title: Crystallization and preliminary X-ray study of the deaminase AmnE from Pseudomonas sp. AP-3.
abstract: The amnE gene from Pseudomonas sp. AP-3 has been verified as encoding a deaminase with 142 amino-acid residues. In order to change the substrate specificity via structure-based protein engineering, the amnE gene, after gene-code optimization, was chemically synthesized and cloned into the expression vector pET-28a. The protein was expressed in Escherichia coli BL21 (DE3) and purified by Ni(2+)-chelating affinity chromatography. Diffraction-quality crystals were obtained using the hanging-drop vapour-diffusion method and diffracted to a resolution of 2.09 Å. The crystals belonged to the orthorhombic space group C2221, with unit-cell parameters a = 63.23, b = 88.93, c = 137.83 Å.
authors.affiliation: Tianjin University of Science and Technology, Tianjin 300222, People's Republic of China.
authors.forename: Dan
authors.initials: D
authors.lastname: Yu
authors.forename: Yongji
authors.initials: Y
authors.lastname: Jiang
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Hou
authors.forename: Shuai
authors.initials: S
authors.lastname: Chen
authors.forename: Guofang
authors.initials: G
authors.lastname: Zhang
authors.forename: Xiang
authors.initials: X
authors.lastname: Liu
authors.forename: Hui
authors.initials: H
authors.lastname: Dong
authors.forename: Bo
authors.initials: B
authors.lastname: Yu
mesh_terms: D000619:Aminohydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013295:Streptococcus mutans / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000619:Aminohydrolases
keywords: AmnE; Pseudomonas sp. AP-3