A xylan hydrolase gene cluster in Prevotella ruminicola B(1)4: sequence relationships, synergistic interactions, and oxygen sensitivity of a novel enzyme with exoxylanase and beta-(1,4)-xylosidase activities

A Gasparic, Jennifer Cynthia Martin, A S Daniel, Harry James Flint

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Two genes concerned with xylan degradation were found to be closely linked in the ruminal anaerobe Prevotella ruminicola B(1)4, being separated by an intergenic region of 75 nucleotides. xynA is shown to encode a family F endoxylanase of 369 amino acids, including a putative amino-terminal signal peptide. xynB encodes an enzyme of 319 amino acids, with no obvious signal peptide, that shows 68% amino acid identity with the xsa product of Bacteroides ovatus and 31% amino acid identity with a beta-xylosidase from Clostridium stercorarium; together, these three enzymes define a new family of beta-(1,4)-glycosidases. The activity of the cloned P. ruminicola xynB gene product, but not that of the xynA gene product, shows considerable sensitivity to oxygen. Studied under anaerobic conditions, the XynB enzyme was found to act as an exoxylanase, releasing xylose from substrates including xylobiose, xylopentaose, and birch wood xylan, but was relatively inactive against oat spelt xylan. A high degree of synergy (up to 10-fold stimulation) was found with respect to the release of reducing sugars from oat spelt xylan when XynB was combined with the XynA endoxylanase from P. ruminicola B(1)4 or with endoxylanases from the cellulolytic rumen anaerobe Ruminococcus flavefaciens 17. Pretreatment with a fungal arabinofuranosidase also stimulated reducing-sugar release from xylans by XynB. In P. ruminicola the XynA and XynB enzymes may act sequentially in the breakdown of xylan.

Original languageEnglish
Pages (from-to)2958-64
Number of pages7
JournalApplied and Environmental Microbiology
Volume61
Issue number8
Publication statusPublished - Aug 1995

Keywords

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • DNA, Bacterial
  • Endo-1,4-beta Xylanases
  • Genes, Bacterial
  • Genetic Linkage
  • Molecular Sequence Data
  • Multigene Family
  • Prevotella
  • Rumen
  • Sequence Homology, Amino Acid
  • Xylans
  • Xylosidases
  • beta-Glucosidase

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