【摘要】：在廢水生物處理過程中微生物代謝產(chǎn)物對(duì)于生物聚集體形成及污染物轉(zhuǎn)化分解具有十分重要的影響。近年來,鐵氧化物強(qiáng)化厭氧產(chǎn)甲烷過程得到了國(guó)內(nèi)外研究學(xué)者的廣泛關(guān)注,但微生物代謝產(chǎn)物在此過程中的作用卻鮮有報(bào)道。因此,本研究通過構(gòu)建針鐵礦-厭氧產(chǎn)甲烷體系,利用不同規(guī)格的透析袋富集微生物代謝產(chǎn)物,進(jìn)而考察反應(yīng)體系甲烷產(chǎn)生效率及微生物群落的變化;以Shewanella oneidensis MR-1為對(duì)象,考察分析不同形態(tài)鐵還原體系內(nèi)微生物代謝產(chǎn)物的產(chǎn)生特征;構(gòu)建產(chǎn)甲烷菌-鐵還原菌-針鐵礦體系,利用iTRAQ蛋白質(zhì)組學(xué)技術(shù)對(duì)添加針鐵礦-產(chǎn)甲烷體系的微生物代謝產(chǎn)物進(jìn)行分析,利用高通量測(cè)序技術(shù)對(duì)微生物群落進(jìn)行了分析表征,得到的主要結(jié)論如下:1)使用透析袋有利于提高針鐵礦和微生物之間的接觸,但孔隙較大的透析(10kDa)對(duì)于產(chǎn)甲烷速率影響不大。而使用孔隙較小的透析袋(2000Da)顯著的提高了SMP濃度和產(chǎn)甲烷速率,SMP及其中的黃素類物質(zhì)可能在促進(jìn)產(chǎn)甲烷過程起到重要的作用。微生物群落分析表明SMP的富集不僅提高了體系內(nèi)具有電化學(xué)活性的微生物相對(duì)豐度,而且也促進(jìn)了乙酸型產(chǎn)甲烷菌Uncultured Methanosaeta sp.的生長(zhǎng)。2)以Shewanella oneidensis MR-1為研究對(duì)象,研究不同導(dǎo)電材料對(duì)鐵還原效率、微生物代謝產(chǎn)物及黃素類物質(zhì)分泌的影響,探究添加針鐵礦的產(chǎn)甲烷體系中可能的鐵還原行為。結(jié)果發(fā)現(xiàn),導(dǎo)電材料對(duì)于S.oneidensis MR-1還原針鐵礦的影響主要由影響電子傳遞效率及菌體活性兩方面組成。針鐵礦刺激S.oneidensis MR-1在培養(yǎng)初期分泌更多的微生物代謝產(chǎn)物,而導(dǎo)電材料的添加降低了S.oneidensis MR-1微生物代謝產(chǎn)物中黃素類物質(zhì)的比例。CNTs有效促進(jìn)了S.oneidensis MR-1對(duì)游離態(tài)鐵Fe-NTA的還原,石墨對(duì)S.oneidensis MR-1還原針鐵礦有一定的促進(jìn)作用。3)高通量分析結(jié)果顯示針鐵礦可以調(diào)節(jié)微生物群落功能。分泌蛋白和微生物群落分析結(jié)果顯示,針鐵礦顯著促進(jìn)了末端產(chǎn)甲烷代謝途徑及胞外電子轉(zhuǎn)移/氧化還原相關(guān)的過程,Firmicutes門、Anaerolinaceae科及Synergistetes門微生物參與了針鐵礦促進(jìn)產(chǎn)甲烷的過程。同時(shí)針鐵礦的添加使得以乙酸鹽為底物的產(chǎn)甲烷體系實(shí)現(xiàn)氫型產(chǎn)甲烷過程。iTRAQ蛋白質(zhì)組學(xué)分析結(jié)果表明SMP蛋白主要來源于微生物自身裂解。
[Abstract]:Microbial metabolites play an important role in the formation of biological aggregates and the transformation and decomposition of pollutants during the biological treatment of wastewater. In recent years, the enhancement of anaerobic methanogenic process by iron oxides has received extensive attention from domestic and foreign researchers, but the role of microbial metabolites in this process is rarely reported. Therefore, through the construction of goethite-anaerobic methanogenic system, the microbial metabolites were enriched with different dialysate bags, and the methane production efficiency and microbial community changes of the reaction system were investigated. Shewanella oneidensis MR-1 was used as the object. The production characteristics of microbial metabolites in different forms of iron reduction system were investigated and analyzed, and the methanogenic bacteria, iron reducing bacteria and goethite system were constructed. ITRAQ proteomics technique was used to analyze the microbial metabolites in the system of pyrite and methane production, and the microbial community was characterized by high-throughput sequencing technique. The main conclusions are as follows: 1) the use of dialysis bags can improve the contact between goethite and microbes, but the larger porosity dialysis (10kDa) has little effect on methanogenic rate. However, the concentration of SMP and methanogenic rate were significantly increased by using dialysate bag (2000Da) with small porosity. SMP and its flavonoids may play an important role in the process of methanogenesis. Microbial community analysis showed that the enrichment of SMP not only increased the relative abundance of microorganisms with electrochemical activity in the system, but also promoted Uncultured Methanosaeta sp. of methanogenic bacteria of acetic acid type. 2) the effects of different conductive materials on iron reduction efficiency, microbial metabolites and the secretion of flavin were studied with Shewanella oneidensis MR-1 as the research object, and the possible iron reduction behavior in methanogenic system with goethite was explored. The results show that the effect of conductive materials on the reduction of goethite by S.oneidensis MR-1 is mainly composed of two aspects which affect the efficiency of electron transport and the activity of bacteria. Goethite stimulated S.oneidensis MR-1 to secrete more microbial metabolites at the early stage of culture, while the addition of conductive materials decreased the ratio of xanthins in S.oneidensis MR-1 microbial metabolites. CNTs effectively promoted the reduction of free iron Fe-NTA by S.oneidensis MR-1. Graphite can promote the reduction of goethite by S.oneidensis MR-1. 3) the results of high throughput analysis show that goethite can regulate the function of microbial community. The results of secretory protein and microbial community analysis showed that goethite significantly promoted end-methanogenic pathway and extracellular electron transfer / redox related processes. Microbes of Firmicutes gate, Anaerolinaceae family and Synergistetes gate were involved in the process of goethite promoting methanogenesis. At the same time, the addition of goethite made the methanogenic system with acetate as the substrate to realize the process of hydrogen methanogenesis. The results of iTRAQ proteomics analysis showed that the SMP protein was mainly derived from microbial self-cleavage.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TQ221.11;X703

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