Plant Molecular Biology

, Volume 26, Issue 3, pp 909–921 | Cite as

Avenacosidase from oat: purification, sequence analysis and biochemical characterization of a new member of the BGA family of β-glucosidases

  • Sabine Gus-Mayer
  • Harald Brunner
  • Hansjörg A. W. Schneider-Poetsch
  • Wolfhart Rüdiger
Research Articles

Abstract

A protein consisting of 60 kDa subunits (As-P60) was isolated from etiolated oat seedlings (Avena sativa L.) and characterized as avenacosidase, a β-glucosidase that belongs to a preformed defence system of oat against fungal infection. The enzyme is highly aggregated; it consists of 300–350 kDa aggregates and multimers thereof. Dissociation by freezing/thawing leads to complete loss of enzyme activity. The specificity of the enzyme was investigated with para-nitrophenyl derivatives which serve as substrates, in decreasing order β-fucoside, β-glucoside, β-galactoside, β-xyloside. The corresponding orthonitrophenyl glycosides are less well accepted. No hydrolysis was found with α-glycosides and β-thioglucoside. An anti-As-P60 antiserum was prepared and used for isolation of a cDNA clone coding for As-P60. A presequence of 55 amino acid residues was deduced from comparison of the cDNA sequence with the N-terminal sequence determined by Edman degradation of the mature protein. The presequence has the characteristics of a stroma-directing signal peptide; localization of As-P60 in plastids of oat seedlings was confirmed by western blotting. The amino acid sequence revealed significant homology (>39% sequence identity) to β-glucosidases that are constituents of a defence mechanism in dicotyledonous plants. 34% sequence identity was even found with mammalian and bacterial β-glucosidases of the BGA family. Avenacosidase extends the occurrence of this family of β-glucosidases to monocotyledonous plants.

Key words

Avena sativa L. avenacosidase β-glucosidases BGA family defence system phytochrome 

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Sabine Gus-Mayer
    • 1
  • Harald Brunner
    • 1
  • Hansjörg A. W. Schneider-Poetsch
    • 2
  • Wolfhart Rüdiger
    • 1
  1. 1.Botanisches InstitutUniversität MünchenMünchenGermany
  2. 2.Botanisches Institut der Universität zu KölnKölnGermany

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