Open AccessCrystallization and preliminary crystallographic analysis of a family 43 β‐ d ‐xylosidase from Geobacillus stearothermophilus T‐6
Author(s) -
Brüx Christian,
Niefind Karsten,
BenDavid Alon,
Leon Maya,
Shoham Gil,
Shoham Yuval,
Schomburg Dietmar
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105036262
Subject(s) - geobacillus stearothermophilus , crystallization , orthorhombic crystal system , crystallography , xylobiose , xylose , chemistry , tetragonal crystal system , recombinant dna , glycoside hydrolase , space group , thermophile , crystal structure , stereochemistry , enzyme , biochemistry , x ray crystallography , diffraction , fermentation , organic chemistry , physics , gene , optics
β‐ 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 β‐ 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, P 2 1 2 1 2 (unit‐cell parameters a = 98.32, b = 99.36, c = 258.64 Å) and P 4 1 2 1 2 (or the enantiomorphic space group P 4 3 2 1 2; unit‐cell parameters a = b = 140.15, c = 233.11 Å), depending on the detergent. Transferring crystals to cryoconditions required a very careful protocol. Orthorhombic crystals diffract to 2.5 Å and tetragonal crystals to 2.2 Å.