STUDIES ON THE ISOLATION OF PENICILLIN ACYLASE FROM ESCHERICHIA-COLI BY AQUEOUS 2-PHASE PARTITIONING

被引：26

作者：

GUAN, Y

WU, XY

TREFFRY, TE

LILLEY, TH

机构：

[1] E CHINA UNIV CHEM TECHNOL, DEPT BIOCHEM, SHANGHAI 200237, PEOPLES R CHINA

[2] UNIV SHEFFIELD, DEPT CHEM, BIOTHERMODYNAM LAB, SHEFFIELD S3 7HF, S YORKSHIRE, ENGLAND

[3] UNIV SHEFFIELD, KREBS INST BIOMOLEC RES, SHEFFIELD S3 7HF, S YORKSHIRE, ENGLAND

[4] UNIV SHEFFIELD, DEPT MOLEC BIOL & TECHNOL, SHEFFIELD S10 2TN, S YORKSHIRE, ENGLAND

来源：

BIOTECHNOLOGY AND BIOENGINEERING | 1992年 / 40卷 / 04期

关键词：

AQUEOUS; 2-PHASE; PENICILLIN ACYLASE; POLYETHYLENE GLYCOL; POLYETHYLENE GLYCOL DERIVATIVES; AFFINITY PARTITIONING; ENZYME RELEASE; EXTRACTION OF INTRACELLULAR ENZYMES;

D O I：

10.1002/bit.260400410

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A method of enzyme release and aqueous two-phase extraction is described for the separation of penicillin acylase from Escherichia coli cells. Butyl acetate, 12% (v/v), treatment combined with freeze-thawing gives up to 70% enzyme release. For polyethylene glycol (PEG) + phosphate two-phase extraction systems the enzyme purity and yield were rather low. Modified PEG, including PEG-ampicillin, PEG-aniline, PEG-phosphate, and PEG-trimethylamine, were synthesized and used in aqueous two-phase systems; PEG-trimethylamine is the most satisfactory. A system containing 12% (w/w) PEG4000, 8% (w/w) of which is PEG-trimethylamine, with 0.7M potassium phosphate at pH 7.2, resulted in the enzyme selective partition being greatly enhanced by charge directed effects. Possible mechanisms for the separation process are discussed.

引用

页码：517 / 524

页数：8

共 30 条

[1]

Albertsson P.-A., 1960, PARTITION CELL PARTI

[2] PARTITION OF PROTEINS IN LIQUID POLYMER-POLYMER 2-PHASE SYSTEMS [J].