title: Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-beta-D-xylanase 10B in complex with xylohexaose.
abstract: The cellulosome of Clostridium thermocellum is a highly organized multi-enzyme complex of cellulases and hemicellulases involved in the hydrolysis of plant cell-wall polysaccharides. The bifunctional multi-modular xylanase Xyn10B is one of the hemicellulase components of the C. thermocellum cellulosome. The enzyme contains an internal glycoside hydrolase family 10 catalytic domain (GH10) and a C-terminal family 1 carbohydrate esterase domain (CE1). The N-terminal moiety of Xyn10B (residues 32-551), comprising a carbohydrate-binding module (CBM22-1) and the GH10 E337A mutant, was crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3(2)21 and contain a dimer in the asymmetric unit. The crystals diffracted to beyond 2.0 A resolution.
authors.affiliation: REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica, Portugal. shabir@dq.fct.unl.pt
authors.forename: Shabir
authors.initials: S
authors.lastname: Najmudin
authors.forename: Benedita A
authors.initials: BA
authors.lastname: Pinheiro
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Romão
authors.forename: José A M
authors.initials: JA
authors.lastname: Prates
authors.forename: Carlos M G A
authors.initials: CM
authors.lastname: Fontes
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D048013:Acetivibrio thermocellus / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D014990:Xylans / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D014990:Xylans; C074759:2-(trimethylsilyl)ethyl-O-(2,3-di-O-benzylxylopyranosyl)-(1-4)-bis(O-(2,3-di-O-benzoylxylopyranosyl)-(1-4)-(2,3-di-O-benzylxylopyranosyl)-(1-4))-2,3-di-O-benzoylxylopyranoside; D043364:Endo-1,4-beta Xylanases