Peptide--(-acetyl-β-glucosaminyl) asparagine amidases (PNGases, -glycanases, EC 3.5.1.52) are indispensable tools in releasing -glycans from glycoproteins. So far, only a limited number of PNGase candidates are available for the structural analysis of glycoproteins and their glycan moieties. Herein, a panel of 13 novel PNGase H candidates (the suffix H refers to the acidic pH optimum of these acidobacterial PNGases) was tested in their recombinant form for their deglycosylation performance. One candidate (originating from the bacterial species ) showed superior properties both in solution-phase and immobilized on amino-, epoxy- and nitrilotriacetate resins when compared to currently acidic available PNGases. The high expression yield compared to a previously described PNGase H, broad substrate specificity, and good storage stability of this novel -glycanase makes it a valuable tool for the analysis of protein glycosylation.