Open AccessExpression, purification, crystallization and preliminary X‐ray diffraction analysis of a ribokinase from the thermohalophile Halothermothrix orenii
Author(s) -
Kori Lokesh D.,
Hofmann Andreas,
Patel Bharat K. C.
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111041091
Subject(s) - orthorhombic crystal system , escherichia coli , crystallization , recombinant dna , resolution (logic) , crystallography , chemistry , diffraction , crystal (programming language) , crystal structure , materials science , gene , biochemistry , physics , optics , organic chemistry , artificial intelligence , computer science , programming language
A ribokinase gene ( rbk ) from the anaerobic halothermophilic bacterium Halothermothrix orenii was cloned and overexpressed in Escherichia coli . The recombinant protein (Ho‐Rbk) was purified using immobilized metal‐ion affinity chromatography and crystals were obtained using the sitting‐drop method. Diffraction data were collected to a resolution of 3.1 Å using synchrotron radiation. The crystals belonged to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 45.6, b = 61.1, c = 220.2, and contained two molecules per asymmetric unit. A molecular‐replacement solution has been found and attempts are currently under way to build a model of the ribokinase. Efforts to improve crystal quality so that higher resolution data can be obtained are also being considered.