Ucon-benzoyl dextran aqueous two-phase systems: Protein purification with phase component recycling

被引：20

作者：

Lu, M ^{[1
]}

Albertsson, PA ^{[1
]}

Johansson, G ^{[1
]}

Tjerneld, F ^{[1
]}

机构：

[1] LUND UNIV,CTR CHEM,DEPT BIOCHEM,S-22100 LUND,SWEDEN

来源：

JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS | 1996年 / 680卷 / 1-2期

关键词：

aqueous two-phase system; partitioning; protein; benzoyl dextran; Ucon;

D O I：

10.1016/0378-4347(95)00484-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Benzoyl dextran with a degree of substitution of 0.18 was synthesized by reacting dextran T500 with benzoyl chloride. A new type of aqueous two-phase system composed of benzoyl dextran as bottom phase polymer and the random copolymer of ethylene oxide and propylene oxide (Ucon 50-KB-5100) as top phase polymer has been formed. The phase diagram for the system Ucon 50-HB-5100-benzoyl dextran with a degree of substitution of 0.18 was determined at room temperature. This two-phase system has been used to purify 3-phosphoglycerate kinase from baker's yeast. The top-phase polymer (Ucon) can be separated from target enzyme by increasing the temperature. The bottom-phase polymer (benzoyl dextran) could be recovered by addition of salt. Yeast homogenate was partitioned in a primary Ucon 50-HB-5100-benzoyl dextran aqueous two-phase system. After phase separation the top phase was removed and temperature-induced phase separation was used for formation of a water phase and a Ucon-rich phase. The benzoyl dextran-enriched bottom phase from the primary system was diluted, and the polymer was separated from water by addition of Na2SO4.

引用

页码：65 / 70

页数：6

共 14 条

[1]

Albertsson P.A., 1986, PARTITION CELL PARTI

[2]

ALRED PA, 1993, J CHROMATOGR, V628, P205

[3] APPLICATION OF TEMPERATURE-INDUCED PHASE PARTITIONING AT AMBIENT-TEMPERATURE FOR ENZYME-PURIFICATION [J].