{Reference Type}: Journal Article
{Title}: Identification and characterization of a novel thermo-stable endo-β-N-acetylglucosaminidase from Rhizomucorpusillus.
{Author}: Nishizawa H;Iwamoto M;Ono Y;
{Journal}: J Biosci Bioeng
{Volume}: 134
{Issue}: 4
{Year}: Oct 2022
{Factor}: 3.185
{DOI}: 10.1016/j.jbiosc.2022.06.013
{Abstract}: Endo-β-N-acetylglucosaminidase (ENGase) is an enzyme that hydrolyzes the chitobiose core of N-glycans and is widely used for glycan analysis on glycoproteins and preparation of precursors for glycosylated compounds. While most of the ENGases that can hydrolyze complex-type glycans are derived from eukaryotes, their production by heterologous expression using Escherichia coli is insufficient, making the production process expensive. From an industrial perspective, there is a need for a less expensive enzyme with higher activity and stability. In this study, we identified a novel ENGase gene from a thermophilic fungus, Rhizomucor pusillus, and named it Endo-Rp. Characterization of the recombinant Endo-Rp showed that the enzyme had maximum hydrolytic activity at 60 °C and hydrolyzed high-mannose-type and biantennary complex-type glycans, but not (2,4)-branched triantennary complex-type or fucosylated glycans. Endo-Rp also hydrolyzed N-glycans attached to RNase B and human transferrin. In summary, we consider Endo-Rp to be a valuable enzyme in various scientific and industrial applications.