Phylogenetic and biochemical characterization of a novel cluster of intracellular fungal α-amylase enzymes

• Published in: Microbiology (Society for General Microbiology) Vol. 153; no. Pt 12; pp. 4003 - 4015
• Main Authors: VAN DER KAAIJ, R. M, JANECEK, S, VAN DER MAAREL, M. J. E. C, DIJKHUIZEN, L
• Format: Journal Article
• Language: English
• Published: Reading Society for General Microbiology 01.12.2007
• Subjects: alpha-Amylases - chemistry; alpha-Amylases - genetics; alpha-Amylases - metabolism; Amino Acid Sequence; Animals; Aspergillus niger - enzymology; Aspergillus niger - genetics; Biological and medical sciences; Cell Wall - metabolism; Fundamental and applied biological sciences. Psychology; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - metabolism; Fungi - chemistry; Fungi - classification; Fungi - enzymology; Fungi - genetics; Glucans - metabolism; Growth, nutrition, metabolism, transports, enzymes. Molecular biology; Histoplasma - enzymology; Histoplasma - genetics; Microbiology; Molecular Sequence Data; Mycology; Phylogeny; Sequence Alignment; Starch - metabolism; Substrate Specificity; Glycosylases; Characterization; Fungi; Glucan; Enzyme; Glycosidases; Oligosaccharide; Hydrolases; Intracellular; Phylogeny; α-Amylase
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/mic.0.2007/008607-0

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.