维生素C二步混菌发酵中L-山梨糖脱氢酶的研究

以氧化葡萄糖酸杆菌和巨大芽孢杆菌混合培养物的无细胞抽提液建立了Vc二步发酵离体实验系统.反应体系中加入山梨糖在pH7.0,35℃下保温24h，2-酮基-L-古龙酸（2KGA）生成.巨大芽孢杆菌胞外活性物质对离体系统的产酸没有影响，一定量的L-山梨糖脱氢酶可促进产酸.从氧化葡萄糖酸杆菌细胞质中分离纯化出L-山梨糖脱氢酶.L-山梨糖脱氢酶酶活与2KGA的形成呈正相关：L-山梨糖的转化是在细胞内进行的;亲缘关系相差甚远的伴生菌均能促进小菌产酸，且“伴生”效率相近;伴生菌通过促进产酸菌生长和提高其L-山梨糖脱氢酶比活力而提高发酵系统中L-山梨糖脱氢酶总活力，并且通过促进产酸菌合成新的RNA而增强其代谢力，从而促进产酸;通过对不同发酵时间L-山梨糖脱氢酶酶活及2KGA累积量的比较表明，此酶可作为生产上2KGA生成的实时监控的指示酶;环境因子通过提高酶活力促进产酸.

额济纳天然绿洲景观格局、动态、演化机制及其健康评价

额济纳天然绿洲位于我国西北干旱内流区黑河流域下游，近年来，随着黑河中、上游地区下泄地表径流量的减少，额济纳天然绿洲面临的生态环境问题日益突出。本文以额济纳天然绿洲景观为研究对象，以景观生态学及生态系统健康学原理为基础，综合运用35技术，基于研究区1987年、1994年和2001年的景观格局、动态变化特征及其演化机制的研究，发展并构建了额济纳天然绿洲景观健康评价指标体系及评价标准，对研究区景观健康问题进行了深入探讨与评价，从而为保护地区景观健康、维持区域生态安全提供科学依据。主要研究结果：①研究区景观属于典型的"基质一廊道一斑块"景观结构特征，景观破碎化程度较高，具有较低的景观多样性和较为粗糙的景观质地特征。②近巧年来研究区水域面积急剧萎缩、绿洲面积明显缩减、荒漠化程度较为严重；景观斑块形状复杂性及景观异质性程度下降，景观质地变得越发粗糙，景观多样性及景观稳定性降低。③上游入境水量的锐减是研究区景观发生退化的根本原因，绿洲人类活动强度的增加、绿洲关键区域的超载和过牧及蒸发度的升高、湿润系数的下降均加剧了研究区景观退化的程度。④基于35项景观变化、生物物理、生态环境及社会经济指标构建了研究区景观健康评价指标体系，运用模糊综合评判方法得出研究区景观健康指数LHI为0.1972，位于健康评价级别中"濒危的"级别偏下的位置。说明近十多年来研究区的景观健康状况较早时期相比趋于恶化，其发展态势表现出严重的不可持续性。

超积累植物筛选及污染土壤植物修复过程研究

超积累植物是植物提取修复的核心，同时也是污染环境修复领域研究的热点与前沿。针对目前已发现的超积累植物数量与对重金属提取种类较少的现状，并根据修复实践的需求，本研究确立了以超积累植物的筛选为主要研究内容并就一些强化措施进行了初步探索，同时根据已报道的植物修复研究的阶段性成果，对污染土壤植物修复的基本概念、原理、方式、强化机制等相关的修复过程进行了系统总结，结果如下：（1）超积累植物应同时具有四个基本特征，即临界含量特征、转移特征、耐性特征和富集系数特征。（2）龙葵和球果掉菜是Cd超积累植物，在Cd投加浓度为259／kg条件下，其茎和叶中Cd含量分别为103.80mg／kg和124.57mg／kg及107.7Omg/kg和150.10mg／kg。植物体内积累的Cd主要分布在茎和叶中。这2种植物不具有同时超积累Pb、Cu或Zn的能力。（3）施加鸡粪后植物对Cd的提取率提高了35.7％～97.0％；采用开花期收获超积累植物的复种方式，植物提取率可以提高到1.43倍和1.75倍。（4）以杂草为筛选对象，将成为筛选超积累植物的一个突破口。（5）盆栽筛选试验具有可操作性强、易于实施等优点，是筛选超积累植物值得尝试的一种方法。（6）现发现的超积累植物几乎均来源于污染区，本研究表明，未污染区也可能存在超积累植物。（7）栽培措施对植物修复效果有较大影响，与现代农业高新技术相结合是植物修复技术成功应用于修复实践的一条捷径。

甲基对硫磷水解酶结构和功能的研究

本研究以本实验室分离的有机磷农药高效降解菌Pseudomonas sp．WBC-3为材料，通过X-射线晶体学研究确定了甲基对硫磷水解酶（MPH）的三维结构，并在结构基础上探讨了甲基对硫磷水解酶的结构与功能的关系。利用生物信息学手段对Pseudomonas sP.WBC-3中的甲基对硫磷水解酶基因进行分析，推测论H的结构基因的编码序列为398-1393 bp，蛋白大小为331个氨基酸，且具有一个由35个氨基酸组成的信号肤。同时发现，MPH是一个不同于具有相似生物学功能的有机磷水解酶（OPH）的蛋白质，而且在PDB库中尚无与MPH同源性较高的蛋白结构。晶体的生长依赖于高纯度的蛋白质的获得。在本研究中，我们采用阳离子交换树脂和凝胶过滤两步纯化获得了纯度达95％以上的MPH溶液，并采用等离子体质谱仪测定MPH为含锌的金属酶。采用悬滴蒸汽扩散法获得了PI和P43212两种晶型的晶体以及硒标记的MPH晶体。在获得单晶以后，最终利用尸43212晶型的晶体通过多波长反常散射法（MAD）解析了MPH的三维结构。MPH的晶体结构为同源二聚体，具有与OPH类似的二价金属离子组成的活性中心：其中一个单体含有两个锌离子，另一个单体含有一个锌离子和一个锅离子；每个单体的两个金属离子通过AsP 151、His 152、His 302、His 147、His 149、His 234和A印255与蛋白质相连，一个H2O分子桥连于两个金属离子，还有一个H2O分子只与一个金属离子配位。在MPH三维结构知识的基础上，利用分子生物学手段对MPH活性中心附近可能的底物结合氨基酸位点进行了研究。通过定点突变获得了针对Trp179，Phe 196和Phe 119三个位点的六个突变体W179F、W179A、F196W、F196A、FllgW和Fll9A，系统比较了突变体与野生酶的催化动力学特点。结果表明Trp 179，Phel％是MPH活性中心的底物结合部位的关键氨基酸。而Phe 119在与底物结合中的作用不明显。

沈阳城市森林生态系统健康与管理研究

本文系统研究了沈阳城市森林的布局与结构、城市森林功能、城市森林病虫害发生与树木健康状况和城市自然资源与社会经济状况等指标对沈阳城市森林生态系统健康与管理的影响。同时一，采用2种生态系统健康评价方法对沈阳城市森林生态系统健康状况进行了评价，并提出了沈阳城市森林生态系统健康管理的对策。研究结果如下：1、截至2004年末，沈阳城市森林植被覆盖率已经达到35％，城市森林林地分布基本合理，但需要进一步加强道路林地、居住区林地和城郊大面积生态林建设。2、沈阳城市森林以乔木为主，乔灌株数比为1.7：1，乔灌的覆盖度比约为7:1。3、沈阳城市森林不同类型林地中植物组成不同。公园林地中有74个属，137个种（变种）；庭院林地中有53个属，104个种（变种）；居住区林地中有45个属，81个种（变种）；道路林地中有43个属，94个种（变种）；运河风景林地中有75个属，142个种（变种）；棋盘山风景林地中有48个属，118个种（变种）。4、公园林地、庭院林地、居住区林地、道路林地和运河风景林地的Shannon一Wiener多样性指数分别为2.78、3.05、3.15、3.18和3.18，均匀度指数分别为0.56、0.66、0.72、0.70和0.64。除了棋盘山风景林地外，沈阳城市森林中栽植总量超过乔木总量5％的乔木树种有7个属，分别为李、柳树、杨树、桧柏、榆树、槐树和银杏，7种树木总量达到了全部乔木总量的82.09％；栽植总量超过灌木总量5％的灌木树种也有7个属，分别为水腊、丁香、李属，小聚、玫瑰、忍冬和连翘，7个属灌木总量达到了全部灌木总量的87.92％。5、公园林地、庭院林地、道路林地和防护林地中OBH＜20cm、20cm＜DBH＜60cm和DBH＞60cm树木的比例分别为：57.9％、40.0％、2.1％，49.2％、47.8％、3.0％，65.3％、33.1％、1.6％和64.6％、34.9％、0.5％，表明沈阳城市森林树木的规格总体上偏小。6、经样方调查和CITYgreen模型计算，沈阳城市森林的生态效益约2.0亿USD／yr.。公园林地、庭院林地和风景林地的景观指标相对较高；道路林地和居住区林地的景观效果一般；防护林地的景观效果较差。7、目前已经发现的沈阳城市森林病害约600余种，虫害约700余种，其中杨树主要病虫害39种，柳树的主要病虫害有33种，榆树和槐树的主要病虫害均为，1种。杨柳树腐烂病、光肩星天牛、天幕毛虫、桃红颈天牛和美国白蛾等是近10年来沈阳城市森林中普遍发生和造成严重危害的主要病虫害。沈阳城市森林主要树木的平均健康指数为2.68，处于一般健康状态。8、沈阳城市森林的土壤和水资源状况均不利于树木的健康生长，沈阳的社会经济发展也有待于进一步提高。9、经过生物指示物法（光肩星天牛为生物指示物）、专家权重法、公众问卷调查和对比研究，沈阳城市森林生态系统总体上处于亚健康状态。10、通过对沈阳城市森林资源、管理状况的调查研究和健康状况的评价，本文提出了沈阳城市森林生态系统健康管理的对策，包括合理规划沈阳城市森林林地布局，增加道路林地、居住区林地和城郊林地的面积和植被覆盖率；调整树木种类组成，避免单一或少数树种的大量栽植，提高生物多样性水平；保护大树和古树；增加城市森林管理资金的投入；应用先进技术，采取科学的病虫害防治和植物养护方法，促进树木的健康生长等。This project systematically studied the urban forest ecosystem health and management in Shenyang. The study explored factors, such as urban forest structure, distribution, pests, aesthetic value, ecological benefit, natural resources and socieo-economic status, that affecting the urban forest ecosystem health and management. Two methods were used to evaluate the ecosystem health. This project also proposed Shenyang's urban forest ecosystem health management strategies. The research results can be summarized as follows: 1. As of the end of 2004, urban forest coverage in Shenyang is about 35%, and is in relatively even patch distribution pattern. However, the street trees and roadside forest patches, residential block forest patches should be enhanced. 2. Trees are the major component of the Shenyang s urban forest, followed by shrubs. The quantity ratio of tree to shrub is about 1.7:1, and the coverage ratio of trees to shrub is about 7:1. 3. Species composition varies by location. There are 74 genera, 137 species (including varieties) in the public parks; 53 genera, 104 species (and var.) in the green spaces of the institution (including school), factory, and company; 45 genera, 81 species (var.) in residential blocks; 43 genera, 94 species (var.) in streets and roadside forest patches; 75 genera, 142 species (var.) in the Canal landscape forest patches; 48 genera, 118 species (var.) in the Qipan Mountain recreation forest. 4. The Shannon-Woener indices varies in parks, in institution, factory, and company yards, in streets and roadside forest patches, in residential blocks.there are 2.78, 3.05, 3.18, 3.15, 3.18, respectively; and the evenness indices are 0.56, 0.66, 0.70, 0.72, 0.64, respectively. Besides the Qipan Mountain forest patches, trees of 7 genera, Prunus spp., Salix spp., Populus spp., Sabina spp., Ulmus spp., Robinia spp. and Ginkgo biloba are of more than 5% the total urban trees, respectively. In fact, trees from these 7 genera are about 82% of all trees in Shenyang's urban forests. In terms of shrubs, species of 7 genera, Ligustrum spp., Syringa spp., Prunus spp., Berberis spp., Rosa spp., Lonicera spp., and Forsythia spp. are more than 5% the total urban shrubs, respectively. 88% of all the shrubs in Shenyang s urban forest are from these 7 genera. 5. The diameter class of DBH<20cm, 20cm60cm of trees in parks, in institution, factory, and company yards, in in streets and roadside forests, and in protective forests are 57.9%, 40.0%, 2.1%; 49.2%, 47.8%, 3.0%; 65.3%, 33.1%, 1.6%; and 64.6%, 34.9%, 0.5%, respectively. The result indicated that the DBH is relatively small in Shenyang s urban forests. 6. Using random plots sampling and CITYgreen model calculation, the ecological benefit of Shenyang's urban is about 0.2 billion US dollars per year. The aesthetic value of parks, institution and company yards, and scenary forest patches is relative high; however, the aesthetic value of protective forest patches is low. 7. There are more than 600 types of diseases and more than 700 kinds of insects in Shenyang's urban forest. Among them, Populus spp., Salix spp., Ulmus spp. and Robinia spp., are attached mainly by 39, 33, 11, and 11, difference types of pests respectively. The rot disease of the Populus spp. and Salix spp., and the insects such as, Anoplophora glabripennis, Malacosoma neustria, Aromia bungii, and Hyphantria cunea are major pests in Shenyang s urban forest for the past 10 years. The trees health condition is relatively poor by having the average health index of 2.68 for major species. 8. The urban soil and water resource are not favorable to tree's growth in Shenyang, and so is the socieo-economic status. 9. Through using bio-indicator, professional and public evaluation, and comparision with other urban forests, it can be concluded that the status of Shenyang s urban forest is not very healthy in general. 10. To improve the urban forest ecosystem health in Shenyang, a better urban forest design need to be implemented to enhance the area and vegetation coverage of the street and roadside forests, the residential block forests, and the surburb forest Species biodiversity need to be enhanced by adjust the species composition of urban forest trees to reduce the single or several tree species populations and to plant more varieties. A better fundings for urban forest health management need to be budgeted. Advanced technologies in tree care and scientific measures to control pests need to be adopted to prove better care for plants and to keep trees grow healthfully.

浑河上游森林变化及其对流域水文的影响

本文以浑河上游流域为研究对象，运用35技术结合野外调查，对研究区1986，1992和2001年三个时段的森林变化及流域出口断面的水文响应进行了研究。研究内容包括研究区近期森林的数量、质量和空间分布状况：研究区森林在各个时段之间变化的数量和具体方式；研究时段内，流域出口断面主要水文要素的变化与定量表达。研究结论将加深人们对该森林流域在上世纪末森林植被变化历程的了解，促进区域生态建设和恢复研究，以实现流域的生态安全。主要结论：1）现阶段研究区森林面积1.57*10~5hm2，森林覆被率达67.04％。森林以阔叶林为主，面积占森林总面积的69.46％，其次是落叶松林，常绿林只占3.17％；对森林质量评价发现研究区森林整体属于良；3）森林的面积权重随海拔增加而增加，森林在研究区平均海拔以下的面积权重是55．52％，平均海拔以上的面积权重是79.64％，差异显著。2）十五年间森林净增加1．8X10'hm，，在1986-1992年间，森林增加0.7*104hm2，1992-2001年间，森林增加1.1*104hm2，两个时期森林变化的速度接近，但变化模式并不相同；根据随机模型模拟结果，按照近期恢复情况，研究区在2010年的覆被率有望达到70％03）森林变化对流域水文要素产生比较明显的影响。流域总蓄水量增加1.5*107t；通过构造径流和泥沙的响应指数发现，1996-2001年和1986-1991年比较，流域每毫米降水在出口断面上产生的径流量减少4.4*104m3，泥沙量减少36.3t。

杂谷脑流域景观格局优化配置研究

景观格局优化理论、方法和技术的研究是景观生态学的一个热点问题，同时也是一个难点。本文以眠江上游典型研究区-杂谷脑流域为研究对象，利用遥感和GIS技术分析该流域从1974年到2000年景观格局的时空变化，并分析了对该流域生态安全存在威胁的主要因素；以空间模拟技术和线性规划为基础建模，从理论和方法上探讨景观格局优化配置问题，以期为该区域的农林复合景观系统的生态安全和可持续发展找出切实可行的格局优化模式；并提出生态敏感度概念，为景观管理优先级提供依据。本文的主要结论如下：（l）杂谷脑流域作为长江上游重要的森林资源输出地，从1974年到2000年景观变化得出，有林地面积减少13％；灌木林地面积增加37％；相应的水源涵养能力下降，土壤侵蚀量增加7.8％；（2）杂谷脑流域陡坡耕作严重，分布在20°-45°坡度上的耕地占耕地总面积的78．2％；干旱河谷分布范围扩大，面积从1974年的217.99km2增加到2000年的237．38kmZ对生态安全造成威胁；（3）产业结构以资源密集型产业和重污染工业为主，结构不合理，第一产业比例大，农民人均纯收入远低于同期全国平均水平，教育水平比较低，这些问题会对杂谷脑流域生态安全产生负向影响；（4）以减少土壤侵蚀为依据，通过空间建模和线性规划方法优化设计杂谷脑流域景观格局，优化结果显示土壤侵蚀量比2000年减少2.35*106吨／年，土壤保持能力与1974年相近，水源涵养能力增强；（5）景观管理对于区域安全必不可少，针对该流域现存问题，建议以提高科学技术水平，调整产业结构和资源利用方式，提高教育水平为突破口。

长白山北坡针叶林下土壤淋洗液的化学性质及土壤特性

为了研究长白山北坡针叶林下土壤淋洗液的化学性质及土壤特性。我们于1987年6-9月，在红松云冷杉林。岳桦云冷杉林标准地，收集了大气降水，林内降水及土壤淋洗液，进行化学分析。结果表明：在红松云冷杉林标准地大气降水经过林冠后产生酸化作用。在红松云冷杉林和岳桦云冷杉林标准地，通过A_0层淋洗液进入土壤A_1层的养分元素（公斤/公顷）为：K 7.4-11.0, Ca 5.7-5.8, Mg 1.1-1.2，有机碳120.4-156.1，全氮1.9-3.4, SO_4-S 1.8-3.6。 A_0+A_1层养分元素的淋失量（公斤/公顷）为：K 1.2-1.6, Ca 2.2-4.6, Mg 0.5-0.9，有机碳35.4-43.9，全氮0.6-0.7, SO_4-S 1.2-2.7。A_0层淋洗液酸度大（pH5.2-5.6）。有机酸含量高（有机碳46-62mg/1）。Fe、Al、Si均从A_1层淋洗下来，并在其下的淀积层（B）淀积。因此，本区针叶林下土壤具有灰化过程的典型特征。土壤分析结果表明：长白山北坡暗针叶带土壤，具有灰化过程，其中杜香落叶松林下土壤具有明显的灰土淀积层，在分类上属灰土。

无介体双室微生物燃料电池产电性能初步研究

本文对无介体双室微生物燃料电池的产电性能进行了初步研究，并根据不同运行阶段产电性能的优劣，对其中微生物的差异性进行了比较分析。全文分为两个部分： 第一部分：以乙酸钠为阳极原料构建双室微生物燃料电池（MFC），研究不同阴极受体、外接电阻、乙酸钠浓度和pH等因素对电池产电性能的影响，研究结果表明：在500mL的阴阳极反应体系中，选用乙酸钠作为阳极底物，质量浓度为6.46 g/L， pH 7.0，接入500Ω外电阻，阴极电子受体选择高锰酸钾的情况下，微生物燃料电池产电性能最好，最大电功率密度达到294.72 mW/m2，库伦效率能达到25.87%。在确定最适外接电阻阻值的同时对MFC内阻进行测定，阻值为871.87Ω。 第二部分：微生物燃料电池运行中，比较以厌氧污泥作为接种源的第一阶段和只接入附着有大量微生物电极的第二阶段的产电性能，得出第二阶段产电性能优于第一阶段，最大电功率密度达到353.57mW/m2，比第一阶段提高58.85 mW/m2；库伦效率为39.35%，增幅达52％左右；针对微生物燃料电池运行过程中，底物CH3COONa可能存在其它的代谢途径，本实验进行了第二阶段产电性能与CH3COONa消耗率关系以及阳极液面上方气体成分和含量的研究，发现第二阶段50h前CH3COONa的大量消耗主要用于微生物的生长，在整个运行过程中，阳极液面上方含有CH4和CO2；对气体测定的同时还发现，振荡能增强电功率密度的输出；通过对电极上和污泥中微生物差异性分析得出，δ-变形菌纲、β-变形菌纲和拟杆菌门的菌种更适应微生物燃料电池的运行环境，能在电极上大量富集，提高电池的产电性能，只接入附着有大量微生物的电极能有效降低热袍菌纲的菌种数量，降低了CH3COONa的无为消耗，有效提高了电池的库伦效率。 Electricity production in the mediator-less two-chambered microbial fuel cell(MFC) was researched. Based on the result in the different operation phase in the MFC, the microbial diversity was analysed. The paper involved two parts: Part 1: A two-chambered microbial fuel cell (MFC) was constructed with high-concentration sodium acetate as fuel in the anode. The influence of different electron acceptors in the cathode, external resistance value, pH value and concentration of sodium acetate on electricity generation in MFC was investigated. The result showed that the maximum power density of 294.72 mW/m2 and the coulombic efficiency of 25.87% was achieved at sodium acetate concentration of 6.46 g/L, pH 7.0, external resistance 500Ωin the anode and when using potassium permanganate as electron acceptor in the cathode. While decided the value of resistor, we found that shaking has effect on electricity production in the MFC. Part 2: Comparing the electricity production in different operation phases when using anaerobic sludge as inoculum in the first phase and microbes in the anodic electrode as inoculum in the second phase, the result showed that electricity production in the second phase was more than that in the first phase, the maximum power density of 353.57 mW/m2 and the coulombic efficiency of 39.35% was achieved, 58.85 mW/m2 and 52％ more than that in the first phase, respectively. According to the fact that CH3COONa might be metabolized in other pathway in the running process in the MFC, we determining the relationship between electricity production and CH3COONa consumption, and the gas content in the anode, we found that CH3COONa was mainly used for microbe growth before 50h, and the anode contained CH4 and CO2. At the same time, we found that shaking could improve power density. The analysis on diversity of microbe in the anodic electrode and anaerobic sludge showed that δ-proteobacterium, β-proteobacterium and Bacteroidetes adapted themselves to the running environment of MFC. The anode could enrich them to improve the electricity production while reduced the quantity of Thermotogales, which were obligately anaerobic organotrophs with a fermentative metabolism, to increase the coulombic efficiency effectively.

自养氨氧化混合种群特性和共存机理初步研究

组特殊自养氨氧化混合种群，表现：无机环境种群生长迅速、生物量高；在一个完全无机的自养生长环境中，不仅保持高氨氧化速率，并出现丰富的异养微生物种群；该种群置于异养、厌氧环境中，迅速表现出产氢特征。对于这样一个特殊的生态体系，研究其共生机理，以及联接这些种群之间的碳源和能源问题，将具有非常重要意义。我们拟从种群特征、细胞表面分泌产物、游离体系产物多糖、蛋白和脂肪酸方面开展研究。 第一部分，自养氨氧化混合种群的基本特征。采用氨氧化培养基，进行种群氨氧化特征研究；采用扫描电镜观察自养混合种群的微观特征；沉降、离心去除微生物种群，分析水相中的总有机碳、糖类等物质；利用LB培养基进行种群的分离、纯化，并采用DGGE手段对微生物种群结构进行分析。结果表明，接入菌种后（2/5000（V/V）），培养液中氨（200mg/L）在3－5天内快速降解；亚硝酸盐与氨氮变化呈负相关趋势，仅有少量硝酸盐含量（< 30mg/L）。氨氧化种群的生物量增长与氨氧化趋势一致，初始生物量7.75 mg/L(蛋白含量)，3-5天后生物量快速增长，并达到最高63.06 mg/L（蛋白含量）。电镜图片显示，种群外包裹一层粘液。离心除去菌体后，检测培养液总有机碳和糖的含量，同样表现出与生物量增长相似的特征，分别由初始的3.73、2.35 mg/L，3－5内天迅速增加，并分别达到最大值35.19、27.45 mg/L。经初步分离、纯化并对纯化菌株进行测序，获得了10株异养微生物分别为布鲁氏菌科苍白杆菌属、纤维单孢菌、类芽孢菌属、黄杆菌属、无色杆菌、鞘脂单胞菌、嗜麦芽寡养单胞菌、噬氢菌属、硫红球菌、假单胞菌；DGGE显示，约有20分条离带，我们对其中的两条优势条带进行切割回收测序，鉴定为欧洲亚硝化单胞菌（Nitrosomonas eur）。 第二部分：混合种群自养-异养菌共生的可能机制。在对微生物种群特征初步分析基础上，针对胞外糖类组分可能被微生物代谢分解，我们重点对微生物细胞蛋白质与糖类进行分析。采用超声结合RIPA裂解液裂解，SDS-PAGE电泳分析混合种群总蛋白种类，并通过氨基酸分析仪及红外光谱法分析氨基酸组成及蛋白红外特征。采用超声破碎结合反复冻融对细胞样品进行处理，提取液采用醇沉、Sevage脱氮白，凝胶过滤方法脱盐和分级分离。对提取物的糖分析包括：紫外扫描，红外光谱，核磁共振，单糖组成分析；扫描电镜观察菌群破裂现象。SDS-PAGE分析结果表明：氨氧化种群不同生长阶段都显示出42kD蛋白表达量很高，d4时42kD蛋白表达已经很强，4-7d内一直持续这种过量表达，直到d8后表达开始减弱。说明42kD蛋白可能与氨氧化密切相关。红外光谱分析显示：细胞提取物的特征峰分布在3427.42cm-1、1718.18 cm-1和1681.72 cm-1、1160.07和1086.74 cm-1，分别对应为OH、 C=O、C－O－C基团，表明具有蛋白的典型特征；氨基酸分析显示蛋白中的Gly,Asp,Ala,Glu含量相对较高。 提取物中胞外多糖分离谱图得到不均一组分，共得到6个收集峰；紫外扫描在201－213 nm处有多糖吸收峰，同样表明多糖成分不均一性；多糖红外光谱特征峰主要分别在3400.49 cm-1、2920.28 cm-1、1154.54和1087.52 cm-1，对应OH、－CH2－ or CH 、C－O－H or C－O－C等多糖特征基团；多糖提取物核磁共振1H d4.3～5.9之间出现强吸收峰，这是1H中，多糖存在的明显证据，1H NMR中，其中O-乙酰基的甲基上的氢信号为d1.1～1.3之间。糖肟全苯甲酸酯衍生物的HPLC测定中，得到单一的单糖峰，由于时间问题，还未进行更深入的试验；电镜图片显示，种群中的细胞有大量的破裂现象。 实验表明，自养氨氧化混合种群显示出快速的氨氧化速率，氨氧化过程生物量和有机质的增加明显。微生物种群包裹粘液层，并分离纯化出大量的异养菌；去除菌体后的游离培养液中存在有机质（包括多糖）说明无机自养生长体系中存在异养菌生长、繁殖的二次碳源；细胞提取物中蛋白条带数目多、种类丰富；细胞多糖提取物具有明显的多糖特征，以及单糖的存在。结合种群的显微特征和游离体系中的有机质的检测结果，我们认为，无机自养生长体系中，种群细胞生长过程中发生的破裂现象可能是导致大量的蛋白、多糖释放到游离胞外，并成为其他异养菌生长的碳源和氮源。这可能是自养体系中，大量异养菌共生的可能机制，至于是什么原因引起种群生长过程中产生的破裂现象，还有待下一步深入研究。 A group of mixed autotrophic ammonia oxidizing populations, having much biological characteristic tested by concerned personnel for pilot test: Performed rapid population growth and obtained high biomass in inorganic environment; Not only maintained a high rate of ammoxidation, promoted a wealth of heterotrophic microbial populations growth in a totally inorganic and autotrophic growth environment; Placed in heterotrophic and anaerobic environment,had the performance characteristics that could rapidly produce hydrogen.For such a special ecological system, Study its symbiotic mechanism and the connection between these populations of carbon and energy issues, will have a very important significance. We intended from the characteristics of the population, the secretion product of cell surface, free substance in the liquid medium like polysaccharide, protein and fatty acids carrying out research. Part I: The basic features of mixed autotrophic ammonia oxidizing populations . Use inorganic liquid medium, processed study for ammonia oxidation characteristics of the population; we used scanning electron microscopy to get micro-features of autotrophic ammonia oxidizing populations .The medium was carried out settlement and centrifugal then removed the microbial populations, after all of that we analysis the water phase for total organic carbon(TOC), carbohydrate and other substances; Solid ammonia oxidizing medium was adopted to separation and purification of population, DGGE means was for structure analysis of microbial population. The results showed that after the inoculum of bacteria (2 / 5000 (V / V)), ammonia in the culture medium (200 mg / L) was rapid degradation in 3-5 days; ammonia and nitrite have the negative correlation between changes in the trend, then only a small amount of nitrate content (<30mg / L). The biomass growth of ammoxidation population in line with the trend of ammonia oxidation, the initial volume of it was 7.75 mg / L (protein content), in 3-5 days upto 63.06 mg / L (protein content). Electron microscope image showed, the populations were wrapped in a layer of mucus, including the a large number ruptted micorbe , Centrifuge to remove bacteria, then detected the medium for total organic carbon and sugar content, result took on the same characteristics with biomass growth, that were from the initial 3.73、2.35 mg / L respectively, in 3-6 days achieved rapid increase in the maximum to 35.19、27.45 mg / L respectively. After initial separation、 purification ,then processed sequencing to strains purified and got the result that there were 10 heterotrophic microorganisms : Brucella Branch pale bacillus, Cellu lomonas, Bacillus species category, a Flavobacterium, colorless Bacteria, Aeromonas sheath fat, little support maltophilia Aeromonas, macrophages species hydrogen, sulphur-MI, Pseudomonas bacteria spores; DGGE display, there were 20 separation bands approximately. Part II: Mixed populations that autotrophic - heterotrophic bacteria symbiotic mechanism. On the basis of preliminary analysis of microbial population characteristics, aiming at extracellular carbohydrate components might be decomposition by microbial, we focused on microbial cell protein and carbohydrate analysis. Using ultrasound combined with RIPA lysis cracking the cells, SDS-PAGE electrophoresis analysis the total protein species of the population, and through the amino acid analyzer studied the compositions of amino acid and infrared spectroscopy analysis of a protein infrared characteristics. Using ultrasound combined with repeatedly freezing and thawing to treated the cell sample, then took the means that alcohol precipitation, deproteinization by Sevage, gel filtration aimed at desalination and grade separation to deal with the lysates . The extraction of sugar analysis included: UV scanning, IR, NMR, single-sugar composition analysis. SDS-PAGE analysis showed that: 42 kD protein expression was very high at different growth stages of mixed autotrophic ammonia oxidizing populations , on the fourth day, 42 kD protein expression had been very strong, 4-7d, it had continued this excessive expression, then started to weaken after 7 days. 42 kD protein that might be closely associated with ammonia oxidation. Infrared spectral analysis showed that: cell extracts with the characteristic that the peak distribution in 3427.42 cm-1、1718.18 cm-1 and 1681.72 cm-1、1160.07 cm-1 and 1086.74 cm-1 corresponding to OH、C = O、C-O-C Groups which had the typical characteristics of protein; and analysis showed that amino acids including Gly, Asp, Ala, Glu ,the content in the protein is relatively high. Exopolysaccharide in the extracts had the separation map that it was uneven, received a total of six collection peaks by the detection mode of phenol-sulphruic acid method ; ultraviolet scan in the 201-213 nm department had polysaccharide absorbing peak, the same ingredients that polysaccharide heterogeneity; infrared polysaccharide spectral characteristics of the main peak at 3400.49 cm-1, 2920.28 cm-1, 1154.54 and 1087.52 cm-1, corresponding OH,-CH2-or CH, C-O-H or C-O-C;and other characteristics of polysaccharide group; 1H NMR of polysaccharide extract appeared absorption peak between d4.3 ~5.9, which is the apparent evidence of polysaccharide, In 1H NMR, the hydrogen signal of one of O-acetyl was between 1.1 to 1.3. The determination of Sugar oxime whole benzoate derivatives by HPLC, there was a single-sugar peak, as a matter of time, yet more in-depth test. Summary: Mixed autotrophic ammonia oxidizing populations show us that it had the ability in ammonia oxidizing and it was great, organic matter and biomass increased significantly in the process of ammonia oxidation. Microbial populations was wrapped up slime layer, the phenomenon of cell breakdown obviously, and there were a lot of separation and purification of the heterotrophic bacteria; a lot of organic matter (including polysaccharides)remined in the medium that removal of cell indicated the inorganic system existed secondary carbon sources that could be used by the heterotrophic bacteria ; there were a large number proteins bands of cell extract, rich variety; cell extracts of polysaccharide had obvious characteristics of polysaccharide, and the existence evidence of single-sugar. Combined population of microscopic characteristics and free of organic matter in the test results, we believe that the health of inorganic system, population growth occurred in the course of the breakdown of the phenomenon is likely to lead to a lot of protein and polysaccharide released into the extracellular free, And other heterotrophic bacteria use them to the growth as carbon and nitrogen. This may be autotrophic system, the large number of heterotrophic bacteria symbiotic mechanism.