Simple fed-batch technique for high cell density cultivation of Escherichia coli

Simple fed-batch technique for high cell density cultivation of Escherichia coli

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1、journalofwtechndogyJournalofBiotechnology39(1995)59-65Simplefed-batchtechniqueforhighcelldensitycultivationofEscherichiacoliD.J.Korz,U.Rinas,K.Hellmuth,E.A.Sanders,W.-D.Deckwer*GBFNationalResearchCenterforBiotechnology,BiochemicalEngineeringDivision,Masch

2、eroderWegI,38124Braunschweig,GermanyReceived31August1994;accepted4November1994AbstractAsimplefed-batchprocessforhighcelldensitycultivationofEscherichiacoliTGlwasdeveloped.Apre-determinedfeedingstrategywaschosentomaintaincarbon-limitedgrowthusingadefinedme

3、dium.Feedingwascarriedouttoincreasethecellmassconcentrationexponentiallyinthebioreactorcontrollingbiomassaccumulationatgrowthrateswhichdonotcausetheformationofaceticacid(II

4、coseorglycerolascarbonsource,respectively.Keywords:Escherichiacoli;Fed-batch;Cultivation,highcelldensity1.Introductiontohighcelldensitiesinfed-batchcultureshavebeenreviewedrecently(Riesenberg,1991;YeeEscherichiacoliisstillthemostimportanthostandBlanch,199

5、2).Thehighestcellconcentra-organismforrecombinantproteinproduction.Totionsofnon-recombinantE.colireportedinthemaximizethevolumetricproductivitiesofbacte-literaturewereobtainedbyfeedingsolidglucoserialculturesitisimportanttogrowE.colitohighviaaspecialappar

6、atusintoapressurizedbioreac-cellconcentrations.Preventingtheaccumulationtor(134g1-lDCW;Matsuietal.,1989)orbyoftoxiclevelsofaceticacidisthemaintasktoremovinginhibitorysubstancesbydialysis(174gachievehighcellconcentrationsinthebioreactor.1-lDCW;MSletal.,199

7、3).Usingfeed-backGrowth-inhibitingacidicby-productsofincom-controlstrategiesforglucosefeedingtomaintainpletesubstrateoxidationsuchasaceticacidarethedissolvedoxygeninacertainrange(CutayarproducedinresponsetooxygenlimitationorandPoillon,1989;Morietal.,1979)

8、andtokeepexcesscarbon.DifferentstrategiestogrowE.colithespecificgrowthrateapproximatelyconstant(Riesenbergetal.,1991)celldensitiesof110-125g1-lDCWhavebeenobtained.Allthesestrategiesrequireconsiderableexper-lCorrespo

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