Open AccessExpression, purification, crystallization and initial crystallographic characterization of the p ‐hydroxybenzoate hydroxylase from Corynebacterium glutamicum
Author(s) -
Kwon SooYoung,
Kang Beom Sik,
Kim GhyungHwa,
Kim KyungJin
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309107046386
Subject(s) - corynebacterium glutamicum , hydroxybenzoate , hydroxylation , crystallization , crystallography , molecule , chemistry , x ray crystallography , stereochemistry , diffraction , enzyme , biochemistry , organic chemistry , gene , physics , optics
p ‐Hydroxybenzoate hydroxylase (PHBH) is an FAD‐dependent monooxygenase that catalyzes the hydroxylation of p ‐hydroxybenzoate (pOHB) to 3,4‐dihydroxybenzoate in an NADPH‐dependent reaction and plays an important role in the biodegradation of aromatic compounds. PHBH from Corynebacterium glutamicum was crystallized using the hanging‐drop vapour‐diffusion method in the presence of NaH 2 PO 4 and K 2 HPO 4 as precipitants. X‐ray diffraction data were collected to a maximum resolution of 2.5 Å on a synchrotron beamline. The crystal belongs to the hexagonal space group P 6 3 22, with unit‐cell parameters a = b = 94.72, c = 359.68 Å, γ = 120°. The asymmetric unit contains two molecules, corresponding to a packing density of 2.65 Å 3 Da −1 . The structure was solved by molecular replacement. Structure refinement is in progress.