Single-step purification of a recombinant thermostable α-amylase after solubilization of the enzyme from insoluble aggregates

被引：16

作者：

Linden, A ^{[1
]}

Niehaus, F ^{[1
]}

Antranikian, G ^{[1
]}

机构：

[1] Tech Univ Hamburg, Dept Tech Microbiol, D-21071 Hamburg, Germany

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2000年 / 737卷 / 1-2期

关键词：

purification; Pyrococcus woesei; Escherichia coli; Halomonas elongata; aggregation; inclusion bodies; solubilization; enzymes; alpha-amylase;

D O I：

10.1016/S0378-4347(99)00364-3

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The expression of the gene encoding a thermostable a-amylase (EC 3.2.1.1) (optimal activity at 100 degrees C) from the hyperthermophilic archaeon Pyrococcus woesei in the mesophilic hosts Escherichia coli and Halomonas elongata resulted in the formation of insoluble aggregates. More than 85% of the recombinant enzyme was present within the cells as insoluble but catalytically active aggregates. The recombinant alpha-amylase was purified to homogeneity in a single step by hydrophobic interaction chromatography on a phenyl superose column after solubilization of the enzyme under nondenaturing conditions. The enzyme was purified 258-fold with a final yield of 54%. (C) 2000 Published by Elsevier Science B.V. All rights reserved.

引用

页码：253 / 259

页数：7

共 22 条

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