Discovery and characterization of a novel extremely acidic bacterial N-glycanase with combined advantages of PNGase F and A |
| |
Authors: | Ting Wang Zhi?P Cai Xiao?Q Gu Hong?Y Ma Ya?M Du Kun Huang Josef Voglmeir Li Liu |
| |
Institution: | *Glycomics and Glycan Bioengineering Research Center, College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, People''s Republic of China;†Department of Plant Pathology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, People''s Republic of China |
| |
Abstract: | Peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidases PNGases (peptide N-glycosidases), N-glycanases, EC 3.5.1.52] are essential tools in the release of N-glycans from glycoproteins. We hereby report the discovery and characterization of a novel bacterial N-glycanase from Terriglobus roseus with an extremely low pH optimum of 2.6, and annotated it therefore as PNGase H+. The gene of PNGase H+ was cloned and the recombinant protein was successfully expressed in Escherichia coli. The recombinant PNGase H+ could liberate high mannose-, hybrid- and complex-type N-glycans including core α1,3-fucosylated oligosaccharides from both glycoproteins and glycopeptides. In addition, PNGase H+ exhibited better release efficiency over N-glycans without core α1,3-fucose compared with PNGase A. The facile expression, non-glycosylated nature, unusual pH optimum and broad substrate specificity of this novel type of N-glycanase makes recombinant PNGase H+ a versatile tool in N-glycan analysis. |
| |
Keywords: | carbohydrate processing core fucosylation glycoprotein structure N-linked glycosylation PNGase Terriglobus |
|
|