萜类生物合成的基因操作

ChinaBiotechnology,2006,26(1):60~64综述*黄瑛**曾庆平(510405)萜类是一组结构迥异的化合物家族,其中很多具有较大的应用价值,如青蒿素和紫杉醇等,它们在多种微生物和植物中合成,但其天然产量低萜类代谢工程通过DNA重组技术改造萜类合成细胞中的代谢途径,以提高萜类最终产量或在不含萜类的生物中合成萜类,为促进有用萜类合成提供了新的机会以萜类化合物生物合成途径的基因转移与表达为切入点,综述了目前在微生物及植物中应用代谢工程提高萜类产量的研究进展萜类基因转移代谢工程Q541:20050721:20051020*(02-03ZP43),(020799),(JQ0205)**:[e-mail],(isoprenoid),(),,,,,,,1(IPP),IPPCoA(MVApathway);(nonMVApathway),5(DXPpathway)(1),IPP,(2)IPPDXP,IPPMVA,DXP2:(1);(2),;(3),2.1DXP(cartenoid),[1,2];,[3,4],[5]Martin[6],2006,26(1):10,(amorpha4,11dienesynthase),,,24g/ml1IPP(a)(b)Fig.1BiosynthesisofIPPandDMAPPviatheMVApathway(a)andthenonMVApathway(b)2IPPFig.2BiosynthesisofterpenoidfromIPP,,,IPP,,Huang[7]78(taxadienesynthase),,70%GGDP,IPPGGDP,,0.5mg/LDXP,2.6Matthews[8],DXSDXS,10.8Reiling[9]GPPGGPP,61ChinaBiotechnologyVo.l26No.120066,,,Martin[10],FPP,500DXP,,Martin[10]5vetispiradiene,IPTG,3(LB2YTM9+2%),trcTrc99ADH5,0.2%M9,;:5vetispiradiene30,37,tRNA,,5,,vetispiradieneKmi[11]3(DH5XL1BlueJM101)DXPDXP,2DXP1.4~2.0,araBADIPTGtrclac2,XLBlueCarter[12]IPP4,,2.2,,(eryosterol)Yamano[13]Misawa[14],Jackson[15],FPP,FPPa,,Polakowski[16]Donald[17]HMGCoA,,Verdoes[18](Xanthophyllomycesdendrorhous),crtYB,,,(crtI),,Shmiada[19](Candidautilis),3crtEcrtBcrtl,,1.1mg/g(),;HMGCoA,1;HMGCoA,4,4.3mg/g;,EMG9,p8ENB.G4ERG9,!∀,,7.8mg/g,,,2.3,,Wallaart[20],,0.2~1.7ng/g(FW);Mann[21]cDNAPds622006,26(1):,;[22][23][24],,FPP,FPP,,FPP,FPS,4,,FPS,FPS[25]Besumbes[26],,,GGPP,30,,,Rosati[27],,,,,Dharmapuri[28],,,,,,3,,,,,,DNA,,,,,;,[29][30],,,[1]LeePC,MijtsBN,SchmidtDannertC.InvestigationoffactorsinfluencingproductionofthemonocycliccarotenoidtoruleneinmetabolicallyengineeredEscherichiacoli.ApplMicrobiolBiotechno,l2004,65:538~546[2]SchmidtDannertC,UmenoD,ArnoldFH.Molecularbreedingofcarotenoidbiosyntheticpathways.NatBiotechno,l2000,18(7):750~753[3]MisawaN,ShimadaH.Metabolicengineeringfortheproductionofcarotenoidsinnoncarotenogenicbacteriaandyeasts.JBiotechno,l1997,59(3):169~181[4]WangCW,OhMK,LiaoJC.EngineeredisopernoidpathwayenhancesastaxanthinproductioninEscherichiacoli.BiotechnolBioeng,2000,62(2):235~241[5]KajiwaraS,FraserPD,KondoK,eta.lExpressionofanexogenousisopentenyldiphosphateisomerasegeneenhancesisoprenoidbiosynthesisinEscherichiacoli.BiochemJ,1997,324:421~426[6]MartinVJ,PiteraDJ,WithersST,eta.lEngineeringamevalonatepathwayinEscherichiacoliforproductionofterpenoids.NatBiotechnology,2003,21(7):796~802[7]HuangQL,RoessnerCA,CroteauR,eta.lEngineeringEscherichiacoliforthesynthesisoftaxadiene,akeyintermediateinthebiosynthesisoftaxo.lBioorgMedChem,2001,9:2237~2242[8]MatthewsPD,WurtzelET.MetabolicengineeringofcarotenoidaccumulationinEscherichiacolibymodulationoftheisoprenoidprecursorpoolwithexpressionofdeoxyxylulosephosphatesynthase.ApplMicrobiolBiotechno,l2000,53:396~400[9]ReilingKK,YoshikuniY,MartinVJ.Monoandditerpene63ChinaBiotechnologyVo.l26No.12006productioninEscherichiacoli.BiotechnolBioeng,2004,87(2):200~212[10]MartinVJ,YoshikuniY,KeaslingJD.TheinvivosynthesisofplantsesquiterpenesbyEscherichiacoli.BiotechnolBioeng,2001,75(5):497~503[11]KimSW,KeaslingJD.MetabolicengineeringofthenonmevalonateisopentenyldiphosphatesynthesispathwayinEscherichiacolienhanceslycopeneproduction.BiotechnolBioeng,2001,72(4):408~415[12]CarterOA,PetersRJ,CroteauR.MonoterpenebiosynthesispathwayconstructioninEscherichiacoli.Phytochemistry,2003,64(2):425~433[13]YamanoS,IshiiT,NakagawaM,eta.lMetabolicengineeringforproductionofbetacaroteneandlycopeneinSaccharomycescerevisiae.BiosciBiotechnolBiochem,1994,58(6):1112~1114[14]MisawaN,ShimadaH.Metabolicengineeringfortheproductionofcarotenoidsinnoncarotenogenicbacteriaandyeasts.JBiotechno,l1997,59(3):169~181[15]JacksonBE,HartWellsEA,MatsudaSP.Metabolicengineeringtoproducesesquiterpenesinyeast.Orglett,2003,5(10):1629~1632[16]PolakowskiT,StahlU,LangC.OverexpressionofacytosolichydroxymethylglutarylCoAreductaseleadstosqualeneaccumulationinyeast.ApplMicrobiolBiotechno,l1998,49:66~71[17]DonaldKA,HamptonRY,FritzIB.Effectsofoverproductionofthecatalyticdomainof3hydroxy3methylglutarylcoenzymeAreductaseonsqualenesynthesisinSaccharomycescerevisiae.ApplEnvironMicrobio,l1997,63(9):3341~3344[18]VerdoesJC,SandmannG,VisserH,eta.lMetabolicengineeringofthecarotenoidbiosyntheticpathwayintheyeastXanthopyhllomycesdendrorhous(Phaffiarhodozyma).ApplEnvironMicrobio,l2003,69(7):3728~3738[19]ShimadaH,KondoK,FraserPD,eta.lIncreasedcarotenoidproductionbythefoodyeastcandidautilisthroughmetabolicengineeringoftheisoprenoidpathway.ApplEnvironMicrobio,l1998,64(7):2676~2680[20]WallaartTE,BouwmeesterHJ,HilleJ,eta.lAmorpha4,11dienesynthase:cloningandfunctionalexpressionofakeyenzymeinthebiosyntheticpathwayofthenovelantimalarialdrugartemisinin.Planta,2001,212(3):460~465[21]MannV,HarkerM,PeckerI,eta.lMetabolicengineeringofastaxanthinproductionintobaccoflowers.NatBiotechno,l2000,18(8):888~892[22]LuckerJ,SchwabW,HautumBV,eta.lIncreasedandalteredfragranceoftobaccoplantsaftermetabolicengineeringusingthreemonoterpenesynthasesfromLemon.PlantPhysio,l2004,134:510~519[23]DucreuxLJ,MorrisW,lHesleyPE,eta.lMetabolicengine