N-Glycosylation patterns in two α-l-arabinofuranosidases from Penicillium canescens belonging to the glycoside hydrolase families 51 and 54

•N-Glycosylation of two Penicillium canescens α-l-arabinofuranosidases was studied.•Enzymes belong to the glycoside hydrolase families 51 and 54 (Abf51A and Abf54A).•N-Linked glycans were found at 5 out of 8 potential N-glycosylation sites in Abf51A.•One out of 3 potential N-glycosylation sites in Abf54A was glycosylated.•Formula of N-linked glycans: (Man)x(GlcNAc)2, where x varied from 7 to 0.

Using MALDI-TOF mass spectrometry (MS) peptide fingerprinting procedure followed by the analysis of MS data with the GlycoMod tool from the ExPASy proteomic site, N-glycosylation of two GH51 and GH54 family α-l-arabinofuranosidases (Abf51A and Abf54A) from Penicillium canescens was studied. Variable N-linked glycans were identified at five out of eight potential N-glycosylation sites in the Abf51A and one out of three potential N-glycosylation sites in the Abf54A. The discriminated glycans represented high-mannose oligosaccharides (Man)x(GlcNAc)2 with a number of Man residues up to 7 or the products of sequential enzymatic trimming of a high-mannose glycan with α-mannosidases and β-N-acetylhexosaminidases. The Abf54A peptide, containing the Asn254 glycosylation site, and one peptide from the Abf51A, containing the Asn163 glycosylation site, were found to exist not only in glycosylated, but also in a native non-modified form.

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