|本期目录/Table of Contents|

[1]邰勇,邓敬轩,任洪艳,等.一株Agrobacterium sp.异养硝化菌的分离鉴定及其氨氧化性能[J].应用与环境生物学报,2020,26(02):325-331.[doi:10.19675/j.cnki.1006-687x.2019.04031]
 TAI Yong,DENG Jingxuan,REN Hongyan,et al.Isolation and identification of a heterotrophic nitrifying strain of Agrobacterium sp. and its ammoxidation performance[J].Chinese Journal of Applied & Environmental Biology,2020,26(02):325-331.[doi:10.19675/j.cnki.1006-687x.2019.04031]
点击复制

一株Agrobacterium sp.异养硝化菌的分离鉴定及其氨氧化性能
分享到:

《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
26卷
期数:
2020年02期
页码:
325-331
栏目:
研究论文
出版日期:
2020-04-25

文章信息/Info

Title:
Isolation and identification of a heterotrophic nitrifying strain of Agrobacterium sp. and its ammoxidation performance
作者:
邰勇邓敬轩任洪艳黄振兴阮文权
1江南大学环境与土木工程学院 无锡 214122 2江苏省厌氧生物技术重点实验室 无锡 214122 3江苏省水处理技术与材料协同创新中心 苏州 215009
Author(s):
TAI Yong1 DENG Jingxuan1 REN Hongyan1 2 3? HUANG Zhengxin1 2 3 & RUAN Wenquan1 2 3
1 School of Environment and Civil Engineering, Jiangnan University, Wuxi 214122, China 2 Jiangsu Key Laboratory of Anaerobic Biotechnology, Wuxi 214122, China 3 Jiangsu Province Water Treatment Technology and Materials Collaborative Innovation Center, Suzhou 215009, China
关键词:
异养硝化菌农杆菌氨氧化氨单加氧酶
Keywords:
heterotrophic nitrifying bacterium Agrobacterium sp. ammonia oxidation ammonia monooxygenase (AMO)
DOI:
10.19675/j.cnki.1006-687x.2019.04031
摘要:
针对传统污水处理工艺中存在的工艺复杂、脱氮效率低等问题,从江苏无锡市桃花山垃圾渗滤液生化反应池活性污泥中富集、分离及筛选出一株异养硝化菌BT1. 通过16S rRNA序列分析,对分离菌株进行鉴定,同时对其异养硝化特性、氨氧化功能基因及氨氧化性能影响因素进行研究. 结果显示:分离到的异养硝化菌为农杆菌属Agrobacterium sp.. 该菌经过32 h培养后,NH4+-N去除率为99.77%;TN去除率为96.99%. 其中,59.62% TN转换为胞内氮,37.37% TN转化为气态氮;检测不到NO3--N和NO2--N的积累. 结合氨单加氧酶基因(amoA)的PCR成功扩增,进一步证明了BT1菌株具有氨氧化能力. 单因子试验结果显示,在温度为30 ℃、C/N为10-15、pH为7.0-9.0、转速为120-160 r/min的条件下,菌株均能去除98.51%以上NH4+-N,体现出良好的氨氧化性能. BT1菌株能够适应较宽的氨氮负荷,在高氨氮浓度(500和1 000 mg/L)下生长良好且NH4+-N去除率均超过64.69%. 本研究表明BT1菌株具有高效的异养硝化性能及优异的氨氮耐受性,具有进一步处理高浓度氨氮废水的应用前景. (图9 参39)
Abstract:
Traditional wastewater treatment processes are complicated and nitrogen removal efficiency is low. In this study, a heterotrophic nitrifying bacterium was enriched, isolated, and screened from activated sludge of a landfill leachate biochemical reaction tank located in Taohuashan, Wuxi City, Jiangsu Province. The strain was identified by morphological observation and 16S rRNA sequence analysis. The heterotrophic nitrification characteristics, ammonia oxidation gene function, and influencing factors of heterotrophic nitrification properties were studied. The results showed that the isolated heterotrophic nitrifying bacterium was Agrobacterium sp. and the NH4+-N removal rate reached 99.77% after 32 h incubation. The total nitrogen (TN) removal rate was 96.99%, of which 59.62% of TN was converted to cell growth as internal nitrogen and 37.37% of TN was converted to gaseous nitrogen. The accumulation of NO3--N and NO2--N was not detected. Moreover, the ammonia monooxygenase gene was successfully amplified. The ability of ammonia oxidation for the BT1 strain was proven. The single factor experiment results showed that the strain could remove more than 98.51% of NH4+-N under conditions of 30 ℃, C/N 10–15, pH 7.0–9.0, and 120–160 r/min, which showed the good ammonia oxidation performance of the strain. The ammonia nitrogen tolerance of the BT1 strain was better and grew well with 500 and 1 000 mg/L ammonia nitrogen. The NH4+-N removal rate exceeded 64.69%. The high-efficiency heterotrophic nitrification performance and excellent ammonia nitrogen tolerance makes the BT1 strain ideal for use in wastewater treatment processes.

参考文献/References:

1 郝明辉, 于鲁冀, 李廷梅, 刘攀龙. 一株异养硝化菌的筛选及生长特性研究[J]. 生物技术通报, 2016, 32 (4): 168-174 [Hao MH, Yu LZ, Li TM, Liu PL. Study on screening and growth characteristics of a heterotrophic nitrifying bacteria [J]. Biotechnol Bull, 2016, 32 (4): 168-174]
2 Yao S. Heterotrophic nitrification and aerobic denitrification at low temperature by a newly isolated bacterium, Acinetobacter sp. HA2 [J]. Bioresour Technol, 2013, 139 (13): 80-86
3 Huang F, Pan L, Lü N, Tang XM. Characterization of novel Bacillus strain N31 from mariculture water capable of halophilic heterotrophic nitrification?—aerobic denitrification [J]. J Biosci Bioeng, 2017, 124 (5): 564-571
4 黄廷林, 何秀秀, 张海涵, 周石磊, 白士远. 水库贫营养异养硝化-好氧反硝化菌Sxf14的脱氮特性[J]. 应用与环境生物学报, 2015, 21 (2): 201-207 [Huang TL, He XX, Zhang HH, Zhou SL, Bai SY. Nitration characteristics of aquaculture heterotrophic nitrification-aerobic denitrifying bacteria Sxf14 [J]. Chin J Appl Environ Biol, 2015, 21 (2): 201-207]
5 丁炜, 朱亮, 徐京, 冯丽娟, 徐向阳. 好氧反硝化菌及其在生物处理与修复中的应用研究进展[J]. 应用与环境生物学报, 2011, 17 (6): 923-929 [Ding W, Zhu L, Xu J, Feng LJ, Xu XX. Advances in aerobic denitrifying bacteria and their application in biological treatment and remediation [J]. Chin J Appl Environ Biol, 2011, 17 (6): 923-929]
6 Zhang XY, Zheng SK, Zhang HY, Duan SP. Autotrophic and heterotrophic nitrification-anoxic denitrification dominated the anoxic/oxic sewage treatment process during optimization for higher loading rate and energy savings [J]. Bioresour Technol, 2018, 263: 84-93
7 葛启隆. 波茨坦短芽孢杆菌异养硝化性能与关键酶活性研究[J]. 太原学院学报 (自然科学版), 2017, 35 (2): 31-35 [Ge QL. Study on heterotrophic nitrification performance and key enzyme activity of Bacillus licheniformis [J]. J Taiyuan Univ (Nat Sci Ed), 2017, 35 (2): 31-35]
8 陈茂霞, 王欢, 周后珍, 李杨, 谭周亮, 李旭东. 异养硝化-好氧反硝化菌HN-02的筛选及其特性[J]. 应用与环境生物学报, 2013, 19 (4): 688-693 [Chen MX, Wang H, Zhou HZ, Li Y, Tan ZL, Li XD. Screening and characteristics of heterotrophic nitrification-aerobic denitrifying bacteria HN-02 [J]. Chin J Appl Environ Biol, 2013, 19 (4): 688-693]
9 刘志培, 刘双江. 硝化作用微生物的分子生物学研究进展[J]. 应用与环境生物学报, 2004, 10 (4): 521-525 [Liu ZP, Liu SJ. Progress in molecular biology of nitrification microorganisms [J]. Chin J Appl Environ Biol, 2004, 10 (4): 521-525]
10 张峰峰, 周可, 谢凤行, 赵琼, 孙海波, 赵玉洁. 一株异养硝化菌AD-28氨氮去除特性研究[J]. 水产科学, 2019, 38 (1): 92-97 [Zhang FF, Zhou K, Xie FX, Zhao Q, Sun HB, Zhao YJ. Study on the removal characteristics of ammonia nitrogen in a heterotrophic nitrifying bacteria AD-28 [J]. J Fish Sci, 2019, 38 (1): 92-97]
11 杨垒. 高效异养硝化细菌的脱氮特性及其处理高氨氮废水研究[D]. 西安: 西安建筑科技大学, 2016 [Yang L. Determination of high-efficiency heterotrophic nitrifying bacteria and its treatment of high ammonia nitrogen wastewater [D]. Xi’an: Xi’an University of Architecture and Technology, 2016]
12 金敏, 王景峰, 孔庆鑫, 赵祖国, 王新为, 谌志强, 陈照立, 邱志刚, 李君文. 好氧异养硝化菌Acinetobacter sp. YY-5的分离鉴定及脱氮机理[J]. 应用与环境生物学报, 2009, 15 (5): 692-697 [Jin M, Wang JF, Kong QX, Zhao ZG, Wang XW, Qi ZQ, Chen ZL, Qiu ZG, Li JW. Isolation, identification and denitrification mechanism of aerobic heterotrophic nitrifying bacteria Acinetobacter sp. YY-5 [J]. Chin J Appl Environ Biol, 2009, 15 (5): 692-697]
13 国家环保局本书编委会. 水和废水监测分析方法[M]. 北京: 中国环境科学出版社, 1989: 252-278 [Editorial Board of the National Environmental Protection Agency. Water and Wastewater Monitoring and Analysis Methods [M]. Beijing: China Environmental Science Press, 1989: 252-278]
14 秦美川. 亚硝酸细菌的快速检测及荧光定量PCR检测试剂盒的创建[D]. 大连: 大连海洋大学, 2016 [Qin MC. Rapid detection of nitrite bacteria and creation of a real-time PCR assay kit [D]. Dalian: Dalian Ocean University, 2016]
15 刘兴, 李连星, 薄香兰, 陈维楚, 周胜杰, 王军, 王立红, 陈成勋. 铜绿假单胞菌YY24的异养硝化-好氧反硝化功能基因的研究[J]. 水产科学, 2018, 37 (4): 45-53 [Liu X, Li LX, Bo XL, Chen WC, Zhou SJ, Wang J, Wang LH, Chen CX. Heterotrophic nitrification-aerobic denitrification functional genes of Pseudomonas aeruginosa YY24 [J]. Fish Sci, 2018, 37 (4): 45-53]
16 康鹏亮, 张海涵, 黄廷林, 陈胜男, 商潘路, 冯稷, 贾竞宇. 湖库沉积物好氧反硝化菌群脱氮特性及种群结构[J]. 环境科学, 2018 (5): 2431-2437 [Kang PL, Zhang HH, Huang TL, Chen SN, Pan SL, Feng J, Jia JY. Nitrogen removal characteristics and population structure of aerobic denitrifying bacteria in lake sediments [J]. Environ Sci, 2018 (5): 2431-2437]
17 王国英, 崔杰, 岳秀萍, 李亚男, 贾子龙. 异养硝化-好氧反硝化菌脱氮同时降解苯酚特性[J]. 中国环境科学, 2015, 35 (9): 2644-2649 [Wang GY, Cui J, Yue XP, Li YN, Jia ZL. Characteristics of denitrification and phenol degradation by heterotrophic nitrification-aerobic denitrifying bacteria [J]. Chin Environ Sci 2015, 35 (9): 2644-2649]
18 游海. 一株异养硝化-好氧反硝化菌的分离,鉴定及降解特性研究[D]. 武汉: 武汉科技大学,2013 [You H. Separation, identification and degradation characteristics of a heterotrophic nitrification-aerobic denitrifying bacteria [D]. Wuhan: Wuhan University of Science and Technology, 2013]
19 Ren YX, Yang L, Liang X. The characteristics of a novel heterotrophic nitrifying and aerobic denitrifying bacterium, Acinetobacter junii YB [J]. Bioresour Technol, 2014, 171: 1-9
20 王洁, 蓝江林, 刘波. 一株异养硝化细菌的分离鉴定和脱氮特性研究[J]. 农业环境科学学报, 2013, 32 (4): 805-810 [Wang J, Lan JL, Liu B. Isolation, Identification and nitrogen removal characteristics of a heterotrophic nitrifying bacteria [J]. J Agro-Environ Sci, 2013, 32 (4): 805-810]
21 Choi KJ, Zhang S, Song JH, Hwang SJ. Aerobic denitrification by a heterotrophic nitrifying-aerobic denitrifying (HN-AD) culture enriched activated sludge [J]. Ksce J Civil Eng, 2017, 21 (6): 2113-2118
22 胡杰, 颜家保, 霍晓琼, 陈美玲, 李超. 低C/N比异养硝化-好氧反硝化菌筛选及硝化特性[J]. 化工进展, 2019 (3): 1567-1572 [Hu J, Yan JB, Huo XQ, Chen ML, Li C. Screening and nitrification characteristics of low C/N ratio heterotrophic nitrification-aerobic denitrification bacteria [J]. Chem Prog, 2019 (3): 1567-1572]
23 刘杰凤, 周天, 刘正辉, 程水明. 一株新型异养硝化细菌的分离鉴定及硝化特性[J]. 环境科学与技术, 2014, 37 (8): 99-103 [Liu JF, Zhou T, Liu ZH, Cheng SM. Isolation, identification and nitrification characteristics of a novel heterotrophic nitrifying bacteria [J]. Environ Sci Technol, 2014, 37 (8): 99-103]
24 连红民, 邱忠平, 何昆明, 周文秀. 一株好氧反硝化-异养硝化菌的筛选及脱氮特性研究[J]. 生物技术通报, 2015, 31 (6): 138-143 [Lian Hm, Qiu ZP, He KM, Zhou WX. Screening and nitrogen removal characteristics of a aerobic denitrification-heterotrophic nitrifying bacteria [J]. Biotechnol Bull, 2015, 31 (6): 138-143]
25 Li D, Liang XH, Jin Y, Wu CD. Isolation and nitrogen removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium, Klebsiella sp. TN-10 [J]. Appl Biochem Biotechnol, 2018 (4): 1-15
26 王兆阳, 陈国耀, 姜珂, 许培雅. 1株耐冷兼性嗜碱好氧反硝化菌的分离鉴定及反硝化特性[J]. 环境科学, 2014 (6): 2341-2348 [Wang ZY, Chen GY, Jiang K, XU PY. Identification and denitrification characteristics of a psychrotolerant facultative basophilic aerobic denitrifier [J]. Environ Sci, 2014 (6): 2341-2348]
27 张婷月, 丁钰, 黄民生.异养硝化-好氧反硝化细菌的筛选及其脱氮性能研究[J]. 华东师范大学学报 (自然科学版), 2018 (6): 22-31+87 [Zhang TY, Ding W, Huang MS. Screening of heterotrophic nitrification-aerobic denitrifying bacteria and its nitrogen removal performance [J]. J East Chin Normal Univ (Nat Sci Ed), 2018 (6): 22-31+87]
28 Qin W, Li WG, Gong XJ, Huang XF, Fan WB, Zhang DY, Yao P, Wang XJ, Song Y. Seasonal-related effects on ammonium removal in activated carbon filter biologically enhanced by heterotrophic nitrifying bacteria for drinking water treatment [J]. Environ Sci Pollut Res, 2017, 24 (24): 1-14
29 Joo HS, Hirai M, Shoda M. Piggery wastewater treatment using Alcaligenes faecalis strain No. 4 with heterotrophic nitrification and aerobic denitrification [J]. Water Res, 2006, 40 (16): 0-3036
30 李焱生. 异养硝化细菌的分离鉴定及荧光标记和关键酶基因研究[D]. 杭州: 浙江工业大学, 2010 [Li YS. Isolation and Identification of Heterotrophic Nitrifying Bacteria and Study on Fluorescent Labeling and Key Enzyme Genes [D]. Hangzhou: Zhejiang University of Technology, 2010]
31 Anthonisen AC, Loehr RC, Prakasam TBS, Srinath EG. Inhibition of nitrification by ammonia and nitrous acid [J]. Water Pollut Control Fed, 1976, 48: 835-852
32 王秀杰, 王维奇, 李军, 王思雨, 张晶, 魏佳, 赵白航. 氨氮对异养硝化菌Acinetobactor sp.活性影响及动力学特性分析[J]. 中国环境科学, 2018 (3): 943-949 [Wang XJ, Wang WQ, Li J, Wang SY, Zhang J, Wei J, Zhao BH. Effects of ammonia nitrogen on the activity of heterotrophic nitrifying bacteria Acinetobactor sp. and its kinetic characteristics [J]. Chin Environ Sci, 2018 (3): 943-949]
33 Xu Y, He T, Li Z, Ye Q, Chen Y, Xie E, Zhang X. Nitrogen removal characteristics of Pseudomonas putida Y-9 capable of heterotrophic nitrification and aerobic denitrification at low temperature [J]. Biomed Res Int, 2017, 2017: 1-7
34 刘万鑫, 杜荣斌, 由翠荣, 付学军, 梁丽琨. 几株异养硝化细菌与好氧反硝化细菌的分离与鉴定[J]. 烟台大学学报 (自然科学与工程版), 2017 (3): 249-254 [Liu WX, Du RB, Yu CR, Fu XJ, Liang LK. Isolation and identification of several heterotrophic nitrifying bacteria and aerobic denitrifying bacteria [J]. J Yantai Univ (Nat Sci Eng Ed), 2017 (3): 249-254]
35 吴建江, 王兆阳, 许培雅. 一株高效异养硝化菌的分离、鉴定及其氨氮去除特性[J]. 中国环境科学, 2013, 33 (7): 1309-1315 [Wu JJ, Wang ZY, Xu PY. Isolation, Identification and ammonia nitrogen removal characteristics of a high-efficiency heterotrophic nitrifying bacteria [J]. Chin Environ Sci, 2013, 33 (7): 1309-1315]
36 Li Y, Wang Y, Fu L, Gao YZ, Zhao HX, Zhou WZ. Aerobic-heterotrophic nitrogen removal through nitrate reduction and ammonium assimilation by marine bacterium Vibrio sp. Y1-5 [J]. Bioresour Technol, 2017, 230 (Complete): 103-111
37 Richardson DJ, Wehrfritz JM, Keech A, Crossman LC, Roldan MD, Sears HJ, Butler CS, Reilly A, Moir JW, Berks BC, Ferguson SJ, Thomson AJ, Spiro S. The diversity of redox proteins involved in bacterial heterotrophic nitrification and aerobic denitrification [J]. Biochem Soc Trans, 1998, 26 (3): 401-408
38 Feng Y, Feng J, Shu Q. Isolation and characterization of heterotrophic nitrifying and aerobic denitrifying\r, Klebsiella pneumoniae\r, and\r, Klebsiella variicola\r, strains from various environments [J]. J Appl Microbiol, 2018, 124 (5): 1195-1211
39 Fu G, Yu T, Huangshen LK, Han JY. The influence of complex fermentation broth on denitrification of saline sewage in constructed wetlands by heterotrophic nitrifying/aerobic denitrifying bacterial communities [J]. Bioresour Technol, 2018, 250: 290-298

相似文献/References:

[1]张培玉,曲洋,杨瑞霞,等.耐盐异养硝化菌驯化方法及分离菌株鉴定[J].应用与环境生物学报,2011,17(01):121.[doi:10.3724/SP.J.1145.2011.00121]
 ZHANG Peiyu,QU Yang,YANG Ruixia,et al.Domestication of Halotolerant Heterotrophic Nitrifying Bacteria and Identification of Two Isolated Strains[J].Chinese Journal of Applied & Environmental Biology,2011,17(02):121.[doi:10.3724/SP.J.1145.2011.00121]

更新日期/Last Update: 2020-04-25