Discovery and characterization of a novel extremely acidic bacterial N-glycanase with combined advantages of PNGase F and A

  • Biosci Rep. 2014 Nov 14;34(6):e00149. doi: 10.1042/BSR20140148.
Ting Wang  1 ,  Zhi P Cai  1 ,  Xiao Q Gu  1 ,  Hong Y Ma  2 ,  Ya M Du  1 ,  Kun Huang  1 ,  Josef Voglmeir  1 ,  Li Liu  1
Affiliations
  • 1. *Glycomics and Glycan Bioengineering Research Center, College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, People's Republic of China.
  • 2. †Department of Plant Pathology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, People's Republic of China.
PMID: 25294009 DOI: 10.1042/BSR20140148
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.

Products