Bhat K M, Sumner I G, Perry B N, Collins M E, Pickersgill R W, Goodenough P W
AFRC Institute of Food Research, Reading Laboratory, Berks, U.K.
Biochem Biophys Res Commun. 1991 Apr 15;176(1):371-7. doi: 10.1016/0006-291x(91)90934-y.
Porcine phospholipaseA2 expressed in E. coli as a fusion protein was isolated, renatured and specifically cleaved by trypsin as described in (1). Active phospholipaseA2, was purified to homogeneity on a column of PBE-94 over a pH region 7.4-4.5. Using this method, several phospholipase A2 mutant enzymes have now been purified in a single step and all behaved identically during chromatofocusing. The method will therefore be extremely useful not only for those interested in understanding the structure-function relationships of phospholipaseA2 but also for preparing the enzyme in large quantities for industrial and pharmaceutical purposes.
如(1)中所述，在大肠杆菌中作为融合蛋白表达的猪磷脂酶A2被分离、复性，并被胰蛋白酶特异性切割。活性磷脂酶A2在PBE - 94柱上于pH 7.4 - 4.5区域被纯化至同质。使用这种方法，现在已经可以一步纯化几种磷脂酶A2突变酶，并且在色谱聚焦过程中它们的行为都相同。因此，该方法不仅对那些有兴趣了解磷脂酶A2结构 - 功能关系的人极为有用，而且对于为工业和制药目的大量制备该酶也非常有用。
