好氧反硝化菌的分离及脱氮特性研究李秀婷

127　　1,2,　1,＊,1,　1,1(1.北京工商大学化学与环境工程学院,北京100048;2.山东阜丰发酵有限公司,山东莒南276600)　:研究了从活性污泥中分离得到好氧反硝化细菌106及其反硝化能力,探讨了培养基碳源、CODCr/N、培养温度及pH对菌株106反硝化作用的影响。结果表明,菌株106能在好氧条件下有效去除培养基中的硝基氮,且其脱氮率达90%以上。其最适培养条件为30℃、摇床转速160r/min、初始pH7.0,分别以琥珀酸钠和硝酸钾为唯一碳源和氮源时,最适CODCr/N为26。该菌在溶解氧为6.0±1.0mg·L-1条件下,培养24h时对NO-3-N的去除率为99.17%。该反硝化细菌106可以在好氧条件下第1d内完成脱氮,具备高效的反硝化性能,且在整个反硝化过程中一直保持低水平的亚硝酸盐浓度。:好氧反硝化菌,筛选,脱氮StudyonisolationanddenitrificationcharacterizationofaerobicdenitrifierLIXiu-ting1,XUGuo-hua2,WANGPing1,＊,LVYue-gang1,ZHAOJin1,LIWen-ting1(1.SchoolofChemicalandEnvironmentalEngineering,BeijingTechnologyandBusinessUniversity,Beijing100048,China;2.ShangdongFufengFermentationCo.,Ltd.,Junan276600,China)Abstract:Abacterialstrain106isolatedfromtheactivatedsludgewasfoundcapableofaerobicdenitrificationandthedenitrifyingcapabilityofthestrainwasstudiedunderaerobiccondition.Theeffectsofcarbonsource,CODCr/Nratio,temperatureandpHondenitrificationoftheaerobicdenitrifierwereinvestigated.Theresultsshowedthatthenitrateintheculturecouldbeefficientlyremovedbystrain106andthenitrogenremovalratewasuptoabove90%.Itshowedthatthenitrite-reducingactivityreachedthehighestat30℃,160r/min,pH7.0andCODCr/Nratio26,usingsodiumsuccinateandpotassiumnitrateasthesolecarbonsourceandnitrogensource,respectively.Whentheconcentrationofdissolvedoxygenwas6.0±1.0mg·L-1,theremovalrateofnitratebystrain106was99.17%in24h.Ourresultsindicatedthatthedenitrifierisolatedcouldremovethenitrogenonthefirstday,nearlycompletedenitrificationcouldbeobtainedevenunderoxicconditionsandtherewasaverylowlevelofnitriteaccumulationinthedenitrificationprocess.Keywords:aerobicdenitrifiers;screening;denitrification:TS201.3　　　　:A　　　　:1002-0306(2010)05-0127-04:2009-12-22　＊通讯联系人:李秀婷(1970-),女,副教授,主要从事酶工程方面的研究。:国家科技支撑计划项目(2007BAK36B07)。　　,,,。,、、、,,,。,、,,,。,,,。,N2O[1]。N2O,。TIN,,,N2O。,,,,。1　1.1　　;DOI:10.13386/j.issn1002-0306.2010.05.054128　　1　106NO-3-N、NO-2-NTIN(h)OD600NO-3-N(mg·L-1)NO-2-N(mg·L-1)NO-3-N(%)TIN(%)00.103121.70.977--240.45059.170.1251.4051.69480.54056.840.29753.3153.44720.38652.190.17257.1357.33960.38145.680.06562.4862.73:。BTB(/L)[2]　20g,KNO31g,KH2PO41g,FeCl2·6H2O0.5g,CaCl2·7H2O0.2g,MgSO4·7H2O1g,8.5g,(BTB)(1%)1mL,1mol/LNaOHpH7.0～7.3,121℃20min,;LB(/L)　KNO31g,KH2PO41g,FeCl2·6H2O0.05g,CaCl2·7H2O0.02g,MgSO4·7H2O1g,8.5g,121℃20min,;DM(/L)　KNO30.72g,KH2PO41g,MgSO4·7H2O1g,2.8g,121℃20min,。1.2　,0.1mLBTB,,30℃2～3d,,。LB,30℃,160r/min24h,10%(DM),30℃,160r/min,。(NO-3-N)(NO-2-N),NO-3-NTIN(NO-3-N+NO-2-N),。NO-3-N:;NO-2-N:N-(1-)-[3]。:,721600nm。1.3　106。、、。,CODCr/N=10,NO-3-N=100mg/L,160r/min,30℃4d,NO-3-NNO-2-N,NO-3-NTIN。,、pH、、。,,CODCr/N26∶1,NO-3-N=100mg/L。150mL,pH7.0,160r/min,30℃,NO-3-NTIN。2　2.1　106BTB22BTB,,,,106(1):10624h,121.7mg/L59.17mg/L;96h10662.48%62.73%。,,0.977mg/L,0.065mg/L,1.00mg/L,N2O。,106,,24h,48h,。,,,,。2.2　106,,、、pH、、DO。,,,[4]。HuangH.K.[5]Citrobacterdiversus,C/NDO。2.2.1　　106、、、4d,1,2。1　1062　106,、、,48h,129　　3　106(℃)(h)OD600NO-2-N(mg·L-1)NO-3-N(mg·L-1)TIN(mg·L-1)NO-3-N(%)TIN(%)2500.1360.090124.8124.9——720.8060.13611.2711.4190.9690.863000.0810.545134.7135.3——720.8070.33810.4010.7392.2892.063500.0860.546140.1140.6——720.9230.67033.5634.2376.0475.664000.1740.612144.2144.9——720.2940.388125.0125.413.3213.42:。。,24h,,。,,。,,,[6]。,NO-3、O2,NO-3O2[7]。。2,,106,TIN57.87%,,TIN52.45%,,,TIN46.17%43.75%。[8]X31、,97.57%97.30%,80%,X31,。106,,,。2.2.2　(CODCr/N)　2106。2,CODCr/N=10,49.65%,CODCr/N26,,93.35%。CODCr/N10～25,CODCr/N,,93.35%,。JooHS[9],,,。2.2.3　pH　3106pH。TINpH,pH5.591.51%pH7.0TIN97.18%;pH;pH8.0,TIN92.72%。106,pH7.0,106,。,pH5.5～8.0,90%,pH7.0,97.18%,106pH,,,。2　106NO-3-NCODCr/NNO-3-N(%)24h48h72h96h1029.4144.4548.1849.651256.5854.1953.3956.891569.8665.0964.6666.661663.3969.2271.471.311964.9475.1074.3075.272065.1475.2779.7481.792268.5880.8983.5784.732565.2889.8890.5890.112661.6691.4691.8893.352863.9488.7092.5292.803069.9173.7384.4083.87:。3　pH1062.2.4　　3106、。,25℃30℃,90.86%92.06%,35℃,75.66%。40℃13.32%,。,30℃,。2.2.5　　,(DO),,[10],130　　NO-3-NNO-2-N,。,。,。4106。100r/min,106,59.03%;,;160r/min,,99.17%;180r/min,91.16%。160r/min106,6.0±1.0mg·L-1。,;,。4　1062.2.6　106　106,106,NO-3-N、NO-2-N,106,5。5,10624h,NO-3-N108.22mg/L2.21mg/L;,,12h0.0969mg/L,,,,。,106,24h,TIN98.05%,30h99.95%,。5　1063　106,,90%。、CODCr/N、pH,10630℃、160r/min、pH7.0,,CODCr/N26。6.0±1.0mg·L-1,24hNO-3-N99.17%。pH,24h,,。,,。[1]BaumannB,SnozziM,ZehnderAJB,etal.DynamicsofdenitrificationactivityofParacoccusdenitrificansincontinuouscultureduringaerobic-anaerobicchanges[J].JBacteriol,1996,178:4367-4374.[2]TakayaN,MariaAntoninaBCS,YasushiS,etal.Aerobicdenitrificationbacteriathatproducelowlevelsofnitrousoxide[J].AppandEnvirMicrobio,2003,69(6):3152-3157.[3]国家环保局《水和废水监测分析方法》编委会.水和废水监测分析方法(第3版)[M].北京:中国环境科学出版社,1989.[4]RobertsonLA,KuenenJG.Thiospaerapantotrophagen.nov.sp.nov.,afacultativelyanaerobic,facultativelyautotrophicsulphurbacterium[J].JournalofGeneralMicrobiology,1983,129(8):2847-2855.[5]HuangHK,TsengSK.NitratereductionbyCitrobacterdiversusunderaerobicenvironment[J].AppliedMicrobiologyandBiotechnology,2001,55(1):90-94.[6]翟茜,汪苹,李秀婷,等.活性污泥中好氧反硝化菌的富集筛选及鉴别[J].环境科学与技术,2007,30(1):11-14.[7]WilsonLP,BouwerEJ.Biodegra