Rapid optimization of enzyme mixtures for deconstruction of diverse pretreatmentbiomass feedstock combinations 英文参考文献.docVIP

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Rapid optimization of enzyme mixtures for deconstruction of diverse pretreatmentbiomass feedstock combinations 英文参考文献.doc

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Rapid optimization of enzyme mixtures for deconstruction of diverse pretreatmentbiomass feedstock combinations 英文参考文献

Banerjeeetal.BiotechnologyforBiofuels2010,3:22 /content/3/1/22 RESEARCH OpenAccess Rapidoptimizationofenzymemixturesfor deconstructionofdiversepretreatment/biomass feedstockcombinations GoutamiBanerjee,SuzanaCar,JohnSScott-Craig,MelissaSBorrusch,JonathanDWalton* Abstract Background:Enzymesforplantcellwalldeconstructionareamajorcostintheproductionofethanolfrom lignocellulosicbiomass.Thegoalofthisresearchwastodevelopoptimizedsyntheticmixturesofenzymesfor multiplepretreatment/substratecombinationsusingourhigh-throughputbiomassdigestionplatform,GENPLAT, whichcombinesroboticliquidhandling,statisticalexperimentaldesignandautomatedGlcandXylassays. Proportionsofsixcorefungalenzymes(CBH1,CBH2,EG1,b-glucosidase,aGH10endo-b1,4-xylanase,andb- xylosidase)wereoptimizedatafixedenzymeloadingof15mg/gglucanforreleaseofGlcandXylfromall combinationsoffivebiomassfeedstocks(cornstover,switchgrass,Miscanthus,drieddistillers’grainsplussolubles [DDGS]andpoplar)subjectedtothreealkalinepretreatments(AFEX,dilutebase[0.25%NaOH]andalkaline peroxide[AP]).A16-componentmixturecomprisingthecoresetplus10accessoryenzymeswasoptimizedfor threepretreatment/substratecombinations.Resultswerecomparedtotheperformanceoftwocommercial enzymes(Accellerase1000andSpezymeCP)atthesameproteinloadings. Results:WhenanalyzedwithGENPLAT,cornstovergavethehighestyieldsofGlcwithcommercialenzymesand withthecoresetwithallpretreatments,whereascornstover,switchgrassandMiscanthusgavecomparableXyl yields.Withcommercialenzymesandwiththecoreset,yieldsofGlcandXylwerehighestforgrassstovers pretreatedbyAPcomparedtoAFEXordilutebase.Cornstover,switchgrassandDDGSpretreatedwithAFEXand digestedwiththecoresetrequiredahigherproportionofendo-b1,4-xylanase(EX3)andalowerproportionof endo-b1,4-glucanase(EG1)comparedtothesamematerialspretreatedwithdilutebaseorAP.Anoptimized enzymemixturecontaining16components(byadditionofa-glucuronidase,aGH11endoxylanase[EX2],Cel5A, Cel61A,Cip1,Cip2,b-mannanase,amyloglucosidase,a-arabinosidase,andCel12Atothecor