorption measurements show that the BET surface area and the Barrett-Joyner-Halenda desorption cumulative pore volume of the foam are 90 m(2) g(-1) and 0.32 cm(3) g(-1) respectively. (C) 2003 Elsevier Inc. All rights reserved.

Shape selectivity in methane dehydroaromatization over Mo/MCM-22 catalysts during a lifetime experiment

Dehydroaromatization of CH4 with 2% CO2 on 6Mo/MCM-22 in a 100-h lifetime test was carried out at 993 K, atmospheric pressure and 1500 mL/gh. The duration of the lifetime test can be divided into an induction stage, stable stage and deactivation stage on the basis of the selectivities of hydrocarbons and coke. The characteristics of deposited coke with different time onstream were studied using TPO and TG techniques. There were two peaks corresponding to two kinds of coke recorded in TPO profiles, and the oxidation temperature of coke shifted to higher values with less hydrogen content with the increase of coke deposits. BET and Benzene-TPD techniques were employed to study the variation of specific surface area of the external and micropore surface versus time onstream. With the accumulation of coke deposits, although the pores became partially blocked and the internal surface decreased, methane could still enter the channel and was converted to benzene with shape selectivity until a critical value of coke deposition was reached.

云雾山本氏针茅群落根系分布特征

为揭示本氏针茅(Stipa bungeana Trin.)群落的生理生态适应机制,采用根系取样器(Φ=9 cm)对宁夏云雾山本氏针茅群落根系分布特征和土壤含水量进行研究。结果表明:本氏针茅群落地下生物量、根长密度、根表面积、比根长均随土壤深度增加而减少,均表现出向表层(0~20 cm)集聚的趋势,且集中分布于0~40 cm土层,最大值均分布在0~20 cm土层,而底层(80~100 cm)最小;所有主要根系分布参数在0~20 cm和20~40 cm土层之间差异显著,以下各层差异不显著;土壤含水量与根生物量和比根长相关性达显著水平(P<0.05),与根表面积、根长密度均呈成正相关;根表面积、根生物量、根长密度和比根长间相关性均达极显著水平(P<0.01)。

黄土丘陵区油松人工林和白桦天然林细根垂直分布及其与土壤养分的关系

在黄土高原子午岭林区,对油松人工林、白桦天然林细根生物量、比根长、根长密度和细根表面积的垂直分布特征,以及这些根系指标与土壤水分、土壤容重、氮素和有机质的关系进行了研究。结果表明,油松人工林细根生物量随土壤深度增加呈单峰曲线,白桦林细根生物量随土壤深度增加呈减少趋势;油松林大部分根系生物量集中分布在0—40 cm土层中,其中0—20 cm土层占37%以上,20—40 cm集中了41%以上;表层土壤(0—20 cm)具有较高的比根长、根长密度和细根表面积,而底层(40—60 cm)的比根长、根长密度和细根表面积最低。油松林土壤全氮和有机质含量垂直变化趋势相似,随土壤深度的增加而降低;硝态氮(NO3--N)均随土壤深度的增加呈单峰曲线变化趋势,而铵态氮(NH4+-N)随土壤深度增加呈先降低后增加的抛物线趋势。白桦林75%的细根生物量集中在0—20 cm土层,比根长、根长密度和细根表面积的垂直分布规律与油松林相似,表层土壤白桦林细根表面积是油松人工林的3.91倍,而20—40 cm土层白桦林细根表面积比油松人工林降低了33%。白桦林土壤全氮、有机质含量、NO3--N和NH4+-N垂直变化趋势与油松林相似。土壤水分、...

Changes in individual plant traits and biomass allocation in alpine meadow with elevation variation on the Qinghai-Tibetan Plateau

Plant traits and individual plant biomass allocation of 57 perennial herbaceous species, belonging to three common functional groups (forbs, grasses and sedges) at subalpine (3700 m ASL), alpine (4300 m ASL) and subnival (>= 5000 m ASL) sites were examined to test the hypothesis that at high altitudes, plants reduce the proportion of aboveground parts and allocate more biomass to belowground parts, especially storage organs, as altitude increases, so as to geminate and resist environmental stress. However, results indicate that some divergence in biomass allocation exists among organs. With increasing altitude, the mean fractions of total biomass allocated to aboveground parts decreased. The mean fractions of total biomass allocation to storage organs at the subalpine site (7%+/- 2% S.E.) were distinct from those at the alpine (23%+/- 6%) and subnival (21%+/- 6%) sites, while the proportions of green leaves at all altitudes remained almost constant. At 4300 m and 5000 m, the mean fractions of flower stems decreased by 45% and 41%, respectively, while fine roots increased by 86% and 102%, respectively. Specific leaf areas and leaf areas of forbs and grasses deceased with rising elevation, while sedges showed opposite trends. For all three functional groups, leaf area ratio and leaf area root mass ratio decreased, while fine root biomass increased at higher altitudes. Biomass allocation patterns of alpine plants were characterized by a reduction in aboveground reproductive organs and enlargement of fine roots, while the proportion of leaves remained stable. It was beneficial for high altitude plants to compensate carbon gain and nutrient uptake under low temperature and limited nutrients by stabilizing biomass investment to photosynthetic structures and increasing the absorption surface area of fine roots. In contrast to forbs and grasses that had high mycorrhizal infection, sedges had higher single leaf area and more root fraction, especially fine roots.

Effects of inoculated Microcoleus vaginatus on the structure and function of biological soil crusts of desert

Microcoleus vaginatus Gom., the dominant species in biological soil crusts (BSCs) in desert regions, plays a significant role in maintaining the BSC structure and function. The BSC quality is commonly assessed by the chlorophyll a content, thickness, and compressive strength. Here, we have studied the effect of different proportions of M. vaginatus, collected from the Gurbantunggut Desert in northwestern China, on the BSC structure and function under laboratory conditions. We found that when M. vaginatus was absent in the BSC, the BSC coverage, quantified by the percentage of BSC area to total land surface area, was low with a chlorophyll a content of 4.77 x 10(-2) mg g(-1) dry soil, a thickness of 0.86 mm, and a compressive strength of 12.21 Pa. By increasing the percentage of M. vaginatus in the BSC, the BSC coverage, chlorophyll a content, crust thickness, and compressive strength all significantly increased (P < 0.01). The maximum chlorophyll a content (13.12 mg g(-1)dry soil), the highest crust thickness, and the compressive strength (1.48 mm and 36.60 Pa, respectively) occurred when the percentage of inoculated M. vaginatus reached 80% with a complex network of filaments under scanning electron microscope. The BSC quality indicated by the above variables, however, declined when the BSC was composed of pure M. vaginatus (monoculture). In addition, we found that secretion of filaments and polymer, which stick sands together in the BSC, increased remarkably with the increase of the dominant species until the percentage of M. vaginatus reached 80%. Our results suggest that not only the dominant species but also the accompanying taxa are critical for maintaining the structure and functions of the BSC and thus the stability of the BSC ecosystems.

Direct electrochemistry of horseradish peroxidase immobilized in calcium carbonate microsphere doped with phospholipids

Protein electrochemistry affords a direct method to study the biological electron transfer processes. However, supplying a biocompatible environment to maintain the native state of protein is all-important and challengeable. Here, we chose vaterite, one of the crystalline polymorphs of calcium carbonate, with highly porous nature and large specific surface area, which was doped with phospholipids, as the matrix to immobilize horseradish peroxidase (HRP). The integrity of HRP was kept during the simple immobilization procedure. By virtue of this organic/inorganic complex matrix, the direct electrochemistry of HRP was realized, and the activity of HRP for catalyzing reduction of O-2 and H2O2 was preserved.