Purification, characterization, and cell wall localization of an alpha-fucosidase that inactivates a xyloglucan oligosaccharin

Plant J. 1993 Mar;3(3):415-26. doi: 10.1046/j.1365-313x.1993.t01-24-00999.x.

Abstract

An alpha-fucosidase that releases fucosyl residues from oligosaccharide fragments of xyloglucan, a plant cell wall hemicellulosic polysaccharide, was purified to homogeneity from pea (Pisum sativum) epicotyls using a combination of cation exchange chromatography and isoelectric focusing. The alpha-fucosidase has a molecular mass of 20 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The alpha-fucosidase has an isoelectric point of 5.5. The substrate specificity of the alpha-fucosidase was determined by high performance anion exchange chromatographic analysis of oligosaccharide substrates and products. The enzyme hydrolyzes the terminal alpha-1,2-fucosidic linkage of oligosaccharides and does not cleave p-nitrophenyl-alpha-L-fucoside. The enzyme does not release measurable amounts of fucosyl residues from large polysaccharides. The subcellular localization of alpha-fucosidase in pea stems and leaves has been studied by immunogold cytochemistry. The alpha-fucosidase accumulates in primary cell walls and is not detectable in the middle lamella or in the cytoplasm of 8-day-old stem tissue and 14-day-old leaf tissue. alpha-Fucosidase activity was readily detected in extracts of 8-day-old stem tissue. No significant alpha-fucosidase activity or immunogold labeling of the alpha-fucosidase was detected in 2- and 4-day-old stem tissue indicating that production of alpha-fucosidase is developmentally regulated.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibody Specificity
  • Carbohydrate Sequence
  • Cell Wall / enzymology*
  • Cell Wall / metabolism
  • Fabaceae / enzymology*
  • Fabaceae / metabolism
  • Fabaceae / ultrastructure
  • Glucans*
  • Immunohistochemistry
  • Molecular Sequence Data
  • Oligosaccharides / isolation & purification
  • Oligosaccharides / metabolism*
  • Plants, Medicinal*
  • Polysaccharides / metabolism*
  • Substrate Specificity
  • Xylans*
  • alpha-L-Fucosidase / immunology
  • alpha-L-Fucosidase / isolation & purification
  • alpha-L-Fucosidase / metabolism*

Substances

  • Glucans
  • Oligosaccharides
  • Polysaccharides
  • Xylans
  • xyloglucan
  • alpha-L-Fucosidase