本文關(guān)鍵詞： 降酚微生物 引物優(yōu)化 微生物群落結(jié)構(gòu) 降解特性　出處：《湖南農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：酚類化合物作為重要的化工原料與副產(chǎn)物,廣泛存在于工業(yè)廢水之中,造成的環(huán)境污染負(fù)荷日益劇增,并逐漸成為當(dāng)前水體的主要污染物之一。在含酚廢水的眾多處理方法中,由于成本低、效率高、操作簡(jiǎn)單、無(wú)二次污染等特點(diǎn),生物法逐漸成為治理含酚工業(yè)廢水的主要方法。為了進(jìn)一步提高生物法的效率和速度,本研究在優(yōu)化降酚微生物分子檢測(cè)技術(shù)的基礎(chǔ)上,研究含酚工業(yè)廢水對(duì)微生物群落結(jié)構(gòu)的影響,并篩選出高濃度高效降酚菌,深入研究其降解特性、影響因素,從而為通過(guò)定向調(diào)控或添加外源降酚微生物,提高含酚工業(yè)廢水治理效率提供理論依據(jù)。主要結(jié)論如下：(1)根據(jù)已知的降酚微生物16S rDNA基因和苯酚羥化酶基因,通過(guò)生物信息學(xué)方法設(shè)計(jì)出4對(duì)降酚微生物16S rDNA引物和3對(duì)苯酚羥化酶基因的引物,并以江西景德鎮(zhèn)某焦化廠活性污泥為對(duì)象,分析了各對(duì)引物的有效檢出率,從而篩選出最優(yōu)引物對(duì)。研究發(fā)現(xiàn),苯酚羥化酶基因的最優(yōu)引物對(duì)是M2; 16S rDNA的最優(yōu)引物是M7。(2)通過(guò)高通量測(cè)序,研究了江西景德鎮(zhèn)某焦化廠厭氧活性污泥和好氧活性污泥的微生物群落結(jié)構(gòu)及季節(jié)對(duì)微生物群落結(jié)構(gòu)的影響。研究發(fā)現(xiàn)焦化廠活性污泥中微生物菌群豐度相對(duì)較低,但多樣性較高。活性污泥中的微生物可分16個(gè)分類單元,其中變形菌門為優(yōu)勢(shì)菌落,占48%,且隨著氣溫升高而降低。從活性污泥中微生物的種屬來(lái)看,Diaphorobacter sp.和Ottowia sp.為優(yōu)勢(shì)菌,分別占20.81%和13.48%。從不同季節(jié)來(lái)看,同季節(jié)的好氧活性污泥與厭氧活性污泥在結(jié)構(gòu)上基本一致,不同季節(jié)優(yōu)勢(shì)菌群不同。其中2月和8月的優(yōu)勢(shì)菌群為Diaphorobacter sp.,占29.38%,而5月和11月的優(yōu)勢(shì)菌群為Ottowia sp.,占17.06%。(3)通過(guò)富集培養(yǎng)分離純化到5株高濃度高效降酚菌,分別命名為pd-A、pd-B、 pd-C、pd-D、pd-E,耐酚能力均在2000 mg/L以上,其中pd-D降酚效果最佳。當(dāng)苯酚初始濃度為1000 mg/L時(shí),48 h內(nèi)pd-D菌株的苯酚降解率達(dá)72.11%。研究發(fā)現(xiàn),pd-D菌株的最佳降解條件為：初始酚濃度1000mg/L、30℃、pH7.0。在該條件下,96h內(nèi)pd-D菌株能將苯酚完全降解,且COD去除率大于80%。(4)研究了以pd-D菌株為陽(yáng)極催化劑,利用微生物燃料電池處理苯酚廢水的新模式。研究表明,當(dāng)在苯酚濃度為1000 mg/L, pd-D菌株的產(chǎn)電周期約7 d,產(chǎn)電電壓達(dá)0.5 V,COD去除明顯升高,苯酚降解時(shí)間縮短了24 h。(5)經(jīng)形態(tài)學(xué)觀察、生理生化實(shí)驗(yàn)及16S rDNA基因序列比對(duì)分析,初步確定pd-D為梭狀芽孢桿菌屬(Clostridium sp.)成員。
[Abstract]:Phenolic compounds, as important chemical raw materials and by-products, widely exist in industrial wastewater, resulting in an increasing environmental pollution load, and gradually become one of the main pollutants in water. Because of the characteristics of low cost, high efficiency, simple operation and no secondary pollution, biological method has gradually become the main method for the treatment of industrial wastewater containing phenol. On the basis of optimizing the molecular detection technology of phenol reducing microorganism, the effect of phenol industrial wastewater on microbial community structure was studied, and the high concentration and high efficiency phenol reducing bacteria were screened out, and its degradation characteristics and influencing factors were studied deeply. It provides a theoretical basis for improving the treatment efficiency of phenol-containing industrial wastewater by regulating or adding exogenous phenol-reducing microorganisms. The main conclusions are as follows: (1) based on the known 16s rDNA gene and phenol-hydroxylase gene of phenol-reducing microorganism, Four pairs of 16s rDNA primers and three pairs of primers for phenol hydroxylase gene were designed by bioinformatics. The effective detection rate of each pair of primers was analyzed by using activated sludge of a coking plant in Jingdezhen, Jiangxi Province. The results showed that the optimal primer pair of phenol hydroxylase gene was M2, and the best primer of 16s rDNA was M7.2) by high-throughput sequencing. The microbial community structure of anaerobic activated sludge and aerobic activated sludge in a coking plant in Jingdezhen, Jiangxi Province, and the effect of season on the microbial community structure were studied. It was found that the abundance of microbial flora in activated sludge of coking plant was relatively low. But the diversity is high. The microorganism in activated sludge can be divided into 16 taxonomic units, among which Proteus is the dominant colony, accounting for 48, and decreases with the increase of temperature. From the species of microorganisms in activated sludge, Diaphorobacter sp. and Ottowia sp. are the dominant bacteria. From different seasons, the structure of aerobic activated sludge and anaerobic activated sludge in the same season was basically the same. In February and August, the dominant flora was Diaphorobacter sp., accounting for 29.38%, while in May and May, the dominant flora was Ottowia sp., which accounted for 17.06. 3) five strains of high concentration high concentration phenol reducing bacteria were isolated and purified by enrichment culture. They were named pd-An pd-B, pd-Ca-pd-Dd-Pd-E, respectively, and the phenolic tolerance was more than 2 000 mg/L. When the initial concentration of phenol was 1000mg / L, the phenol degradation rate of pd-D strain reached 72.11. The optimum degradation conditions of pd-D strain were found as follows: initial phenol concentration 1000mg / L ~ 30 鈩，

本文編號(hào)：1508260