972 resultados para ETHANOL FERMENTATION


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The TiO2-supported zeolite with core/shell heterostructure was fabricated by coating aluminosilicate zeolite (ASZ) on the TiO2 inoculating seed via in situ hydrothermal synthesis. The catalysts were characterized by transmission electron microscope (TEM), X-ray diffraction (XRD), nitrogen physisorption (BET), and Fourier transform infrared spectroscopy (FT-IR). The surface acidity of the catalysts was measured by pyridine-TPD method. The catalytic performance of the catalysts for ethanol dehydration to ethylene was also investigated. The results show that the TiO2-supported zeolite composite catalyst with core/shell heterostructure exhibits prominent conversion efficiency for ethanol dehydration to ethylene.

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TiO2/4A zeolite composite catalysts were prepared by coating TiO2 on 4A zeolite via liquid phase deposition. The TiO 2/4A zeolite composite catalysts wtih higher surface weak acidity and lower mediate strong acidity exhibit much better catalytic performance on ethanol dehydration to ethylene compared with 4A zeolite. It is suggested that the TiO2 promoter could improve the effective Lewis acidity of composite catalyst which consequently enhanced the catalytic performance.

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The hydrogen production from the organic fraction of municipal solid waste (OFMSW) by anaerobic mixed culture fermentation was investigated using batch experiments at 37 degrees C. Seven varieties of typical individual components of OFMSW including rice, potato, lettuce, lean meat, oil, fat and banyan leaves were selected to estimate the hydrogen production potential. Experimental results showed that the boiling treated anaerobic sludge was effective mixed inoculum for fermentative hydrogen production from OFMSW. Mechanism of fermentative hydrogen production indicates that, among the OFMSW, carbohydrates is the most optimal substrate for fermentative hydrogen production compared with proteins, lipids and lignocelluloses. This conclusion was also substantiated by experimental results of this study. The hydrogen production potentials of rice, potato and lettuce were 134 mL/g-VS, 106 mL/g-VS, and 50 mL/g-VS respectively. The hydrogen percentages of the total gas produced from rice, potato and lettuce were 57-70%, 41-55% and 37-67%. 2008 International Association for Hydrogen Energy.

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The novel hexagon SnO2 nanosheets are successfully synthesized in ethanol/water solution by hydrothermal process. The samples are characterized by X-ray diffraction (XRD), infrared ray (IR) and transmission electron microscopy (TEM). By changing the reaction conditions, the size and the morphology can be controlled. Comparison experiments show that when the temperature increased from 140 degrees C to 180 degrees C, the edge length of the hexagon nanoparticles increases from 300-450 nm to 700-900 nm. On the other hand, by adjusting the ratios of water to ethanol from 2 to 0.5, SnO2 nanoparticles with different morphologies of triangle and sphere are obtained. When the concentration of NaOH is increased from 0.15 M to 0.30 M, a hollow ring structure can be obtained. (c) 2006 Elsevier B.V. All rights reserved.

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Two biosensors for fermentation process control have been introduced, which were developed in our lab recently. One is an enzyme electrode-based on-line monitoring system for glutamate fermentation process control and the other is an H+-ISFET-based ENFET for penicillin G fermentation process control.

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We present in this paper results obtained from a parabolic flight campaign regarding ethanol sessile drop evaporation under reduced gravity conditions. Drops are created using a syringe pump by means of injection through a PTFE (polytetrafluoroethylene) substrate. The drops are recorded using a video camera and an infrared camera to observe the thermal motion inside the drop and on the heating substrate. The experimental set-up presented in this paper enables the simultaneous visualization and access to the heat flux density that is transferred to the drop using a heat flux meter placed between the heating block and the PTFE substrate. We evidence original thermal spreading phenomena during the ethanol drop creation on a heated PTFE substrate. The drop exhibits specific behaviour which is discussed here. This work is performed in the frame of a French-Chinese collaboration (project IMPACHT) for future experiments in a Chinese scientific satellite.

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本文从四川绵竹酒厂、成都市龙泉长安垃圾填埋场以及四川大学荷花池底的厌氧污泥中先后分离得到63株厌氧产氢菌,其中H-8、H-61、HC-10等16株产氢细菌产氢能力较高,HC-10的产氢能力最高,最大产氢量和最大产氢速率分别达到2840 ml H2/L培养基和25.39 mmol H2/g drycell·h,对HC-10进行生理生化鉴定和分子生物学鉴定,判定其为clostridium sp.,对HC-10的产氢条件进行了研究,结果表明,该菌的最适生长温度为35 ℃,最适生长初始pH为7,以葡萄糖为最佳碳源,以蛋白胨为最佳氮源,不利用无机氮源,其产氢发酵液相产物以乙醇和乙酸为主,其发酵类型属于乙醇型发酵。此外,以酒糟废液作为底物,进行了菌株HC-10的生物强化试验,研究表明,投加了HC-10的强化系统其产氢量比对照高出40.32%。 同时为了获得厌氧产氢菌的高效突变株,分别以产氢菌H-8和H-61为原始菌株进行微波诱变处理,对微波诱变参数进行了优化,考察了突变株的遗传稳定性、产氢特性及耐酸性。菌株H-8经过微波诱变得到5株高产氢突变株HW7、HW33、HW181、HW184、HW195,经多次传代表明HW195是稳定的高产突变株。突变株HW195具有较好的耐酸性,在pH值为2.8时仍能生长。通过间歇发酵实验,其最大产氢量和最大产氢速率分别达到2460 mL/L培养基和27.97 mmol H2/g drycell·h,比原始菌分别提高了50.75%和41.7%。菌株H-61经过微波诱变后选育得到的突变株HW-18,其最大产氢量和最大产氢速率分别达到2190 mL/L培养基和25.86 mmol H2/g drycell·h,比原始菌分别提高了23.03%和31.00%。 为了对比各种诱变方式对产氢菌产氢能力的影响,以厌氧产氢菌H-61为原始菌株,先后经亚硝基胍(NTG)、紫外(UV)诱变,选育得到1株高产突变株HCM-23。在葡萄糖浓度为10 g/L的条件下,其产氢量为3024 mL/L培养基,比原始菌株提高了69.89%;其最大产氢速率为33.19 mmol H2/g drycell·h,比原始菌株提高了68.14%。经过多次传代实验,稳定性良好。其发酵末端产物以乙醇和乙酸为主,属于典型乙醇型发酵。其最适产氢初始pH为6.5,最适生长温度为36 ℃,以蔗糖为最佳碳源。与原始菌株相比,突变株HCM-23的产氢特性发生了改变,如生长延滞期延长,可利用无机氮源等。 From anaerobic activated sludge, 16 strains of hydrogen producing bacteria were newly isolated. One of them named as HC-10 had the highest hydrogen producing capability, under the batch fermentative hydrogen production condition, the maximal hydrogen yield and hydrogen production rate was 2840 mL/L culture and 25.39 mmol H2/g drycell·h. It was identified as clostridium sp.HC-10 by 16S rDNA sequence analysis. Various parameters for hydrogen production, including substrates, initial pH and temperature, have been studied. The optimum condition for hydrogen producing of strain HC-10 were achieved as: initial pH 7.0, temperature 35 ℃, glucose as the favorite substrate, Moreover, using distiller's solubles wastewater as substrate, HC-10 strain was added in the biohydrogen producing system to research the bioaugmentation effection. The results showed that the hydrogen production of bioaugmentation system was 40.32% higher than the noaugmentation system. An anaerobic, hydrogen producing strain H-8 was irradiated by microwave to optimize the microwave mutagenesis condition, and to test the heredity, hydrogen-producing potential and aciduric of the mutants. An aciduric mutant named as HW195 with steady hydrogen-producing capability was obtained, which can grow at pH 2.8. Its capability of hydrogen production was tested in the batch culture experiments. The maximum hydrogen yield and hydrogen production rate was 2460 mL/L culture and 29.97 mmol H2/g drycell·h, which was 50.7% and 41.7% higher than those of the initial strain, respectively. When used the strain H-61 as original strain, a mutant named as HW18 was obtained. The maximum hydrogen yield and hydrogen production rate was 2190 mL/L culture and 25.86 mmol H2/g drycell·h, which was 23.03% and 31.00% higher than those of the initial strain, respectively. The results demonstrated that microwave mutagenesis could be used in the field of hydrogen producing microorganism. The hydrogen producing strain H-61 was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production capability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production capability. which was tested in the batch culture experiments. With the condition of 10 g/L glucose, its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L culture and 33.19 mmol H2/g drycell·h, 69.89%and 68.14% higher than that of the original strain, respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type, while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation were studied, including time, carbon source, nitrogen source, glucose concentration, glucose utilization, initial pH and incubation temperature had been studied, indicated the optimum condition of hydrogen production for the mutant HCM-23 as initial pH6.5, temperature 36 ℃, and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different, such as, the growth lag phase and the utilization of inorganic nitrogen source, etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

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本文对不同菌种(酵母菌和运动发酵单胞菌)快速生产燃料乙醇的条件进行了研究,实现了鲜甘薯快速转化为燃料乙醇。全文分为两部分: 第一部分:酵母菌快速生产燃料乙醇的条件研究。通过单因素试验,酵母菌快速生产燃料乙醇的条件为:发酵方式采用边糖化边发酵(SSF),蒸煮温度为85 ℃,料水比2:1(初始糖浓度 210 g/kg),糖化酶用量0.75 AGU/g 鲜甘薯,接种量10%(v/w)。在最优条件下,经过24 h发酵,乙醇浓度可达97.44 g/kg, 发酵效率为92%,发酵强度为4.06 g/kg/h。由于采用了低温蒸煮和SSF,可以大大节约能耗,从而降低乙醇生产的成本。同时,利用摇瓶优化的条件,进行了10 L,100 L,500 L发酵罐的放大试验,由于发酵罐初期可以人为通氧,使菌体能迅速积累,发酵时间缩短2 h,发酵效率在90%以上。 第二部分:运动发酵单胞菌快速生产燃料乙醇条件研究。通过单因素试验和正交试验获得了发酵的最佳参数:初始pH值6.0-7.0,硫酸铵5.0 g/kg,糖化酶量1.6 AUG/kg淀粉,初始糖浓度200 g/kg,接种量12.5%(v/w)。经过21 h发酵,乙醇浓度为95.15 g/kg,发酵效率可达94%。同时对不灭菌发酵也进行了研究,发酵效率可达92%。为鲜甘薯运动发酵单胞菌燃料乙醇的工业化生产打下基础。 对发酵结束后的残糖进行了研究。通过薄层层析和葡萄氧化酶测定证明:无论是酵母菌还是运动发酵单胞菌发酵结束后的发酵液中都不含葡萄糖。经过HPLC进一步分析残糖说明:发酵液中已没有葡萄糖成分;经糖化酶水解后仍没有葡萄糖出现;但经酸水解后又出现了葡萄糖,说明结束后的残糖是一些低聚糖结构。有关残糖的结构需要进一步研究。可以通过开发高效的低聚糖水解酶来降低发酵液的残糖,提高原料的利用率。 A new technology for rapid production fuel ethanol from fresh sweet potato by different microorganisms (Saccharomyces cerevisiae and Zymomonas mobilis) was gained in this research. The paper involved two parts: Part 1: The study on fuel ethanol rapid production from fresh sweet potato by Saccharomyces cerevisiae. The following parameters of Saccharomyces cerevisiae was investigated by a series of experiments: fermentation models, cooking temperature, initial sugar concentration and glucoamylase dosage. The results showed that SSF (simultaneous saccharification and fermentation) not only reduced the fermentation time (from 30 to 24h) but also enhanced the ethanol concentration (from 73.56 to 95.96 g/kg). With low-temperature-cooking (85 ℃) using SSF, the Saccharomyces cerevisiae was able to produce ethanol 97.44 g/kg which the fermentation yield could reach to 92% and ethanol productivity 4.06 g/kg/h from sweet potato enzymatic hydrolysis. Furthermore, the savings in energy by carrying out the cooking (85 ℃) and saccharification (30 ℃) step at low temperature had been realized. The results were also verified in 10 L, 100 L and 500 L fermentor. The fermentation yield was no less than 90%. The fermentation time of fermenter was shorter than Erlenmeyer flask. This may be that the aeration in the early fermentation period is available, which lead to the rapidly commutations of biomass. Part 2: The technology of ethanol rapid production with simultaneous saccharification and fermentation ( SSF ) by Zymomonas mobilis,using fresh sweet potato as raw material was studied. The effects of various factors on the yield of ethanol were investigated by the single factor and the orthogonal experiments. As a result, the optimal technical conditions were obtained from those experiments:initial pH value 6.0-7.0, nitride 5.0 g/kg,(NH4)2SO4, glucoamylase 1.6 AUG/kg starch, inoculums concentration 12.5% (v/w). The Zymomonas mobilis was able to produce ethanol 95.15 g/kg, with 94% of the theoretical yield, from fresh sweet potato after 24 h fermentation. The fermentation efficiency of non-sterilized was also reach to 92%. We also analyzed the final fermentation residual sugars of Saccharomyces cerevisiae and Zymomonas mobilis. When the residual sugars were analyzed by thin-layer chromatogram and glucose oxidase, there was no glucose. The analysis of reducing sugars by HPLC showed that there was no glucose existed in the fermentation liquor. However, the glucose appeared after being hydrolyzed by acid. It is indicated that the residual sugars in the final fermentation liquor were the configuration of oligosaccharide, which was linked by the special glycosidic bonds. It was feasible for reducing residual sugars to develope the enzyme that can degradation the oligosaccharide.

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红发夫酵母分离于北美西部高山地区和日本一些岛屿上落叶树的渗出液中,因其所产主要色素为在水产养殖、食品和医药工业有广阔应用前景的虾青素而成为研究的热点。本论文对红发夫酵母Phaffia rhodozyma 的生长特性、培养参数与培养基组分对生长和虾青素积累的影响及其优化、虾青素合成的调节控制、虾青素的提取测定及红发夫酵母耐高温菌种的诱变进行了系统的研究。 虾青素是红发夫酵母的胞内色素,要对其进行分析首先要对红发夫酵母进行破壁处理,实验发现二甲亚砜是最有效的破壁溶剂,用氯仿和丙酮可以有效地把类胡萝卜素从二甲亚砜破壁后的红发夫酵母细胞中提取出来。 在固定摇床转速为200 rpm,温度为20 ℃的条件下,当种龄为36 h,以10%的接种量接入装液量为30 mL的250 mL三角瓶,初始pH为5.5时最有利于红发夫酵母的生长及类胡萝卜素的合成。 本实验中红发夫酵母最佳利用碳、氮源分别为蔗糖和蛋白胨,但蛋白胨价格昂贵,不适宜作单一氮源,因此使用硫酸铵和酵母膏作为复合氮源。 本论文采用了BP神经网络结合遗传算法的方法来优化红发夫酵母的发酵培养基,得到红发夫酵母发酵培养基的最佳配比为:蔗糖45.10 g/L、硫酸铵3.00 g/L、硫酸镁0.80 g/L、磷酸二氢钾1.40 g/L、酵母膏3.00 g/L、氯化钙0.50 g/L,使用优化后的培养基发酵类胡萝卜素产量达到8.20 mg/L,干重达到9.47 g/L,类胡萝卜素的产量比起始培养基提高了95.90%,干重提高了89.40%。 从代谢途径出发对红发夫酵母合成虾青素调控调控,选择谷氨酸、乙醇、VB1作为添加剂,通过正交试验设计得出三者添加水平分别为0.2 g/L,0.1% (V/V),10 mg/L时,类胡萝卜素产量提高了25.73%,达到了10.31mg/L。 通过上述优化培养,本论文中红发夫酵母的虾青素产量从1.33 mg/L提高到9.12 mg/L,产量提高了6.86倍;总类胡萝卜素产量从4.23 mg/L提高到10.31 mg/L,产量提高了2.44倍;细胞干重从5.00 g/L提高到11.35 g/L,提高了2.27倍,总体提高效果显著。 红发夫酵母属于中低温菌,本论文采用紫外复合诱变的方式,通过高温筛选,得到一株能在35 ℃下能生长的突变株,但所产类胡萝卜素中虾青素所占比例很小,可能是诱变改变了红发夫酵母的代谢途径,阻断了虾青素的合成。 Phaffia rhodozyma is a heterobasidiomyceteous yeast that was originally isolated from the slime fluxes of brich tree wounds in mountain regions of northern Japan and southern Alaska. Phaffia rhodozyma produces astaxanthin as its principal carotenoid pigment, which has potential applications in acquaculture, food and pharmaceutical industry. This paper researched ways to break cell, analysis of astaxanthin, characteristics of growth, culture parameters and the effects of components of medium on growth and astaxanthin formation , optimization of culture medium, control of astaxanthin synthesis and mutagenesis of Phaffia rhodozyma. It is necessary to disrupt the yeast cell for extracting astaxanthin considering the yeast accumulating carotenoids in cell. Dimethyisulphoxide was the most effective solvent for breaking the yeast cell; acetone and chloroform were effective to extract carotenoids out of the disrupted cell. The optimum pH for growth and carotenoids synthesis is 5.5, the optimum medium volume is 30 mL (in 250 mL flask), the optimum culture time of inoculum is 36 h, the optimum inoculum concentration is 10%. The research on culture medium showed: sucrose is the best one of 6 carbon sources for growth and astaxanthin synthesis. Peptone is the best nitrogen source for growth and astaxanthin synthesis. Uniform Design was used for trial design of the formula medium components, then back-propagation neural network was established to modeling the relationships between the carotenoid yield and the concentration of medium components. Genetic algorithm (GA) was used for global optimization of the model. The optimum combination of the medium was obtained: sucrose 45.10 g/L, ammonium sulfate 3.00 g/L, magnesium sulfate 0.80 g/L, potassium dihydrogen phosphate 1.40 g/L, yeast extract 3.00 g/L, calcium chloride 0.50 g/L. The yield of carotenoid reached 8.20 mg/L, which was 95.90% higher than that of the original medium. Glu, VB1 and ethanol were selected as fermentation addictives, after Orthogonal Test, the carotenoid contents increased by 25.73% when adding 0.16 g/L Glu, VB1 10 mg/L and ethanol 0.1% (V/V). After the above optimization, the astaxanthin content increased 6.86 folds, which is 9.12 mg/L. The carotenoids content increased 2.44 folds, which is 10.31 mg/L. The biomass increased 2.27 folds, which is 11.35 g/L. Phaffia rhodozyma grows in the mild temperature range of 0 to 27 ℃, in this work, a thermotolerant mutant was selected through UV-irradiation. It can grows at 35 ℃, and showed increased carotenoid content. The optimal growth temperature for this mutant is 30 ℃. But the mutant can only produce carotenoids with little astaxanthin accumulation.

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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.