Identifying and characterising a plant GH1 β‐glucosidase that exhibits hydrolytic activity on N ‐linked glucopyranoside

β‐Glucosidases (Bgls) catalyse the hydrolysis of β‐D‐glycosidic bonds and are widely exploited as industrial biocatalysts. While their activity on O ‐ and S ‐linked glycopyranosides is well established, hydrolysis of N ‐linked glycopyranosides by Bgls has not previously been demonstrated. Here, we screened a eukaryotic Bgl library for activity on methyl anthranilate‐ N ‐β‐D‐glucopyranoside (MANT‐ N ‐glucose), a novel N ‐glucopyranoside we recently produced biocatalytically. We identify ZmGlu1 as capable of hydrolysing this substrate, albeit with substantially reduced catalytic efficiency relative to native O ‐glucopyranoside substrates. Structural modelling of the enzyme–substrate complex reveals interactions likely underlying this reduced activity. These findings establish N ‐glycopyranosides as Bgl substrates and lay the groundwork for engineering Bgls with broader glycosidic bond specificity.