Mode of action of glycoside hydrolase family 5 glucuronoxylan xylanohydrolase from Erwinia chrysanthemi

FEBS J. 2007 Apr;274(7):1666-77. doi: 10.1111/j.1742-4658.2007.05710.x.

Abstract

The mode of action of xylanase A from a phytopathogenic bacterium, Erwinia chrysanthemi, classified in glycoside hydrolase family 5, was investigated on xylooligosaccharides and polysaccharides using TLC, MALDI-TOF MS and enzyme treatment with exoglycosidases. The hydrolytic action of xylanase A was found to be absolutely dependent on the presence of 4-O-methyl-D-glucuronosyl (MeGlcA) side residues in both oligosaccharides and polysaccharides. Neutral linear beta-1,4-xylooligosaccharides and esterified aldouronic acids were resistant towards enzymatic action. Aldouronic acids of the structure MeGlcA(3)Xyl(3) (aldotetraouronic acid), MeGlcA(3)Xyl(4) (aldopentaouronic acid) and MeGlcA(3)Xyl(5) (aldohexaouronic acid) were cleaved with the enzyme to give xylose from the reducing end and products shorter by one xylopyranosyl residue: MeGlcA(2)Xyl(2), MeGlcA(2)Xyl(3) and MeGlcA(2)Xyl(4). As a rule, the enzyme attacked the second glycosidic linkage following the MeGlcA branch towards the reducing end. Depending on the distribution of MeGlcA residues on the glucuronoxylan main chain, the enzyme generated series of shorter and longer aldouronic acids of backbone polymerization degree 3-14, in which the MeGlcA is linked exclusively to the second xylopyranosyl residue from the reducing end. Upon incubation with beta-xylosidase, all acidic hydrolysis products of acidic oligosaccharides and hardwood glucuronoxylans were converted to aldotriouronic acid, MeGlcA(2)Xyl(2). In agreement with this mode of action, xylose and unsubstituted oligosaccharides were essentially absent in the hydrolysates. The E. chrysanthemi xylanase A thus appears to be an excellent biocatalyst for the production of large acidic oligosaccharides from glucuronoxylans as well as an invaluable tool for determination of the distribution of MeGlcA residues along the main chain of this major plant hemicellulose.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism
  • Catalysis
  • Dickeya chrysanthemi / enzymology*
  • Glycoside Hydrolases / chemistry
  • Glycoside Hydrolases / metabolism
  • Hydrolysis
  • Kinetics
  • Models, Molecular
  • Molecular Structure
  • Oligosaccharides, Branched-Chain / chemistry*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Substrate Specificity
  • Xylans / chemistry
  • Xylans / metabolism
  • Xylosidases / chemistry*
  • Xylosidases / metabolism

Substances

  • Bacterial Proteins
  • Oligosaccharides, Branched-Chain
  • Xylans
  • 4-O-methyl glucuronoxylan
  • Glycoside Hydrolases
  • Xylosidases
  • glucuronoxylan xylanohydrolase
  • alpha-glucuronidase