生物发酵工程

陈国强,微生物发酵工程前沿,微生物发酵工程是工业生物技术的基础,什么是“工业生物技术”？,IndustrialMicrobiologyandFermentationTechnology,IndustrialMicrobiologyisthedisciplinethatusesmicroorganisms,usuallygrownonalargescale,toproducevaluablecommercialproductsorcarryoutimportantchemicaltransformations.FermentationTechnologyisthetechnologytogrowcellsinalargescalewithhighefficiency,italsoincludesproductrecoveryprocesses.,MicrobialProducts:MicrobialCells,Enzymes,PharmaceuticalProducts,SpecialtyChemicalsandFoodAdditives,Flowsheetfordevelopinganindustrialmicrobialfermentationprocess,StrainselectionLaboratoryprocessdevelopmentPilotScaleupIndustrialScaleupDownstreamprocessdevelopmentProductpackagingtechniquesOthercommercialconsiderationExamples,StrainSelection,PurchasefromCultureCollectionsScreeningofnaturecircumstancesGeneticengineeringMutationsCellbiologytechniques,InternationalCultureCollections,Domestically,strainscanbepurchasedfrom:CGMCCorChinaGeneralMicrobiologicalCultureCollectionCenter中国微生物菌种保藏管理委员会普通微生物中心，北京中关村,Screeningofnaturecircumstances,GeneticEngineering,Incorporationintoartificialplasmidsofgenesfromawidevarietyofsourceshasmadepossiblethetransferofgeneticmaterialacrossvirtuallyanyspeciesbarrier,VarioushighvalueaddedproductshavebeenproducedfromGeneticengineeringmethods,Mutation,Viachemicalorphysical,andbiologicalmeans,CellBiologyTechniques,Protoplastfusion(promotehighfrequenciesofgeneticrecombinants)removingthecellwallwithlyticenzymesinthepresenceofosmoticstabilizers.Inthepresenceoffusogenicagentsuchaspolyethyleneglycol(PEG),protoplastsareinducedtofuseandformtransienthybridsordiploids.Regenerationofviablecellsfromthefusedprotoplasts.,Stagesneededfortransferringanindustrialprocessfromthelaboratorytothecommercialfermentor,ShakeflashExperimentsLabscalefermentor(5-10L)Pilotscalefermentor(300-3000L)Commercialfermentor(10,000-500,000L),LaboratoryProcessDevelopmentShakeFlaskExperiments,Optimizationofconditionsforcellgrowthandproductformationusingshakeflaskexperiments:1.pH2.Temperature3.Dissolvedoxygen(DO)4.Substratechoice5.Maximalandoptimalsubstrateconcentration6.Others,LaboratoryProcessDevelopmentFermentorExperiments,AgitationCoolingandheatingAirinletandoutletpHcontrolNutrientadditionInoculationViewingport,AseriesofFermentorsusedforLabScaleProcessDevelopment,LaboratoryProcessDevelopmentLabscalefermentorexperiments,BatchprocessFed-batchprocessContinuousprocessSemi-continuousprocess,FermentationProcess:BatchProcess,Allthenutrientsneededforcellgrowthwillonlybeaddedonceatthebeginningoffermentation.,Fermentationprocess:Fed-batchprocess,Duringthefermentation,additionalnutrientswillbeaddedinabatchwaytopromotethecellgrowthorproductformationandtoavoidnutrientdeficiency.,Fermentationprocess:Continuousprocess,Fermentationprocess:Semi-continuousprocess,Similartocontinuousprocess,withtheadditionofnutrientsandoutflowoffermentationbrothinacontinuousandbatchway.,Processcontrolandmonitoring,Processparameterstobemonitors,Sugarconsumption,pH,Temperature,Fermentationtime(h),Agitation,CellDryWeight,Product,Computersoftwareshavebeendevelopedtomonitorandchangetheprocessonline,PilotScaleUp,Scaleup:Thetransferofaprocessfromsmall-scalelaboratoryequipmenttolarge-scalecommercialequipmentPilotexperimentTotestthefeasibilityofthelabscalefermentationprocessinasemi-industrialscalePilotfermentorsnormallyhaveasizerangingfrom100Lto10,000L,dependingontheproductstobemassproducedlater.,Fermentorsizesforvariouspurposes,Problemsemergingduringthescaleup,Asthesizeoftheequipmentisincreased,thesurface-volumeratiochangesLargefermentorhasmuchmorevolumeforagivensurfacearea,itisobviouslymoredifficulttomixthebigtankthanthesmallflaskInscaleupstudiesonaerobicfermentations,oxygenrateinthefermentorisbestkeptconstantasthesizeofthefermentorisincreased.HowtokeepDOconstant?IncreasestirringrateIncreaseairpressureUsepureoxygenIncreaseairinlet,IndustrialScaleup,Totransferthepilotscaleresultsintoacommerciallyfeasibleproductionsetting.Fermentorsizesrangefrom100Lto500,000L,dependingonproducts.,IndustrialFermentationSetting,Flowsheetfordevelopinganindustrialmicrobialfermentationprocess,StrainselectionLaboratoryprocessdevelopmentPilotScaleupIndustrialScaleupDownstreamprocessdevelopmentProductpackagingtechniquesOthercommercialconsiderationExamples,DownstreamProcessDevelopment,Chromatography,Filter(pressfilter),DownstreamProcessDevelopment,Continuouscentrifuges,UnitOperationsinDownstreamSeparation,MicrofiltersUltrafiltrationGelchromatographyReverseosmosisDialysisElectrodialysisIonexchangeDistillation/freezedrySolventextractionFoamandbubblefractionationUltracentrifugationCentrifugationLiquidcyclonesGravitysedimentation,Size,Diffusivity,Ionexchange,Vapourtemperature,pressure,Solubility,Surfaceactivity,Density,Processingfermentationbroth,Fermentation,Diluteslurry,CentrifugationFiltrationSedimentation,Clarifiedliquor,Filtration,Brightliquor,Concentratedslurry,Dewatering,Drying,DryBiomass,Centrifugation:Continuouscentrifugation,Disc-Centrifuges,Solids-retainingcentrifuge,Nozzlewithpressurizeddischargeofconcentrate,Nozzlewithperipheralnozzles,Periodicallysolids-ejectingcentrifuge,Periodicallysolids-ejectingcentrifugewithaxialchannels,ProductPackagingTechniques,SterilePackagingTechniquesforMedicalApplicationsandFoodPreservationPyrogeneFreeandSterilePackagingforInjectionPurposeFreezeDryPackagingforFoodsorMedicinesDewateredPackaging(suchasDryYeast)NormalFoodPackaging(suchasNa-glutamate)SaltyPackagingPreservationChemicalsOrdinaryPackaging,Strengthsandweaknessesofbiotechnologicalprocesses,StrengthsRelianceonrenewablefeedstocksVersatilitywithdifferentfeedstocksFood,feedanddrugapplicationsFinechemicalstobulkchemicalapplicationsLowtemperatureOperationinaqueousmediaSeveralreactionsachievedinasinglefermentationstepHighlevelofautomationStereospecificityComplexmoleculesconvertedand/orproduced“Benign”effluentsproduced,WeaknessesFeedstocksareoxidizedandunsuitableformanyapplicationsFeedstockcostsfluctuateSterilizationisamajorcostProductoftenindiluteaqueoussolutionsProductrecoverycostlyEquipmentcostshighReactionsslowleadingtopoorvolumetricproductivityComplicatedreactionconditionsMainlybatchoperationHighcell(catalyst)regenerationcostsHighBODwastes,OtherCommercialConsiderationCostEvaluation,OutlineoftotalcapitalinvestmentFixedcapitalDirectcosts(Land,sitedevelopment,buildings,processing,services)Indirectcosts(Engineering,construction,Contingency,Fees)Start-upcostsWorkingcapitalInventory,accountsreceivable,accountpayable,BiotechnologyandBioengineeringTowardsindustrialchemistry,生物工程发展趋势,Biotechnologymajorsectors,PharmaAgricultureEnvironmentChemistryEnergy,GreenChemistry,Newpharmacompoundsandmaterials,devices,Controlsystems,genetherapy,NewmaterialsNewcompoundsNewprocesses,BiomassBiofuelsBioelectricity,BiodegradationEnd-of-pipesolutionIntegratedprocesses,Agro-efficiencyFoodqualityAgrochemicals,Biochemicalengineeringisnolongeranacademicchallenge,US:mostnewprofessorshipsarebiomolecularlyorientedEurope:movinginsamedirectionLongsearchestofindnewprofessorsofbiochemicalengineeringMostcomesfromindustry(Patents,fewpublications)Solution:movetowardsbiomolecularfocusMetabolicengineeringComplexcontrolsystemstore-engineereucaryoticcellsTissueengineering,BiochemicalengineeringtobiomolecularengineeringExamplesfromtheETHZurich,DepartmentofProcessTechnologyFromProcessTechnologytoBioprocessMolecularDevelopmentProfessorshipofBioprocessTechnologyMajorbiomolecularorientationDepartmentofChemicalTechnologyFromChemicalTechnologytoEucaryoticMolecularTechnologyProfessorshipofBiochemicalTechnologyMajoreucaryoticbiomolecularsciencesandtechnologyorientationDepartmentofBiologyFromBiotechnologytoSystemsapproachesProfessorshipofSystemsBiologyProfessorshipofMetabolicEngineering,BacktoWhiteBiotech:basedonbiomolecularratherthanmacroengineering,BiocatalysismoleculardesignandbuildingattheatomiclevelfornewmoleculesProcessimprovementsmilderreactionconditions,fewersideproducts,NewmaterialsNewcompoundsNewprocesses,BioprocessesToday,Highvaluecompounds:typicallypharmaproductsproteins:therapeuticantibodiespeptidehormones:insulinsmallmolecules:drugsordrugintermediatesMediumvaluecompoundsantibioticsvitaminsaminoacidsBulkcompoundshighfructosecornsyrup(glucoseisomerase)acrylamide(MitsubishiRayon)polylacticacid(NatureWork)propanediol(Dupont),MichaelHoppertandFrankMayer,Am.Sci.87,vol6(1999),Biosynthesisbywholecells,Whendobioprocessesbecomepractical?,BiocatalystRelevantenzymesystems:wheretofindthese?Sufficientactivities:howtoachievethis?Biosystemstability:howtomaintainthesystem?BioprocessMultiphasesystems:essentialforapolarproductsBatchorContinuousDown-streamprocessing,Challengesforabiocatalysisb