European Journal of Plant Pathology

, Volume 101, Issue 3, pp 291–299 | Cite as

Purification and partial characterization of xylanase from the fungal maize pathogenHelminthosporium turcicum (Pass)

  • Yeshitila Degefu
  • Richard Fagerström
  • Nisse Kalkkinen
Article

Abstract

The fungal pathogenHelminthosporium turcicum was found to secret xylanase when grown on minimal medium containing xylans, wheat straw or isolated maize cell walls. The highest xylanase activity occurre when the fungus was grown on maize cell walls. When glucose was added to this medium xylanase activity was suppressed. The xylanase enzyme was purified from the culture filtrate by subsequent anion exchange chromatography, cation exchange chromatography and gel filtration. The purified xylanase gave a single band on sodium dodecyl sulphate polyacrylamide gel electrophoresis corresponding to an apparent molecular weight of 22.5 kDa. It is determined to have a pI of 7.4, specific activity of 11300 nanokatals mg−1, pH optimum between pH 5.5 and 6.5 and optimal temperature between 50 °C and 60 °C. The half-life of the enzyme at pH 6.0 and 50 °C was found to be 35 min. For primary structure comparison with other xylanases, the protein was digested with trypsin and the resulting peptides were separated by reversed phase chromatography and selected peptides were sequenced. The determined amino acid sequence showed high homology with xylanase fromCochliobolus carbonum and three other fungal xylanases.

Key words

cell wall degrading enzymes Helminthosporium turcicum xylanase 

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Yeshitila Degefu
    • 1
  • Richard Fagerström
    • 2
  • Nisse Kalkkinen
    • 3
  1. 1.Department of Plant Biology, Plant and Forest PathologyUniversity of HelsinkiFinland
  2. 2.Alko Research LaboratoriesAlko LTDHelsinkiFinland
  3. 3.Institute of Biotechnology, Protein Chemistry LaboratoryUniversity of HelsinkiFinland

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