苯酚降解细菌的分离与选育研究

凌琪,汤利华,陶勇,王莉,丁媛媛(安徽建筑工业学院环境工程系,安徽合肥230022):采用焦化废水处理厂的活性污泥为菌源,以苯酚为唯一碳源进行苯酚降解细菌的分离与选育研究,结果从焦化废水处理厂的活性污泥中驯化筛选出了5株具有高效降解苯酚能力的菌株;通过生理生化反应及外观特征观察,对5株苯酚降解菌株的初步鉴定结果为:phen-8为产碱杆菌属(Alcaligenes),phen-9为微球菌属(Micrococcus),phen-24为芽孢杆菌属(Bacillus),phen-78phen-103为假单胞菌属(Pseudomonas);经物化诱变处理后得到1株生长更快并对苯酚具有更高降解率的诱变菌株(phen-24),该菌株在30e下,于pH值为7.0苯酚浓度为400mg/L的唯一碳源培养基中的生长状况较好,培养9h后苯酚几乎被完全降解:焦化废水;苯酚;生物降解;诱变;分离与选育:X703:C:1000-4602(2007)11-0078-05StudyonIsolationandScreeningofPheno-ldegradingBacteriaLINGQ,iTANGL-ihua,TAOYong,WANGL,iDINGYuan-yuan(DepartmentofEnvironmentalEngineering,AnhuiInstituteofArchitecture&Industry,Hefei230022,China)Abstract:Thestudyonisolationandscreeningofpheno-ldegradingbacteriawasconductedusingtheactivatedsludgeofacokingwastewatertreatmentplantasbacteriasourceandphenolassinglecarbonsource.Fivekindsofstrainswithhighdegradationabilitiesofphenolwereacclimatedandscreenedoutfromtheactivatedsludgeofthecokewastewatertreatmentplan.tThephysiologica,lbiochemicalandap-pearanceidentificationsofthefivestrainsshowthatphen-8belongstoAlcaligenes,phen-9toMicrococcus,phen-24toBacillus,phen-78andphen-103toPseudomonas.Thefiveoriginalstrainsarerepeatedlytrea-tedwithUVandNTGmutagenesis,whichscreensoutphen-24mutantstrain,withthehighestpheno-lde-gradingability.Thebestgrowthconditionsforthephen-24mutantstrainaretemperatureof30e,pHof7.0,phenolconcentrationof400mg/L.Within9hours,phenolisalmostcompletelydegraded.Keywords:cokingwastewater;pheno;lbiodegradation;mutagenesis;isolationandscreening:(2006KJ108B);/0(01013024);(07021012)[12],5~25mg/L[3][4],,[56]#78#第23卷第11期2007年6月中国给水排水CHINAWATER&WASTEWATERVo.l23No.11Jun.2007,,,,,phen-24,,1材料与方法11111111,11112¹,15W,253.7nmº(NTG)NTG:NTG,))),1B9,NTG1mg/mL100Lg/mL11113¹:5g/L,10g/L,5g/L,18g/L,pH7.4~7.6º:5g/L,10g/L,5g/L,20g/L,400mg/L»:5g/L,5g/L,1g/L,20g/L,K2HPO41g/L,pH7.4~7.6¼:NH4NO31.0g/L,KH2PO40.5g/L,K2HPO40.5g/L,MgSO4#7H2O0.2g/L,CaCl20.1g/L,NaCl0.2g/LMnSO4#H2O10%FeCl2,2004006008001000mg/L1121121124h,5mL101001000,0.1mL,30e2d,,,15,24h4e11212,0.1mL,30e2d2004006008001000mg/L15mL(100mL),30e200r/min24h,,511213¹3mL6cm,,,30cm15W253.7nm2min10~106,0.1mL,,30e2d,,5~6,ºNTG100Lg/mLNTG(15mL),30e30min,,2,0.1mL,30e2d,0.1mL(),30e2d,1121410mm,,600nm(OD600),#79#凌琪,等:苯酚降解细菌的分离与选育研究第23卷第11期112154-,()7500r/min10min,100LL10mL,5mL0.5mol/L100LL,pH6.8pH7.2,50LL2%4-,50LL,15min,510nm11216,[7]2结果与讨论211,5,11Tab.1Resultsofphysiologicalandbiochemicaltestforscreenedstrainsphen-8phen-9phen-24phen-78phen-103G-G+G+G-G--++++--+-------+--+++-+--++/----+--++-----+----+-++++++++(Alcaligenes)(Micrococcus)(Bacillus)(Pseudomonas)(Pseudomonas)2125,22Tab.2Phenoldegradationratebeforeandaftermutagenesisofstrains%200mg/L400mg/L600mg/L800mg/L1000mg/Lphen-871.374.674.475.977.479.534.435.510.411.5phen-975.388.285.189.129.934.25.96.20.91.2phen-2480.010085.510032.945.57.09.30.62.1phen-7872.277.475.177.634.637.46.66.80.61.4phen-10368.872.175.480.531.738.67.78.61.71.92,,,,phen-24213phen-2421311phen-24pH7.0400mg/L,200r/min24h,(OD600),11,25~35e,phen-24,30e,OD6000.679,#80#第23卷第11期中国给水排水~7.582.5%;pH4.0,OD6000.202,17.4%pH,phen-24,pH11.0,OD6000.019,,pH,phen-24,2131330epH7.0,phen-2424h,OD600,33,phen-24,400mg/L,OD600,100%,;400mg/L,,;800mg/L,,3,500mg/L,,24h95%,1000mg/L;1000mg/L,,(400mg/L),phen-243phen-24Fig.3Effectofphenolconcentrationongrowthanddegradationcapacityofphen-24mutantstrain21314phen-242%pH7.0400mg/L,30e200r/min24h,OD600,44phen-24Fig.4Phenoldegradationcurveandgrowthcurveofphen-24mutantstrain4,phen-246h,;6~9h,OD600#81#凌琪,等:苯酚降解细菌的分离与选育研究第23卷第11期,phen-24;9~10h,phen-24;,4h,,,9h100%11h,(),phen-24,OD600,phen-243结论¹15,5,phen-8(Al-caligenes),phen-9(Micrococcus),phen-24(Bacillus),phen-78phen-103(Pseudomonas)º5,1(phen-24)»phen-24pH7.0400mg/L,30e9h,:[1]WatanabeK,TeramotoM,HarayamaS.Anoutbreakofnon-flocculatingcatabolicpopulationscausedthebreak-downofapheno-ldigestingactivated-sludgeprocess[J].ApplEnvironMicrobio,l1999,65(7):2813-2819.[2]PaulaM,vanSchie,YoungLY.Isolationandcharacter-izationofpheno-ldegradingdenitrifyingbacteria[J].ApplEnvironMicrobio,l1998,64(7):2432-2438.[3]ChungTP,TsengHY,JuangRS.Masstransfereffectandintermediatedetectionforphenoldegradationinim-mobilizedPseudomonasputidasystems[J].ProcessBio-chem,2003,38(10):1497-1507.[4],,.[J].,2001,83(2):36-37.[5]JeongJJ,KimJH,KimCK,etal.3-and4-alkylphenoldegradationpathwayinPseudomonassp.strainKL28:ge-neticorganizationofthelapgeneclusterandsubstratespecificitiesofphenolhydroxylaseandcatechol2,3-diox-ygenase[J].Microbiology,2003,149(11):3265-3277.[6]TeramotoM,OhnishiK,HarayamaS,etal.AnAraC/Xy-lSfamilymemberatahighlevelinahierarchyofregula-torsforpheno-lmetabolizingenzymesinComamonastes-tosteroniR5[J].JBacterio,l2002,184(14):3941-3946.[7],.[M].:,2001.:13955105084E-mail:[e-mail]:2006-12-08#82#第23卷第11期中国给水排水