PURIFICATION AND PROPERTIES OF HUMAN GLUCOSE-6-PHOSPHATE-DEHYDROGENASE MADE IN ESCHERICHIA-COLI

被引：51

作者：

BAUTISTA, JM ^{[1
]}

MASON, PJ ^{[1
]}

LUZZATTO, L ^{[1
]}

机构：

[1] ROYAL POSTGRAD MED SCH, DEPT HAEMATOL, DUNCANE RD, LONDON W12 ONN, ENGLAND

来源：

BIOCHIMICA ET BIOPHYSICA ACTA | 1992年 / 1119卷 / 01期

关键词：

GLUCOSE-6-PHOSPHATE DEHYDROGENASE; EXPRESSION; PURIFICATION; (ESCHERICHIA-COLI);

D O I：

10.1016/0167-4838(92)90237-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The cDNA for the X-chromosome encoded human glucose-6-phosphate dehydrogenase (G6PD) has been expressed in E. coli and the enzyme purified to homogeneity, using a simple one-step fractionation on 2'5'-ADP-Sepharose. By selecting one of several different expression vectors and by optimizing culture conditions a yield of more than 10 mg of pure enzyme per liter of culture is obtained reproducibly. When the recombinant enzyme and authentic G6PD purified from normal human red cells were compared, they proved to be indistinguishable by the following criteria: electrophoretic mobility in both native and denaturing conditions, the K(m) values for glucose 6-phosphate and NADP and the K(i) value for NADPH. The recombinant enzyme, unlike the red cell enzyme, retained 100% activity when stored at 4-degrees-C for over 1 year.

引用

页码：74 / 80

页数：7

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