次级代谢产物菌种改良的策略和手段

21420027JournalofWuxiUniversityofLightIndustryVol.21No.4Jul.2002:1009-038X(2002)04-0428-06:2002-03-25;:2002-04-20.:(1944-),,,,.,,,(,214036):,,,.:;;;:Q591:AStrategyandMeansUtilizedfortheImprovementofSecondaryMetabolite2ProducingStrainsZHANGXing2yuan,PANJun2hua,ZENGMei2juan,LINai2qiang(TheKeyLaboratoryofIndustrialBiotechnologyofMinistryofEducation,SouthernYangtzeUniversity,Wuxi214036,China)Abstract:Startingwithantibiotics,anintroductiontostrategyandmeansutilizedfortheimprove2mentofsecondarymetabolites2producingstrainswasgiven.Basedonthetheoryofmetabolicengineer2ing,afurtheradvanceswasmadeonthefive2wordstrategywhichwaseffectiveindesignedbreedingofprimarymetabolites,andadiscussionwasmadeintheprospectforitsapplicationindesignedbreed2ingfortheoverproductionofsecondarymetabolites.Keywords:designedbreeding;metabolicengineering;secondarymetabolism;antibiotics,,.,,..,,,.(),.,,.DNA,.[13];;,.,+.,,.,,.,.,,,,.,(),.,.,,,..,..[4].1(),,.,.,,,.1.1,.,,,,.,,.,,,.1.2+(),.,,(A).,[5,6],.:,;,;,,;,;,.,.,4,;45(,).,[7].,..,,.,,.[9,10].9244:2(),,.,.,,,[7].1().,.,.1Fig.1Strategyforimprovementofantibiotic2producingstrains2.12.1.1(),.,,(buildingblocks,),.,Streptomycescattleya,29.3kbDNA[1].Streptomyceslividans,CStreptomyceslac2tamgens,23.,,,.2.1.2()(),.,,.,.,(,).,.,,,,.,03421,.,,,;,[11].tetracenomycinStreptomycesglaucescens2.,,[12].(2).,,TcmD330,tetracenomycin30%.2.1.3().[13]:,,;(),.,.,100mmol/L.,.,,,.,,(pH).,,..().,ABC(ATPbindingcassettetrans2porters)[14].,,.[15].,;,,.2.2,.,.(activator).,(repressor)[16].,.A2Streptomycesgriseus[17]virginiaebutano2lidesStreptomycesvirginiae[18].A2virginiaebutanolides,,,[19].,.,.,,.,,,().,,(timeprofile).,,,,,.2.2.1()1)(pathwayspecificregula21344:tor).(acti2vatorprotein),.(),.,.,,.,,.,srmR(),(Strepto2mycesambofaciens)[20].,100g/mL500g/mL.ccaRStreptomycesclavuligerusC[21,22].(clavulanicacid)1[21].2)(globalregulator),(pleiotropicregulator).StreptomycescoelicolorabsA[23]().absA,actinorhodinunde2cylprodigiosin.S.lividanscutRS[24],2actinorhodin3.S.lividansasfR2,63.,actinorhodinundecyl2prodigiosin[25].2.2.2(),(transcriptionalrepressor),:S.coelicolor(methylenomycin)mmyRactinorhodinact2orf[26];S.glaucescenC(tetracenomycinC)tcmR[27];Streptomycespeucetius(daunorubicin)dnrO[28];Strep2tomycesvenezuelaejadomycinBjadR2[29].MmyR,S.coelicolor[16].,.,,.3,.,,.:[1]CHENCW,LINHF,LUOCL,etal.CloningandexpressionofaDNAsequenceconferringcephamycinCproduction[J].Bio/Technology,1988,6:1222-1224.[2]ISOGAIT,FUKAGAWAM,ARAMORII,etal.Constructionofa72aminocephalosporanicacid(7ACA)biosyntheticoperonanddirectproductionof7ACAinAcremoniumchrysogenum[J].Bio/Technology,1991,(9):188-191.[3]CRAWFORDL,STEPANAM,MCADAPC,etal.Productionofcephalosporinintermediatesbyfeedingadipicacidtore2combinantPenicilliumchrysogenumstrainsexpressingringexpansionactivity[J].Bio/Technology,1995,13:58-62.[4],.[J].,2002,27(4):252-256.[5].[J].,1999,26(2):147-149.23421[6].[J].,2001,20(4):447-449.[7],.C1a[J].,1998,23(6):410-414.[8]ZHANGXY.Threebasicviewpointsofindustrialfermentation[A].Foodof21stCentury(4thInternationalConferenceofFoodScienceandTechnology)[C].Beijing:ChinaLightIndustryPress,2000.[9]CRAMERIR,DAVIESJE.Increasedproductionofaminoglycosidesassociatedwithamplifiedantibiotictesistancegenes[J].JAntibiot,1986,39:128-35.[10]WEISBLUMB.Methodofincreasingtheantibioticyieldofproducingorganisms[P].U.S.Patent4376823,1983203201.[11]THOMASDI,COVEJH,BAUMBERGS,etal.Plasmideffectsonsecondarymetaboliteproductionbyastreptomycetesynthesizingananthelminticmacrolide[J].JGenMirobiol,1991,137:2331-2337.[12]DECKERH,SUMMERSRG,HUTCHINSONCR.OverproductionoftheacylcarrierproteincomponentofatypepolyketidesynthasestimulatesproductionoftetracenomycinbiosyntheticintermediatesinStreptomycesglaucescens[J].JAn2tibiot,1994,47:54-63.[13]CUNDLIFFEE.Howantibiotic2producingorganismsavoidsuicide[J].AnnuRevMicrobiol,1989,43:207-233.[14]MENDEZC,SALASJA.ABCtransportersinantibiotic2producingactinomycetes[J].FEMSMicrobiolLett,1998,158:1-8.[15]LEWISK.Multidrugresistancepumpsinbacteria:variationsonatheme[J].TrendsBiochemSci,1994,19:119-123.[16]CHATERKF,BRUTONCJ.Resistance,regulatoryandproductiongenesfortheantibioticmethylenomycinareclustered[J].EmboJ,1985,4:1893-1897.[17]HORINOUCHIS,BEPPUT.A2factorasamicrobialhormonethatcontrolscellulardifferentiationandsecondarymetabolisminStreptomycesgriseus[J].MolMicrobiol,1994,12:859-864.[18]YAMADAY,NIHIRAT,SAKUDAS.Butyrolactoneautoregulators,inducersofvirginiamycinbiosynthesis[A].Biotechnol2ogyofAntibiotics,2nded[C].NewYork:MarcelDekker,1997.63-79.[19]BEPPUT.Signaltransductionandsecondarymetabolism:prospectsforcontrollingproductivity[J].TrendsBiotechnol,1995,132:264-269.[20]GEISTLICHM,LOSICKR,TURNERJR,etal.CharacterizationofanovelregulatorygenegoverningtheexpressionofapolyketidesynthasegeneinStreptomycesambofaciens[J].MolMicrobiol,1992,6:2019-2029.[21]PEREZ2LLARENAFJ,LIRASP,RODRIGUEZ2GARCIAA,etal.Aregulatorygene(ccaR)requiredforcephamycinandclavulanicacidproductioninStreptomycesclavuligerus:amplificationresultsinoverproductionofbothbeta2lactamcompounds[J].JBacteriol,199