Purification, crystallization and preliminary X-ray diffraction analysis of thermostable nitrile hydratase: research letter
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Date
2004
Journal Title
Journal ISSN
Volume Title
Publisher
Academy of Science of South Africa (ASSAf)
Abstract
Microbial nitrile hydratases are important industrial enzymes
that catalyse the conversion of nitriles to the corresponding
amides. Bacillus strain RAPc8 nitrile hydratase has recently been
cloned and functionally expressed in E. coli. Here, the purification,
crystallization and preliminary X-ray diffraction data of this nitrile
hydratase are described. The heterotetrameric enzyme was
crystallized using the hanging-drop vapour-diffusion method.
Crystals produced in the presence of 30% PEG 400, 0.1 M MES
(pH 6.5) and 0.1 M magnesium chloride were selected for X-ray
diffraction studies. A data set complete to 2.5 Å was collected under
cryoconditions at the in-house X-ray source at the University of the
Western Cape. The space group was determined to be primitive
tetragonal (P41212) with unit cell dimensions a = 106.61 Å, b =
106.61 Å, c=83.23 Å, = = =90°; with one dimer per asymmetric
unit. Solution of the three-dimensional structure via molecular
replacement is in progress.
Description
Keywords
X-ray diffraction analysis, Thermostable protein, Nitrile hydratase, Bacillus pallidus, Bacillus strain RAPc8
Citation
Tsekoa, T.L., et al. (2004). Purification, crystallization and preliminary X-ray diffraction analysis of thermostable nitrile hydratase: research letter. South African Journal of Science, 100: 488-490