Expression, purification, crystallization and preliminary X-ray diffraction analysis of Thermotoga neapolitana beta-glucosidase B.

Abstract

Beta-glucosidases belong to families 1, 3 and 9 of the glycoside hydrolases and act on cello-oligosaccharides. Family 1 and 3 enzymes are retaining and are reported to have transglycosylation activity, which can be used to produce oligosaccharides and glycoconjugates. Family 3 enzymes are less well characterized than their family 1 homologues and to date only two crystal structures have been solved. Here, the expression, purification, crystallization and X-ray diffraction data of a family 3 beta-glucosidase from the hyperthermophilic bacterium Thermotoga neapolitana are reported. Crystals of selenomethionine-substituted protein have also been grown. The crystals belong to space group C222(1), with unit-cell parameters a = 74.9, b = 127.0, c = 175.2 A. Native data have been collected to 2.4 A resolution and the structure has been solved to 2.7 A using the selenomethionine MAD method. Model building and refinement of the structure are under way.