生物工程下游技术-第五章ppt课件.ppt

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1、SeparationandpurificationofproductsChapter5Celldisruption,proteinrecoveryandseparation第五章细胞破碎、蛋白质复性和固液分离Inthischapter,Weconsiderthemethodsforcellrupturemostlikelytobeusefulatlargerscales.Thesemethodsareconvenientlysplitintotwogroups,describedas“chemical”and“mechanical”.Intheche

2、micalmethods,thecellmembraneisrupturedbyosmoticpressure,dissolvedbydetergents,orenfeebledbyorganicsolvents.Thesechemicalcasesareoftengentlesothattheproductsarenotirreversiblydenaturedevenwhilethecellsareruptured.Scale-upiseasy:Ifwewanttotreat10timesthebiomass,weneedtouse10times

3、asmuchchemical.Themechanicalmethodsaredominatedbyhomogenizationorballmillgrinding.Inbothcases,cellsarebrokenbyhighmechanicalshear.Heatgenerationisaproblem.TABLE1.ChemicalCellDisintegrationTechniquesMethodTechniquePrincipleStressCostExamplesChemicalOsmoticshockOsmoticruptureofme

4、mbraneGentleCheapRuptureofredbloodcellsEnzymedigestionCellwalldigested,providingdisruptionGentleExpensiveEgglysozymeSolubilizationDetergentssolubilizecellmembraneGentleModerate-expensiveBilesaltsactingonE.coliLipiddissolutionOrganicsolventdissolvesincellwallModerateCheapToluene

5、disruptionofyeastAlkalitreatmentSaponificationoflipidssolubilizesmembraneHarshCheap方法技术原理效果成本举例化学法渗透冲击渗透压破坏细胞温和便宜血红细胞的破坏酶消化法细胞壁被消化，使细胞破碎温和昂贵增溶法表面活性剂溶解细胞壁温和适中胆盐作用于大肠杆菌脂溶法有机溶剂溶解细胞壁并使之失稳适中便宜甲苯破碎酵母细胞碱处理法碱的皂化作用使细胞壁融解剧烈便宜表1.细胞化学破碎法MethodTechniquePrincipleProductCostExamplesMechanical

6、Homogenization(bladetype)CellschoppedinWaringblenderModerateModerateAnimaltissueandcellsGrindingCellsrupturedbygrindingwithabrasivesModerateCheapUltrasonicationCellsbrokenwithultrasoniccavitationHarshExpensiveCellsuspensionsonasmallscaleHomogenization(orificetype)Cellsforcedthr

7、oughsmallholearebrokenbyshearHarshModerateLargescaletreatmentofcellsuspensions,CrushinginballmillCellscrushedbetweenglassorsteelballsHarshCheapLargescaletreatmentofcellsuspensionsandplantcellsTABLE2.MechanicalCellDisintegrationTechniques方法技术原理效果成本举例机械法匀浆法（片型）细胞被搅拌器劈碎适中适中动物组织及动物

8、细胞研磨法细胞被研磨物磨碎适中便宜超声波法用超声波的空穴作用使细胞破碎适中昂贵细胞悬浮液小规模处理匀浆法（孔