Overexpression, purification and crystallization of the two C-terminal domains of the bifunctional cellulase ctCel9D-Cel44A from Clostridium thermocellum

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 61; no. 12; pp. 1043 - 1045
• Main Authors: Najmudin, Shabir, Guerreiro, Catarina I. P. D., Ferreira, Luís M. A., Romão, Maria J. C., Fontes, Carlos M. G. A., Prates, José A. M.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.12.2005; International Union of Crystallography
• Subjects: Bacterial Proteins - chemistry; carbohydrate-binding module; Catalytic Domain; Cell Wall - metabolism; cellulase; Cellulase - chemistry; Chromatography, Gel; Clostridium thermocellum; Clostridium thermocellum - metabolism; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; Crystallization Communications; Crystallography, X-Ray; CRYSTALS; Electrophoresis, Polyacrylamide Gel; Escherichia coli - metabolism; glycoside hydrolase; Hydrolysis; MOLECULES; Polysaccharides - metabolism; Protein Binding; Protein Structure, Tertiary; Recombinant Proteins - chemistry; SPACE GROUPS; X-RAY DIFFRACTION
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309105035670

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 P43212, containing a single molecule in the asymmetric unit. Native and seleno‐l‐methionine‐derivative crystals diffracted to 2.1 and 2.8 Å, respectively.