离子型稀土矿山集液沟区域耐盐异养硝化细菌的筛选及其脱氮特征
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摘要
为筛选处理高盐含氨氮矿冶废水的耐盐微生物,从赣南离子型稀土矿山集液沟区域采集样品,利用选择性培养基富集培养分离耐盐异养硝化细菌,并考察碳源、pH、盐度等因素对其脱氮性能的影响.结果表明,分离得到一株耐盐异养硝化细菌X1,经16S rDNA鉴定为肠杆菌属Enterobacter sp.X1,确定该菌适宜的脱氮条件为:以蔗糖作为碳源,pH值为5,盐度≤15%.菌株X1的NH_4~+-N去除率可达到60%以上.研究结果可为高盐含氨氮矿冶废水的微生物处理提供菌株和数据参考.
To screen the salt tolerant microorganism with high salinity and ammonia nitrogen mining and metallurgy wastewater, the samples were collected from the collecting groove region of ion-type rare earth mines in south of Jiangxi province, and the salt tolerant heterotrophic nitrifying bacteria were cultured and isolated by selective medium. Apart from that, the effect of carbon source, p H, and salinity on its nitrogen removal performance was investigated. The results show that a salt tolerant heterotrophic nitrifying bacterium X1 is isolated and identified as Enterobacter sp. X1 by 16 S r DNA analysis. The suitable condition for nitrogen removal is determined as follows: the sucrose is the carbon source, the p H value equal to 5, and the salinity less than or equal to 15 %. The removal rate of NH_4~+-N of strain X1 could reach above 60 %. These results could provide a reference for the microbial treatment of wastewater with high salt and ammonia nitrogen.
To screen the salt tolerant microorganism with high salinity and ammonia nitrogen mining and metallurgy wastewater, the samples were collected from the collecting groove region of ion-type rare earth mines in south of Jiangxi province, and the salt tolerant heterotrophic nitrifying bacteria were cultured and isolated by selective medium. Apart from that, the effect of carbon source, p H, and salinity on its nitrogen removal performance was investigated. The results show that a salt tolerant heterotrophic nitrifying bacterium X1 is isolated and identified as Enterobacter sp. X1 by 16 S r DNA analysis. The suitable condition for nitrogen removal is determined as follows: the sucrose is the carbon source, the p H value equal to 5, and the salinity less than or equal to 15 %. The removal rate of NH_4~+-N of strain X1 could reach above 60 %. These results could provide a reference for the microbial treatment of wastewater with high salt and ammonia nitrogen.
引文
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[13]邱廷省,伍红强,方夕辉,等.风化壳淋积型稀土矿提取除杂技术现状及进展[J].稀土,2012,33(4):81-85.
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[15]LEI Y,WANG Y,LIU H,et al.A novel heterotrophic nitrifying and aerobic denitrifying bacterium,Zobellella taiwanensis DN-7,can remove high-strength ammonium[J].Appl Microbiol Biotechnol,2016,100(9):4219-4229.
[16]FAN L,CHEN J,LIU Q,et al.Exploration of three heterotrophic nitrifying strains from a tilapia pond for their characteristics of inorganic nitrogen use and application in aquaculture water[J].Journal of Biosci Bioeng,2015,119(3):303-309.
[17]SHODA M,Y ISHIKAWA.Heterotrophic nitrification and aerobic denitrification of high-strength ammonium in anaerobically digested sludge by Alcaligenes faecalis strain No.4[J].Journal of Biosci Bioeng,2014,117(6):737-741.
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[19]KUSHNER D J.Life in high salt and solute concentrations,in Microbial Life in Extreme Environments[M].London:Academic Press,1978.
[20]JIN R,LIU T,LIU G,et al.Simultaneous heterotrophic nitrification and aerobic denitrification by the marine origin bacterium Pseudomonas sp.ADN-42[J].Appl Biochem Biotechnol,2015,175(4):2000-2011.
[21]DUAN J,FANG H,SU B,et al.Characterization of a halophilic heterotrophic nitrification-aerobic denitrification bacterium and its application on treatment of saline wastewater[J].Bioresour Technol,2015,179:421-428.
[22]YOU J,LIU J F,YANG S,et al.Low-temperature-active and salttolerant beta-mannanase from a newly isolated Enterobacter sp.strain N18[J].Journal of Biosci Bioeng,2016,121(2):140-146.
[23]KIM K,JANG Y J,LEE S M,et al.Alleviation of salt stress by enterobacter sp.EJ01 in tomato and Arabidopsis is accompanied by up-regulation of conserved salinity responsive factors in plants[J].Mol Cells,2014,37(2):109-117.
[24]苟莎,黄钧.异养硝化细菌脱氮特性及研究进展[J].微生物学通报,2009(2):255-260.
[25]刘杰凤,周天,刘正辉,等.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.
[26]王弘宇,马放,杨开,等.两株异养硝化细菌的氨氮去除特性[J].中国环境科学,2009,29(1):47-52.
[27]RICHARDSON D J,WEHRFRITZ J M,KEECH A,et al.The diversity of redox proteins involved in bacterial heterotrophic nitrification and aerobic denitrification[J].Biochem Soc Trans,1998,26(3):p.401-408.
[28]金敏,王景峰,庆鑫,等.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.
[29]何霞,吕剑,何义亮,等.异养硝化机理的研究进展[J].微生物学报,2006,46(5):844-847.
[2]HE T,LI Z,SUN Q,et al.Heterotrophic nitrification and aerobic denitrification by Pseudomonas tolaasii Y-11 without nitrite accumulation during nitrogen conversion[J].Bioresour Technol,2016,200(1):493-499.
[3]JI B,YANG K,WANG H,et al.Aerobic denitrification by Pseudomonas stutzeri C3 incapable of heterotrophic nitrification[J].Bioprocess Biosyst Eng,2015,38(2):407-409.
[4]LI C,YANG J,WANG X,et al.Removal of nitrogen by heterotrophic nitrification-aerobic denitrification of a phosphate accumulating bacterium Pseudomonas stutzeri YG-24[J].Bioresour Technol,2015,182:18-25.
[5]SUN Y,LI A,ZHANG X,et al.Regulation of dissolved oxygen from accumulated nitrite during the heterotrophic nitrification and aerobic denitrification of Pseudomonas stutzeri T13[J].Appl Microbiol Biotechnol,2015,99(7):3243-3248.
[6]DAUM M,ZIMMER W,PAPEN H,et al.Physiological and molecular biological characterization of ammonia oxidation of the heterotrophic nitrifier Pseudomonas putida[J].Curr Microbiol,1998,37(4):281-288.
[7]SU J J,YEH K S,TSENG P W,et al.A strain of Pseudomonas sp.isolated from piggery wastewater treatment systems with heterotrophic nitrification capability in Taiwan[J].Curr Microbiol,2006,53(1):77-81.
[8]PAL R R,KHARDENAVIS A A,PUROHIT H J.Identification and monitoring of nitrification and denitrification genes in Klebsiella pneumoniae EGD-HP19-C for its ability to perform heterotrophic nitrification and aerobic denitrification[J].Funct Integr Genomics,2015,15(1):63-76.
[9]MOIR J W,CROSSMAN L C,SPIRO S,et al.The purification of ammonia monooxygenase from Paracoccus denitrificans[J].FEBS Lett,1996,387(1):71-74.
[10]JOO H S,HIRAI M,SHODA M.Characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis No.4[J].J Biosci Bioeng,2005,100(2):184-191.
[11]KIM J K,PARK K J,CHO K S,et al.Aerobic nitrification-denitrification by heterotrophic Bacillus strains[J].Bioresour Technol,2005,96(17):1897-906.
[12]李春.原地浸矿新工艺在离子型稀土矿的推广应用[J].有色金属科学与工程,2011,2(1):63-67.
[13]邱廷省,伍红强,方夕辉,等.风化壳淋积型稀土矿提取除杂技术现状及进展[J].稀土,2012,33(4):81-85.
[14]DEREEPER A,GUIGNON V,BLANC G,et al.Phylogeny.fr:robust phylogenetic analysis for the non-specialist[J].Nucleic Acids Res,2008,36:465-469.
[15]LEI Y,WANG Y,LIU H,et al.A novel heterotrophic nitrifying and aerobic denitrifying bacterium,Zobellella taiwanensis DN-7,can remove high-strength ammonium[J].Appl Microbiol Biotechnol,2016,100(9):4219-4229.
[16]FAN L,CHEN J,LIU Q,et al.Exploration of three heterotrophic nitrifying strains from a tilapia pond for their characteristics of inorganic nitrogen use and application in aquaculture water[J].Journal of Biosci Bioeng,2015,119(3):303-309.
[17]SHODA M,Y ISHIKAWA.Heterotrophic nitrification and aerobic denitrification of high-strength ammonium in anaerobically digested sludge by Alcaligenes faecalis strain No.4[J].Journal of Biosci Bioeng,2014,117(6):737-741.
[18]KUNDU P,PRAMANIK A,DASGUPTA A,et al.Simultaneous heterotrophic nitrification and aerobic denitrification by Chryseobacterium sp.R31 isolated from abattoir wastewater[J].Biomed Res Int,2014,ID,436056:1-12.
[19]KUSHNER D J.Life in high salt and solute concentrations,in Microbial Life in Extreme Environments[M].London:Academic Press,1978.
[20]JIN R,LIU T,LIU G,et al.Simultaneous heterotrophic nitrification and aerobic denitrification by the marine origin bacterium Pseudomonas sp.ADN-42[J].Appl Biochem Biotechnol,2015,175(4):2000-2011.
[21]DUAN J,FANG H,SU B,et al.Characterization of a halophilic heterotrophic nitrification-aerobic denitrification bacterium and its application on treatment of saline wastewater[J].Bioresour Technol,2015,179:421-428.
[22]YOU J,LIU J F,YANG S,et al.Low-temperature-active and salttolerant beta-mannanase from a newly isolated Enterobacter sp.strain N18[J].Journal of Biosci Bioeng,2016,121(2):140-146.
[23]KIM K,JANG Y J,LEE S M,et al.Alleviation of salt stress by enterobacter sp.EJ01 in tomato and Arabidopsis is accompanied by up-regulation of conserved salinity responsive factors in plants[J].Mol Cells,2014,37(2):109-117.
[24]苟莎,黄钧.异养硝化细菌脱氮特性及研究进展[J].微生物学通报,2009(2):255-260.
[25]刘杰凤,周天,刘正辉,等.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.
[26]王弘宇,马放,杨开,等.两株异养硝化细菌的氨氮去除特性[J].中国环境科学,2009,29(1):47-52.
[27]RICHARDSON D J,WEHRFRITZ J M,KEECH A,et al.The diversity of redox proteins involved in bacterial heterotrophic nitrification and aerobic denitrification[J].Biochem Soc Trans,1998,26(3):p.401-408.
[28]金敏,王景峰,庆鑫,等.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.
[29]何霞,吕剑,何义亮,等.异养硝化机理的研究进展[J].微生物学报,2006,46(5):844-847.