AUTHOR=Guo Rui-Rui , Comamala Gerard , Yang Huan-Huan , Gramlich Marius , Du Ya-Min , Wang Ting , Zeck Anne , Rand Kasper Dyrberg , Liu Li , Voglmeir Josef 

TITLE=Discovery of Highly Active Recombinant PNGase H+ Variants Through the Rational Exploration of Unstudied Acidobacterial Genomes

JOURNAL=Frontiers in Bioengineering and Biotechnology

VOLUME=8

YEAR=2020

URL=https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2020.00741

DOI=10.3389/fbioe.2020.00741

ISSN=2296-4185

ABSTRACT=<p>Peptide-<italic>N</italic><sup>4</sup>-(<italic>N</italic>-acetyl-β-glucosaminyl) asparagine amidases (PNGases, <italic>N</italic>-glycanases, EC 3.5.1.52) are indispensable tools in releasing <italic>N</italic>-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<sup>+</sup> candidates (the suffix H<sup>+</sup> 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 <italic>Dyella japonica</italic>) 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<sup>+</sup>, broad substrate specificity, and good storage stability of this novel <italic>N</italic>-glycanase makes it a valuable tool for the analysis of protein glycosylation.</p>