高温好氧反硝化菌的分离鉴定及其反硝化性能研究

32120111ENVIRONMENTALSCIENCEVol.32No.1Jan.201113123*1.5100062.5100063.5100061TAD1.0.67～0.89μm×1.03～1.41μm16SrDNAChelatococcussp..TAD150℃、24h63.79mg/L0.46mg/L99.12%.pHORP.TAD1pH7.0～9.02.1～7.2mg/L.16SrDNAX172X703A0250-3301201101-0259-072010-01-282010-04-1320777019200820086501986～[e-mail]*[e-mail]rificationCharacteristicsofaThermophilicAerobicDenitrifierZHANGMiao13HUANGShao-bin1231.CollegeofEnvironmentalScienceandEngineeringSouthChinaUniversityofTechnologyGuangzhou510006China2.StateKeyLaboratoryofPulpandPaperEngineeringSouthChinaUniversityofTechnologyGuangzhou510006China3.TheKeyLaboratoryofEnvironmentalProtectionandEco-RemediationofGuangdongRegularHigherEducationInstitutionsGuangzhou510006ChinaAbstractAbacterialstrainTAD1withhighnitrogenremovalefficiencywasisolatedfrombiofilmofthebiotricklingfilterofacoal-firedpowerplantbythermophilicdomestication.ThisbacteriumwasGramnegativeshortrodwiththesizeof0.67-0.89μm×1.03-1.41μm.ItwasidentifiedasChelatococcussp.accordingtoitsphysiologicalpropertiesandtheanalysisofits16SrDNAgene.Studiedonitsfunctionofaerobicdenitrificationatthetemperatureof50℃theresultsshowedthatnitrateintheculturemediawasefficientlyremovedfrom63.79mg/Lto0.46mg/Landthenitrogenremovalefficiencywasupto99.12%in24hoursandnonitritewasobservedduringtheincubationthemajorendproductofthedenitrificationwasnitrogen.DuringthedenitrificationofTAD1thepHintheculturemediumgraduallyincreaseswhiletheoxidation-reductionpotentialgraduallydecreases.ThefactorsaffectingaerobicdenitrificationbystrainTAD1werealsodiscussedindicatingthatthemostsuitablepHvalueforaerobicdenitrificationwas7.0-9.0andtheDOwas2.1-7.2mg/L.Keywordsthermophilicaerobicdenitrifier16SrDNAsequenceanalysisdenitrifyingcharacteristicsbiologicalnitrogenremoval0.1mg/L.Pochana1SBR95%30%2.3～7Takaya81Pseudomonassp.straink50DO38μmol/L0.03μmol/min.91S.maltophiliastrainX041DO3.80～5.21mg/L10h26.81mg/L0..310①②32③OH－pH.11～131450℃..150℃TAD1.、.11.1DMg/LNH4Cl0.3Na2HPO4·7H2O7.9KH2PO41.5MgSO4·7H2O0.14.0KNO30.62mLpH7.0～7.5.g/LEDTA50.0ZnSO42.2CaCl25.5MnCl2·4H2O5.06FeSO4·7H2O5.0NH46Mo7O2·4H2O1.1CuSO4·5H2O1.57CoCl2·6H2O1.61.1.2NOx15.1.2.190mL..30℃160r/min48h10%100mL35℃48h10%100mL50℃.24h.50℃3d1mL.pH.24hDM3..1.2.2100mLDM450℃160r/min24h..124h80%TAD1.1.31.3.1DM50℃12h.1.3.216TAD1.1.3.316SrDNAPCRPCR.16SrDNAF275′-AGAGTTTGATCCTGGCTCAG-3′R15225′-AAGGAGGTGATCCAGCCGCA-3′.PCR50μL94℃4min94℃30s54℃30s72℃2min3072℃10min.GenBankBlast.1.410%100mL、50℃0621160r/min4hNO－3-N、NO－2-N、N2、、pH、ORPCOD.1.5NO－3-NGB7480-87NO－2-NGB7493-87N-1--N2GC-2104He25mL/min32℃85℃75℃721480nmpHORPS-25pHCODDOYSIDO.216SrDNANJTAD1Fig.2PhylogenetictreesofstrainTAD1andrelatedbacteriabasedonthepartial16SrDNAsequenceswithNeighbor-Joining22.12.1.1TAD10.67～0.89μm×1.03～1.41μm.TAD11.2.1.2TAD1TAD1ONPG、、MR、V-P、、、.1TAD1Fig.1ElectronandmicroscopyphotographofstrainTAD12.1.316SrDNA1385bpGenBankTAD1Chelatococcusdaeguensis99%ChelatococcusdaeguensisstrainK10699%.MEGANeighbor-Joining16SrDNA2.Chelatococcussp.ChelatococcusdaeguensisstrainTAD1.2.2C.daeguensisTAD1C.daeguensisTAD11623224hNO－3-N、NO－2-N、N2、pH、ORPCOD3～6.3TAD1Fig.3GrowthcurveofaerobicdenitrificationofstrainTAD14TAD1Fig.4AerobicdenitrificationperformanceofstrainTAD130～4h4h12h.463.79mg/L0.46mg/L99.12%TAD1.TAD15TAD1pHORPFig.5CurvesofpHandORPinthedenitrificationprocessofstrainTAD16TAD1CODFig.6CurveofCODinthedenitrificationprocessofstrainTAD1.TAD1.5pHORP.pH17.ORP.64～12hCOD..2621.18～20.2.3C.daeguensisTAD1TAD1.21.TAD1..29113.69mg/L.50℃pH7.0160r/min.7NO－3-N113.69mg/L1.21mg/L98.94%NO－2-N113.58mg/L85.31mg/L24.89%.TAD1..TAD1..7Fig.7Aerobicdenitrificationprocessofnitrateandnitrite2.4pHC.daeguensisTAD1pH50℃160r/min8.8pHTAD1Fig.8EffectsofbeginningpHonaerobicdenitrificationbystrainTAD18pH.pHTAD1pH42%.pH.pH799.02%pH898.81%pH998.14%pH1010.47%.pH7～9TAD1pH6pH≥10.pHpH22.TAD1pH7.0～9.0.2.5C.daeguensisTAD1DO.C.daeguensisTAD1DO23DO24hDO.50℃.9.92.1～11.3362329DOTAD1Fig.9EffectsofDOonaerobicdenitrificationbystrainTAD1mg/L78.14mg/L50℃24hTAD1.7.2mg/L99.08%.TAD1..DODO.DO、.24.Huang18CitrohucterdiversusDODO2～6mg/L.Patureau19MicrovirgulaaerodenitrificansDO4.5mg/LDO4.5mg/LDO.Su25PseudomonasstutzeriSU2O292%90h99.24%.C.daeguensisTAD12.1～11.3mg/L.311TAD1、16SrDNAChelatococcussp..1.2C.daeguensisTAD150℃、24h99.12%.TAD1.3C.daeguensisTAD1pH7.0～9.0DODO2.1～11.3mg/L.1PochanaKKellerJ.StudyoffactorsaffectingsimultaneousnitrificationanddenitrificationSNDJ.WaterScienceandTechnology199939661-68.2RandallCWBarnardJLStenselHD.DesignandretrofitofwastewatertreatmentplantsforbiologicalnutrientremovalM.LancasterPATechnomicPublishingCompany1992.3.-J.200930123614-3618.4.Acinetobactersp.YY-5J.2009155692-697.5.J.200425175-78.6./N2OJ.20093072105-2111.7KimJBChoKSJeongSKetal.Identificationandcharacterizationofapigment-producingdenitrifyingbacteriumJ.BiotechnologyandBioprocessEngineering200813217-223.8TakayaNCatalan-SakairiMABSakaguchiYetal.AerobicdenitrifyingbacteriathatproducelowlevelsofnitrousoxideJ.AppliedandMicrobiologyBiotechnology20036963152-3157.9.J.200626113718-3724.10WangPLiXTXiangM