一株耐冷异养硝化菌的分离鉴定及脱氮特性
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摘要
利用异养硝化培养基,冬季从南四湖人工湿地底泥中分离筛选到一株具有较高脱氮性能的菌株Z01.经形态特征、生理生化特性以及16S r DNA序列分析,初步鉴定该菌株为假单胞菌属(Pseudomonas sp.),命名为Pseudomonas sp.Z01.实验探究得到菌株Z01的最佳初始投菌量为7%,最适温度、pH和盐度分别为15℃、6.0～8.0和0.3%,最佳碳源为葡萄糖.在最适生长条件下,通过氨氮去除试验发现该菌株对模拟污水中氨氮的去除率有一定提高,是异养硝化培养基中氨氮去除效率的1.1倍.此外,该菌株对模拟废水中总氮和COD也具有较好的去除效果.研究表明,菌株Z01在冬季污水处理中硝化脱氮具有良好的应用前景和很大的发展空间.
A heterotrophic nitrifying bacteria strain Z01 was isolated from the sediment of Lake Nansi constructed wetland in winter,and it was studied for its capacity of nitrification and nitrogen removal. Methods of morphological,physiological and biochemical were used to identify strain Z01 as well as 16 S r DNA analysis. It is affiliated to be Pseudomonas sp.,which named Pseudomonas sp. Z01. These data showed that the optimum dosage,temperature,pH and salinity of microbial strains Z01 were 7%,15℃,6.0-8.0 and 0. 3%,as sucrose as optimum carbon. Under the optimum removal efficiency of ammonia nitrogen from simulated wastewater by strain 10 was 1.1 times higher than that of heterotrophic nitrification medium. Moreover,it was better on the removal performance of the total nitrogen and COD in the simulated wastewater. Therefore,strain Z01 has a good application prospect and great development space in nitrification denitrification of sewage treatment in winter.
A heterotrophic nitrifying bacteria strain Z01 was isolated from the sediment of Lake Nansi constructed wetland in winter,and it was studied for its capacity of nitrification and nitrogen removal. Methods of morphological,physiological and biochemical were used to identify strain Z01 as well as 16 S r DNA analysis. It is affiliated to be Pseudomonas sp.,which named Pseudomonas sp. Z01. These data showed that the optimum dosage,temperature,pH and salinity of microbial strains Z01 were 7%,15℃,6.0-8.0 and 0. 3%,as sucrose as optimum carbon. Under the optimum removal efficiency of ammonia nitrogen from simulated wastewater by strain 10 was 1.1 times higher than that of heterotrophic nitrification medium. Moreover,it was better on the removal performance of the total nitrogen and COD in the simulated wastewater. Therefore,strain Z01 has a good application prospect and great development space in nitrification denitrification of sewage treatment in winter.
引文
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[8] Lin Y,Kong HN,He YL et al. Isolation and characterization of heterotrophic nitrifying bacteria. Environmental Science,2006,27(2):324-328.[林燕,孔海南,何义亮等.异养硝化细菌的分离及其硝化特性实验研究.环境科学,2006,27(2):324-328.]
[9] Zhao B,An Q,He YL et al. N2O and N2production during heterotrophic nitrification by Alcaligenes faecalis strain NR.Bioresource Technology,2012,116:379-385.
[10] Kundu P,Pramanik A,Mitra S et al. Heterotrophic nitrification by Achromobacter xylosoxidans S18 isolated from a smallscale slaughterhouse wastewater. Bioprocess and Biosystems Engineering,2012,35(5):721-728.
[11] Li B,Zhang QF,Dou SH et al. Screening of a psychrotrophic oil-degrading strain LHB16 and its degradation characteristics. Biotechnology,2010,20(5):83-85.[李兵,张庆芳,窦少华等.低温石油降解菌LHB16的筛选及降解特性研究.生物技术,2010,20(5):83-85.]
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[14] Chevalier P. Nitrogen and phosphorus removal by high latitude mat-forming cyanobacteria for potential use in tertiary wastewater treatment. Applied Phycology,2000,12(2):105-113.
[15] Xu Q,Zhu GB,Du GC. Separation and identification of an efficient psychrotrophic bacteria and research on its degradation characteristics. Chinese Journal of Environmental Engineering,2010,4(2):377-382.[徐巧,诸葛斌,堵国成.一株高效耐冷菌的分离鉴定及去除特性研究.环境工程学报,2010,4(2):377-382.]
[16] Zou YX. Enhance ammonia removal effect in constructed wetland at low temperature[Dissertation]. Changchun:Jilin University,2014.[邹雨璇.低温条件下人工湿地氨氮强化脱氮机理的研究[学位论文].长春:吉林大学,2014.]
[17] Cheng Y,Li QF,Fei YT et al. Screening and nitrogen removing characteristics of heterotrophic nitrification-aerobic denitrification bacteria SLWX2 from sea water. Environmental Science,2016,37(7):2681-2688.[成钰,李秋芬,费聿涛等.海水异养硝化-好氧反硝化芽孢杆菌SLWX2的筛选及脱氮特性.环境科学,2016,37(7):2681-2688.]
[18] Zhou DQ ed. Microbiology experiment course:The third edition. Beijing:Higher Education Press,2013.[周德庆.微生物学实验教程:第三版.北京:高等教育出版社,2013.]
[19] Dong XZ,Cai MY eds. Manual of identification in common bacteria system. Beijing:Science Press,2001:364-379.[东秀珠,蔡妙英.常见细菌系统鉴定手册.北京:科学出版社,2001:364-379.]
[20] Editorial board of"Water and wastewater monitoring and analysis method",Ministry of Environmental Protection of the People's Republic of China ed. Monitoring and analysis methods of water and wastewater:fourth edition. Beijing:China Environmental Science Press,2002.[国家环境保护总局《水和废水监测分析方法》编委会.水和废水监测分析方法:第4版.北京:中国环境科学出版社,2002.]
[21] Pang D,Peng ZR,He WH et al. Screening,identification and immobilized optimization of heterotrophic nitrifying bacteria. Genomics and Applied Biology,2018,37(6):2430-2437.[庞朵,彭自然,何文辉等.异养高效硝化细菌的筛选鉴定与固定化最优化.基因组学与应用生物学,2018,37(6):2430-2437.]
[22] He TX,Ni JP,Li ZL et al. Heterotrophic nitrification and aerobic denitrification of the hypothermia aerobic denitrification bacterium:Arthrobacter arilaitensis. Environmental Science,2016,37(3):296-302.[何腾霞,倪九派,李振轮等. 1株Arthrobacter arilaitensis菌的耐冷异养硝化和好氧反硝化作用.环境科学,2016,37(3):296-302.]
[23] Buchanan RE,Gibbons NE eds. Bergey's manual of systematic bacteriology:Eight edition. Baltimore:The Williams&Wilkins Company,1974.
[24] Joo HS,Hirai M,Shoda M. Characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis no. 4. Journal of Bioscience and Bioengineering,2005,100(2):184-191.
[25] Wang HY,Ma F,Yang K et al. Ammonia removal by two strains of heterotrophic nitrifying bacteria. China Environmental Science,2009,29(1):47-52.[王弘宇,马放,杨开等.两株异养硝化细菌的氨氮去除特性.中国环境科学,2009,29(1):47-52.]
[26] Gupta AB. Thiosphaera pantotropha a sulphur bacterium capable of simultaneous heterotrophic nitrification and aerobic denitrification. Enzyme and Miicrobial Technology,1997,21(8):589-595.
[27] Tang MZ,Li TT,Wang YN et al. Identification and characterization of an efficient psychrotrophic bacterial strain isolated from artificial wetlands. Acta Scientiae Circumstantiae,2013,33(3):708-714.[唐美珍,李婷婷,王艳娜等.人工湿地中一株高效低温菌的分离鉴定与去除特性研究.环境科学学报,2013,33(3):708-714.]
[28] Zhu F,Wang S,Zhou PJ. Molecular mechanisms of cold-adapted enzymes to cold environment and their application in biotechnology industry. Acta Microbiologica Sinaca,2002,42(5):640-664.[朱非,王珊,周培瑾.低温酶冷适应的分子机制及其在生物技术中的应用.微生物学报,2002,42(5):640-644.]
[29] Liu FF,Zhou DP,Wu SH et al. Isolation and identification of heterotrophic nitrifiers from cultivation wastewater. Journal of Agro-Environment Science,2010,29(11):2232-2237.[刘芳芳,周德平,吴淑杭等.养殖废水中异养硝化细菌的分离筛选和鉴定.农业环境科学学报,2010,29(11):2232-2237.]
[30] Li XL. Screening of cryogenic nitrification bacteria and its preliminary application in a low temperature wastewater treatment[Dissertation]. Qingdao:Qingdao University,2017.[李晓亮.低温硝化细菌的筛选及其在低温污水处理中的初步应用研究[学位论文].青岛:青岛大学,2017.]
[31] Xin YF,Qu XH,Yuan MD et al. Isolation and identification of a nitrifying and aerobic denitrifying Acinetobacter sp. YF14and its denitrification activity. Acta Microbiologica Sinica,2011,51(12):1646-1654.[辛玉峰,曲晓华,袁梦东等.一株异养硝化-反硝化不动杆菌的分离鉴定及脱氮活性.微生物学报,2011,51(12):1646-1654.]
[2] Bai SN,Xu TY,Liu Y et al. Separation and selection of heterotrophic nitrifying bacteria from lawn soil and their nitrification products characteristics. Acta Horticulturae Sinica,2018,45(7):1338-1346.[白三女,许天月,刘英等.草坪土壤异养硝化菌的分离筛选及硝化产物特征分析.园艺学报,2018,45(7):1338-1346.]
[3] Yao S,Ni JR,Ma T et al. Heterotrophic nitrification and aerobic denitrification at low temperature by a newly isolated bacterium Acinetobacter sp. HA2. Bioresource Technology,2013,139:80-86.
[4] Wang J,Lan JL,Liu B. Isolation and denitrification characteristics of a heterotrophic nitrification bacterium. Journal of Agro-Environment Science,2013,32(4):805-810.[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.农业环境科学学报,2013,32(4):805-810.]
[5] Tian FR,Zhang BB,Yang ZL et al. Isolation,identification of heterotrophic nitrifiers and optimization of their ammonia nitrogen removal efficiency. Environmental Engineering,2014,32(5):64-68.[田凤蓉,张彬彬,杨志林等.高效脱氨除臭异养硝化菌的筛选鉴定及脱氨性能研究.环境工程,2014,32(5):64-68.]
[6] Li XL,Zhang L,Zhang PY et al. Screening,biodiversity,and growth characteristics of cold-adapted nitrobacteria isolated from Arctic Ocean sediments. Chin J Appl Environ Biol,2017,23(2):294-300.[李晓亮,张良,张培玉等.北极海洋沉积物低温硝化细菌的筛选、多样性与生长特性.应用与环境生物学报,2017,23(2):294-300.]
[7] Wang TY,Wei HF,Hu ZQ et al. Isolation and identification of a heterotrophic nitrifying and aerobic denitrifying strain and its denitrification characteristics. Acta Scientiae Circumstantiae,2017,37(3):945-953.[王田野,魏荷芬,胡子全等.一株异养硝化好氧反硝化菌的筛选鉴定及其脱氮特性.环境科学学报,2017,37(3):945-953.]
[8] Lin Y,Kong HN,He YL et al. Isolation and characterization of heterotrophic nitrifying bacteria. Environmental Science,2006,27(2):324-328.[林燕,孔海南,何义亮等.异养硝化细菌的分离及其硝化特性实验研究.环境科学,2006,27(2):324-328.]
[9] Zhao B,An Q,He YL et al. N2O and N2production during heterotrophic nitrification by Alcaligenes faecalis strain NR.Bioresource Technology,2012,116:379-385.
[10] Kundu P,Pramanik A,Mitra S et al. Heterotrophic nitrification by Achromobacter xylosoxidans S18 isolated from a smallscale slaughterhouse wastewater. Bioprocess and Biosystems Engineering,2012,35(5):721-728.
[11] Li B,Zhang QF,Dou SH et al. Screening of a psychrotrophic oil-degrading strain LHB16 and its degradation characteristics. Biotechnology,2010,20(5):83-85.[李兵,张庆芳,窦少华等.低温石油降解菌LHB16的筛选及降解特性研究.生物技术,2010,20(5):83-85.]
[12] Yan FJ,Tian XL,Dong SL et al. Isolation,identification and characterization of a low-temperature degrading bacterium.Periodical of Ocean University of China,2011,41(3):24-30.[闫法军,田相利,董双林等.一株低温去除菌的分离鉴定及其去除特性研究.中国海洋大学学报,2011,41(3):24-30.]
[13] Zhao Y. Separation and identification of psychrotrophic ammonia degradation bacteria and their applications in wastewater treatment[Dissertation]. Lanzhou:Lanzhou Jiaotong University,2013.[赵燕.耐冷氨氮降解菌的分离、鉴定及其在废水处理中的应用研究[学位论文].兰州:兰州交通大学,2013.]
[14] Chevalier P. Nitrogen and phosphorus removal by high latitude mat-forming cyanobacteria for potential use in tertiary wastewater treatment. Applied Phycology,2000,12(2):105-113.
[15] Xu Q,Zhu GB,Du GC. Separation and identification of an efficient psychrotrophic bacteria and research on its degradation characteristics. Chinese Journal of Environmental Engineering,2010,4(2):377-382.[徐巧,诸葛斌,堵国成.一株高效耐冷菌的分离鉴定及去除特性研究.环境工程学报,2010,4(2):377-382.]
[16] Zou YX. Enhance ammonia removal effect in constructed wetland at low temperature[Dissertation]. Changchun:Jilin University,2014.[邹雨璇.低温条件下人工湿地氨氮强化脱氮机理的研究[学位论文].长春:吉林大学,2014.]
[17] Cheng Y,Li QF,Fei YT et al. Screening and nitrogen removing characteristics of heterotrophic nitrification-aerobic denitrification bacteria SLWX2 from sea water. Environmental Science,2016,37(7):2681-2688.[成钰,李秋芬,费聿涛等.海水异养硝化-好氧反硝化芽孢杆菌SLWX2的筛选及脱氮特性.环境科学,2016,37(7):2681-2688.]
[18] Zhou DQ ed. Microbiology experiment course:The third edition. Beijing:Higher Education Press,2013.[周德庆.微生物学实验教程:第三版.北京:高等教育出版社,2013.]
[19] Dong XZ,Cai MY eds. Manual of identification in common bacteria system. Beijing:Science Press,2001:364-379.[东秀珠,蔡妙英.常见细菌系统鉴定手册.北京:科学出版社,2001:364-379.]
[20] Editorial board of"Water and wastewater monitoring and analysis method",Ministry of Environmental Protection of the People's Republic of China ed. Monitoring and analysis methods of water and wastewater:fourth edition. Beijing:China Environmental Science Press,2002.[国家环境保护总局《水和废水监测分析方法》编委会.水和废水监测分析方法:第4版.北京:中国环境科学出版社,2002.]
[21] Pang D,Peng ZR,He WH et al. Screening,identification and immobilized optimization of heterotrophic nitrifying bacteria. Genomics and Applied Biology,2018,37(6):2430-2437.[庞朵,彭自然,何文辉等.异养高效硝化细菌的筛选鉴定与固定化最优化.基因组学与应用生物学,2018,37(6):2430-2437.]
[22] He TX,Ni JP,Li ZL et al. Heterotrophic nitrification and aerobic denitrification of the hypothermia aerobic denitrification bacterium:Arthrobacter arilaitensis. Environmental Science,2016,37(3):296-302.[何腾霞,倪九派,李振轮等. 1株Arthrobacter arilaitensis菌的耐冷异养硝化和好氧反硝化作用.环境科学,2016,37(3):296-302.]
[23] Buchanan RE,Gibbons NE eds. Bergey's manual of systematic bacteriology:Eight edition. Baltimore:The Williams&Wilkins Company,1974.
[24] Joo HS,Hirai M,Shoda M. Characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis no. 4. Journal of Bioscience and Bioengineering,2005,100(2):184-191.
[25] Wang HY,Ma F,Yang K et al. Ammonia removal by two strains of heterotrophic nitrifying bacteria. China Environmental Science,2009,29(1):47-52.[王弘宇,马放,杨开等.两株异养硝化细菌的氨氮去除特性.中国环境科学,2009,29(1):47-52.]
[26] Gupta AB. Thiosphaera pantotropha a sulphur bacterium capable of simultaneous heterotrophic nitrification and aerobic denitrification. Enzyme and Miicrobial Technology,1997,21(8):589-595.
[27] Tang MZ,Li TT,Wang YN et al. Identification and characterization of an efficient psychrotrophic bacterial strain isolated from artificial wetlands. Acta Scientiae Circumstantiae,2013,33(3):708-714.[唐美珍,李婷婷,王艳娜等.人工湿地中一株高效低温菌的分离鉴定与去除特性研究.环境科学学报,2013,33(3):708-714.]
[28] Zhu F,Wang S,Zhou PJ. Molecular mechanisms of cold-adapted enzymes to cold environment and their application in biotechnology industry. Acta Microbiologica Sinaca,2002,42(5):640-664.[朱非,王珊,周培瑾.低温酶冷适应的分子机制及其在生物技术中的应用.微生物学报,2002,42(5):640-644.]
[29] Liu FF,Zhou DP,Wu SH et al. Isolation and identification of heterotrophic nitrifiers from cultivation wastewater. Journal of Agro-Environment Science,2010,29(11):2232-2237.[刘芳芳,周德平,吴淑杭等.养殖废水中异养硝化细菌的分离筛选和鉴定.农业环境科学学报,2010,29(11):2232-2237.]
[30] Li XL. Screening of cryogenic nitrification bacteria and its preliminary application in a low temperature wastewater treatment[Dissertation]. Qingdao:Qingdao University,2017.[李晓亮.低温硝化细菌的筛选及其在低温污水处理中的初步应用研究[学位论文].青岛:青岛大学,2017.]
[31] Xin YF,Qu XH,Yuan MD et al. Isolation and identification of a nitrifying and aerobic denitrifying Acinetobacter sp. YF14and its denitrification activity. Acta Microbiologica Sinica,2011,51(12):1646-1654.[辛玉峰,曲晓华,袁梦东等.一株异养硝化-反硝化不动杆菌的分离鉴定及脱氮活性.微生物学报,2011,51(12):1646-1654.]