Improving heterologous polyketide production in Escherichia coli by overexpression of an S-adenosylmethionine synthetase gene

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1、ApplMicrobiolBiotechnol(2007)77:367–373DOI10.1007/s00253-007-1172-9APPLIEDGENETICSANDMOLECULARBIOTECHNOLOGYImprovingheterologouspolyketideproductioninEscherichiacolibyoverexpressionofanS-adenosylmethioninesynthetasegeneYongWang&BrettA.Boghigian&BlaineA.PfeiferReceived:13June2007/R

2、evised:12August2007/Accepted:16August2007/Publishedonline:18September2007#Springer-Verlag2007AbstractAnS-adenosylmethioninesynthetasegene(metK)IntroductionfromStreptomycesspectabiliswasclonedintoanexpressionplasmidunderthecontrolofaninducibleT7promoterandPolyketidesarenaturalprodu

3、ctsproducedbybacteria,fungi,introducedintoastrainofEscherichiacoli(BAP1(pBP130/andplantsandrepresentastructurallydiverseclassofpBP144))capableofproducingthepolyketideproductcompoundswithabroadrangeofactivities(antibacterial,6-deoxyerythronolideB(6-dEB).ThemetKcoexpressionantifunga

4、ls,anticanceragents,immunosuppressants).OverinBAP1(pBP130/pBP144)improvedthespecificproductionthepast5years,effortshavefocusedongeneratingsuch−1
1of6-dEBfrom10.86to20.08mglOD600.Inanefforttoimportantcompoundsthroughtechnicallyconvenientmicro-probethereasonforthisimprovement,aserie

5、sofgenebialhosts(Mutkaetal.2006;Pfeiferetal.2001,2003;deletionandexpressionexperimentswereconductedbasedWatanabeetal.2003).HostssuchasEscherichiacoliofferonametKmetabolicpathwaythatbranchesbetweennumerousadvantages(mostnotably,fastergrowthratesandpropionyl-CoA(a6-dEBprecursor)anda

6、utoinducercom-well-establishedgeneticmanipulationprotocols)whencom-pounds.Thedeletionandexpressionstudiessuggestedthatparedtomostnativepolyketideproducingmicroorganisms,theautoinducerpathwayhadalargerimpactonimprovedand,therefore,severalpolyketideproductshavenowbeen6-dEBbiosynthes

7、is.Supportingtheseresultswereexperi-generatedthroughE.coli.Inoneexample,metabolicmentsdemonstratingthepositiveeffectconditionedmediaengineeringwasusedtosupplytheprecursors(propionyl-(thesuspectedlocationoftheautoinducercompounds)hadCoAand2S-methylmalonyl-CoA)neededfortheproduction

8、on6-dEBproduction.Takentogether,t