Substrate specificity of novel GH16 endo-β-(1→3)-galactanases acting on linear and branched β-(1→3)-galactooligosaccharides

J Biotechnol. 2019 Jan 20:290:44-52. doi: 10.1016/j.jbiotec.2018.12.006. Epub 2018 Dec 18.

Abstract

Arabinogalactan proteins are proteoglycans located in the plant cell wall. Most arabinogalactan proteins are composed of carbohydrate moieties of β-(1→3)-galactan main chains with β-(1→6)-galactan side chains terminated by other glycans. In this study, three novel endo-β-(1→3)-galactanases were identified and the substrate specificity was further studied using well-defined galactan oligomers. Linear and branched β-(1→3)-linked galactans, which resemble the carbohydrate core of the arabinogalactan protein, were used for the characterization of endo-β-(1→3)-galactanases. The identified enzymes required at least three consecutive galactose residues for activity. Non-substituted regions were preferred, but substituents in the -2 and +2 and in some cases also -1 and +1 subsites were tolerated to some extent, depending on the branching pattern, however at a significantly lower rate/frequency.

Keywords: Glycosyl hydrolase (GH) family 16; Gum arabic; Type II arabinogalactan; β-(1→3)-Galactan; β-(1→3)-Galactanases.

MeSH terms

  • Aspergillus oryzae / genetics
  • Carbohydrate Conformation
  • Cloning, Molecular
  • Galactose / chemistry
  • Galactose / metabolism*
  • Oligosaccharides / chemistry
  • Oligosaccharides / metabolism*
  • Plant Proteins* / chemistry
  • Plant Proteins* / genetics
  • Plant Proteins* / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • beta-Galactosidase* / chemistry
  • beta-Galactosidase* / genetics
  • beta-Galactosidase* / metabolism

Substances

  • Oligosaccharides
  • Plant Proteins
  • Recombinant Proteins
  • beta-Galactosidase
  • Galactose