好氧反硝化菌T7用于废水脱氮特性的研究汤丽娟

1　3　　　　　　Vol.1　No.3200811EnvironmentandEcologyintheThreeGorgesNov.2008T7:2008-10-15:(50378095):(1983-),,,,,E-mail:[e-mail].汤丽娟,罗固源,许晓毅,林晓东,陈燕云(,400045)　:(SND)OGO,T7,,,,PCR16SrDNA,16SrDNAT7(Rhodococcus)。、,C/N5。,(PVA),,,。:,,PVA。　　:;;;;PVA　　:X703　　　　　　　　　:A　　:1674-2842(2008)03-0024-04StudyonNitrogenRemovalCharacteristicsofanAerobicDenitrifyingBacteriumRhodococcusStrainT7TANGLi-juan,LUOGu-yuan,XUXiao-yi,LINXiao-dong,CHENYan-yun(KeyLaboratoryofThreeGorgesReservoirRegionsEco-EnvironmentMinistryofEducation,ChongqingUniversity,Chongqing400045,China)Abstract:Onestrainofaerobicdenitrifyingbacterium,namedasT7,withhighactivityofnitrogenremovalwasiso-latedfromOGOreactorpossessingsimultaneousnitrificationanddenitrification(SND)phenomenon,T7isGram-positive,rod-likeandpinkandidentifiedasRhodococcusaccordingtoitsPCRamplificationand16SrDNAsequencehomologycomparison.Itcouldgrowwellwithpotassiumnitrateasonlynitrogensourceandsodiumcitrateasonlycarbonsource,andoptimumratioofC/Nwasfive.Theamplificationcultureofthisbacteriumwasusedinsyntheti-calnitratewastewatertreatment,andpolyvinylalcohol(PVA)wasselectedasentrapmentcarriertoimmobilizeT7.Theexperimentalresultsindicatethatthebacteriumcanmetabolizenitratewhensufficientoxygenexists,andcantreatdifferentconcentrationsofnitrogen-containingwastewater.Besides,themodeofdroppingbacteriaimmobilizedbypolyvinylalcohol(PVA)canprovideadvantageconditionsforengineeringapplicationofaerobicdenitrifyingbacte-ria.Keywords:simultaneousnitrificationanddenitrification(SND);nitrogenremoval;aerobicdenitrifying;amplificationculture;polyvinylalcohol(PVA)immobilization　　,。、、、。,,,[1-3]。,30%[1]。。[4-5],。,,(BTB)pH,DOI:10.14068/j.ceia.2008.03.004(SND)OGO,16SrDNA,Rhodococcus,T7。T7,,,。1　1.1　DM(g·L-1):5.0g;KNO32.0g;K2HPO41.0g;KH2PO41.0g;MgSO40.2g;2ml;1000ml;2%;1%(BTB)1ml,pH7.0～7.2。(g·L-1):(ED-TA)50g;EnSO42.2g;CaCl25.5g;CuSO4·5H2O1.57g;MnCl2·4H2O5.06g;FeSO4·7H2O5.0g;COCl2·6H2O1.61g;pH7.0。1.2　、KNO3(NaNO2)、K2HPO4、KH2PO4、MgSO4、。1。1　　　　　CODcrHACH-COD()NH+4-NNO-2-NN—(1-)—NO-3-NTN—DOYS15100DOOD600nm　　＊:用铬酸盐标准法GB11914进行校正2　2.1　T7T7DM,。30℃5d,,(O2,Ar),。()NO-3-NNO-2-N。,3。2.2　T7T724h5%BTBDM,30℃、120r/min96h,3h1,600nm,。3。2.3　、T7、,。:(0.2%),、、、、、;:(0.5%),、、、、、。:(0.5%),(0.2%),1、5、10、15、20、256。5%,30℃,120r/min24h,600nm,。2.4　T7T75%,4000r/min25min,,。PVA1080℃,,30℃。PVA,,,,,11.5℃24h。T710L,[6]。,。:30℃,DO(5.0±0.5)mg·L-1,pH7.0～7.2,C/N5。2.5　2.5.1　形态学鉴定T7。2.5.2　16SrDNA同源性比较[7]DNA,16SrDNA:8F:AGAGTTTGATC-CTGGCTCAG(E.coli16SrRNA8-253:T727),1495R:CTACGGCTACCTT-GTTACGA(E.coli16SrRNA1514-1495),16SrDNA。PCR,,341。3　3.1　T7T75d133.00mg/L50.47mg/L,62.05%,0.6mg/L。1。1　T73.2　T7T72,,T74:,,,。12h,24h,,72h。,。,,,,。3.3　、、T7、、DM2。2,:T7,,、,。:T7,、,2　T7,,。:T75,5。,,Rhodococcussp.[8]。、、5。2　T7、、OD600OD600C/NOD6001.0660.33210.2140.0831.31550.4120.4130.441100.3441.2590.181150.1761.2030.298200.1030.1781.235250.1553.4　3　T7NO-3-N/(mg·L-1)PVA/(mg·L-1)/%/(mg·L-1)/%10.08691.400.10889.20101.81281.882.17178.2910040.3659.6450.0649.94500420.1815.96450.759.851000939.206.08970.922.914　T7NO-2-N/(mg·L-1)PVA/(mg·L-1)/%/(mg·L-1)/%10.09490.600.11588.50103.61663.845.29147.0910089.7210.2895.394.61500473.285.34492.031.591000979.082.09991.500.8526　　　　　　　　　　　　　　　　　　　　　　　　　　　　　13、4:。1mg·L-1,,85%。,,,PVA。,,。3.5　3.5.1　形态学鉴定T7、,BTBDM。3　T7(×100)3.5.2　16SrDNA同源性比较T7PCR、16SrDNA,NCBI()BLAST,:T7Rhodococcus99%,T7Rhodococcus。4　4.1　OGO,T7。、,5。16SrDNAT7Rhodococcus,Rhodococcussp.。4.2　T7,,,。4.3　T7,PVA,。:[1]　,,　.[J].,2002,21(9):527-529.[2]　LukowT,DiekmannH.Aerobicdenitrificationbyanewlyi-solatedheterotrophicbacteriumstrainT[J].BiotechnologyLetters,1997,11(9):1157-1159.[3]　OtaniY,HasegawaK,HanakiK.Comparisonofaerobicde-nitrifyingactivityamongthreeculturalspecieswithvariouscarbonsources[J].WaterSci.Technol,2004,50(8):15-22.[4]　FretteL,GejlsbjergB,WestermannP.Aerobicdenitrifiersi-solatedfromanaltermatingactivatedsludgesystem[J].FEMSMicrobiologyEcology,1997,24:363-370.[5]　PaiSL,ChongNM,ChenCH.Potentialapplicationsofaer-obicdenitrifyingbacteriabioagensinwastewatertreatment[J].BioresourceTechnology,1999,68:179-185.[6]　.[J].,1998,18(2):189-191.[7]　SambrookJ,RussellDW.Molecularcloning:alaboratorymanual3rded[M].3rdedition.NewYork:ColdSpringHar-borLaboratoryPress,2001:27-30.[8]　,,,.[J].(),2004,25(1):75-78.(23)　　。,。:[1]　.GB7479—87[S].[2]　.《》[M].4.:,2002.[3]　.[M].2.:,2005.273:T7