2细胞的分离与胞内产物的溶解.docVIP

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 In some cases, the products of interest are not in the broth but are in the biomass. In particular, most proteins produced in quantity by genetically manipulated bacteria are not excreted into the broth, but are precipitated within the cell. Lipids and some antibiotics are also trapped in the biomass. In a few cases, like baker抯 yeast, the desired product is the cell mass itself. In a few others, desired products like steroids can be extracted without rupturing the cells. In many cases, the product is trapped in the biomass: It is intracellular. Releasing this trapped material usually involves rupturing the cell wall, and is the subject of this short chapter. The methods of cell rupture have largely been developed in biochemistry, and hence are of small scale. Their application to the larger scale operations implied by genetic engineering is speculative. The equipment which is used is not designed for biotechnology. Some equipment is borrowed from the dairy industry, where it was developed for the homogenization of milk; other equipment is taken from the paint industry, where it is used for the size reduction of pigments. 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 At this point, we pause briefly to explore the physical structure of microbial membranes and the complexity of the problems which we face. At present, knowledge of this general structure does not provide a direct guide to methods of cell rupture. In the future, it may. As a result, we feel that a synopsis has merit. In this synopsis, we emphasize Gram-negative procaryotes. 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 化学法 The chemical methods of cell rupture listed in Table 4.0-1 are dominated by osmotic shock, detergent so1ubilization, and lipid dissoluti