optimization of peptide linkers

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1、ApplMicrobiolBiotechnol(2008)79:579–587DOI10.1007/s00253-008-1468-4BIOTECHNOLOGICALLYRELEVANTENZYMESANDPROTEINSBifunctionalenhancementofaβ-glucanase-xylanasefusionenzymebyoptimizationofpeptidelinkersPingLu&Ming-GuangFengReceived:30January2008/Revised:20March2008/Accepted:23M

2、arch2008/Publishedonline:16April2008#Springer-Verlag2008AbstractTheflexiblepeptides(GGGGS)n(n≤3),theα-andcanbeconvertedintoanimalnutritionandfuelethanolbyhelicalpeptides(EAAAK)n(n≤3)andtwootherpeptidesmicrobialenzymes(Dienetal.2003;Levasseuretal.2005).wereusedaslinkerstocons

3、tructbifunctionalfusionsofHydrolysisoftheendospermcellwallsrequiresactionsofβ-glucanase(Glu)andxylanase(Xyl)forimproveddifferentenzymestodegradethepolysaccharidesintocatalyticefficienciesofbothmoieties.EightGlu-Xylfusionutilizableoligo-ormonosaccharides.Sincefibrolyticenzyme

4、sconstructedwithdifferentlinkerswereallenzymestendtoevolvetowardbi-ormultifunctionalexpressedastheproteinsofca.46kDainEscherichiacolidirectionsformoreefficientbiodegradationofplantfibreBL21anddisplayedtheactivitiesofbothβ-glucanaseand(Xueetal.1992;Flintetal.1993),aninteresti

5、nconstructionxylanase.Comparedtoallthecharacterizedfusionswithofbi-ormultifunctionalfusionsoftheenzymesforthetheparentalenzymes,thecatalyticefficienciesoftheGludegradationofcerealpolysaccharideshasbeenincreasing.andXylmoietieswereequivalentto304–426%and82–Plentyoffusionenzym

6、eshavebeendevelopedbymeans143%oftheparentalones,respectively.Thepeptidelinkerofend-to-endfusiontechnique(Trujilloetal.1997;Hong(GGGGS)2resultedinthebestfusion,whosecatalyticetal.2006).Withthistechnique,individualgenesencodingefficiencyhadanetincreaseof326%fortheGluandofintac

7、tfunctionalproteinsarefusedintoachimericgenethat43%fortheXyl.Thetwomoietiesofafusionwiththelinkermayencodealargerproteintoperformtwoormore(EAAAK)3alsoshowednetincreasesof262and31%infunctions(DoiandYanagawa1999).Previousstudieshavecatalyticefficiency.Ourresultshighlight,forth

8、efirsttime,shownthatproperlinkerpeptidescanbeinsertedbetweentheenhancedbifu