Expression and Characterization of a Rice β-Xylosidase with Xylooligosaccharide Hydrolysis and Transglycosylation Activities

Plant β-xylosidases are less well characterized for hemicellulose degradation than their microbial counterparts. To address this, a broadly expressed rice (<i>Oryza sativa</i>) glycoside hydrolase family 3 (GH3) β-xylosidase designated OsXyl1 was expressed in heterologous <i>Pichia pastoris</i>. OsXyl1 showed maximal enzyme activity at pH 4.0 and 60 °C. It was relatively stable at 30-50 °C. It hydrolyzed 4NP-β-<i>d</i>-xylopyranoside (4NPXyl) and β-1,4-linked xylooligosaccharides (XOS) with degrees of polymerization (DP) of 2-6. OsXyl1 hydrolylsis of 4NPXyl was much more rapid and specific than that of other 4NP glycosides with an apparent <i>k</i>_cat/<i>K</i>_m value of 19.0 mM^-1 s^-1. OsXyl1 had similar specificity toward XOS having DP values of 2-5 with apparent <i>k</i>_cat/<i>K</i>_m values of 2.6-4.2 mM^-1 s^-1. OsXyl1 was also efficient at transglycosylating short alcohols with 4NPXyl and XOS xylosyl donors. Therefore, rice OsXyl1 β-xylosidase may function in recycling of xylans in plant cell wall recycling and it may be applied for transglycosylation of alcohol acceptors.