Recombinant Production and Characterization of a Novel ?-L-Fucosidase from Bifidobacterium castoris

alpha-L-fucosidases (EC 3.2.1.51) are of particular interest due to their ability to cleave terminal alpha-L-fucose residues from glycoconjugates, a property associated with numerous biological and therapeutic effects. They have also been investigated for their potential use in glycan remodeling, disease biomarker analysis, and particularly as therapeutic agents in the context of fucosidosis, a rare lysosomal storage disorder, caused by a deficiency in alpha-L-fucosidase activity. However, limitations in enzyme availability, stability, and substrate specificity highlight the need for novel and more efficient enzyme sources. Bifidobacterium castoris (B. castor is) is a newly identified species first discovered in the beaver gut microbiota in 2019. Phylogenetic studies have revealed its advanced metabolic capacity, and genomic analyses have demonstrated its extensive carbohydrate metabolism potential. This research article focuses on the recombinant production and biochemical characterization of a novel alpha-L-fucosidase from B. castoris LMG (Laboratorium voor Microbiologie Gent) 30937, predicted to belong to glycoside hydrolase family 29 (GH29) according to Universal Protein Resource (UniProt) annotation. Under optimized reaction conditions the recombinant alpha-L-fucosidase exhibited a specific activity of 0.264 U/mg to pNP-Fuc (4-Nitrophenyl-alpha-L-fucopyranoside). The results indicate that the enzyme is active in the pH range of 3.0-8.0 and temperatures of 24-42 degrees C, but its optimum conditions are the slightly acidic pH of 5.5 and the elevated temperature of 42 degrees C. This profile suggests that the enzyme is adapted to acidic intestinal-like environments. This novel enzyme expands the GH29 alpha-L-fucosidase repertoire and offers a promising new candidate for future biotechnological applications.