Overexpression, purification and crystallization of the two C-terminal domains of the bifunctional cellulase ctCel9D-Cel44A from Clostridium thermocellum.
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.
Najmudin
Shabir
S
REQUIMTE, Departamento de Química, FCT-UNL, 2829-516 Caparica, Portugal.
Guerreiro
Catarina I P D
CI
Ferreira
Luís M A
LM
Romão
Maria J C
MJ
Fontes
Carlos M G A
CM
Prates
José A M
JA
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
D001426:Bacterial Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; C101903:CelJ protein, Clostridium thermocellum; D002480:Cellulase