厌氧氨氧化过程指纹研究

The anaerobic ammonia oxidation (Anammox) is one of hotspots in the research field of environmental science and microbiology. More than ten papers on Anammox have been published in the international journal Nature or Science, and dozens of Anammox plants have been put into use all over the world. However, no pure culture of anaerobic ammonia oxidation bacteria (AnAOB) has been achieved yet. The quality of AnAOB mixed culture is not so good as expected. It has been reported that cytochrome c is one of the important components of AnAOB, an essential prosthetic group of Anammox enzymes, a main factor for the red colour of AnAOB. So far, however, no information is available about the coupling relationship between the activity, cytochrome c and red colour of AnAOB. In this project, the performance of Anammox reactor will be investigated. The component, community and function of AnAOB mixed culture will be analyzed. The colour characteristic of AnAOB mixed culture in different forms will be observed. The coupling relationship between the activity, cytochrome c and red colour of AnAOB mixed culture will be studied. The fingerprints for Anammox process will be created for the effective control and real time optimization. So it is significant to support the project for the development of Anammox technology.

厌氧氨氧化（Anammox）是环境和微生物领域的研究热点，已有10余篇论文发表于《Nature》和《Science》上，并有多个工程用于废水脱氮。但厌氧氨氧化菌（AnAOB）生长极慢，迄今未能纯培，菌种（混培物）质控困难。细胞色素c（Cyt c）是AnAOB的关键成分、Anammox酶的必要辅基、AnAOB呈现红色的主要致因。然而，人们对AnAOB活性、Cyt c含量、红色特征的耦联性知之甚少。在典型工况下，试验Anammox反应器的运行性能，检测AnAOB混培物的组成、结构和功能性状，研究AnAOB混培物各级结构的显色特征，探索AnAOB混培物功能活性、Cyt c[Fe(II)/Fe(III)]含量、显色特征的耦联关系，并以AnAOB混培物的红色特征构建Anammox反应器状态指纹（对应于运行性能的颜色指标），可指导Anammox过程有效控制和实时优化，推动Anammox技术持续发展。

作为极具潜力的新型废水生物脱氮技术，厌氧氨氧化（Anammox）工艺已成为环境工程领域的研究热点，但其存在种质易变、直观判据缺乏的难题，严重限制了工程化应用。本项目基于Anammox颗粒污泥床反应器，研创了典型过程状态指纹，可为Anammox工艺的运行调控提供理论依据和技术参数。主要结论如下：① 测定Anammox污泥反应动力学曲线，根据氨氮动力学特征参数（Ks-55.86 mg N/L，Ki-412.22 mg N/L，Kmax-151.75 mg N/L）划分了Anammox过程典型状态：饥饿状态（0-Ks, q/qmax<50%）、饱足状态（Ks-Kmax, q/qmax>50%）、耐受状态（Kmax-Ki, q/qmax>50%）和中毒状态（>Ki, q/qmax<50%）。② 通过长期稳定运行，分别建立了各典型状态的Anammox脱氮系统，测得了各自的稳态效能参数：饥饿、饱足、耐受和中毒状态下的容积去除速率分别为0.69、4.97、9.65、13.73 kg N/(m3·d)，污泥比活性分别为188.94±26.58、313.29±53.27、397.50±50.93、198.60±3.12 mg N/(g VSS·d)。③ 探究Anammox混培物的组效关系，创建了对应的颜色指纹。建立了各典型状态的色度值聚类中心（a*, b*）；色度值与污泥比活性之间存在极显著的相关关系（r=0.940, p<0.01）；通过色度值，以细胞色素c的浓度变化和价态比例变化关联了厌氧氨氧化菌联氨脱氢酶量及酶活。⑤ 探究厌氧氨氧化颗粒污泥形效关系，创建了对应的形态指纹。各典型工况下颗粒污泥体积平均粒径依次为1970、2187、2351、2566 μm，颗粒污泥表面凹凸度依次为0.931、0.916、0.882、0.870；通过相对光亮度（LSCE*/ LSCI*），以颗粒菌群、EPS和孔隙分布关联了颗粒污泥的粒径、凹凸度和传质通量。⑥ 探究厌氧氨氧化颗粒污泥床构效关系，创建了对应的质地指纹。污泥床自下而上颗粒污泥体积平均粒径由2135.18 μm递减至1470.50 μm，粒径分布变异系数由0.42逐级增加到0.83；光亮度L*可以表征颗粒污泥粒径及污泥床高度；由污泥床沿程密度差主导的颗粒污泥跳选分层运动是质地特征的成因。