【作者】 邢德峰；

【导师】 任南琪；

【作者基本信息】 哈尔滨工业大学 ， 环境工程， 2006， 博士

【摘要】 有机废水发酵法生物制氢技术具有能源和环保的双重功效,已经成为环境生物技术领域的研究热点之一。虽然,发酵法生物制氢技术有着广阔的应用前景,但是该技术还处于研发阶段。提高系统的产氢效率和运行稳定性,降低生产成本,仍然是生物制氢技术实现工业化进程中面临的根本问题。利用活性污泥作为菌种发酵产氢时,反应系统中的微生物群落结构对产氢效能具有决定性的影响。因此,利用现代分子生物学手段阐明微生物群落结构与产氢效能的关系,确定重要的功能菌群,建立产氢群落的快速分子检测系统,实现高效产氢群落的定向构建和优化,对实现发酵法生物制氢技术的工业化具有重要的理论意义和应用价值。目前,这些内容还没有被系统性地研究。目前所报道的产氢细菌都因不具备凝集能力,而造成大量流失,降低了系统的产氢效能。而通过载体固定化技术必然大幅度增加运行成本。所以,分离具有高效产氢并能形成颗粒生长的自凝集细菌,通过生物强化作用实现高效产氢群落结构的构建,对实现发酵法生物制氢工业化更具有现实意义和重要的应用价值。针对目前发酵法生物制氢技术工业化进程中存在的问题,进行了自凝集产氢细菌的分离和筛选,并结合DGGE、FISH、克隆文库、Real-time PCR、酶电泳等分子生物学手段,重点探讨了微生物群落结构和产氢效能的关系,深入研究了乙醇型产氢发酵的形成机制,开发了产氢细菌的快速检测方法,初步探讨了生物强化技术定向优化群落结构。论文取得了一些创新性的研究成果。(1)发现并建立了产乙醇杆菌属(Ethanoligenens)。根据系统进化关系、独特的化能异养特性、G+C(mol%)含量和生理生化特征,证实所分离的菌株YUAN-3~T和菌株X-29代表了与最相近的柔嫩梭菌(Clostridium leptum)亚群现有属完全不同的一个新属。分类名为Ethanoligenens harbinense gen. nov., sp. nov.,哈尔滨产乙醇杆菌(Ethanoligenens harbinense)为模式种,菌株YUAN-3~T(=JCM 12961T= CGMCC 1.5033~T)为模式菌株。菌株YUAN-3~T振荡培养时可形成凝集颗粒,直径一般为0.5~5mm,最大时可达到1.5cm,培养液澄清;最大比产氢率为2.81molH2/mol-glucose,最大产氢速率为27.6mmolH2/g-drycell·h;是目前唯一报道的自凝集产氢细菌。(2)揭示了不同产氢发酵类型微生物群落结构和演替规律。DGGE分析更多还原

【Abstract】 Fermentative bio-hydrogen production of organic wastewater has been attracting attention as an environmentally friendly process that does not consume fossil fuels, and become one of the research focuses in environmental biotechnology. Although the fermentative bio-hydrogen production has a wide potential, this technique is still under the research and development. There is a long way to the industrial production. Many important problems which haven’t been solved well restrict the industrialization process. The enhancement of the hydrogen production rate and the stable operation of the system and reducing the production cost are essential to realize the industrialization of fermentative bio-hydrogen production.When activated sludge as inoculums produces hydrogen, microbial community structure plays a key role for hydrogen producing in the reactors. Thereforeelucidating relationship of structure and function of microbial communities by modern molecular biological techniques, determining important functional populations for hydrogen producing, developing a quick molecular detection system of hydrogen-producing community, and realizing directional establishment and optimization of community with high efficient hydrogen production will make an important theoretical significance and practical value. At present, these contents have not been investigated yet. High efficient hydrogen-producing populations are essence of enhancing hydrogen production of reactors. Increases of hydrogen production efficiency and stability of operation are favored by putting high efficient hydrogen-producing bacteria. However, almost reported hydrogen-producing bacteria haven’t the aggregative ability, and running out resulted in depression of hydrogen production efficiency in reactors. Carrier immobilization of hydrogen-producing bacteria increases the cost of reactor operation. So isolating auto-aggregative bacteria with high efficient hydrogen production and forming granule, realizing establishment of community structure with high efficient hydrogen production by bioaugmentation will make an important practical significance and value for realizing of industrialization of fermentative bio-hydrogen production.更多还原