Richardson P, Hoare M, Dunnill P
SERC Centre for Biochemical Engineering, Department of Chemical and Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom.
Biotechnol Bioeng. 1990 Aug 5;36(4):354-66. doi: 10.1002/bit.260360406.
A biochemical engineering framework for optimizing the design and operation of fractional protein precipitation has been developed. The method utilizes a fractionation diagram to represent the purification of a product protein relative to total contaminating protein. The purification factor for a single or double-cut fractional precipitation is obtained as the gradient of an appropriate operating tie-line. A computer algorithm has been devised to maximize the tie-line gradient for a given yield enabling a plot of optimum purification factor versus yield to be constructed. The recovery of the enzyme alcohol dehydrogenase from clarified bakers homogenate using saturated ammonium sulphate has been examined. Fractionation and purification versus yield diagrams were used to investigate the effects of such process parameters as pH, temperature, and initial total protein concentration on fractionation efficiency. The results are discussed in terms of the underlying solubility and mixing phenomena and the industrial application of fractional precipitation.
已开发出一种用于优化分级蛋白质沉淀设计与操作的生化工程框架。该方法利用分级图来表示产物蛋白相对于总污染蛋白的纯化情况。单次或双次分级沉淀的纯化因子可作为适当操作连接线的斜率获得。已设计出一种计算机算法，以使给定产率下的连接线斜率最大化，从而构建出最佳纯化因子与产率的关系图。已研究了使用饱和硫酸铵从澄清的面包师匀浆中回收醇脱氢酶的情况。利用分级和纯化与产率关系图来研究诸如pH、温度和初始总蛋白浓度等工艺参数对分级效率的影响。根据潜在的溶解度和混合现象以及分级沉淀的工业应用对结果进行了讨论。
