微生物燃料电池产电菌株的筛选及群落结构研究

发布时间：2018-03-19 20:22

本文选题：微生物燃料电池　切入点：产电微生物　出处：《浙江海洋大学》2017年硕士论文　论文类型：学位论文

【摘要】：微生物燃料电池(microbial fuel cell,简称MFC)是利用微生物氧化有机或无机底物进行产电的新兴技术。本研究样品取自浙江省海水增养殖基地养殖废水储水池的沉积物(M01、M02、M03、M04),对其进行微生物产电菌株的筛选及群落结构分析。使用厌氧试管对产电菌富集培养并对特征菌进行分离纯化,共纯化11株兼性厌氧菌,分别记为ME1~ME11。16S rDNA分子鉴定结果及系统发育分析表明ME1属于过氧微杆菌(Microbacterium paraoxydans),ME2和ME3同属于Shewanella haliotis,ME4为Sunxiuqinia elliptica,ME6、ME8、ME10同属于Burkholderia contaminansz,ME7属于高地芽孢杆菌(Bacillus altitudinis),ME9属于Exiguobacterium sp.,ME10属于梭菌目(Clostridiales),未能确定到属。为了研究沉积物与以其作为接种物制作的微生物燃料电池中微生物群落结构变化,首先对沉积物中微生物基因组进行高通量测序,发现4个样品中有24个门类的细菌,其中变形菌门为优势类群,其次为拟杆菌门、绿弯菌门,样品之间群落结构没有明显差别,所含细菌类群以及相应丰度基本一致;3个门类的真菌,分别为子囊菌门(Ascomycota)、担子菌门(Basidiomycota)、接合菌门(Zygomycota),样品间群落结构和相对丰度存在差异性;3个门类古菌,分别是泉古菌门(Crenarchaeota)、广古菌门(Euryarchaeota)以及Parvarchaeota门,其中泉古菌门为优势类群,样品间群落结构和相对丰度差异不明显。以养殖废水沉积物(M01、M02、M03)为接种物,阳极室添加乙酸钠作为电子供体,阴极室添加铁氰化钾作为电子受体,构建双室型(H型)微生物燃料电池(01、02、03)筛选产电菌。MFC运行45~60天后,测其电压分别为5.043、4.172、4.428 mV,然后对阳极生物膜和阳极室底泥微生物进行高通量测序。结果显示阳极底泥细菌分为9个门,分别为变形菌门(Proteobacteria)、浮霉菌门(Planctomycetes)、芽单胞菌门(Gemmatimonadetes)、厚壁菌门(Firmicutes)、绿弯菌门(Chloroflexi)、绿菌门(Chlorobi)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)。其中变形菌门相对丰度分别为73.20%、66.47%、69.96%,为优势菌群。其次为绿弯菌门、拟杆菌门。阳极生物膜细菌分为6个门,分别为变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、绿弯菌门(Chloroflexi)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)。其中变形菌门相对丰度都在97%以上,是优势菌群。阳极阳极生物膜和阳极底泥中古菌都分为3个门,分别为泉古菌门(Crenarchaeota)、广古菌门(Euryarchaeota)以及Parvarchaeota,并且都是以泉古菌门为优势类群,但是阳极底泥中各门类古菌的相对丰度都比阳极生物膜上的高1~2个数量级。阳极底泥和阳极生物膜中真菌群落分为3个门,分别为子囊菌门(Ascomycota)、担子菌门(Basidiomycota)以及接合菌门(Zygomycota),子囊菌门为优势菌群。通过比较种泥沉积物与阳极生物膜和阳极底泥中微生物群落结构变化,发现MFC运行后希万氏菌属(Shewanella)相对丰度显著增高,阳极生物膜和阳极底泥中微生物群落多样性明显降低,验证了MFC对阳极微生物群落结构具有选择性。
[Abstract]:Microbial fuel cell (microbial fuel cell, referred to as MFC) is a new technology of electricity production by microbial oxidation of organic or inorganic substrates. The study samples were collected from the sediments of Zhejiang province marine aquaculture base of aquaculture wastewater storage tank (M01, M02, M03, M04), analysis of the microbial strains and electricity production community structure. Use anaerobic tube enriched electricigens and characteristics of bacteria were isolated and purified by CO purification of 11 strains of facultative anaerobic bacteria, respectively rDNA and ME1~ME11.16S identification results of molecular phylogenetic analysis showed that ME1 belongs to Microbacterium peroxy (Microbacterium paraoxydans), ME2 and ME3 belong to Shewanella Haliotis, ME4 Sunxiuqinia elliptica ME6, ME8, Burkholderia, ME10 belong to contaminansz, ME7 belong to the highlands (Bacillus altitudinis), Bacillus ME9 belongs to Exiguobacterium sp., ME10 belongs to clostridiales (Clostridiales ), failed to confirm to the genus. In order to study the sediment and the changes of microbial community structure of microbial fuel cell inoculum production in the first high-throughput sequencing of microbial genomes in sediments, found that there are 24 Categories of bacteria in 4 samples, including Proteobacteria were the dominant species, followed by Bacteroidetes. Chloroflexi, there was no difference in community structure between samples, including bacterial taxa and the corresponding abundances are consistent; 3 kinds of fungi were Ascomycota, Basidiomycota (Ascomycota) (Basidiomycota), Zygomycota (Zygomycota), the sample between the community structure and relative abundance differences; 3 categories of archaea, namely Crenarchaeota (Crenarchaeota), Euryarchaeota (Euryarchaeota) and Parvarchaeota, which Crenarchaeota were the dominant species, community structure and relative abundance between samples in order to keep the difference is not obvious. Aquaculture wastewater sediment (M01, M02, M03) as inoculum, the anode chamber of adding sodium acetate as an electron donor, the cathode chamber added potassium ferricyanide as electron acceptor, the construction of double chamber type (H type) microbial fuel cell (01,02,03) screening of exoelectrogens.MFC operation for 45~60 days, the electric pressure are respectively 5.043,4.172,4.428 and mV. High throughput sequencing of the anode biofilm and the anode chamber. The results showed that the sediment microbial anode sediment bacteria are divided into 9, respectively, Proteobacteria (Proteobacteria), planctomycetes (Planctomycetes), gemmatimonadetes (Gemmatimonadetes), Firmicutes (Firmicutes), Chloroflexi (Chloroflexi), green (Chlorobi), bacterial phyla Bacteroidetes (Bacteroidetes), actinobacteria (Actinobacteria), acidobacteria (Acidobacteria). The relative abundance of Proteobacteria were 73.20%, 66.47%, 69.96%, as the dominant bacteria. Followed by Chloroflexi, Bacteroidetes. The anode biofilm bacteria are divided into 6, respectively (Proteobacteria) Proteobacteria, Firmicutes (Firmicutes), Chloroflexi (Chloroflexi), Bacteroidetes (Bacteroidetes), actinobacteria (Actinobacteria), acidobacteria (Acidobacteria). The relative abundance of bacterial strain in more than 97%, is the dominant bacteria. The biofilm anode and anode sediment archaeal are divided into 3, respectively (Crenarchaeota) Crenarchaeota and Euryarchaeota (Euryarchaeota) and Parvarchaeota, and are based on the Crenarchaeota were the dominant groups, but the relative abundance of anode mud in the various categories of ancient bacteria are higher than 1~2 orders of magnitude on the anode biofilm. The anode and the anode biofilm in the sediment fungal community is divided into 3, respectively (Ascomycota) Ascomycota, Basidiomycota (Basidiomycota) and Zygomycota Ascomycota (Zygomycota), as the dominant bacteria. By comparison Changes in microbial community structure and sediment from the anode biofilm and the anode in the sediment, found Xiwanshi genus of MFC after the operation (Shewanella) significantly increased the relative abundance and diversity of microbial communities in the sediment and the anode biofilm anode decreased obviously, verify that MFC is selective to the anode microbial community structure.

【学位授予单位】：浙江海洋大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X714;TM911.45;X172

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