title: Crystallization and preliminary crystallographic analysis of a family 43 beta-D-xylosidase from Geobacillus stearothermophilus T-6.
abstract: Beta-D-Xylosidases (EC 3.2.1.37) are hemicellulases that cleave single xylose units from the nonreducing end of xylooligomers. In this study, the crystallization and preliminary X-ray analysis of a beta-D-xylosidase from Geobacillus stearothermophilus T-6 (XynB3), a family 43 glycoside hydrolase, is described. XynB3 is a 535-amino-acid protein with a calculated molecular weight of 61 891 Da. Purified recombinant native and catalytic inactive mutant proteins were crystallized and cocrystallized with xylobiose in two different space groups, P2(1)2(1)2 (unit-cell parameters a = 98.32, b = 99.36, c = 258.64 A) and P4(1)2(1)2 (or the enantiomorphic space group P4(3)2(1)2; unit-cell parameters a = b = 140.15, c = 233.11 A), depending on the detergent. Transferring crystals to cryoconditions required a very careful protocol. Orthorhombic crystals diffract to 2.5 A and tetragonal crystals to 2.2 A.
authors.affiliation: Institute for Biochemistry, University of Cologne, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Brüx
authors.forename: Karsten
authors.initials: K
authors.lastname: Niefind
authors.forename: Alon
authors.initials: A
authors.lastname: Ben-David
authors.forename: Maya
authors.initials: M
authors.lastname: Leon
authors.forename: Gil
authors.initials: G
authors.lastname: Shoham
authors.forename: Yuval
authors.initials: Y
authors.lastname: Shoham
authors.forename: Dietmar
authors.initials: D
authors.lastname: Schomburg
mesh_terms: D002021:Buffers; D002384:Catalysis; D015925:Cryopreservation; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004187:Disaccharides / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000737:chemistry; D008970:Molecular Weight; D009154:Mutation; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction; D014995:Xylosidases / Q000737:chemistry*
chemical_list: D002021:Buffers; D004187:Disaccharides; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; D014995:Xylosidases; C026579:exo-1,4-beta-D-xylosidase; C004173:xylobiose