Affinity partitioning of glucose-6-phosphate dehydrogenase and hexokinase in aqueous two-phase systems with free triazine dye ligands

被引：22

作者：

Xu, Y

Vitolo, M

Albuquerque, CN

Pessoa, A

机构：

[1] Univ Sao Paulo, Biochem Pharmaceut Technol Dept, FCF, BR-05508900 Sao Paulo, Brazil

[2] Hebei Inst Technol, Dept Chem Engn, Tangshan 063009, Hebei, Peoples R China

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2002年 / 780卷 / 01期

基金：

巴西圣保罗研究基金会;

关键词：

partitioning; aqueous two-phase systems; glucose-6-phosphate dehydrogenase; hexokinase; enzymes; triazine dyes;

D O I：

10.1016/S1570-0232(02)00409-9

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The partitioning of glucose-6-phosphate dehydrogenase (G6PDH) (E.C. 1.1.1.49) and hexokinase (E.C. 2.7.1.1) in polyethylene glycol (PEG)-hydroxypropyl starch (PES) and PEG-phosphate aqueous two-phase systems was investigated with free triazine dyes, Cibacron Blue F3GA and Procion Red HE3B, as their affinity ligands. It was found that the free reactive triazine dyes, not bound to phase-forming polymers, preferentially partitioned in the top-PEG phase in the PEG-salt and PEG-PES systems. The effect of various parameters such as type and concentration of affinity ligands, pH of the system, molecular mass of PEG and phase composition on partitioning of the enzymes was estimated. Phosphate is a key factor affecting the enzyme partitioning in the PEG-PES system. Cibacron F3GA changed the partition coefficient of G6PDH from 0.73 to 1.59. (C) 2002 Elsevier Science B.V. All rights reserved.

引用

页码：53 / 60

页数：8

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