Overproduction, purification, crystallization and preliminary X-ray characterization of a novel carbohydrate-binding module of endoglucanase Cel5A from Eubacterium cellulosolvens

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Apr 1;67(Pt 4):491-3. doi: 10.1107/S1744309111004246. Epub 2011 Mar 25.

Abstract

The anaerobic cellulolytic rumen bacterium Eubacterium cellulosolvens produces a large array of cellulases and hemicellulases that are responsible for the hydrolysis of plant cell-wall polysaccharides. One of these enzymes, endoglucanase Cel5A, comprises two tandemly repeated novel carbohydrate-binding modules (CBMs) and two catalytic domains belonging to glycoside hydrolase family 5 joined by flexible linker sequences. The novel CBM located at the N-terminus of the endoglucanase has been crystallized. The crystals belonged to the hexagonal space group P6(1)22 and contained a single molecule in the asymmetric unit. The structure of the L-selenomethionine derivative has been solved by a MAD experiment on crystals that diffracted to 1.75 Å resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbohydrate Metabolism
  • Cellulase / chemistry*
  • Cellulase / isolation & purification
  • Cellulase / metabolism
  • Crystallization
  • Crystallography, X-Ray
  • Eubacterium / enzymology*
  • Protein Binding

Substances

  • endoglucanase Cel5A
  • Cellulase