Purification of a D-hydantoinase using a laboratory-scale Streamline phenyl column as the initial step

被引：10

作者：

Abendroth, J ^{[1
]}

Chatterjee, S ^{[1
]}

Schomburg, D ^{[1
]}

机构：

[1] Univ Cologne, Inst Biochem, D-50674 Cologne, Germany

来源：

JOURNAL OF CHROMATOGRAPHY B | 2000年 / 737卷 / 1-2期

关键词：

purification; enzymes; D-hydantoinase;

D O I：

10.1016/S0378-4347(99)00443-0

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A D-hydantoinase from Thermus sp. was overexpressed in Escherichia coli and purified to homogeneity for subsequent crystallization. The purification was performed with hydrophobic interaction chromatography as the capture step followed by anion-exchange chromatography and gel permeation chromatography as intermediate purification and polishing steps, respectively. The hydrophobic interaction step was done in fluidized bed mode in a laboratory-scale Streamline column made from conventional laboratory equipment. The whole purification protocol could be finished within one day. The purified enzyme crystallizes. The crystals are suitable for X-ray protein structure analysis and diffract to at least 2.3 Angstrom resolution. Complete data sets have been measured up to 2.6 Angstrom resolution. The X-ray structure is currently being solved. (C) 2000 Published by Elsevier Science BN. All rights reserved.

引用

页码：187 / 194

页数：8

共 12 条

[1]

ABENDROTH J, 1999, 5 WORKSH CRYST MACR

[2] Dynamic light scattering in evaluating crystallizability of macromolecules [J].