title: Cloning, purification, crystallization and preliminary X-ray diffraction analysis of nitrile hydratase from the themophilic Bacillus smithii SC-J05-1.
abstract: 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 alpha- and beta-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.
authors.affiliation: Sumitomo Chemical Co. Ltd Environmental Health Science Laboratory, 3-1-98 Kasugade-naka, Konohanaku, Osaka 554-8558, Japan. shinji-horai@ds-pharma.co.jp
authors.forename: Shinji
authors.initials: S
authors.lastname: Hourai
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ishii
authors.forename: Misao
authors.initials: M
authors.lastname: Miki
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Takashima
authors.forename: Satoshi
authors.initials: S
authors.lastname: Mitsuda
authors.forename: Kazunori
authors.initials: K
authors.lastname: Yanagi
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D002845:Chromatography; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006836:Hydro-Lyases / Q000737:chemistry*; D008027:Light; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012542:Scattering, Radiation; D012685:Sepharose / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D012685:Sepharose; D006836:Hydro-Lyases; C048831:nitrile hydratase