Cloning, purification, crystallization and preliminary X-ray diffraction analysis of nitrile hydratase from the themophilic Bacillus smithii SC-­J05-1

Nitrile hydratase (NHase) converts nitriles to the corresponding amides and is recognized as having important industrial applications. Purification, cloning, crystallization and initial crystallographic studies of the NHase from Bacillus smithii SC‐J05‐1 (Bs NHase) were conducted to analyze the acti...

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Bibliographic Details
Published in:Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2005-11, Vol.61 (11), p.974-977
Main Authors: Hourai, Shinji, Ishii, Takeshi, Miki, Misao, Takashima, Yoshiki, Mitsuda, Satoshi, Yanagi, Kazunori
Format: Article
Language:English
Subjects:
Bacillus - enzymology
Bacillus smithii
Chromatography
Cloning, Molecular
CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY
CRYSTALLIZATION
Crystallization Communications
Crystallography, X-Ray
CRYSTALS
DIFFUSION
Electrophoresis, Polyacrylamide Gel
Hydro-Lyases - chemistry
Light
MAGNESIUM IONS
NHase
Protein Conformation
Protein Structure, Secondary
Protein Structure, Tertiary
Scattering, Radiation
Sepharose - chemistry
SPECIFICITY
STABILITY
X-RAY DIFFRACTION
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Summary:Nitrile hydratase (NHase) converts nitriles to the corresponding amides and is recognized as having important industrial applications. Purification, cloning, crystallization and initial crystallographic studies of the NHase from Bacillus smithii SC‐J05‐1 (Bs NHase) were conducted to analyze the activity, specificity and thermal stability of this hydrolytic enzyme. Bs NHase was purified to homogeneity from microbial cells of B. smithii SC‐J05‐1 and the nucleotide sequences of both the α‐ and β‐subunits were determined. Purified Bs NHase was used for crystallization and several crystal forms were obtained by the vapour‐diffusion method. Microseeding and the addition of magnesium ions were essential components to obtain crystals suitable for X‐ray diffraction analysis.
ISSN:1744-3091
1744-3091
DOI:10.1107/S1744309105030939