Background: Peptide: N-glycanase is a deglycosylation enzyme releasing N-glycan from
glycoproteins. Although glycan specificity analysis of this enzyme has been reported, recognition
requirements for the peptide sequence have not been precisely elucidated.
Objective: In this study, we carried out peptide specificity analysis of several peptide:N-glycanases.
Methods: Using synthetic chitobiose-pentapeptide substrates having a systematic series of amino
acid sequences composed of hydrophobic leucine and hydrophilic serine, we examined the peptide
specificities of peptide: N-glycanases comprising yeast cytoplasmic PNGase, bacterial PNGase F,
and plant PNGase A by ultra-performance liquid chromatography combined with electrospray ionization
Results: We found that each of the PNGases had higher activity for the more hydrophobic (leucinerich)
chitobiose-pentapeptides, although the sensitivities of the PNGases for hydrophobicity varied.
Cytoplasmic PNGase showed broad specificity. In contrast, PNGase A showed moderate specificity.
PNGase F showed the highest specificity.
Conclusion: PNGases from different origins had similar but significantly independent peptide