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

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Dec 1;61(Pt 12):1043-5. doi: 10.1107/S1744309105035670. Epub 2005 Nov 5.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Catalytic Domain
  • Cell Wall / metabolism
  • Cellulase / chemistry*
  • Chromatography, Gel
  • Clostridium thermocellum / metabolism*
  • Crystallography, X-Ray
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / metabolism
  • Hydrolysis
  • Polysaccharides / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry

Substances

  • Bacterial Proteins
  • Polysaccharides
  • Recombinant Proteins
  • CelJ protein, Clostridium thermocellum
  • Cellulase