title: Crystallization and preliminary crystallographic studies of the recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114.
abstract: Dihydropyrimidinases are involved in the reductive pathway of pyrimidine degradation, catalysing the hydrolysis of 5,6-dihydrouracil and 5,6-dihydrothymine to the corresponding N-carbamoyl beta-amino acids. This enzyme has often been referred to as hydantoinase owing to its industrial application in the production of optically pure amino acids starting from racemic mixtures of 5-monosubstituted hydantoins. Recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114 (SmelDhp) has been expressed, purified and crystallized. Crystallization was performed using the counter-diffusion method with capillaries of 0.3 mm inner diameter. Crystals of SmelDhp suitable for data collection and structure determination were grown in the presence of agarose at 0.1%(w/v) in order to ensure mass transport controlled by diffusion. X-ray data were collected to a resolution of 1.85 A. The crystal belongs to the orthorhombic space group C222(1), with unit-cell parameters a = 124.89, b = 126.28, c = 196.10 A and two molecules in the asymmetric unit. A molecular-replacement solution has been determined and refinement is in progress.
authors.affiliation: Departamento de Química Física, Bioquímica y Química Inorgánica, Area de Bioquímica y Biología Molecular, Edificio CITE I, Universidad de Almería, Spain.
authors.forename: Sergio
authors.initials: S
authors.lastname: Martínez-Rodríguez
authors.forename: Luis Antonio
authors.initials: LA
authors.lastname: González-Ramírez
authors.forename: Josefa María
authors.initials: JM
authors.lastname: Clemente-Jiménez
authors.forename: Felipe
authors.initials: F
authors.lastname: Rodríguez-Vico
authors.forename: Francisco Javier
authors.initials: FJ
authors.lastname: Las Heras-Vázquez
authors.forename: Jose A
authors.initials: JA
authors.lastname: Gavira
authors.forename: Juan Manuel
authors.initials: JM
authors.lastname: García-Ruíz
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D011994:Recombinant Proteins / Q000737:chemistry; D016962:Sinorhizobium meliloti / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D000581:Amidohydrolases; C020047:dihydropyrimidinase