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1、第二讲细胞破碎2学时一、通过本章学习应掌握的内容1、细胞破碎的意义2、常用的细胞破碎方法3、渗透压计算方法4、了解机械破碎法所用设备5、渗透压的计算二、细胞破碎的目的Insomecases,theproductsofinterestarenotinthebrothbutareinthebiomass.Inparticular,mostproteinsproducedinquantitybygeneticallymanipulatedbacteriaarenotexcretedintothebroth,bu
2、tareprecipitatedwithinthecell.Lipidsandsomeantibioticsarealsotrappedinthebiomass.Inafewcases,likebaker’syeast,thedesiredproductisthecellmassitself.Inafewothers,desiredproductslikesteroidscanbeextractedwithoutrupturingthecells.Inmanycases,theproductistrapp
3、edinthebiomass:Itisintracellular.Releasingthistrappedmaterialusuallyinvolvesrupturingthecellwall,andisthesubjectofthisshortchapter.Themethodsofcellrupturehavelargelybeendevelopedinbiochemistry,andhenceareofsmallscale.Theirapplicationtothelargerscaleoperat
4、ionsimpliedbygeneticengineeringisspeculative.Theequipmentwhichisusedisnotdesignedforbiotechnology.Someequipmentisborrowedfromthedairyindustry,whereitwasdevelopedforthehomogenizationofmilk;otherequipmentistakenfromthepaintindustry,whereitisusedforthesizere
5、ductionofpigments.由于有许多生化物质存在于细胞内部,必须在纯化以前将细胞破碎,使胞内物质释放到液相中,然后方可进行提取三、细胞膜Atthispoint,wepausebrieflytoexplorethephysicalstructureofmicrobialmembranesandthecomplexityoftheproblemswhichweface.Atpresent,knowledgeofthisgeneralstructuredoesnotprovideadirectguid
6、etomethodsofcellrupture.Inthefuture,itmay.Asaresult,wefeelthatasynopsishasmerit.Inthissynopsis,weemphasizeGram-negativeprocaryotes.本节主要探寻细胞膜的物质结构及我们所面对的几个复杂问题。现在所有的关于细胞膜结构的基本知识,并不能为细胞破碎方法提供基本的引导。也许将来可以。正因如此,对此进行提纲切领的介绍很有必要。主要强调的是革兰氏阴性原核生物。其细胞结构中没有细胞核:基因物质
7、位于单链DNA上。典型的生物是大肠杆菌,是生物技术研究的主体。通过这种细胞生产了很多细胞重组的产物三、细胞破碎的主要方法1、化学法ThechemicalmethodsofcellrupturelistedinTable4.0-1aredominatedbyosmoticshock,detergentso1ubilization,andlipiddissolution.Thesethreemethodsaredescribedinthefollowingparagraphs.Beforethisdescri
8、ption,wewanttogivesomebriefgeneralizationsabouttheothertwomethods:enzymedigestionandalkalitreatment.渗透冲击:无论动物还是植物细胞,胞内物质都是由膜组织包裹起来的,因此,破坏细胞膜系统是胞内物质释放的关键步骤通常细胞膜系统强度较差,易受渗透压冲击而破碎Thesimplestofthethreemajorchemicalmetho