Novel xylan-binding properties of an engineered family 4 carbohydrate-binding module

Biochem J. 2007 Sep 1;406(2):209-14. doi: 10.1042/BJ20070128.

Abstract

Molecular engineering of ligand-binding proteins is commonly used for identification of variants that display novel specificities. Using this approach to introduce novel specificities into CBMs (carbohydrate-binding modules) has not been extensively explored. Here, we report the engineering of a CBM, CBM4-2 from the Rhodothermus marinus xylanase Xyn10A, and the identification of the X-2 variant. As compared with the wild-type protein, this engineered module displays higher specificity for the polysaccharide xylan, and a lower preference for binding xylo-oligomers rather than binding the natural decorated polysaccharide. The mode of binding of X-2 differs from other xylan-specific CBMs in that it only has one aromatic residue in the binding site that can make hydrophobic interactions with the sugar rings of the ligand. The evolution of CBM4-2 has thus generated a xylan-binding module with different binding properties to those displayed by CBMs available in Nature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / chemistry
  • Amides / metabolism
  • Amino Acid Sequence
  • Endo-1,4-beta Xylanases / chemistry*
  • Endo-1,4-beta Xylanases / classification
  • Endo-1,4-beta Xylanases / genetics
  • Endo-1,4-beta Xylanases / metabolism*
  • Ligands
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation / genetics
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Binding
  • Protein Structure, Tertiary
  • Rhodothermus / enzymology
  • Rhodothermus / genetics
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Solubility
  • Structural Homology, Protein
  • Thermodynamics
  • Xylans / chemistry*
  • Xylans / metabolism*
  • beta-Glucans / metabolism

Substances

  • Amides
  • Ligands
  • Xylans
  • beta-Glucans
  • Endo-1,4-beta Xylanases