title: Overexpression, purification and crystallization of the two C-terminal domains of the bifunctional cellulase ctCel9D-Cel44A from Clostridium thermocellum.
abstract: Clostridium thermocellum produces a highly organized multi-enzyme complex of cellulases and hemicellulases for the hydrolysis of plant cell-wall polysaccharides, which is termed the cellulosome. The bifunctional multi-modular cellulase ctCel9D-Cel44A is one of the largest components of the C. thermocellum cellulosome. The enzyme contains two internal catalytic domains belonging to glycoside hydrolase families 9 and 44. The C-terminus of this cellulase, comprising a polycystic kidney-disease module (PKD) and a carbohydrate-binding module (CBM44), has been crystallized. The crystals belong to the tetragonal space group P4(3)2(1)2, containing a single molecule in the asymmetric unit. Native and seleno-L-methionine-derivative crystals diffracted to 2.1 and 2.8 A, respectively.
authors.affiliation: REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica, Portugal.
authors.forename: Shabir
authors.initials: S
authors.lastname: Najmudin
authors.forename: Catarina I P D
authors.initials: CI
authors.lastname: Guerreiro
authors.forename: Luís M A
authors.initials: LM
authors.lastname: Ferreira
authors.forename: Maria J C
authors.initials: MJ
authors.lastname: Romão
authors.forename: Carlos M G A
authors.initials: CM
authors.lastname: Fontes
authors.forename: José A M
authors.initials: JA
authors.lastname: Prates
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D020134:Catalytic Domain; D002473:Cell Wall / Q000378:metabolism; D002480:Cellulase / Q000737:chemistry*; D002850:Chromatography, Gel; D048013:Acetivibrio thermocellus / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006868:Hydrolysis; D011134:Polysaccharides / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; C101903:CelJ protein, Clostridium thermocellum; D002480:Cellulase