Fazi A, Magnani M, Accorsi A, Ninfali P, Fornaini G
Istituto di Chimica Biologica, Università degli Studi di Urbino, Italy.
Prep Biochem. 1988;18(2):153-63. doi: 10.1080/00327488808062518.
A procedure for the simultaneous purification to homogeneity of hexokinase, phosphoglucomutase 1 and 2, aldolase, phosphoglucose isomerase and glucose-6-phosphate dehydrogenase from human origin has been developed. Human placenta homogenate was first chromatographed on DE-52 column which retains hexokinase and glucose-6-phosphate dehydrogenase while the other enzymes are recovered in the unabsorbed protein fraction. The other steps in the purification involve Matrex gel and specific affinity chromatography for the DE-52 retained enzymes and phosphocellulose and Matrex gel chromatography for the other enzymes. All the enzymes mentioned were obtained in one week, with recoveries from 14 percent for glucose-6-phosphate dehydrogenase to 75 percent for hexokinase. Thus, the procedures utilized seem to be useful in obtaining large amounts of enzymes in a a homogeneous form from an easily available human tissue.
已开发出一种从人源同时纯化己糖激酶、磷酸葡萄糖变位酶1和2、醛缩酶、磷酸葡萄糖异构酶和葡萄糖-6-磷酸脱氢酶至均一性的方法。人胎盘匀浆首先在DE-52柱上进行层析，该柱保留己糖激酶和葡萄糖-6-磷酸脱氢酶，而其他酶则在未吸附的蛋白质部分中回收。纯化的其他步骤包括针对DE-52保留的酶进行Matrex凝胶和特异性亲和层析，以及针对其他酶进行磷酸纤维素和Matrex凝胶层析。所有提及的酶在一周内获得，回收率从葡萄糖-6-磷酸脱氢酶的14%到己糖激酶的75%不等。因此，所采用的方法似乎有助于从易于获取的人体组织中以均一形式获得大量酶。
