1株高效产絮凝剂菌株的分离及其在冷轧含油废水处理中的应用雷国元

32920119ENVIRONMENTALSCIENCEVol．32No．9Sep．201114300811T-3、16SrDNAKlebsiellasp．．pH、、、．、pH925℃．、、Fourier—OH—COO－．．CaCl2、、4g·L－1、10%1g·L－1pH7、COD4819mg·L－1、28456.8mg·L－1、395010mg·L－1、218.4mg·L－1、1.3699.79%、92.32%99.97%．、Ca2+Ca2+．X703．1A0250-3301201109-2716-082010-09-292011-01-21505780511964～、[e-mail]tBio-flocculant-producingStrainandItsApplicationintheTreatmentofCold-rollingWasteOilyWaterLEIGuo-yuanDINGCui-pingYANGJia-xuanHubeiKeyLaboratoryforEfficientUtilizationandAgglomerationofMetallurgicMineralResourcesWuhanUniversityofScienceandTechnologyWuhan430081ChinaAbstractAnexcellentstraindesignatedasT-3whichproducesbio-flocculantswasisolatedfromsoilsamplesandidentifiedasKlebsiellasp．speciesbasedontheanalysisofmorphologyphysiologyandbiochemistryand16SrDNAsequencesmeasurement．TheeffectsofcultureconditionssuchaspHvaluestemperaturecarbonsourcesandnitrogensourcesonbio-flocculantsproductionbyT-3strainwerestudied．TheexperimentresultsshowthatT-3strainhasbetteradaptabilitytocarbonsourcesandnitrogensourcesandhighercapacityofbio-flocculantswasobtainedwhentheinitialpHvalueofcultureandtemperaturewere9and25℃respectively．BasedonthecolorimetricreactionsofproteinsandpolysaccharidesubstanceultravioletscanninganalysisandFourierTransformInfraredSpectroscopyanalysisitisfoundthatthebio-flocculantsproducedbyT-3straincontains—OHand—COO－groupsandbelongstoanionictypeflocculant．Moreoverthemaincomponentispolysaccharides．Thetreatmentofoilycold-rollingwastewaterbythebio-flocculantwasinvestigatedandthebetterresultwasobtained．WhenthedosagesofCaCl2bio-flocculantsandpolyaluminiumchloridewere4g·L－110%volumefractionand1g·L－1respectivelyandthepHvaluewas7.0theoilconcentrationCODandturbidityweredecreasedto10mg·L－1218.4mg·L－1and1.36from4819mg·L－128456.8mg·L－1and3950withtheremovalefficienciesof99.79%92.32%and99.97%respectively．TheinteractionbetweenflocculantandoilydropletsisachievedbytheinteractionofVanderWaalsforcehydrogenbondandthebridgedcoordinationofCa2+inwhichthebridgedcoordinationofCa2+isthedominant．Keywordsbio-flocculantsoilywastewatercold-rollingmechanismwastewatertreatment1．2～7、8～10、．．MBF、、11．、、、．12～14DOI:10.13227/j.hjkx.2011.09.00891．、、1T-3、Zeta．11.1．16、18、20．．．COD20000～30000mg·L－11000～10000mg·L－11710～17400pH4～6MS2000G0.1～4.5μmd0.50.616μmZetaCDZeta－30mV．、．1.21.2.115、1T-3．20g、KH2PO42g、K2HPO45g、NH42SO40.2g、0.5g、0.5g、NaCl0.1g、MgSO4·7H2O0.2g、pH7.0、1000mL．250mL100mL3%25℃130r/min．1.2.2、1516、．1.2.316SrDNA16SrRNA27F5'-AGAGTTTGATCCTGGCTCAG-3'1492R5'-CGGTTACCTTGTTACGACTTC-3'PCR．50μL10×PCRbuffer5μLPfu1μLdNTPs10mmol/L2μL27F2μL1492R2μLDNA1μLddH2O37μL．PCR94℃5min3594℃30s56℃30s72℃2min72℃10min．PCR1.2%DNADNA．16SrDNABlastGenBank16SrDNAClustalWGenBank16SrDNAMega4.1NeighbourJoiningNJ16SrDNAbootstrap1000．1.31.3.1T-3、、、pH．1.3.2pH50mLCaCl2CaCl20.2gT-310mLpH5min．MBFT-350mLCaCl2CaCl20.2gpH7.05min．50mL10mLCaCl2pH7.05min．50mLCaCl2CaCl20.2gpH7.05min．1.41.4.1．0.5%2.5μm100mL0.6mL5717232min-550nmD550EaEa%=D550－D550D550×100%1Et．50mLCaCl2pH7.05min．EtEt%=－×100%21.4.2MBFT-315℃11000r/min10min35℃4000r/min10min．1.4.317．1.4.40.6%2min5minD550．1.4.5MBFT-3、Molish、、、MBFT-3FourierBRUKERVERTEX70．1.4.6、COD、18TDT-1．22.1、1T-395%．2.2T-3T-32～4μm．1．1T-3Table1IdentificationofstrainT-3－－+M．R．+－－－V．P．+2.3T-316SrDNAPCR1500bp16SrDNA1．1435bp．1T-316SrDNAFig．1ElectrophoresisofPCRproductsof16SrDNAgeneofstrainT-32.4T-316SrDNAGenBankBlast16SrDNAGenBankKlebsiellasp．16SrDNA99%Klebsiellasp．．216SrDNA．2.5T-3T-33．8172912T-3Fig．2PhylogentictreeofstrainT-33T3Fig．3GrowthcurveofT3strainandflocculatingrateduringthegrowthstage．．2.6T-3、、pH24．、、．pHpH9～10．pH9～10pH6～7．pHpH．．15、25、35℃94.36%、94.53%、87.12%2、T-3Table2EffectsofcarbonandnitrogensourcesonMBFT-3productionEa/%Ea/%96.20NH42SO493.6795.9793.5596.5596.32D-96.7895.5288.2696.094pHT-3Fig．4EffectoftheinitialpHvalueofcultureonflocculantsproductionbystrainT-325℃．2.7MBFT-32.7.1pHMBFT-3pHZeta、19．pH．MBFT-3pH5．pH5～7pHpH7～890%．2.7.2MBFT-3．6．．16%96.94%．2.7.3CaCl2MBFT-320．CaCl2CaCl27．9172325pHFig．5EffectofthepHvalueonthetreatmentofoilywastewaterbybio-flocculant6MBFT-3Fig．6EffectofMBFT-3dosageonthetreatmentofoilywastewater＜3.2g·L－1CaCl2＞3.2g·L－1100%CaCl24g·L－1．2.7.4MBFT-3MBFT-3MBFT-38．6%、0.6g·L－12010%1g·L－12.0．．2.7.5CaCl24g·L－17CaCl2Fig．7EffectofCaCl2dosageonthetreatmentofoilywastewater8MBFT-3Fig．8EffectofthecooperationofMBFT-3andPAConthetreatmentofoilywastewater10%1g·L－1pH7．3．3．3MBFT-3Table3ResultofoilywastewatertreatmentbyMBFT-3/%/mg·L－148191099.7939501.3699.97COD/mg·L－128456.8218.492.323MBFT-33.1MBFT-34．02729117．3.2、MBFT-3、5．MBFT-3/．4MBFT-3Table4HeatstabilityofMBFT-3/℃253545556580Ea/%92.9194.6495.5995.7996.9396.935MBFT-3、Table5ColorreactionofcarbohydrateandproteinonMBFT-3Molish3.3MBFT-3MBFT-3．9．、280nm260nm．．9MBFT-3Fig．9UltravioletspectrumofMBFT-33.4MBFT-3FouirFouir10．10．①3600～3400cm－13439cm－1—OHN—H21．②3000～2900cm－12929cm－1C—H22．③1720～1600cm－11640cm－1—COO－23．④1462cm－1—CH2—1392cm－1C—H2—CH321．⑤1307cm－1—COO－PO11．⑥1101cm－1C—O24．⑦859cm－1．⑧542cm－111．10MBFT-3FourierFig．10FouriertransactioninfraredspectraofMBFT-3MBFT-3．MBFT-3．4MBFT-3、、、Butter-fiel