Molecular characterization of acidic peptide:N-glycanase from the dimorphic yeast Yarrowia lipolytica

In this study, we identified and characterized a PNGase A-like enzyme, PNGase Yl, in the dimorphic yeast Yarrowia lipolytica. The corresponding gene was cloned and recombinantly expressed in Pichia pastoris. The purified enzyme cleaved glycans from glycopeptides with the maximum activity at pH 5. No metal ions were required for full activity, and rather it was repressed by three metal ions (Fe3+, Cu2+ and Zn2+). Using glycopeptide substrates, PNGase Yl was shown to release various types of N-glycans including high-mannose and complex-type glycans as well as glycans containing core-linked α(1,3)-fucose that are frequently found in plants and insects. Moreover, in comparison with PNGase A, PNGase Yl was able to cleave with higher efficiency the glycans from some denatured glycoproteins. Taken together, our results suggest that PNGase Yl, the first biochemically characterized yeast PNGase A homologue, can be developed through protein engineering as a useful deglycosylation tool for N-glycosylation study.

Source: Journal of Biochemistry - January 5, 2015 Category: Biochemistry Authors: Lee, K. J., Gil, J. Y., Kim, S.-Y., Kwon, O., Ko, K., Kim, D.-I., Kim, D. K., Kim, H. H., Oh, D.-B. Tags: Regular Papers Source Type: research