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.
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Similar content being viewed by others Explore related subjectsDiscover the latest articles and news from researchers in related subjects, suggested using machine learning. Author information Authors and AffiliationsBiotechnology Group, Danish Institute of Agricultural Sciences, 1871 Copenhagen, Denmark, , , , ,
M. Skjøt
Molecular Biotechnology, Enzyme Research, Novo Nordisk A/S, 2880 Bagsværd, Denmark, , , , ,
S. Kauppinen, L. Kofod, C. Fuglsang, H. Dalbøge & L. Andersen
Department of Plant Biology, Plant Biochemistry Laboratory, The Royal Veterinary and Agricultural University, 1871 Copenhagen, Denmark, , , , ,
M. Pauly
Electronic Publication
About this article Cite this articleSkjøt, .M., Kauppinen, .S., Kofod, .L. et al. Functional cloning of an endo-arabinanase from Aspergillus aculeatus and its heterologous expression in A. oryzae and tobacco. Mol Gen Genomics 265, 913–921 (2001). https://doi.org/10.1007/s004380100489
Received: 12 October 2000
Accepted: 16 March 2001
Issue Date: July 2001
DOI: https://doi.org/10.1007/s004380100489
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