Expression, purification, crystallization and preliminary crystallographic analysis of a GH20 ?-N-acetylglucosaminidase from the marine bacterium Vibrio harveyi.

ABSTRACT: Vibrio harveyi ?-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 (GlcNAc? or diNAG) used for soaking was cleaved by the active enzyme, leaving only a single GlcNAc molecule bound to the protein.

SUBMITTER: Meekrathok P 

PROVIDER: S-EPMC4388178 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature