title: Expression, crystallization and preliminary crystallographic data analysis of VioD, a hydroxylase in the violacein-biosynthesis pathway.
abstract: Violacein, a natural purple secondary metabolite, is sequentially biosynthesized by five enzymes in the following pathway: VioA-VioB-VioE-VioD-VioC. VioD, a flavin-dependent oxygenase, catalyzes the hydroxylation of the intermediate product prodeoxyviolaceinic acid (PVA) at the 5-position of one indole ring to yield proviolacein. In vitro biochemical data have revealed this process, but the catalytic mechanism still remains largely unclear. Here, the cloning, expression, purification, crystallization and diffraction of VioD are reported. Crystals of VioD diffracted to 1.7 Å resolution and belonged to space group P31, with unit-cell parameters a = b = 90.0, c = 94.5 Å, α = β = 90, γ = 120°. Solvent-content calculation and molecular-replacement results suggest the presence of two molecules of VioD in the asymmetric unit.
authors: Department of Microbiology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Tingting
authors: T
authors: Ran
authors: Department of Microbiology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Mengxiao
authors: M
authors: Gao
authors: Department of Microbiology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Qiaoe
authors: Q
authors: Wei
authors: Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, People's Republic of China.
authors: Jianhua
authors: J
authors: He
authors: Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, People's Republic of China.
authors: Lin
authors: L
authors: Tang
authors: Department of Microbiology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Weiwu
authors: W
authors: Wang
authors: Department of Microbiology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Dongqing
authors: D
authors: Xu
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D053898:Biosynthetic Pathways; D002846:Chromatography, Affinity; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D007211:Indoles / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000737:chemistry*; D008969:Molecular Sequence Data; D020560:Proteobacteria / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D007211:Indoles; D006899:Mixed Function Oxygenases; C063155:violacein
keywords: VioD; violacein