Preparation of ginsenoside Rd using a novel α-L-arabinofuranosidase BpAbf51A from Bacillus pumilus

α-L-Arabinofuranosidase has been widely used in the fields of enhancing pasta production quality, fruit juice clarification, enhancing wine flavor, increasing feed utilization rate, and developing special drug ingredients, playing a pivotal role in the food processing, feed, and medical care industries. In this study, a novel α-L-arabinofuranosidase (BpAbf51A), belonging to glycoside hydrolase family 51 (GH51), was cloned from the Bacillus pumilus strain 145 and expressed in Escherichia coli BL21 (DE3), with a molecular weight of approximately 56.0 kDa. BpAbf51A comprises two characteristic domains of the GH51 family: an N-terminal (β/α)₈-barrel catalytic domain and a C-terminal jelly-roll domain. The enzyme exhibits high substrate specificity for p-nitrophenyl-α-L-arabinofuranoside and demonstrates optimal catalytic activity at 50°C and pH 8.0, suggesting its potential for industrial applications under moderate conditions. Notably, BpAbf51A specifically hydrolyzes the arabinofuranosyl moiety at the C-20 position of ginsenoside Rc to produce ginsenoside Rd. Molecular docking and two-dimensional interaction diagrams further revealed that the key amino acid residues, Ser213 and Asn214 of BpAbf51A, form strong hydrogen bonds with ginsenoside Rc. In this study, a novel α-L-arabinofuranosidase, BpAbf51A, has demonstrated significant potential for industrial applications in the production of rare saponins and other glycoside-based natural products, providing new research directions for the development of efficient biocatalysts.