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Microbiology 153 (2007), 4003-4015; DOI  10.1099/mic.0.2007/008607-0
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Microbiology 153 (2007), 4003-4015; DOI  10.1099/mic.0.2007/008607-0
© 2007 Society for General Microbiology

Phylogenetic and biochemical characterization of a novel cluster of intracellular fungal {alpha}-amylase enzymes

R. M. van der Kaaij1,2, S. Janecek3, M. J. E. C. van der Maarel1,2,4 and L. Dijkhuizen1,2

1 Microbial Physiology Research Group, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, The Netherlands
2 Centre for Carbohydrate Bioprocessing, TNO-University of Groningen, Haren, The Netherlands
3 Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Slovakia
4 Business Unit Food and Biotechnology Innovations, TNO Quality of Life, Groningen, The Netherlands

Correspondence
M. J. E. C. van der Maarel
m.j.e.c.van.der.maarel{at}rug.nl

Currently known fungal {alpha}-amylases are well-characterized extracellular enzymes that are classified into glycoside hydrolase subfamily GH13_1. This study describes the identification, and phylogenetic and biochemical analysis of novel intracellular fungal {alpha}-amylases. The phylogenetic analysis shows that they cluster in the recently identified subfamily GH13_5 and display very low similarity to fungal {alpha}-amylases of family GH13_1. Homologues of these intracellular enzymes are present in the genome sequences of all filamentous fungi studied, including ascomycetes and basidiomycetes. One of the enzymes belonging to this new group, Amy1p from Histoplasma capsulatum, has recently been functionally linked to the formation of cell wall {alpha}-glucan. To study the biochemical characteristics of this novel cluster of {alpha}-amylases, we overexpressed and purified a homologue from Aspergillus niger, AmyD, and studied its activity product profile with starch and related substrates. AmyD has a relatively low hydrolysing activity on starch (2.2 U mg–1), producing mainly maltotriose. A possible function of these enzymes in relation to cell wall {alpha}-glucan synthesis is discussed.

Abbreviations: GH, glycoside hydrolase; GPI, glycosylphosphatidylinositol






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