基于丙氨酸为底物的厌氧氨氧化过程研究
发布时间：2018-07-22 11:29
【摘要】：厌氧氨氧化(Anaerobic Ammonium Oxidation,ANMMOX)工艺因其效率高、能耗低、污泥产率低而备受关注。但是由于厌氧氨氧化菌(Anaerobic Ammonia-Oxidizing Bacteria,AAOB)生长较为缓慢,对环境因素敏感,限制了ANMMOX工艺的进一步推广应用。本课题通过研究丙氨酸作用下的厌氧氨氧化过程,探讨了丙氨酸被AAOB利用的可能性。试验中利用以厌氧氨氧化菌为优势菌群的厌氧氨氧化富集培养物作为研究对象,考察了不同丙氨酸浓度以及不同底物在短期与长期过程中对厌氧氨氧化过程的影响。采用一些常规的分析方法测定水中含氮化合物的含量评估系统的脱氮效果,采用现代分子生物学技术q PCR技术考察不同底物条件下厌氧氨氧化菌的变化情况。研究结果表明,短期培养过程中,在不同底物的作用下厌氧氨氧化过程受到的影响不同。当丙氨酸作为唯一底物时,体系中未发生厌氧氨氧化反应;当利用丙氨酸作为电子供体以及碳源和利用丙氨酸作为电子供体时,厌氧氨氧化活性较差,厌氧氨氧化过程受到的影响较大;当利用丙氨酸作为碳源和在不缺乏电子供体以及碳源的体系中投加丙氨酸时,厌氧氨氧化过程受到的影响较小。长期培养过程中,丙氨酸对于厌氧氨氧化过程的影响较大。当仅存在丙氨酸一种底物时,体系中未发生厌氧氨氧化反应,厌氧氨氧化活性被抑制,厌氧氨氧化菌不能直接利用丙氨酸;在利用丙氨酸同时作为碳源与电子供体和利用丙氨酸作为电子供体这两种条件下,丙氨酸对于厌氧氨氧化过程影响较大。氮去除负荷较低,分别为0.09 kg N/(m3·d),0.10 kg N/(m3·d)。平均比厌氧氨氧化活性分别为0.005 kgNH_4~+-N/(kg VSS·d),0.023 kg NH_4~+-N/(kg VSS·d),厌氧氨氧化活性受到抑制,利用丙氨酸同时作为碳源与电子供体和利用丙氨酸作为电子供体均不利于厌氧氨氧化过程的进行;在利用丙氨酸作为碳源的条件下,氮去除负荷为0.19 kg N/(m3·d),总氮去除率为50%,平均比厌氧氨氧化活性为0.03 kg NH_4~+-N/(kgVSS·d),厌氧氨氧化活性较低,利用丙氨酸作为碳源不利于厌氧氨氧化过程的进行;在不缺乏电子供体以及碳源的体系中投加丙氨酸的条件下,氮去除负荷为0.21 kg N/(m3·d),去除率达到55%。平均比厌氧氨氧化活性为0.08 kg NH_4~+-N/(kg VSS·d),厌氧氨氧化过程受到的影响最小。在不同底物作用下,△NO_2~-/△NH_4~+,△NO_3~-/△NH_4~+比值均偏离理论比值1.32,0.26,以在不缺乏电子供体以及碳源的体系中投加丙氨酸的条件下的偏离程度最低,厌氧氨氧菌在体系中不再是唯一的优势菌群。q PCR结果表明,在不同底物作用下厌氧氨氧化菌16S r DNA基因拷贝数数量均减少。丙氨酸被体系内的异养菌利用,使得丙氨酸的去除率均达到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%.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：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%.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X703
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