嗜热脂肪芽孢杆菌6-磷酸葡萄糖异构酶两种同工酶的表达、纯化及结晶

Expression, purification, and crystallization of two isozymes of 6-phosphoglucose isomerase of Bacillus stearothermophilus.

作者信息

Hsiao C D, Chou C C, Hsiao Y Y, Sun Y J, Meng M

机构信息

Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, 11529, Republic of China.

出版信息

J Struct Biol. 1997 Nov;120(2):196-200. doi: 10.1006/jsbi.1997.3913.

DOI:10.1006/jsbi.1997.3913

PMID:9417984

Abstract

Two isozymes of 6-phosphoglucose isomerase (phosphoglucose isomerase A & phosphoglucose isomerase B), isolated from Bacillus stearothermophilus, have been overexpressed in Escherichia coli strain DF2145 and purified to homogeneity. Crystals of both isozymes have been obtained by the vapor diffusion method. The crystals of phosphoglucose isomerase A have unit cell dimensions a = b = 132.0 A, c = 183.6 A, and diffract to about 2.8 A resolution. An analysis of the reflection data indicates that the crystal system is hexagonal, space group P6122 or P6522. The crystals of phosphoglucose isomerase B complexed with 6-phosphogluconate belong to the orthorhombic space group I222 (or I212121), with cell dimensions a = 75.1 A, b = 95.7 A, c = 171.5 A, and diffract to a resolution of 2.3 A. These crystals promise to yield more detail for the substrate recognition and higher resolution structures of 6-phosphoglucose isomerase.

摘要

从嗜热脂肪芽孢杆菌中分离出的6-磷酸葡萄糖异构酶的两种同工酶（磷酸葡萄糖异构酶A和磷酸葡萄糖异构酶B）已在大肠杆菌DF2145菌株中过表达并纯化至同质。两种同工酶的晶体均通过气相扩散法获得。磷酸葡萄糖异构酶A的晶体晶胞参数为a = b = 132.0 Å，c = 183.6 Å，衍射分辨率约为2.8 Å。对衍射数据的分析表明，晶体系统为六方晶系，空间群为P6122或P6522。与6-磷酸葡萄糖酸复合的磷酸葡萄糖异构酶B的晶体属于正交空间群I222（或I212121），晶胞参数为a = 75.1 Å，b = 95.7 Å，c = 171.5 Å，衍射分辨率为2.3 Å。这些晶体有望为6-磷酸葡萄糖异构酶的底物识别和更高分辨率结构提供更多细节。