Expression, purification, crystallization and initial crystallographic characterization of the p-hydroxybenzoate hydroxylase from Corynebacterium glutamicum

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 63; no. 11; pp. 944 - 946
• Main Authors: Kwon, Soo-Young, Kang, Beom Sik, Kim, Ghyung-Hwa, Kim, Kyung-Jin
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Blackwell Publishing Ltd 01.11.2007; International Union of Crystallography
• Subjects: 4-Hydroxybenzoate-3-Monooxygenase - biosynthesis; 4-Hydroxybenzoate-3-Monooxygenase - chemistry; 4-Hydroxybenzoate-3-Monooxygenase - isolation & purification; BIODEGRADATION; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; Corynebacterium glutamicum; Corynebacterium glutamicum - enzymology; CRYSTALLIZATION; Crystallization Communications; Crystallography, X-Ray; CRYSTALS; DENSITY; DIFFUSION; FAD-dependent monooxygenases; MOLECULES; OXYGEN; p-hydroxybenzoate hydroxylase; RESOLUTION; SPACE GROUPS; STOWING; SYNCHROTRONS; X-RAY DIFFRACTION
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309107046386

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 NaH2PO4 and K2HPO4 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 P6322, 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 Å3 Da−1. The structure was solved by molecular replacement. Structure refinement is in progress.