Aims: This study focused on the cloning, expression and characterization of recombinant alpha-L-arabinosidases from Bifidobacterium longum H-1. Methods and Results: alpha-L-Arabinofuranosidase (AfuB-H1) and bifunctional alpha-L-arabinopyranosidase/beta-D-galactosidase (Apy-H1) from B. longum H-1 were identified by Southern blotting, and their recombinant enzymes were overexpressed in Escherichia coli BL21 (DE3). Recombinant AfuB-H1 (rAfuB-H1) was purified by single-step Ni(2+)-affinity column chromatography, whereas recombinant Apy-H1 (rApy-H1) was purified by serial Q-HP and Ni(2+)-affinity column chromatography. Enzymatic properties and substrate specificities of the two enzymes were assessed, and their kinetic constants were calculated. According to the results, rAfuB-H1 hydrolysed p-nitrophenyl-alpha-L-arabinofuranoside (pNP-alpha L-Af) and ginsenoside Rc, but did not hydrolyse p-nitrophenyl-alpha-L-arabinopyranoside (pNP-alpha L-Ap). On the other hand, rApy-H1 hydrolysed pNP-alpha L-Ap, p-nitrophenyl-beta-D-galactopyranoside (pNP-beta D-Ga) and ginsenoside Rb2. Conclusions: Ginsenoside-metabolizing bifidobacterial rAfuB-H1 and rApy-H1 were successfully cloned, expressed, and characterized. rAfuB-H1 specifically recognized the alpha-L-arabinofuranoside, whereas rApy-H1 had dual functions, that is, it could hydrolyse both beta-D-galactopyranoside and alpha-L-arabinopyranoside. Significance and Impact of the Study: These findings suggest that the biochemical properties and substrate specificities of these recombinant enzymes differ from those of previously identified alpha-L-arabinosidases from Bifidobacterium breve K-110 and Clostridium cellulovorans.