大肠杆菌β-葡萄糖醛酸酶的表达与纯化

Expression and purification of Escherichia coli beta-glucuronidase.

作者信息

Aich S, Delbaere L T, Chen R

机构信息

Department of Biochemistry, College of Medicine, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5, Canada.

出版信息

Protein Expr Purif. 2001 Jun;22(1):75-81. doi: 10.1006/prep.2001.1401.

DOI:10.1006/prep.2001.1401

PMID:11388802

Abstract

A strong and constitutive expression vector of Escherichia coli beta-glucuronidase with the isocitrate dehydrogenase promoter has been developed for producing a large amount of recombinant protein. More than 95% pure enzyme was obtained by a four step purification procedure-ammonium sulfate precipitation, DEAE ion-exchange chromatography, Superose 12 gel filtration, and hydroxyapatite steric ion-exchange chromatography. The overexpressed gene can produce 23 mg of pure enzyme from one liter of bacterial culture.

摘要

一种带有异柠檬酸脱氢酶启动子的大肠杆菌β-葡萄糖醛酸酶的高效组成型表达载体已被开发出来，用于大量生产重组蛋白。通过硫酸铵沉淀、DEAE离子交换色谱、Superose 12凝胶过滤和羟基磷灰石空间离子交换色谱这四步纯化程序，获得了纯度超过95%的酶。过表达的基因从一升细菌培养物中可产生23毫克纯酶。