Crystallization and preliminary X-ray crystallographic analysis of a novel α-L-arabinofuranosidase (CtGH43) from Clostridium thermocellum ATCC 27405

Acta Crystallogr F Struct Biol Commun. 2014 May;70(Pt 5):616-8. doi: 10.1107/S2053230X14006402. Epub 2014 Apr 15.

Abstract

The truncated carbohydrate-active enzyme belonging to family 43 glycoside hydrolase from Clostridium thermocellum (CtGH43) is an α-L-arabinofuranosidase that in combination with endoxylanase leads to complete breakdown of L-arabinosyl-substituted xylans. The recombinant enzyme CtGH43 from C. thermocellum was overexpressed in Escherichia coli and purified by immobilized metal-ion affinity chromatography. The recombinant CtGH43 has a molecular mass of 35.86 kDa. Preliminary structural characterization was carried out on CtGH43 crystallized from different conditions, which gave either cube-shaped or brick-shaped crystals. These diffracted to a resolution of 1.65 Å for the cubic form and 1.1 Å for the monoclinic form. Molecular replacement was used to solve the CtGH43 structure.

Keywords: C. thermocellum; CtGH43; glycoside hydrolase; α-l-arabinofuranosidase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Clostridium thermocellum / enzymology*
  • Clostridium thermocellum / genetics*
  • Crystallization
  • Crystallography, X-Ray
  • Glycoside Hydrolases / chemistry*
  • Glycoside Hydrolases / genetics*
  • Glycoside Hydrolases / isolation & purification
  • Molecular Sequence Data

Substances

  • Glycoside Hydrolases
  • alpha-N-arabinofuranosidase