抗水解液中抑制物的秸稈乙醇發(fā)酵菌劑構建及其功能研究.pdf

1、中國農業(yè)大學博士學位論文抗水解液中抑制物的秸稈乙醇發(fā)酵菌劑構建及其功能研究姓名：路鵬申請學位級別：博士專業(yè)：生態(tài)學指導教師：李國學20070601：雪查些鑾主曼圭蘭蘭笙蘭竺：：圣蘭：ABSTRACTChinabecomingpureoilimportcountryfrom1993andisalargefossilfuelconsumptionareaoftheworldTothecountrydevelopmentinthefuture

2、，energyproblemisoneimportantinhibitingfactorAndenergysecuritypolicybecomingbigproblemtoChinagovernmentUselignioceIlulosicmaterialtoproduceethanolasreproducibleenergyfromfermentationmethodbymicrobialstrainsmayrevivethepro

3、blemNaturalcellulosemateriolisonekindofmacromoleculesubstancewhichhasundisslovehighpolymerizationcharactersandnoteasytodecomposedbymicrobialorganism，Howtoimprovethehydrolyzingrateofcellulosematerialandethanolconversingra

4、tefromhydrolyzatebymicrobialorganismismainobiectionofthisresearchThisresearchanalyzedhydrolyzingresultsandregulationoforiginalcellulosematerialbyseveralpretreatmentmethodsusingsinglefactordesignexperimentsAndthenuseortho

5、gonalexperimenttoimprovinghydrolyzingefficencyandacquiredtheoptimummodeofdilutedacidpretreatmentmethodComparedtheresultofalkaljpretreatmentanddilutedacidpretreatmentexperiment，wefindthealkalipretreatmentathightemperature

6、(100。C、maydecreasedthereducingsugarobviouslyAlkalipretreatmentisunfitfororiginalcellulosematerialpretreatmentalonecomparedwithdilutedacidpretreatmentmethodWeselectandconstructedanewethanolfelillentationmicrobialmixturein

7、severalstepsFirstlyweselectonegroupofmicrobialorganismsystemtofermentedhydrolysatetoethanoldirectlyThismicrobialsystemcanconvertthetomplicatedhydrolyzatetoethanol，glycerolandseveralothermetabolitesbuttheethanolconcentrat

8、ionistwo10wtonextextractionstepSecondlyweselectandacqukedpurecultureofthe4mainfermentationmicrobialstrainsinthemicrobialorganismsystemThirdlyusethe4sirainsandoneTrichodermakon證UOudemstraincombineonenewmicrobialstrainsgro

9、upbyorthogonalexperimentThemicrobialorganismgrouphashigherethanolconverteffectcomparedwiththeoldmicrobia】organismsystemEthanolcoucentIationinthefermentationsolutionreachedto1219￡／LanditCaBalsoenduretheinhibitorsinthehydr

10、olyzateunder10％solidcontent，05％sulfuricacidconcentrationand10礦C120。CpretleatmenttemperaturepretreatmentconditinnsandneedupdetoxificationMicrobialstrainSaccharomyceskluyveriinthemicrobialsystemhasprominentresistanceeffect

11、tofuffuraJinhibitorButitnothasobviouslyeffecttesistaceticacidinfermentationprocessTheaceticacidresistanceofmicrobialstraindropprominentlywhentheoriginalsugarconcentrationabove7％Ethan01productionprominentlydecreasedwhenac

12、eticacidconcentrationabove2mg／rMBut，whenfermentationatlowerpriginalsugarconcentration，ethanolproductiondroppeddownprominentlyataceticacidconcentrationexceeded4mg／m1ToexploringtotheresistancemechanismofSaccharomyceskluyve

13、ristraintofuffuralinhibitorweculturedthestrainunderseveraldifferentfufloralconcentrationsconditionfor18handdetecttheproteinspotchangeonthe2DPAGECompared453proteinspotswhichdistinguishedfrom2D—PAGE，wefindithas26upregulate

14、dproteinspots，19down—regulatedproteinspotsand4inducedproteinspotsFinally7proteinsidentifiedbyMALDITOFMSmethodinthismicrobialstrainThisthesisexploreethanolfermentationpasswayuseorigina】cellulosematerialbyresearchthestrawp

15、retreatmentprocessandhydrolyzatefermentationconditions(includingconstructthemicrobialorganismsystemmicrobialstainsgroupoptimizingandmetabolizeproductionanalyzing)wefindthe4inducedproteinsunderfurfuralinhibitorintimidated

16、conditionsbyusingproteomicsmethod，Itprovideconveniencetoanalyzinginhibitorresistancemechanismofmicrobialstrain，andalsoprovideconveniencetoidentifytheresistingeontr01geneandrecombinethemicrobialstrainKeywords：COITtstalk，p