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1、MetabolicengineeringofplantalkaloidbiosynthesisFumihikoSato*²³,TakashiHashimoto³§,AkiraHachiya§,Ken-ichiTamura*,Kum-BooChoi*,TakashiMorishige*,HidekiFujimoto*,andYasuyukiYamada§*DivisionofAppliedLifeSciences,GraduateSchoolofAgriculture,and²DivisionofIntegratedLifeScience,GraduateSchoolofBio
2、studies,KyotoUniversity,Kyoto606-8502,Japan;and§NaraInstituteofScienceandTechnology,Ikoma630-0101,JapanThiscontributionispartofthespecialseriesofInauguralArticlesbymembersoftheNationalAcademyofScienceselectedonApril27,1999.ContributedbyYasuyukiYamada,November3,2000Plantalkaloids,oneofthelar
3、gestgroupsofnaturalproducts,providescine),whicharefoundmainlyinHyoscyamus,Duboisia,Atropa,manypharmacologicallyactivecompounds.SeveralgenesintheandScopoliaspecies,togetherwiththeirsemisyntheticderivatives,biosyntheticpathwaysforscopolamine,nicotine,andberberinehaveareusedasparasympatholytic
4、sthatcompetitivelyantagonizeace-beencloned,makingthemetabolicengineeringofthesealkaloidstylcholine.Boththetropaneringmoietyofthetropanealkaloidspossible.Expressionoftwobranching-pointenzymeswasengi-andthepyrrolidineringofnicotinearederivedfromputrescinebyneered:putrescineN-methyltransferase
5、(PMT)intransgenicplantsofwayofN-methylputrescine(MP)(Fig.1).BecauseputrescineisAtropabelladonnaandNicotianasylvestrisand(S)-scoulerine9-O-metabolizedtopolyaminessuchasspermidineandspermine,themethyltransferase(SMT)inculturedcellsofCoptisjaponicaandN-methylationofputrescinecatalyzedbyputresc
6、ineN-methyltrans-Eschscholziacalifornica.OverexpressionofPMTincreasedthenico-ferase(PMT)isthefirstcommittedstepinthebiosynthesisofthesetinecontentinN.sylvestris,whereassuppressionofendogenousPMTalkaloids.activityseverelydecreasedthenicotinecontentandinducedabnor-Isoquinolinealkaloidsaresome
7、ofthemajormetabolitessuc-malmorphologies.EctopicexpressionofSMTcausedtheaccumula-cessfullyproducedbyplantcellculture.Sofar,about60havebeentionofbenzylisoquinolinealkaloidsinE.californica.Theprospectsisolatedfromplantcellcultures(ref.4andreferencestherein
