基于丙氨酸為底物的厭氧氨氧化過(guò)程研究
發(fā)布時(shí)間：2018-07-22 11:29
【摘要】：厭氧氨氧化(Anaerobic Ammonium Oxidation,ANMMOX)工藝因其效率高、能耗低、污泥產(chǎn)率低而備受關(guān)注。但是由于厭氧氨氧化菌(Anaerobic Ammonia-Oxidizing Bacteria,AAOB)生長(zhǎng)較為緩慢,對(duì)環(huán)境因素敏感,限制了ANMMOX工藝的進(jìn)一步推廣應(yīng)用。本課題通過(guò)研究丙氨酸作用下的厭氧氨氧化過(guò)程,探討了丙氨酸被AAOB利用的可能性。試驗(yàn)中利用以厭氧氨氧化菌為優(yōu)勢(shì)菌群的厭氧氨氧化富集培養(yǎng)物作為研究對(duì)象,考察了不同丙氨酸濃度以及不同底物在短期與長(zhǎng)期過(guò)程中對(duì)厭氧氨氧化過(guò)程的影響。采用一些常規(guī)的分析方法測(cè)定水中含氮化合物的含量評(píng)估系統(tǒng)的脫氮效果,采用現(xiàn)代分子生物學(xué)技術(shù)q PCR技術(shù)考察不同底物條件下厭氧氨氧化菌的變化情況。研究結(jié)果表明,短期培養(yǎng)過(guò)程中,在不同底物的作用下厭氧氨氧化過(guò)程受到的影響不同。當(dāng)丙氨酸作為唯一底物時(shí),體系中未發(fā)生厭氧氨氧化反應(yīng);當(dāng)利用丙氨酸作為電子供體以及碳源和利用丙氨酸作為電子供體時(shí),厭氧氨氧化活性較差,厭氧氨氧化過(guò)程受到的影響較大;當(dāng)利用丙氨酸作為碳源和在不缺乏電子供體以及碳源的體系中投加丙氨酸時(shí),厭氧氨氧化過(guò)程受到的影響較小。長(zhǎng)期培養(yǎng)過(guò)程中,丙氨酸對(duì)于厭氧氨氧化過(guò)程的影響較大。當(dāng)僅存在丙氨酸一種底物時(shí),體系中未發(fā)生厭氧氨氧化反應(yīng),厭氧氨氧化活性被抑制,厭氧氨氧化菌不能直接利用丙氨酸;在利用丙氨酸同時(shí)作為碳源與電子供體和利用丙氨酸作為電子供體這兩種條件下,丙氨酸對(duì)于厭氧氨氧化過(guò)程影響較大。氮去除負(fù)荷較低,分別為0.09 kg N/(m3·d),0.10 kg N/(m3·d)。平均比厭氧氨氧化活性分別為0.005 kgNH_4~+-N/(kg VSS·d),0.023 kg NH_4~+-N/(kg VSS·d),厭氧氨氧化活性受到抑制,利用丙氨酸同時(shí)作為碳源與電子供體和利用丙氨酸作為電子供體均不利于厭氧氨氧化過(guò)程的進(jìn)行;在利用丙氨酸作為碳源的條件下,氮去除負(fù)荷為0.19 kg N/(m3·d),總氮去除率為50%,平均比厭氧氨氧化活性為0.03 kg NH_4~+-N/(kgVSS·d),厭氧氨氧化活性較低,利用丙氨酸作為碳源不利于厭氧氨氧化過(guò)程的進(jìn)行;在不缺乏電子供體以及碳源的體系中投加丙氨酸的條件下,氮去除負(fù)荷為0.21 kg N/(m3·d),去除率達(dá)到55%。平均比厭氧氨氧化活性為0.08 kg NH_4~+-N/(kg VSS·d),厭氧氨氧化過(guò)程受到的影響最小。在不同底物作用下,△NO_2~-/△NH_4~+,△NO_3~-/△NH_4~+比值均偏離理論比值1.32,0.26,以在不缺乏電子供體以及碳源的體系中投加丙氨酸的條件下的偏離程度最低,厭氧氨氧菌在體系中不再是唯一的優(yōu)勢(shì)菌群。q PCR結(jié)果表明,在不同底物作用下厭氧氨氧化菌16S r DNA基因拷貝數(shù)數(shù)量均減少。丙氨酸被體系內(nèi)的異養(yǎng)菌利用,使得丙氨酸的去除率均達(dá)到96%以上。
[Abstract]:Anaerobic ammonia oxidation (Anaerobic Ammonium oxidation / ANMMOX) process has attracted much attention due to its high efficiency, low energy consumption and low sludge yield. However, the growth of Anaerobic Ammonia-Oxidizing BacteriaAAOB (AAOB) was slow and sensitive to environmental factors, which limited the further application of ANMMOX process. In this paper, the possibility of using alanine by AAOB was discussed by studying the anammox process of alanine. The effects of different alanine concentrations and substrates on the anaerobic ammonia oxidation process in the short and long term were investigated by using the enrichment culture of anaerobic ammonia oxidation with anaerobic ammonia oxidizing bacteria as the dominant bacteria. Some conventional analytical methods were used to determine the denitrification effect of the evaluation system for the content of nitrogen-containing compounds in water. The changes of anaerobic ammonia-oxidizing bacteria under different substrate conditions were investigated by using the modern molecular biological technique q-PCR. The results showed that the process of anaerobic ammoxidation was affected by different substrates in the short-term culture. When alanine is the sole substrate, there is no anammoxidation reaction in the system, but when alanine is used as electron donor and carbon source and alanine is used as electron donor, the anaerobic ammonia oxidation activity is poor. When alanine was used as carbon source and alanine was added in the system without electron donor and carbon source, the anammox process was less affected. During the long-term culture, alanine has a great effect on the anaerobic ammoxidation process. When there is only one kind of alanine substrate, there is no anaerobic ammonia oxidation reaction in the system, the anaerobic ammonia oxidation activity is inhibited, and the anaerobic ammonia oxidizing bacteria can not utilize alanine directly. Under the conditions of using alanine as carbon source and electron donor and alanine as electron donor, alanine has a great influence on anaerobic ammoxidation process. Nitrogen removal load was lower, 0.09 kg N / (m 3 d) 0.10 kg N / (m 3 d).), respectively. The average specific anaerobic ammonia oxidation activity was 0.005 kg NH _ 4 ~ -N / (kg VSS _ d) 0.023 kg NH _ 4 ~ -N / (kg VSS d),) anaerobic ammonia oxidation activity was inhibited. The use of alanine as carbon source and electron donor and alanine as electron donor were not conducive to the anaerobic ammonia oxidation process. Under the condition of using alanine as carbon source, the nitrogen removal load was 0.19 kg N / (m 3 d), total nitrogen removal rate was 50%), and the average anaerobic ammonia oxidation activity was 0.03 kg NH 4 ~ -N / (kg VSS d),). Using alanine as carbon source is not conducive to anaerobic ammoxidation, and nitrogen removal load is 0.21 kg / (m ~ 3 d),) under the condition that alanine is not added to the system without electron donor and carbon source, and the removal rate of nitrogen reaches 55 kg / (m ~ 3 d),). The average specific anaerobic ammonia oxidation activity was 0.08 kg NH _ 4 ~ -N / (kg VSS d),). Under the action of different substrates, the ratios of no _ 2O _ (- / NH _ 4 ~) and no _ 3O _ (- / NH _ 4 ~) all deviated from the theoretical ratio of 1.32 ~ 0.26, and the lowest deviation was found when alanine was added to the system without electron donor and carbon source. The results of QPCR showed that the copies of 16s r DNA gene of anaerobic ammonia-oxidizing bacteria decreased under different substrate. Alanine was used by heterotrophic bacteria in the system and the removal rate of alanine was over 96%.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：X703
[Abstract]:Anaerobic ammonia oxidation (Anaerobic Ammonium oxidation / ANMMOX) process has attracted much attention due to its high efficiency, low energy consumption and low sludge yield. However, the growth of Anaerobic Ammonia-Oxidizing BacteriaAAOB (AAOB) was slow and sensitive to environmental factors, which limited the further application of ANMMOX process. In this paper, the possibility of using alanine by AAOB was discussed by studying the anammox process of alanine. The effects of different alanine concentrations and substrates on the anaerobic ammonia oxidation process in the short and long term were investigated by using the enrichment culture of anaerobic ammonia oxidation with anaerobic ammonia oxidizing bacteria as the dominant bacteria. Some conventional analytical methods were used to determine the denitrification effect of the evaluation system for the content of nitrogen-containing compounds in water. The changes of anaerobic ammonia-oxidizing bacteria under different substrate conditions were investigated by using the modern molecular biological technique q-PCR. The results showed that the process of anaerobic ammoxidation was affected by different substrates in the short-term culture. When alanine is the sole substrate, there is no anammoxidation reaction in the system, but when alanine is used as electron donor and carbon source and alanine is used as electron donor, the anaerobic ammonia oxidation activity is poor. When alanine was used as carbon source and alanine was added in the system without electron donor and carbon source, the anammox process was less affected. During the long-term culture, alanine has a great effect on the anaerobic ammoxidation process. When there is only one kind of alanine substrate, there is no anaerobic ammonia oxidation reaction in the system, the anaerobic ammonia oxidation activity is inhibited, and the anaerobic ammonia oxidizing bacteria can not utilize alanine directly. Under the conditions of using alanine as carbon source and electron donor and alanine as electron donor, alanine has a great influence on anaerobic ammoxidation process. Nitrogen removal load was lower, 0.09 kg N / (m 3 d) 0.10 kg N / (m 3 d).), respectively. The average specific anaerobic ammonia oxidation activity was 0.005 kg NH _ 4 ~ -N / (kg VSS _ d) 0.023 kg NH _ 4 ~ -N / (kg VSS d),) anaerobic ammonia oxidation activity was inhibited. The use of alanine as carbon source and electron donor and alanine as electron donor were not conducive to the anaerobic ammonia oxidation process. Under the condition of using alanine as carbon source, the nitrogen removal load was 0.19 kg N / (m 3 d), total nitrogen removal rate was 50%), and the average anaerobic ammonia oxidation activity was 0.03 kg NH 4 ~ -N / (kg VSS d),). Using alanine as carbon source is not conducive to anaerobic ammoxidation, and nitrogen removal load is 0.21 kg / (m ~ 3 d),) under the condition that alanine is not added to the system without electron donor and carbon source, and the removal rate of nitrogen reaches 55 kg / (m ~ 3 d),). The average specific anaerobic ammonia oxidation activity was 0.08 kg NH _ 4 ~ -N / (kg VSS d),). Under the action of different substrates, the ratios of no _ 2O _ (- / NH _ 4 ~) and no _ 3O _ (- / NH _ 4 ~) all deviated from the theoretical ratio of 1.32 ~ 0.26, and the lowest deviation was found when alanine was added to the system without electron donor and carbon source. The results of QPCR showed that the copies of 16s r DNA gene of anaerobic ammonia-oxidizing bacteria decreased under different substrate. Alanine was used by heterotrophic bacteria in the system and the removal rate of alanine was over 96%.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：X703
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