Enterococcus faecalis α1-2-mannosidase (EfMan-I): an efficient catalyst for glycoprotein N-glycan modification

While multiple alpha 1-2-mannosidases are necessary for glycoprotein N-glycan maturation in vertebrates, a single bacterial alpha 1-2-mannosidase can be sufficient to cleave all alpha 1-2-linked mannose residues in host glycoprotein N-glycans. We report here the characterization and crystal structure of a new alpha 1-2-mannosidase (EfMan-I) from Enterococcus faecalis, a Gram-positive opportunistic human pathogen. EfMan-I catalyzes the cleavage of alpha 1-2-mannose from not only oligomannoses but also high-mannose-type N-glycans on glycoproteins. Its 2.15 angstrom resolution crystal structure reveals a two-domain enzyme fold similar to other CAZy GH92 mannosidases. An unexpected potassium ion was observed bridging two domains near the active site. These findings support EfMan-I as an effective catalyst for in vitro N-glycan modification of glycoproteins with high-mannose-type N-glycans.