Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-beta-D-xylanase 10B in complex with xylohexaose

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Aug 1;64(Pt 8):715-8. doi: 10.1107/S1744309108019696. Epub 2008 Jul 5.

Abstract

The cellulosome of Clostridium thermocellum is a highly organized multi-enzyme complex of cellulases and hemicellulases involved in the hydrolysis of plant cell-wall polysaccharides. The bifunctional multi-modular xylanase Xyn10B is one of the hemicellulase components of the C. thermocellum cellulosome. The enzyme contains an internal glycoside hydrolase family 10 catalytic domain (GH10) and a C-terminal family 1 carbohydrate esterase domain (CE1). The N-terminal moiety of Xyn10B (residues 32-551), comprising a carbohydrate-binding module (CBM22-1) and the GH10 E337A mutant, was crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3(2)21 and contain a dimer in the asymmetric unit. The crystals diffracted to beyond 2.0 A resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Clostridium thermocellum / enzymology*
  • Crystallization
  • Crystallography, X-Ray
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Endo-1,4-beta Xylanases / chemistry*
  • Endo-1,4-beta Xylanases / genetics
  • Endo-1,4-beta Xylanases / isolation & purification
  • Mutagenesis, Site-Directed
  • Protein Conformation
  • Xylans / chemistry*

Substances

  • DNA Primers
  • Xylans
  • 2-(trimethylsilyl)ethyl-O-(2,3-di-O-benzylxylopyranosyl)-(1-4)-bis(O-(2,3-di-O-benzoylxylopyranosyl)-(1-4)-(2,3-di-O-benzylxylopyranosyl)-(1-4))-2,3-di-O-benzoylxylopyranoside
  • Endo-1,4-beta Xylanases