非粮原料燃料乙醇发酵关键技术的研究

生物质燃料乙醇是一种高度清洁的交通液体燃料，是减少温室气体排放，缓解大气污染的最佳技术选择。以非粮原料生产燃料乙醇可以在进行能源生产的同时保证粮食安全，有利于产业的可持续发展。在众多的非粮原料中，甘薯是我国开发潜力最大的生物质能源作物之一。我国占世界甘薯种植总面积和产量的90％。同时，甘薯的单位面积燃料乙醇产量远大于玉米和小麦。其成本是目前酒精中最低廉的，因此利用甘薯生产乙醇是发展生物质燃料乙醇的首要选择。目前采用薯类全原料主要采用分批发酵生产乙醇，其技术水平低，发酵强度低，一般在0.7－2.5g/（L•h），乙醇浓度低，甘薯发酵乙醇为6－8％(v/v)，能耗高，环境负荷大，污染严重。针对上述问题，本文从菌株选育、原料预处理、中试放大、残糖成分分析等方面进行研究。 为了研究乙醇发酵生产规模扩大过程中，大型发酵罐底部高压条件下，CO2对酵母乙醇发酵的影响，我们通过CO2 加压的方法进行模拟试验，研究结果表明，发酵时间随压强的升高而逐渐延长，高压CO2 对乙醇发酵效率影响不大，在0.3 MPa 以下时，发酵效率均可达到90%以上。高压CO2 对发酵的抑制作用是高压和CO2 这两个因素联合作用的结果。高压CO2 条件下，酵母胞外酶和胞内重要酶类的酶活均表现出特征性。0.2 MPa 下，酶活性的变化趋势和0.1 MPa 条件下的较为一致。而0.3 MPa 下的酶活变化趋势与0.4 MPa 下的酶活更为接近。通过全基因表达分析发现在CO2 压力为0.3 MPa 下，乙醇发酵途径中多个基因表达量下调，同时海藻糖合成酶和热激蛋白基因表达量上调。 筛选耐高温的乙醇酵母菌株能够解决糖化温度和发酵温度不协调的矛盾，实现真正意义上的边糖化边发酵。高温发酵还能够降低发酵时的冷却成本，实现乙醇的周年生产。本研究筛选出一株高温发酵菌株Y-H1，进而我们对该菌株的胞外酶和胞内乙醇代谢重要酶类的酶活性进行了分析。结果表明Y-H1 能够在40 ℃条件下正常进行乙醇发酵，发酵33h，最终乙醇浓度达到10.7%（w/w），发酵效率达到90%以上。同时发酵液最终pH 在3.5 左右，显示菌株具有一定的耐酸性能力。同时观察到40 ℃下，菌株的胞外酶和胞内乙醇代谢重要酶类的酶活性发生了变化，乙醇发酵途径中关键酶基因表达下调，而海藻糖合成酶与热激蛋白基因表达量上调，这些结果为进一步研究酵母菌耐热调控机理提供了依据。 糖蜜是一种大规模工业生产乙醇的理想原料，本研究利用选育高浓度乙醇发酵菌株结合配套的发酵稳定剂，研究了糖蜜高浓度乙醇发酵情况。结果表明采用冷酸沉淀预处理糖蜜溶液，采用分批补料的发酵方式，乙醇浓度最高达到了10.26% (w/w)，发酵时间为42 h。同时观察到在糖蜜发酵中，乙醛含量与乙醇浓度存在一定的相关性。 快速乙醇发酵对于缩短乙醇生产周期、降低乙醇生产成本、减少原料腐烂损失具有重要意义。本研究诱变和筛选得到了一株快速乙醇发酵菌株10232B。在优化后的发酵条件下，采用10L 发酵罐进行分批乙醇发酵，经过18h,乙醇的最终浓度达到88.5g/L,发酵效率93.6%，平均乙醇生产速度达到4.92 g/L/h。此菌株在保持较高乙醇生产浓度的同时，拥有快速生产乙醇的能力，适合作为快速乙醇发酵生产菌种。 由于鲜甘薯具有粘度大的特点，传统液化糖化处理很难在短时间内充分糖化原料；高粘度的醪液也难以进行管道输送，容易堵塞管路；同时，也会降低后续的乙醇发酵效率。 本文采用了快速粘度分析法对鲜甘薯糊化粘度特性进行了分析，进而对预处理条件进行了研究，在最佳预处理条件下，糖化2h 后，醪液葡萄糖值最高可达99.3，粘度4.5×104 mPa.s，而采用传统糖化工艺，醪液DE 值仅为85.8，粘度大于1.0×105 mPa.s。 此预处理方法也可用于快速糖化不加水的醪液。后续的乙醇发酵试验表明，通过此预处理方法获得的糖化醪液对乙醇发酵无负面影响。 在前期已实现了实验室水平的鲜甘薯燃料乙醇快速乙醇发酵基础上，进一步将发酵规模扩大到500L，在中试水平上对甘薯乙醇发酵进行了研究。结果表明在500L 中试规模，采用边糖化边发酵（SSF）工艺，在料液比为3∶1，发酵醪液最高粘度为6×104mPa.s 条件下，发酵37h，乙醇浓度达到了12.7%（v/v），发酵效率91%，发酵强度为2.7 g/（L•h）。与目前国内的薯类乙醇发酵生产技术水平具有明显的优越性。 为研究甘薯、木薯乙醇发酵中残糖的组成，采用了高效液相色谱—蒸发光散射检测法，对乙醇发酵残糖进行了分析。结果表明，甘薯、木薯乙醇发酵残糖均为寡聚糖，主要由葡萄糖、木糖、半乳糖、阿拉伯糖和甘露糖构成。随着发酵时间延长，寡聚糖中的葡萄糖、半乳糖、甘露糖可被缓慢的水解释放。提高糖化酶量仅在一定程度上降低残糖，过量的糖化酶反而会导致残糖增加。同时发现3, 5-二硝基水杨酸法不能准确测定甘薯、木薯乙醇发酵中的残总糖含量。进一步筛选了两株残糖降解菌株，对甘薯乙醇发酵残糖的降解利用率均达到了40%以上，而且还能显著降低发酵醪液粘度。经形态学和rRNA ITS 序列分析，确定这两株菌分别属于为木霉属和曲霉属黑曲霉组。 通过对以甘薯原料为代表的非粮原料发酵技术研究开发，以期形成乙醇转化率高，能耗低，生产效率高、季节适应性好，原料适应性广，经济性强，符合清洁生产机制的燃料乙醇高效转化技术，为具有我国特色的燃料乙醇发展模式提供技术支持。 Sweet potato is one of the major feedstock for the fuel ethanol production in China. The planting area and the yield in China take 90% of the world. Sweet potato is an efficient kind of energy crops. The energy outcome per area is higher than corn or wheat. And the manufacture cost of ethanol is the lowest, compared with corn and wheat. So sweet potato is the favorable crop for the bioethanol production in China. However, the low-level fermentation technology restricts the development of ethanol production by sweet potato, including slow ethanol production rate, low ethanol concentration and high energy cost. To solve these problems, we conducted research on the strain breeding, pretreatment, pilot fermentation test and residual saccharides analysis. To study the impact of hyperbaric condition at bottom of the large fermentor on yeast fermentation, high pressure carbon dioxide (CO2) was adopted to simulate the situation. The results showed that the fermentation was prolonged with the increasing pressure. The pressure of CO2 had little impact on the ethanol yield which could reach 90% under the pressure below 0.3 MPa. The inhibition was combined by the high pressure and CO2. Under the high CO2 pressure, the extracellular and important intracellular enzyme activities were different from those under normal state. The changes under 0.1 MPa and 0.2 MPa were similar. The changes under 0.3 MPa were closer to those under 0.4 MPa. The application of thermotolerance yeast could solve the problem of the inconsistent temperature between fermentation and saccharificaton and fulfill the real simultaneous saccharification and fermentation. And it could reduce the cooling cost. A thermotolerance strain Y-H1 was isolated in our research. It gave high ethanol concentration of 10.7%（w/w）at 40 ℃ for 33 h. The ethanol yield efficiency was over 90%. At 40 ℃, the extracellular and important intracellular enzyme activities of Y-H1 showed the difference with normal state, which may indicate its physiological changes at the high temperature. Molasses is another feedstock for industrial ethanol production. By our ethanol-tolerance strain and the regulation reagents, the fermentation with high ethanol concentration was investigated. In fed-batch mode combined with cold acid deposition, the highest ethanol concentration was 10.26% (w/w) for 42h. The aldehyde concentration in fermentation was found to be related to ethanol concentration. The development of a rapid ethanol fermentation strain of Zymomonas mobilis is essential for reducing the cost of ethanol production and for the timely utilization of fresh material that is easily decayed in the Chinese bioethanol industry. A mutant Z. mobilis strain, 10232B, was generated by UV mutagenesis. Under these optimized conditions, fermentation of the mutant Z. mobilis 10232B strain was completed in just 18 h with a high ethanol production rate, at an average of 4.92 gL-1h-1 per batch. The final maximum ethanol concentration was 88.5 gL-1, with an ethanol yield efficiency of 93.6%. This result illustrated the potential use of the mutant Z. mobilis 10232B strain in rapid ethanol fermentation in order to help reduce the cost of industrial ethanol production. As fresh sweet potato syrup shows high viscosity, it is hard to be fully converted to glucose by enzymes in the traditional saccharification process. The high-viscosity syrup is difficult to be transmitted in pipes, which may be easily blocked. Meanwhile it could also reduce the later ethanol fermentation efficiency. To solve these problems, effects of the pretreatment conditions were investigated. The highest dextrose equivalent value of 99.3 and the lowest viscosity of 4.5×104 mPa.s were obtained by the most favorable pretreatment conditions, while those of 85.8 and over 1.0×105 mPa.s was produced by traditional treatment conditions. The pretreatment could also be applied on the material syrup without adding water. The later experiments showed that the pretreated syrup had no negative effect on the ethanol fermentation and exhibited lower viscosity. The fuel ethanol rapid production from fresh sweet potato was enlarged in the 500L pilot scale after its fulfillment on the laboratory level. The optimal ratio of material to water was 3 to 1 in 500L fermentor. With low-temperature-cooking (85 ℃) using SSF, the Saccharomyces cerevisiae was able to produce ethanol 97.44 g/kg for 37h, which reached 92% of theoretical yield. The average ethanol production rate was 4.06 g/kg/h. And the maximum viscosity of syrup reached 6×104mPa.s. The results showed its superiority over current industrial ethanol fermentation. The compositions of the residual saccharides in the ethanol fermentation by sweet potato and cassava were analyzed by high performance liquid chromatography coupled with evaporative light-scattering detector. The results showed that all the residual saccharides were oligosaccharides, mainly composed of glucose, xylose, galactose, arabinose and mannose. The glucose, galactose and mannose could be slowly hydrolyzed from oligosaccharides in syrup during a long period. To increase the glucoamylase dosage could lower the residual saccharides to a certain extent. However, excess glucoamylase dosage led to more residual saccharides. And the method of 3, 5-dinitrosalicylic acid could not accurately quantify the residual total saccharides content. Two residual saccharides degrading strains were isolated, which could utilize 40% of total residual saccharide and lower the syrup viscosity. With the analysis of morphology and internal transcribed spacer sequence, they were finally identified as species of Trichoderma and Aspergillus niger.

Remediation of Persistent Organic Pollutants (POPs) in Two Different Soils

Persistent organic pollutants (POPs) are a set of chemicals that are toxic, persist in the environment for long periods of time, and biomagnify as they move up through the food chain. The most widely used method of POP destruction is incineration, which is expensive and could result in undesirable by-products. An alternative bioremediation technology, which is cheaper and environ-mentally friendly, was tested during this experiment. Two different soil types containing high and low organic matter (OM) were spiked with 100 mg/kg each of pyrene and Aroclor 1248 and planted with three different species of grasses. The objective of the study was to determine residue recovery levels (availability) and potential effectiveness of these plant species for the remediation of POPs. The results showed that recovery levels were highly dependent on the soil organic matter content—very low in all treatments with the high OM content soil compared to recoveries in the low OM soil. This indicates that availability, and, hence, biodegradability of the contaminants is dependent on the organic matter content of the soil. Moreover, the degree of availability was also significantly different for the two classes of chemicals. The polyaromatic hydrocarbon (PAH) recovery (availability) was extremely low in the high organic matter content soil compared to that of the polychlorinated biphenyls (PCBs). In both soil types, all of the plant species treatments showed significantly greater PCB biodegradation compared to the unplanted controls. Planting did not have any significant effect on the transformation of the PAHs in both soil types; however, planting with switchgrass was the best remedial option for both soil types contaminated with PCB.

黄土高原不同密度刺槐(Robinia pseudoacia)林地水分生产力与土壤干燥化效应模拟

应用WinEPIC模型模拟研究了1957~2001年期间黄土高原半湿润区长武和半干旱区延安不同密度刺槐(Robinia pseudoacia)林地水分生产力演变规律和深层土壤干燥化效应。结果:(1)长武和延安高密度(6000株/hm2)、中高密度(4500株/hm2)、中低密度(3000株/hm2)和低密度(1500株/hm2)等4种处理刺槐林地逐年生物量模拟值均呈现快速增加、达到最大值后又逐年波动性降低的变化趋势,林地密度越高早期逐年生物量越高,后期逐年生物量差异缩小,两地均以低密度处理逐年生物量平均值和累积生物量模拟值最高;(2)4种密度处理45年生刺槐林地年均耗水量值基本相等,长武和延安分别为603mm和566mm,但生长前期年耗水量明显高于后期,并高于同期年降水量,林地密度越高前期耗水量越高,中期以后各密度处理耗水量基本接近且波动趋势基本一致,林地密度越高干旱胁迫程度越重;(3)模拟生长初期,4种密度处理刺槐林地0~10m土层逐月土壤有效含水量均呈现强烈的波动性降低趋势,长武各密度处理刺槐林地分别在7~23年生、延安分别在7~17年生之后逐月土壤有效含水量均在0~200mm较低水平上随降水量变化而波动..

黄土高原典型土壤有机碳和微生物碳分布特征的研究

以阐明黄土高原典型区域土壤有机碳(SOC)含量和储量及微生物碳(Mc)含量随土壤类型、土层和土地利用方式变异规律为目的,研究了从北向南依次分布的干润砂质新成土(神木)、黄土正常新成土(延安)和土垫旱耕人为土(杨凌)等典型土壤的SOC含量和储量及Mc含量的变化特征。结果表明,不同土壤类型、不同土层SOC和Mc含量存在显著差异。同一土壤类型SOC和Mc含量在0～60cm随土层深度增加下降很明显,60～120cm土层有轻微下降,120cm土层以下低而稳定,同层次土壤从南到北,SOC、Mc和SOC储量含量显著下降,均以土垫旱耕人为土最高,黄土正常新成土次之,干润砂质新成土最低,且差异显著(P<0.05);0～200cm土层SOC总储量也沿土垫旱耕人为土(102.23±30.12t/hm2)、黄土正常新成土(67.78±9.23t/hm2)、干润砂质新成土(27.07±4.59t/hm2)依次下降;土垫旱耕人为土、黄土正常新成土和干润砂质新成土在100～200cm土层SOC累积量分别是0～100cm土层的65%、74%和58%,因此在研究黄土高原SOC贮量时必需考虑深层贮量的贡献。Mc随土壤类型的变化趋势与SOC基本相...

人工苜蓿草地土壤理化性状变化

为了明确西北旱地种植苜蓿对土壤物理性状、水分和养分的影响,对其进行了较为详尽地探讨。种植苜蓿能增加土壤团聚体含量,提高土壤通气透水能力。苜蓿生长发育时需要大量消耗土壤水分,连续多年种植苜蓿会导致土壤干燥化;种植苜蓿能提高土壤有机质和氮素水平,降低土壤磷、钾含量。施肥既能平衡土壤养分又能增加产草量。

施氮和不同栽培模式对半湿润农田生态系统冬小麦根系特征的影响

在陕北黄土丘陵沟壑区对清耕和问作沙打旺(Astragalus adsurgens)6年、4年、2年仁用杏园0～500cm土层土壤水分、有机质、全氮、全磷、速效氮、速效磷、速效钾含量及树体生长状况进行了测定,结果表明,间作沙打旺,除了在秋季提高了0～100cm土层土壤水分以外,在春季、夏季和秋季均显著降低了0～500cm土层土壤水分,且间作沙打旺年限越长,降低越显著。间作沙打旺提高了0～100cm土层土壤有机质和全氮含量,间作年限越长效果越显著,但对100cm土层以下的土壤有机质和全氮无显著影响。间作沙打旺对土壤全磷含量无显著影响。间作沙打旺显著降低了土壤速效养分含量,其中速效氮降低深度达500cm土层,速效磷、速效钾达300cm土层,且间作年限越长,降低越显著。间作沙打旺显著削弱了树体的长势,降低了坐果率和杏仁产量。间作沙打旺存在着与仁用杏争水争肥的矛盾,黄土丘陵沟壑区仁用杏园不宜间作沙打旺。

密度对三种豆科牧草生产力和水分利用率的影响

研究播种密度对3种豆科牧草生产力和水分利用效率的影响,以探讨其在草地农业系统中的合理优化模式。结果表明:沙打旺(Astragelus adsurgens)在高密区第2年生产力最高(18321 kg.hm-2),第3年开始下滑;苜蓿(Medicago sativa)在中、高密区生产力的增幅和增量差别不显著,以第3年最高(22563和22108kg.hm-2);胡枝子(Lespedeza daurica)在中密区第3年最高(7856 kg.hm-2);加大密度使沙打旺提早进入生长盛期,第2年即达到高生产力和较高水分利用效率;苜蓿以中密区(20~30株/m2)效果最好;达乌里胡枝子的生产力和水分利用率偏低,在黄土塬区草地农业中没有优势。

黄土丘陵典型地区植被建设中有关问题的研究Ⅱ、立地条件类型划分及小流域造林种草布局模式

以土壤水分条件及地形等因素为依据 ,进行了立地条件类型的划分 ,共划分了 3个类型组和 9个类型 ,并提出了小流域林草布局的典型模式。根据立地条件类型划分和小流域布局模式可以看出 ,延安研究区有条件发展乔木林 ,但并不是到处都可发展乔木林 ,因此 ,依据立地条件类型进行造林种草的合理布局进行植被建设是取得成功的必要措施。

陕北黄土高原植被区划及与林草建设的关系

在陕北黄土高原 ,根据植被的地理分布、植物区系背景、环境条件的地理变化及造林种草实践 ,从南至北可划分为落叶阔叶林区、森林草原区和干草原、沙化草原区。以延安一线为界 ,南部为森林地区 ,北部为草原地区 ;再以长城沿线为界 ,南部为森林草原区 ,北部为干草原、沙化草原区。延安一线和长城沿线为两条重要的生态分界线 ,就是这两条生态线把陕北黄土高原划分为森林、森林草原及干草原、沙化草原三个植被区。植被区划是造林种草的理论依据之一 ,陕北黄土高原的种树种草应遵循这两条生态线 ,使其与植被分区相符合

宁夏南部山区不同耕作方式对冬小麦产量及水分利用效率的影响

为探讨在宁南山区推广保护性耕作技术的可行性,2005~2006年在宁夏彭阳县就垄沟、传统、覆膜3种不同的耕作方式的适应性进行试验研究,对比分析了3种耕作方式下冬小麦的生长状况、产量及农田土壤含水量变化的关系。结果表明:覆膜耕作可增加土壤蓄水保墒性能,提高水分利用效率,增产增效明显。对三种耕作方式下作物产量进行分析,覆膜耕作比传统耕作增产46%;垄沟耕作比传统耕作减产71%。覆膜耕作水分利用效率比传统耕作提高了33%;垄沟耕作由于在越冬期垄上冬小麦大面积冻死,不适宜在宁南山区推广。

密度对刺槐幼苗生物量及异速生长模式的影响

刺槐(Robinia pseudoacacia)具有较强的生态适应性和抗逆能力,是黄土高原植被恢复和生态重建的主要优良树种之一(程积民等,2002;陈云明等,2002)。黄土高原地区刺槐人工林密度偏大,且多为低产林(余新晓等,1996;韩芯莲等,1996;2003)。王百田等(2005)认为黄土高原刺槐单木总生物量及各部分生物量都是密度的幂函数关系,但低密度林分和高密度林分的总生物量都比中间密度林分的总生物量高。种植密度不仅影响刺槐林地生物量,而且刺槐林地出现土壤干层的部分原因就是栽植密度过大(韩芯莲等,2003),其林地生产力的维持以及可持续生长成为林草植被建设中亟待解决的问题。植物密度对种群个体数量和生长的调控可以维持种群对资源的合理利用,保持种群的稳定性(Watkinson,1980)。因生长资源的强制分配,种植密度能引起植株个体间相互作用,密度增加导致植物种内竞争产生,使种群中单株生长量和生物量发生改变(李博等,2000)。竞争的实质是植物对地上光照和地下养分资源的利用,在共享资源有限的情况下竞争会导致植物个体生长量和存活率的降低(李博等,1998)。种内竞争对植物个体生长的影响,与植物“自疏法则”的...

辽西油松纯林可持续经营途径探讨

Under the direction of Sustainable Development, some key issues of forestry in Western Liaoning Province were analyzed in this paper. "Planting Broad-leaved Trees and Accelerating Conifers" (PBTAC) was put forward as the effective approach to sustainable management on the Pure Plantation of Pinus tabulaeformis and the unique path of forestry sustainable development in Western Liaoning Province. The concept, aim, and notices of "PBTAC" were also analyzed and discussed.

陕北丘陵沟壑区果树适地适栽与节水灌溉

根据陕北丘陵沟壑区川地、坡地以及不同坡向、不同坡位的气候条件 ,提出果树的最佳适栽地块。通过对陕北丘陵沟壑区果园土壤含水量的分析 ,提出了适合于该区的果树节水灌溉方法和灌溉量

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干旱是威胁宁南山区胡麻生产持续发展的首要限制因子。采用地膜覆盖穴播种植技术 ,探索了旱地胡麻覆膜穴播种植的增产机理及其增产效应。结果表明 ,覆膜穴播可以提高土壤地温 ,保持土壤水分 ,促进土壤速效养分的充分释放和有效利用。又因穴播种植 ,胡麻籽粒顶土合力增强 ,出苗率高达 79.4%～ 96.2 % ,籽粒产量提高 37.9%～ 46.2 %。与此同时 ,作者于胡麻覆膜穴播种植后 ,在胡麻的关键生育期枞形后期—现蕾初期 ,进行了膜上节水补灌试验。结果表明 ,节水补灌 30 0～ 450 m3 /hm2可提高籽粒产量 30 %～ 44% ,水分生产效率达0 .30～ 0 .32 kg/(hm2 · mm) ,水分生产效率提高 8%～ 1 4 .9%。这一新的种植方式 ,效益显著 ,应在干旱半干旱地区的胡麻产地大面积推广应用

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自 1 98 4年开始的长期不同轮作与施肥试验表明 :种植作物可提高土壤供氮能力 6.88%～ 36.58%,以种植苜蓿提高幅度最大 ,是裸地的 1 .37倍。长期施用氮肥、磷肥、有机肥及三者配施可提高土壤供氮能力 9.8%～ 1 33.98%,其中以施氮、磷及有机肥玉米连作提高幅度最大。不同轮作系统中土壤供氮能力 :粮草 3年轮作 >粮草 8年轮作 >粮豆 3年轮作 >粮饲豆 4年轮作 ;种植作物可有效增加土壤有机氮水平 ,提高剖面土壤供氮能力。施用有机肥可有效地增加深层土壤供氮潜力并在 1 0 0 cm处出现迅速减小的现象