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Molecular Genetics and Genomics

, Volume 265, Issue 5, pp 913–921 | Cite as

Functional cloning of an endo-arabinanase from Aspergillus aculeatus and its heterologous expression in A. oryzae and tobacco

  •  M. Skjøt
  •  S. Kauppinen
  •  L. Kofod
  •  C. Fuglsang
  •  M. Pauly
  •  H. Dalbøge
  •  L. Andersen
Original Paper

Abstract.

Functional cloning in yeast has been used to isolate full-length cDNAs encoding an endo-α-1,5-L-arabinanase from the filamentous fungus Aspergillus aculeatus. Screening of a cDNA library constructed in a yeast expression vector for transformants that hydrolysed AZCL-arabinan identified 44 Saccharomyces cerevisiae clones all harbouring the same arabinanase-encoding cDNA. The cloned cDNA was expressed in A. oryzae and the recombinant enzyme was purified and characterized. The mode of action of the enzyme was studied by analysis of the digestion pattern towards debranched arabinan. The digestion profile obtained strongly suggests that the enzyme is an endo-arabinanase. In addition, the feasibility using Nicotiana tabacum as an alternative host for arabinanase expression was investigated.

Endo-arabinanase Aspergillus aculeatus Expression cloning Molecular farming 

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Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  •  M. Skjøt
    • 1
  •  S. Kauppinen
    • 2
  •  L. Kofod
    • 2
  •  C. Fuglsang
    • 2
  •  M. Pauly
    • 3
  •  H. Dalbøge
    • 2
  •  L. Andersen
    • 2
  1. 1.Biotechnology Group, Danish Institute of Agricultural Sciences, 1871 Copenhagen, Denmark
  2. 2.Molecular Biotechnology, Enzyme Research, Novo Nordisk A/S, 2880 Bagsværd, Denmark
  3. 3.Department of Plant Biology, Plant Biochemistry Laboratory, The Royal Veterinary and Agricultural University, 1871 Copenhagen, Denmark

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