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1、ResearchArticlepubs.acs.org/synthbioExploringLysineRiboswitchforMetabolicFluxControlandImprovementofL‑LysineSynthesisinCorynebacteriumglutamicumLi-BangZhouandAn-PingZeng*InstituteofBioprocessandBiosystemsEngineering,HamburgUniversityofTechnologyDenickestrasse15,D-21073Hamburg,Germany
2、*SSupportingInformationABSTRACT:Riboswitch,aregulatorypartofanmRNAmoleculethatcanspecificallybindametaboliteandregulategeneexpression,isattractiveforengineeringbiologicalsystems,especiallyforthecontrolofmetabolicfluxesinindustrialmicroorganisms.Here,wedemonstratetheuseoflysineriboswitc
3、handintracellularL-lysineasasignaltocontrolthecompetingbutessentialmetabolicby-pathwaysoflysinebiosynthesis.Tothisend,wefirstexaminedthenaturallysineriboswitchesofEschericiacoli(ECRS)andBacillussubtilis(BSRS)tocontroltheexpressionofcitratesynthase(gltA)andthusthemetabolicfluxinthetrica
4、rboxylicacid(TCA)cycleinE.coli.ECRSandBSRSwerethensuccessfullyusedtocontrolthegltAgeneandTCAcycleactivityinalysineproducingstrainCorynebacteriumglutamicumLP917,respectively.ComparedwiththestrainLP917,thegrowthofbothlysineriboswitch-gltAmutantswasslower,suggestingareducedTCAcycleactiv
5、ity.Thelysineproductionwas63%higherinthemutantECRS-gltAand38%higherinthemutantBSRS-gltA,indicatingahighermetabolicfluxintothelysinesynthesispathway.Thisisthefirstreportonusinganaminoacidriboswitchforimprovementoflysinebiosynthesis.Thelysineriboswitchescanbeeasilyadaptedtodynamicallycon
6、trolotheressentialbutcompetingmetabolicpathwaysorevenbeengineeredasan“on-switch”toenhancethemetabolicfluxesofdesiredmetabolicpathways.KEYWORDS:dynamicmetaboliccontrol,lysineriboswtch,lysinebiosynthesis,citratesynthase,C.glutamicumMetabolicfluxcontrolisamajorissueindevelopinghighlylysin
7、ebiosynthesisinCorynebacteriumglutamicumbyusingproductivemicroorganismsforbioproduction.Inlysineasanintracellularsignal.particular,thecontrolofmetabolicpathwaysthatareessentialC.glutamicumisaGram-positivebacteriumwidelyusedfor6−8forcellgrowthbutnotdesiredfortheproductionofaspecialpro
8、ducingaminoa
