title: Expression, purification, crystallization and preliminary crystallographic analysis of a GH20 β-N-acetylglucosaminidase from the marine bacterium Vibrio harveyi.
abstract: Vibrio harveyi β-N-acetylglucosaminidase (VhGlcNAcase) is a new member of the GH20 glycoside hydrolase family responsible for the complete degradation of chitin fragments, with N-acetylglucosamine (GlcNAc) monomers as the final products. In this study, the crystallization and preliminary crystallographic data of wild-type VhGlcNAcase and its catalytically inactive mutant D437A in the absence and the presence of substrate are reported. Crystals of wild-type VhGlcNAcase were grown in 0.1 M sodium acetate pH 4.6, 1.4 M sodium malonate, while crystals of the D437A mutant were obtained in 0.1 M bis-tris pH 7.5, 0.1 M sodium acetate, 20% PEG 3350. X-ray data from the wild-type and the mutant crystals were collected at a synchrotron-radiation light source and were complete to a resolution of 2.5 Å. All crystals were composed of the same type of dimer, with the substrate N,N'-diacetylglucosamine (GlcNAc₂ or diNAG) used for soaking was cleaved by the active enzyme, leaving only a single GlcNAc molecule bound to the protein.
authors: Biochemistry-Electrochemistry Research Unit, School of Biochemistry, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
authors: Piyanat
authors: P
authors: Meekrathok
authors: Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.
authors: Marco
authors: M
authors: Bürger
authors: Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.
authors: Arthur T
authors: AT
authors: Porfetye
authors: Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.
authors: Ingrid R
authors: IR
authors: Vetter
authors: Biochemistry-Electrochemistry Research Unit, School of Biochemistry, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
authors: Wipa
authors: W
authors: Suginta
mesh_terms: D000118:Acetylglucosaminidase / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D014733:Vibrio / Q000201:enzymology* / Q000235:genetics
chemical_list: D000118:Acetylglucosaminidase
keywords: GH20 glycoside hydrolase family; Vibrio harveyi; β-N-acetylglucosaminidase