Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-beta-D-xylanase 10B in complex with xylohexaose.
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.
Najmudin
Shabir
S
REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica, Portugal. shabir@dq.fct.unl.pt
Pinheiro
Benedita A
BA
Romão
Maria J
MJ
Prates
José A M
JA
Fontes
Carlos M G A
CM
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*
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