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(BYS1),,,,3(,210095):BYS120h1gL-195%,0314molL-1NaClGibbons16SrDNA(GenBankAccessionNo1AY0662217),(Halomonassp1),pHNaCl715,30110molL-1:;;:S15614:A:10002030(2003)02005504Onephenylaceticacid2degradingmoderatehalophilicbacterium(BYS1)isolatedfromsludgeHONGQing,HEJian,LIUZhi,ZHANGGuo2shun,LIShun2peng3(KeyLaboratoryofMicrobiologicalEngineeringAgriculturalEnvironment,MinistryofAgriculture,NanjingAgricUniv,Nanjing210095,China)Abstract:Onebacteriumstrainwasisolatedfromtheactivatedsludgeproducedinthetreatmentofwastewaterfromonephenylaceticacidmanufacturer.ItcouldusephenylaceticacidassolecarbonsourceandgrowunderNaClconcentrationof0314molL-1inGibbonsmedium.Itsdegradingratewashigherthan95%withtheinitialphenylaceticacidconcentrationbeing1gL-1.ItwasidentifiedasHalomonassp1accordingtoitsphysiologicalandbiochemicalcharactersandanalysisofits16SrDNAse2quence(GenBankAccessionNo1AY0662217).ItsbestdegradingefficiencywasachievedwithpH,temperatureandNaClbeing712,30and110molL-1,respectively.Keywords:phenylaceticacid;degradation;moderatehalophilicbacterium,(xenobiotics),,,,,,,[1]Woolard[2](GreatSaltLake)NaCl,NaCl310molL-1,BYS1,1111:20020918:863(20011AA214121);(,:2000121):(1970),3Correspondingauthor:(1942),,Tel:02543963142003,26(2):5558JournalofNanjingAgriculturalUniversity112Gibbons[3](gL-1):5,10,5,3,KCl2,MgSO47H2O20,NaCl;pH7(gL-1):M63[4],KH2PO41316,KOH412,(NH4)2SO42,Fe2SO4,MgSO47H2O0125,1,NaCl;pH711310g,1gL-1,,30,180rmin-1;3d10%,NaCl210molL-1,NaCl210molL-1,,114,Na2SO4,25818nm,,11511511[5]1151216SrDNA[6]SDS,(CTAB),BYS1DNA[3]BYS116SrDNA(fD1):5CCGAATTCGTCGACAACAGAGTTTGATCCTGGCTAG3,(rP2):5CCCGGGATCCAAGCTTACGGCTACCTTGTTACGACTT3;BYS1DNA,50L:1L,dNTP(215mmolL-1)4L,(1mmolL-1)1L,10Buffer5L,Mg2+(210mmolL-1)3L,Taq(5UL-1)015L,3514LPCR:955min;941min,521min,723min,30;7210minPCR,BamHIEcoRI;pUC19,DH5,;,,(116kb);TAKARABLASTGenBank16SrDNA2211,,11,,BYS1212BYS1(1gL-1),5%BYS1,4h,1,25818nm,4h,16h,20h1BYS1Fig11DegradingofphenylaceticacidbyBYS12BYS1NaClFig12GrowthofBYS1underdifferentNaClconcentrations6526213NaClGibbonsNaCl,24h,,5000rmin-110min,,OD6102,0314molL-1NaCl,,NaCl110molL-1214,pHNaCl,20h,,BYS1,2035,85%,30100%pH,pH510,pH54518%,pH105816%,pH715,100%NaCl,016112molL-185%,110molL-1100%3BYS1(1000)Fig13ThemorphologyofBYS14BYS1Fig14ThecolonyofBYS121521511,(3),(018110)(115215)m,,,,,,(4),,,,,NO-3,,0314molL-1NaClGibbons2151216SrDNABYS1DNA,16SrD2NAPCR,116kb(5)TAKARAGenBank,AY06622171,BYS116SrDNA16SrDNA,,16SrDNA95%,[7],,,(Halomonassp1)[8]5BYS116SrDNAPCRFig15Electrophoresismapofcloningprocedureof16SrDNAfromBYS1M:DNAmarkerDL15000;A:TotalchromosomeDNAofBYS1;B:pUC19;C:pUC19ligatedwith16SrDNAfragment;D:pUC19+16SrDNAfragmentdigestedwithBamHIandEcoRI;E:16SrDNAfragment752:(BYS1)1BYS116SrDNABLASTTable1BLASTanalysisof16SrDNAsequencesshowingtheclassificationofstrainBYS1BacteriumforalignmentGenBankAccessionNo1IdentityHalomonasvenustaAJ306894111495/1498(99180%)HalomonasaquamarinaAJ306888111458/1498(97133%)HalomonasmeridianaAJ306891111457/1498(97126%)Halomonassp.MBIC1142AB022662111436/1475(97136%)Halomonassp.MIBHP1H25AB016053111450/1499(96173%)Halomonassp.NIBHP1H22AB016052111454/1504(96168%)Halomonassp.NIBHP1H14AB016051111452/1503(96161%)Halomonassp.NIBHP1H13AB016051111423/1477(96134%):[1]GalinskiEA.Osmoadaptioninbacteria[J].AdvMicroPhysiol,1995,37:273328.[2]WoolardCR,IrvineRL.Biologicaltreatmentofhypersalinewastewaterbyabiofilmofhalophilicbacteria[C].AbstractsoftheAnnualWaterEnvironmentalFederationConference,NewYork,1992.[3]SegalSN,GibbonsNE.EffectofsomemetalionsonthegrowthofHalobacteriumcuirubrum[J].CanJMicrobiol,1960,6:165169.[4]LarsenPL,SydneLK,Landfald,etal.OsmoregulationinEscherichiacolibyaccumulationoforganicosmolytes:betaines,glutamicacid,andtrehalose[J].ArchMicrobiol,1987,147:17.[5],.[M].:,20011370410.[6]F,R,RE.[M].,.:,199913940.[7].[M].:,1997.17.[8]HoltJG,KriegNR,PeterHA.BergeysofDeterminativeBacteriology[M].9thed.Baltimore:TheWilliamsandWilkinsCompa2ny,1993184,142.:8526