生物分离工程-双语版-6培训资料.ppt

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1、生物分离工程-双语版-6Bioseparationsteps1Purificationsteps:(1)Removalofinsolubles(2)Isolationofproduct(3)Purification(4)Polishing2Twotechniquesinthischapter(1)Reversemicellesextraction(2)Aqueoustwo-phasesystems3Fornewtechniquesshouldbe:(1)Screen(2)Develop(3)Analyze(4)

2、Reject4Shouldnotbe:ignoreorminimizeimportanceofmoreestablishedanddevelopedtechniquesIIliquid-liquidextractionAdvantage(1)Large-scaleprocesses(2)Highthroughputs(3)Easytooperate(4)FlexiblemilderconditionsANophasechangeBEnergy-savingprocessesAReversemicellesext

3、raction1Reversiblemicellesareaggregatesofsurfactantmoleculesinorganicsolvents(1)Innercoreofwater(2)Innerlayermadeupofsurfactantmoleculewithpolarionicgroups(3)Externallayercomposedofahydrocarbonchain2W0:themoleratioofthewatertothesurfactant3Affinityreversemic

4、ellespartition4Proteinspartitioningfactors(1)Volumeofthetwophases(2)Polarsolvent(3)Ionicstrength(4)pH(5)surfactants蛋白质在反胶束中的溶解反胶束萃取蛋白质示意图Example6.1Thermodynamicanalysisofactivityandstabilityofglobularproteins1Materials:lysozyme,ribonucleaseA,cytochromeC2Met

5、hod:scanningcalorimetry(DSC)3W0action(1)GlubularproteinsexhibitamaximumofactivityatW08-12(2)ProteinsstabilitywillbedecreaseatmorethanoptimumW0(3)Amonolayerofwateraroundproteinmoleculeissufficienttopreservetheactivityandstabilityofprotein.(4)Conformationalfle

6、xibilityofglubularproteinsdecreaseinreversemicellesMethodsofproteinssolubilizedinreversemecilles1Dryproteinspowderaddedtoasolutioncontainingreversemicelles(1)Sizeofreversemicellesshouldroughlybethesizeofproteinsmolecule(2)Energeticsisunfavorable(3)Minimalene

7、rgybarrierleadtoaminimumofproteinstructuralchangesasproteinsenterthereversemicelles2Proteinsolutioninjectedintoanorganicsolution(1)lesserdegreeofproteinstructuralchanges(2)micellesizedoesnotaffectproteinsolubilization3Phase–transfermethod(1)organicandaqueous

8、phaseareinequilibrium(2)proteinstransferabovePIasexpectedwhenpositivechargedsurfactants(3)proteinstransferbelowPIishinderedwhennegativechargedsurfactants蛋白质溶解方式Moreinformationrequirementsaboutre