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1、Chapter4CellDisruption第四章细胞破碎※4.1 CellMembranes细胞膜※4.0Summary概述※4.2 ChemicalMethods化学破碎方法※4.3mechanicaldisruption机械破碎※4.0Summary概述Bioseparationsusuallybeginwiththeseparationofbiomassfrombroth.TheseparationcommonlyusesfiltrationorcentrifugationasdescribedinChapters2an
2、d3.Inmanycases,thedesiredproductisinthebroth.Antibiotics,arecommonlyinthebroth;soareextracellularenzymes,manypolysaccharides,andmostaminoacids.Inallthesecases,theseparatedbrothcanbetreatedtoisolateandpurifytheproduct,asabyproduct.生物分离的第一步是将生物机体从发酵液中分离,通常使用过滤和离心等方法,这在第
3、二、三章中已有陈述。大多数情况下,抗生素,胞外酶,一些多糖,及氨基酸等目标产物存在于在发酵液中。在上述过程中,需被分离的发酵液可被看作一种副产物来处理,以此分离和纯化产物。Insomecases,theproductsofinterestarenotinthebrothbutareinthebiomass.Inparticular,mostproteinsproducedinquantitybygeneticallymanipulatedbacteriaarenotexcretedintothebroth,butareprecip
4、itatedwithinthecell.Lipidsandsomeantibioticsarealsotrappedinthebiomass.Inafewcases,likebaker’syeast,thedesiredproductisthecellmassitself.Inafewothers,desiredproductslikesteroidscanbeextractedwithoutrupturingthecells.Inmanycases,theproductistrappedinthebiomass:Itisintr
5、acellular.有些目标产物不在发酵液中,而是存在于生物体中。尤其是由基因工程菌产生的大多数蛋白质不会被分泌到发酵液中,而是在细胞内沉积。脂类物质和一些抗生素也是包含在生物体中。还有一些目标产物就是细胞本身,如面包酵母。还有,产物如类固醇不必通过细胞破碎提取。大多数情况下,产物还是包裹在生物体内,属胞内产物。Releasingthistrappedmaterialusuallyinvolvesrupturingthecellwall,andisthesubjectofthisshortchapter.Themethodsofc
6、ellrupturehavelargelybeendevelopedinbiochemistry,andhenceareofsmallscale.Theirapplicationtothelargerscaleoperationsimpliedbygeneticengineeringisspeculative.Theequipmentwhichisusedisnotdesignedforbiotechnology.使胞内产物释放出来一般需要破碎细胞壁,这也是这一章的主题。细胞破碎的方法在生物化学领域中得到了很广泛的运用,但多数在小
7、规模生产中,在大规模生产尤其是基因工程中应用极少。Inthischapter,webegininSection4.1byreviewingthestructureofthecellwall.Wethenconsiderthemethodsforcellrupturemostlikelytobeusefulatlargerscales.Thesemethods,listedinTable4.0-1,areconvenientlysplitintotwogroups,describedas“chemical”and“mechanica
8、l.”InthechemicalmethodsdetailedinSection4.2,thecellmembraneisrupturedbyosmoticpressure,dissolvedbydetergents,orenfeebledbyor