發(fā)布時間：2018-05-31 19:53

  本文選題：菌株篩選 + 產(chǎn)氫��； 參考：《太原理工大學(xué)》2015年碩士論文

【摘要】：氫氣熱值高，燃燒產(chǎn)物清潔無污染，被認(rèn)為是21世紀(jì)最具應(yīng)用價值的可再生能源之一。與傳統(tǒng)的電化學(xué)、熱化學(xué)制氫方法相比，生物制氫有著節(jié)約能源、不消耗礦物資源、可持續(xù)等優(yōu)點(diǎn)，其在研究與應(yīng)用中的地位越來越受到重視，許多國家都投入大量的人力物力對其進(jìn)行研究。在生物制氫研究中，光合細(xì)菌光發(fā)酵制氫因具有產(chǎn)氫效率高、底物轉(zhuǎn)化效率高、對環(huán)境無污染等突出的優(yōu)點(diǎn)，而日益受到人們的廣泛關(guān)注。 本實(shí)驗利用光照富集以及雙層平板劃線法從污水廠二沉池的活性污泥中分離純化得到一株光合細(xì)菌，結(jié)合菌落特征以及16SrDNA基因序列分析等對其進(jìn)行了分類學(xué)鑒定，確定該菌株為沼澤紅假單胞菌（Rhodopseudomonas palustris），命名為Rhodopseudomonas palustris.PB-Z。 以所篩選的光合細(xì)菌PB-Z為研究對象，對其生長條件進(jìn)行初步優(yōu)化，并研究了PB-Z在不同氮源、碳源、氮源濃度、碳源濃度、初始pH值、鎂離子濃度下的產(chǎn)氫特性。實(shí)驗結(jié)果表明：（1）厭氧光照是PB-Z的最佳生長模式；乙酸鈉為PB-Z生長的最佳碳源；在光照強(qiáng)度為4500lux、初始pH為7時，PB-Z的生長活性最高。（2）以尿素為氮源時，PB-Z有著最高的產(chǎn)氫活性，累積產(chǎn)氫量為91.2±4.5ml，尿素為PB-Z的最佳產(chǎn)氫氮源；在尿素濃度為10mmol/L時，PB-Z的產(chǎn)氫量最大，，為101.1±3.4ml。（3）PB-Z產(chǎn)氫的最佳碳源為葡萄糖，當(dāng)葡萄糖濃度為70mmol/L時，PB-Z有著最大的產(chǎn)氫量與底物轉(zhuǎn)化率，分別為123±3ml、69.7±1.9%。PB-Z的最大產(chǎn)氫活性出現(xiàn)在PB-Z的對數(shù)生長期。（4）PB-Z產(chǎn)氫的最佳初始pH為7，PB-Z在利用葡萄糖產(chǎn)氫的過程中，會產(chǎn)生有機(jī)酸使培養(yǎng)基變?yōu)樗嵝�，通過外源調(diào)節(jié)培養(yǎng)基的pH能夠顯著提高光合細(xì)菌PB-Z的產(chǎn)氫量以及底物轉(zhuǎn)化率，產(chǎn)氫量由124.7±2.7ml（對照組）提高到221.7±8ml，底物轉(zhuǎn)化率由68.1%（對照組）提高到91.2%。（5）光合細(xì)菌PB-Z產(chǎn)氫的最適鎂離子濃度為5mmol/L。PB-Z中含有多種色素，在一定的鎂離子濃度范圍內(nèi)，PB-Z的色素合成狀況良好，但是過高濃度的鎂離子會對PB-Z的色素合成造成不利影響，光合色素合成受到抑制，PB-Z的產(chǎn)氫量出現(xiàn)顯著地下降。為探究光合細(xì)菌PB-Z產(chǎn)氫的最適條件，根據(jù)單因素實(shí)驗的結(jié)果，以鎂離子濃度、pH、反應(yīng)時間為實(shí)驗因素，以光合細(xì)菌PB-Z產(chǎn)氫量為響應(yīng)值，采用響應(yīng)面分析法對實(shí)驗結(jié)果進(jìn)行模擬及分析，建立了產(chǎn)氫量與3個產(chǎn)氫影響因素關(guān)系的二次多項式數(shù)學(xué)模型。結(jié)果表明，鎂離子濃度、pH值、反應(yīng)時間與產(chǎn)氫量存在顯著的相關(guān)性，確定PB-Z的最佳產(chǎn)氫條件為：鎂離子濃度4.77mmol/L，初始pH7.05，反應(yīng)時間77.5h，在此實(shí)驗條件下，實(shí)際產(chǎn)氫量為137.8ml，與預(yù)測值相比偏差僅4.8%。說明響應(yīng)曲面法優(yōu)化PB-Z的產(chǎn)氫條件是可行的。
[Abstract]:Because of its high calorific value and clean combustion products, hydrogen is considered as one of the most valuable renewable energy sources in the 21st century. Compared with the traditional electrochemical and thermochemical hydrogen production methods, biological hydrogen production has the advantages of saving energy, not consuming mineral resources, and being sustainable, so it has been paid more and more attention in the research and application. Many countries have invested a lot of manpower and material resources to study it. In the research of biological hydrogen production, photo-fermenting hydrogen production by photosynthetic bacteria has attracted more and more attention for its advantages of high efficiency of hydrogen production, high conversion efficiency of substrate and no pollution to the environment. In this experiment, a photosynthetic bacteria was isolated and purified from activated sludge of secondary settling tank of wastewater treatment plant by the method of light enrichment and double layer plate drawing. The bacteria was identified by taxonomy combining with colony characteristics and 16SrDNA gene sequence analysis. The strain was identified as Rhodoppseudomonas palustris, named Rhodopseudomonas palustris.PB-Z. The growth conditions of photosynthetic bacteria (PB-Z) were preliminarily optimized, and the hydrogen production characteristics of PB-Z under different nitrogen sources, carbon sources, nitrogen source concentrations, carbon source concentrations, initial pH values and magnesium ion concentrations were studied. The results show that anaerobic illumination is the best growth mode for PB-Z, sodium acetate is the best carbon source for PB-Z growth, and the growth activity of PB-Z is the highest when the light intensity is 4500luxand and the initial pH is 7) when urea is used as nitrogen source, PB-Z has the highest hydrogen production activity. The accumulative hydrogen production was 91.2 鹵4.5 ml, urea was the best hydrogen and nitrogen source for PB-Z, the hydrogen production of PB-Z was the largest when urea concentration was 10mmol/L, and the optimum carbon source of hydrogen production was glucose when the concentration of urea was 101.1 鹵3.4ml.(3)PB-Z. When the concentration of glucose was 70mmol/L, PB-Z had the largest hydrogen production and substrate conversion. The maximum hydrogen production activity of 69.7 鹵1.9%.PB-Z (123 鹵3 ml) appeared in the logarithmic growth period of PB-Z. The optimum initial pH of hydrogen production of PB-Z was 7g PB-Z. In the process of hydrogen production from glucose, organic acids were produced to make the medium acidic. The hydrogen production and substrate conversion of photosynthetic bacteria PB-Z could be significantly increased by adjusting the pH of the medium. Hydrogen production increased from 124.7 鹵2.7 ml (control group) to 221.7 鹵8 ml, and substrate conversion rate increased from 68.1 ml (control group) to 91.22%. 5) the optimum magnesium ion concentration of photosynthetic bacteria PB-Z for hydrogen production was that 5mmol/L.PB-Z contained many kinds of pigments, and the pigment synthesis of PB-Z was good in a certain range of mg ~ (2 +) concentration. However, too high concentration of magnesium ions had a negative effect on the synthesis of PB-Z pigments, and the hydrogen production of PB-Z decreased significantly when the synthesis of photosynthetic pigments was inhibited. In order to explore the optimum conditions for hydrogen production of photosynthetic bacteria (PB-Z), according to the results of single factor experiments, the pH of mg ~ (2 +) and reaction time were taken as experimental factors, and the hydrogen production of photosynthetic bacteria (PB-Z) was taken as response value. The response surface analysis method is used to simulate and analyze the experimental results, and a quadratic polynomial mathematical model of the relationship between hydrogen production and three factors affecting hydrogen production is established. The results showed that there was a significant correlation between mg ~ (2 +) concentration, pH value, reaction time and hydrogen production. The optimum hydrogen production conditions of PB-Z were determined as follows: mg ~ (2 +) concentration 4.77 mmol / L, initial pH 7.05, reaction time 77.5 h. The actual hydrogen production is 137.8 ml, the deviation is only 4.8% compared with the predicted value. It is feasible to optimize the hydrogen production conditions of PB-Z by response surface method.
【學(xué)位授予單位】：太原理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：TQ116.2;Q93

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