PARTIAL-PURIFICATION OF GLUCOSE-6-PHOSPHATE-DEHYDROGENASE AND PHOSPHOFRUCTOKINASE FROM RAT ERYTHROCYTE HEMOLYSATE BY PARTITIONING IN AQUEOUS 2-PHASE SYSTEMS

被引：20

作者：

DELGADO, C ^{[1
]}

TEJEDOR, MC ^{[1
]}

LUQUE, J ^{[1
]}

机构：

[1] UNIV ALCALA DE HENARES, DEPT BIOQUIM & BIOL MOLEC, E-28871 ALCALA DE HENARES, SPAIN

来源：

JOURNAL OF CHROMATOGRAPHY | 1990年 / 498卷 / 01期

关键词：

D O I：

10.1016/S0021-9673(01)84244-8

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Glucose 6-phosphate dehydrogenase shows a high partition coefficient in poly(ethylene glycol)-dextran aqueous two-phase systems in comparison with those for 6-phosphogluconate dehydrogenase, phosphofructokinase and the bulk of proteins present in rat erythrocyte haemolysates. As a consequence, fractions highly enriched in glucose 6-phosphate dehydrogenase can be obtained after multiple partitions in the above systems with a counter-current distribution procedure. Phosphofructokinase shows a high affinity for Cibacron Blue and, as a result, the enzyme can be extracted in the top phase of poly(ethylene glycol)-dextran systems containing Cibacron Bluepoly(ethylene glycol) (affinity systems). The efficiency for the purification of the enzymes by partitioning is increased up to 10-fold when enzyme-rich fractions, obtained by precipitation with poly(ethylene glycol), are used instead of original haemolysate. The recovery of enzyme activities is near 100% in both instances. © 1990.

引用

页码：159 / 168

页数：10

共 21 条

[1]

Albertsson P. -<Angstrom>., 1985, PARTITION CELL PARTI

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3]

Brooks D. E., 1985, PARTITIONING AQUEOUS, P11

[4]

FLANAGAN SD, 1975, J BIOL CHEM, V250, P1484

[5] PROTEIN RECOVERY USING 2-PHASE SYSTEMS [J].

HUSTEDT, H ;

KRONER, KH ;

MENGE, U ;

KULA, MR .

TRENDS IN BIOTECHNOLOGY, 1985, 3 (06) :139-144

[6]

Hustedt H., 1985, PARTITIONING AQUEOUS, P529

[7] PREPARATION OF CIBACRON BLUE F3G-A (POLYETHYLENE-GLYCOL) IN LARGE-SCALE FOR USE IN AFFINITY PARTITIONING [J].

JOHANSSON, G ;

JOELSSON, M .

BIOTECHNOLOGY AND BIOENGINEERING, 1985, 27 (05) :621-625

[8] AFFINITY PARTITIONING OF PHOSPHOFRUCTOKINASE FROM BAKERS-YEAST USING POLYMER-BOUND CIBACRON BLUE F3G-A [J].

JOHANSSON, G ;

KOPPERSCHLAGER, G ;

ALBERTSSON, PA .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1983, 131 (03) :589-594

[9]

JOHANSSON G, 1984, J CHROMATOGR, V291, P175, DOI 10.1016/S0021-9673(00)95018-0

[10] COUNTERCURRENT DISTRIBUTION OF YEAST ENZYMES WITH POLYMER-BOUND TRIAZINE DYE AFFINITY LIGANDS [J].

JOHANSSON, G ;

ANDERSSON, M ;

AKERLUND, HE .

JOURNAL OF CHROMATOGRAPHY, 1984, 298 (03) :483-493

← 1 2 3 →