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1、CHAPTER1TheEssenceofMetabolicEngineeringTheconceptofmetabolicpathwaymanipulationforthepurposeofendowingmicroorganismswithdesirablepropertiesisaveryoldoneindeed.Wehavemanyoutstandingexamplesofthisstrategyintheareasofaminoacids,antibiotics,solvents,andvitaminproduct
2、ion.Thesemethodsrelyheavilyontheuseofchemicalmutagensandcreativeselectiontechniquestoidentifysuperiorstrainsforachievingacertainobjective.Despitewidespreadacceptanceandimpressivesuccesses,thegeneticandmetabolicprofilesofmutantstrainswerepoorlycharacterizedandmutag
3、enesisremainedaran-domprocesswheresciencewascomplementedwithelementsofart.Thedevelopmentofmolecularbiologicaltechniquesfordeoxyribonucleicacid(DNA)recombinationintroducedanewdimensiontopathwaymanipu-lation.Geneticengineeringallowedtheprecisemodificationofspecifice
4、nzymaticreaction(s)inmetabolicpathwaysand,hence,theconstructionofwell-definedgeneticbackgrounds.ShortlyafterthefeasibilityofrecombinantDNAtechnologywasestablished,varioustermswerecoinedtorepresentthe2MetabolicEngineeringpotentialapplicationsofthistechnologytodirec
5、tedpathwaymodification.Someofthetermssuggestedweremolecularbreeding(Kelloggetal.,1981),invitroevolution(Timmisetal.,1988),(microbialormetabolic)pathwayengineering(MacQuitty,1988;Tongetal.,1991),cellularengineering(Nerem,1991),andmetabolicengineering(Stephanopoulos
6、andVallino,1991;Bailey,1991).Althoughtheexactdefinitionvariesfromauthortoauthor,allconveysimilarmeaningswithrespecttothegeneralgoalsandmeansofmetabolicengineering.Herewedefinemetabolicengineeringasthedirectedimprovementofproductformationorcellularpropertiesthrough
7、themodificationofspecificbiochemicalreaction(s)ortheintroductionofnewone(s)withtheuseofrecombinantDNAtechnology.Anessentialcharacteristicoftheprecedingdefinitionisthespecificityoftheparticularbiochemicalreaction(s)targetedformodificationortobenewlyintroduced.Onces
8、uchreactiontargetsareidentified,establishedmolecularbiologicaltechniquesareappliedinordertoamplify,inhibitordelete,transfer,orderegulatethecorrespond-in
