好氧同时硝化反硝化脱氮微生物的混合培养堵国成

　第21卷第5期2002年9月　　　　　　　无锡轻工大学学报JournalofWuxiUniversityofLightIndustry　　　　　　Vol.21　No.5Sep.　2002　:1009-038X(2002)05-0515-04　　:2002-05-02;　:2002-06-26.:(1965-),,,,.好氧同时硝化—反硝化脱氮微生物的混合培养堵国成,　耿金菊,　陈坚(江南大学工业生物技术教育部重点实验室,江苏无锡214036)　:筛选和分离得到的多株脱氮微生物,能在完全好氧条件下将氨氮转化为NO2-,随即在好氧反硝化菌的作用下还原为N2排放,整个生物脱氮过程历时较短,30h内对200mg/L的氨氮去除率达99%,而且无中间产物NO2-的积累.混合脱氮微生物菌群生长的适宜pH范围为7～10,探索实现混合脱氮微生物菌群高密度培养的pH:发酵前期补酸控制pH≤8,发酵中后期不控制pH值,可缩短菌体的生长周期,提高菌体的氨氮降解速率,细胞质量浓度达3.9g/L,比自然pH条件下提高了62.5%.:脱氮微生物;硝化—反硝化;好氧脱氮;混合培养:Q939:AMixedCultureofDenitrifyingBacteriaforAerobicSimultaneousNitrification-DenitrificationDUGuo-cheng,　GENGJin-ju,　CHENJian(KeyLabofIndustrialBiotechnology,MinistryofEducation,SouthernYangtzeUniversity,Wuxi214036,China)Abstract:NH4+wasconvertedtoNO2-underaerobicconditionsbythescreenedbacteria,theintermediateNO2-wasthenreducedtoN2byaerobicdenitrifyingbacteria,andnoNO2-wasaccumulated.200mg/LofNH4+canberemovedwithin30hwitharemovalrateof99%.TheappropriatepHforthegrowthofmixedbacteriawas7～10.pHcontrolpolicyforthehighdensitymixedculturein5Lfermentorwasproposed.BycontrollingpHvaluelowerthan8intheearlystageandwithoutpHcontrollingafterwards,cellgrowthperiodwasshortenedandNH4+removalratecouldbeimproved,drycellweightreached3.9g/L,increased62.5%higherthanthatundernaturalpHconditions.Keywords:denitrifyingbacteria;nitrification-denitrification;aerobicdenitrifying;mixedculture　　,.,,.,,2080,—.Pseudomonasspp,Al-caligenusfaecalisThiosphaerapantotropha[1～3],(NO2-)(NO3-)(N2).,,,:(1),NO2-NO3-,—;(2),OH-,pH;(3),,.,—.pH—.,,pH,pH,.,5LpH,pH,pH,.1　材料与方法1.1　:(Aerobicdenitrify-ingBacteria),.(g/L):NaHCO31.75,CH3COONa0.854,(NH4)2SO41.0,NaCl1.0,FeSO40.4,K2HPO41.0,MgSO40.25.NaNO2NaNO3.1.2　:,50mL500mL,30℃,200r/min.:5L(KF-5L)3L,10%,30℃,400r/min,1L/(L·min),pH.1.3　:[4].:5mL11000r/min10min,,0.1mol/LHCl,105℃,(DCW).:-N,N-[5].:[4].2　结果与讨论2.1　,(NH4+—N)、(NO2-—N(NO3-—N).NO2-—NNO3-—N,NH4+—N,NO2-—NNO3-—N,.,,,,.2.2　NO2-NO3-,,,—,,,,.,,.NH4+、NO2-NO3-,、NH4+—N(r)、NO2-—NNO3-—N1(a,b,cd).(a)516　　　　　　　　　　　　　　　无　锡　轻　工　大　学　学　报　　　　　　　　　　　第21卷(b)NH4+—N(c)NO2-—N1　NH4+、NO2-NO3-Fig.1　ParameterchangeswiththeadditionofNH4+、NO2-andNO3-　　1(d),NO3-,NO3-,NO3-;NH4+—N,NO3-.2(a),.1(c),NO2-,NO2-,12hNO2-0.05g/L0.003g/L,()1.7g/L,NO2-,[6].NO2-NH4+,NH4+—N,NO2-,30hNO2-100%.NO2-—N,,,[7].NO2-,NH4+—N,NO2-—NNO3-—N,—.NH4+、NO2-NO3-,(),—(Aerobicshortcutsimultaneousnitrification—denitrification),NH4+—NNO2-—N,,NO2-—NN2.2:2　Fig.2　Sketchofbio-denitrificationprocess　　,,,[8]:(1)25%,;(2)40%,;(3);(4),NO2-NO3-63%.—517第5期堵国成等:好氧同时硝化—反硝化脱氮微生物的混合培养.,.2.3　5LpH5LpH=7(pH),pH=8(pH),pH≤7pH≤8pH.3(ab),pH,pH,.(a)(b)3　5LpHFig.3　TimecoursesofDCWandtheremovalrateofammonia-nitrogenunderdifferentpHconditionsin5Lfermentor　　3(a)3(b),,,pH.pH,pH(4).pH8,,20h,pH10h.3.9g/L,62.5%,.pH≤7,,pH,pHpH,,.—,pH,pH78,.,pH≤8,.4　5LpHFig.4　TimecourseofpHchangesin5Lfermentor3　结　论(),,NH4+—NNO2-—N,,NO2-—NN2.pH≤8,pH,,,3.9g/L,pH62.5%.:[1]ROBERTSONLA,VANNIELEWJ,TORREMANSRAM.Simultaneousnitrificationanddenitrificationinaerobicchemo-statculturesofThiosphaerapantotropha[J].Applied&EnvironmentalMicrobiology,1988,54(1):2812-2818.(下转第532页)518　　　　　　　　　　　　　　　无　锡　轻　工　大　学　学　报　　　　　　　　　　　第21卷2.4　SBAGlu6,SBAGlu125,200,SBA.7,SBAGlu,125,100%,SBA,.7　SBAGluTab.7　ThetestofcalibratingvalueafteremendationinthedeterminationofGlufromthedesaltingliquorusingSBAmethodNo.SBA/u1/(μg/L)SBA/u2/%172.039.3111.4100.3266.849.1116.0100.0　　:SBA,100,,NH4+,SBA.3　结　论SBA[4],NH4+,;91%,80%～83%,SBA120～125,L-.,:108～110,、、、120～125.:[1].[M].:,1995.570-587.[2].[M].:,1999.427.[3]SMITHTJ,MATHIASLJ.Newmethacrylatederivativesbasedonpyroglutamicacid[J].PolymerPreprints,2000,41(2):1254-1255.[4].———[A].[C].:,2001.79-83.(责任编辑:杨萌)(上接第518页)[2]OTTES,SCHALKJ,KUENENJG.HydroxylamineoxidationandsubsequentnitrousoxideproductionbytheheterotrophicammoniaoxidizerAlcaligenesfaecalis[J].AppliedMicrobiology&Biotechnology,1999,51(2):255-261.[3]GUPTAAB.Thiosphaerapantotropha:Asulphurbacteriumcapableofsimultaneousheterotrophicnitrificationandaerobicdenitrification[J].Enzyme&MicrobialTechnology,1997,21(8):589-595.[4]《》.()[M].:,1989.[5],,.—N,N—.[J],1992,11(3):75-77.[6]YOOH,AHNKH,LEEHJ,etal.Nitrogenremovalfromsyntheticwastewaterbysimultaneousnitrificationanddenitrifica-tion(SND)vianitriteinanintermittently-aeratedreactor[J].WaterResearch,1999,33(1):145-154.[7]MULLEREB,STOUTHAMERAH,VANVERSEVELDHW.SimultaneousNH3oxidationandN2productionatreducedO2tensionsbysewagesludgesubculturedwithchemolithotrophicmedium[J].Biodegradation,1995,6(4):339-349.[8]JETTENMSM.Towardsamoresustainablemunicipalwastewatertreatmentsystem[J].WaterScienceTechnology,1997,35(9):171-180.(责任编辑:杨萌)532　　　　　　　　　　　　　　　无　锡　轻　工　大　学　学　报　　　　　　　　　　　第21卷