无溶剂系统中固定化脂肪酶催化废油脂转酯生产生物柴油

:02539837(2004)11090306:903~908:20040225.:,,1971,.:.Tel:[phone];Email:[e-mail]:(2003C33102).吴虹,宗敏华,娄文勇(华南理工大学生物工程系,广东广州510640):Novozym435.,./1,66U/g,35~40150r/min,..614h,/1,30h886%.300h,.:,,,,,:O643/Q55:ATransesterificationofWasteOiltoBiodieselinSolventFreeSystemCatalyzedbyImmobilizedLipaseWUHong,ZONGMinhua*,LOUWenyong(DepartmentofBiotechnology,SouthChinaUniversityofTechnology,Guangzhou510640,Guangdong,China)Abstract:ThetransesterificationofwasteoiltobiodieselinasolventfreesystemcatalyzedbyanimmobilizedlipaseNovozym435wasexplored.Thebyproductglycerolwasfoundtobeadsorbedonthesupportandthestabilityoftheimmobilizedlipasewasenhancedbyrinsingtheenzymewithacetone.Theoptimalreactionconditionssuchasmolarratioofmethanoltooil,enzymeamount,temperatureandshakingratewere1,66U/g,35~40and150r/min,respectively.Wateradditionwasnotfavorableforthereaction.Athreestepmethanolysisprotocolwasusedtoprotecttheimmobilizedlipasefrominactivationbymethanol.Onemolarequivalentofmethanolwassuccessivelyaddedatreactiontimeof0,6and14h,bywhichamethylestercontentof886%inthereactionmixturecouldbeachievedaftertotalreactiontimeof30h.Acetonewasusedtoremoveglycerolfromtheenzymesurfaceateachstep.Theenzymeshowednoappreciablelossinactivityafterbeingcontinuouslyoperatedfor300h.Keywords:lipase,immobilizedenzyme,wasteoil,transesterification,biodiesel,solventfreesystem,,.,[1].,,.,[2],,(75%)..2511200411Vol.25No.11ChineseJournalofCatalysisNovember2004,,,.,[3~9].,,,!.,,,!.,,,.,.,,Shimada[6,7],..,.Shimada[7],Novozym435,.,,,,.,[9,10],.Dossat[10],RhizomucormeiheiLipozyme∀R,.,;,,,,.LipozymeTLIM()..Shimada[7]Novozym435,,,,.11.1Novozym435Candidaantarctica,,164U/g(U1min1mol),NovoNordiskIndustries.,Sigma...1.2,,,.GB/T173761998,GB553485,GB/T55301998GB552885.,.19%(36),500mg/kg,2003,8402.1.350ml10g,,.01ml,10000r/min10min,,,.1.4HP4890D(3398A,)HP5(053mm#15m).:250,280,1801min,08/min186,1min,20/min280;,25ml/min;1∃100;1l.,4223,904256072,7636,76808386min.1%.22.1,1mol3mol,.,.,,2448h10g,72h,882%.,,,.4,754%.1Fig1InfraredspectraofthesampleobtainedbywashingtheimmobilizedlipaseNovozym435withwateraftertransesterificationofwasteoil(1)andglycerol(2),,,,,,276g.(1(1)),(1(2)).,1463cm-1,,(10%).Shimada[7].,[1,9,10],,.,,4,884%..2/Fig2Effectofmethanol/oilmolarratioontheenzymatictransesterificationofwasteoil(Reactionconditions:10g(00119mol)wasteoil,66UNovozym435,35,150r/min,12h.Theconversionisexpressedasthepercentageofmethanolconsumedfortheformationofesterwhenthemolarratioofmethanol/oilislessthan3,andastheratioofmethylestertooilwhenthemolarratioofmethanol/oilismorethan3.)2.2//(2).(/1),12h95%,/;/1~15,12h/;/15,12h/.,,,,90511:!.,/23/1,12h133%36%.,/15.[7,11,12],/1.2.3,,,,,,[13].3.,,.66U,.33U66U,24h5h;66U,.,24h,.66U,5h298%.,66U.3Fig3Effectoflipaseamountontheenzymatictransesterificationofwasteoil(Reactionconditions:10gwasteoil,038gmethanol,35,150r/min.)2.4.,,,,.,.,.4,,Novozym435,.4Fig4EffectofwatercontentontheenzymatictransesterificationofwasteoilReactiontime:(1)1h,(2)5h(Reactionconditions:10gwasteoil,038gmethanol,66UNovozym435,35,150r/min.)2.5,,,.,,,;,,,.5,,,50~60,,.,,30%.Shimada[11].,35~40.906255Fig5EffectoftemperatureontheenzymatictransesterificationofwasteoilReactiontime:(1)1h,(2)5h,(3)12h(Reactionconditions:10gwasteoil,038gmethanol,66UNovozym435,150r/min.)2.6,,.6,150r/min,;150r/min,,.,150r/min.6Fig6EffectofshakingrateontheenzymatictransesterificationofwasteoilReactiontime:(1)1h,(2)5h(Reactionconditions:10gwasteoil,038gmethanol,66UNovozym435,35.)2.77Novozym435..1;6h,90%,,mol;14h90%,,,16h.30h,,886%.7Fig7Timecourseoftheenzymatictransesterificationofwasteoil(Reactionconditions:10gwasteoil,038gmethanol,66UNovozym435,35,150r/min.Atreactiontimeof6and14h,theenzymewasrinsedwithacetoneandthen038gmethanolwasadded,respectively.)8Novozym435Fig8OperationalstabilityofNovozym435inthetransesterificationofwasteoil(Reactionconditions:10gwasteoil,038gmethanol,66UNovozym435,35,150r/min.Atreactiontimeof6and14h,theenzymewasrinsedwithacetoneandthen038gmethanolwasadded,respectively.Thethreestepsservedasonebatchwithtotalreactiontimebeingof30h.)90711:Novozym435,8.,Novozym43510(300h),.1SoumanouMM,BornscheuerUT.EnzymeMicrobTechnol,2003,33(1):972,,,,.(TanTW,WangF,DengL,XuJL,WangLJ.ModernChemInd),2002,22(2):43NelsonLA,FogliaTA,MarmerWN.JAmOilChemSoc,1996,73(9):11914FukudaH,KondoA,NodaH.JBiosciBioeng,2001,92(5):4055KseO,TterM,AksoyHA.BioresourTechnol,2002,83(2):1256WatanabeY,ShimadaY,SugiharaA,TominagaY.JAmOilChemSoc,2001,78(7):7037ShimadaY,WatanabeY,SugiharaA,TominagaY.JMolCatalB,2002,17(35):1338LaraPV,ParkEY.EnzymeMicrobTechnol,2004,34(34):2709DuW,XuYY,LiuDH.BiotechnolApplBiochem,2003,38(2):10310DossatV,CombesD,MartyA.EnzymeMicrobTechnol,1999,25(35):19411ShimadaY,WatanabeY,SamukawaT,SugiharaA,NodaH,FukudaH,TominagaY.JAmOilChemSoc,1999,76(7):78912KaiedaM,SamukawaT,KondoA,FukudaH.JBiosciBioeng,2001,91(1):1213,,,,.(LiuSL,ZongMH,YuSW,ZhangXY,XingM.ChinJCatal),2003,24(1):7(EdLYX)90825