Members of the genus
Bifidobacterium
are commonly found in the human gut and are known to utilize complex carbohydrates that are indigestible by the human host. Members of the
Bifidobacterium longum
subsp.
longum
taxon can metabolize various plant-derived carbohydrates common to the human diet. To metabolize such polysaccharides, which include arabinoxylan, bifidobacteria need to encode appropriate carbohydrate-active enzymes in their genome. In the current study, we describe two GH43 family enzymes, denoted here as AxuA and AxuB, which are encoded by
B. longum
subsp.
longum
NCIMB 8809 and are shown to be required for cereal-derived arabinoxylan metabolism by this strain. Based on the observed hydrolytic activity of AxuA and AxuB, assessed by employing various synthetic and natural substrates, and based on
in silico
analyses, it is proposed that both AxuA and AxuB represent extracellular α-L-arabinofuranosidases with distinct substrate preferences. The variable presence of the
axuA
and
axuB
genes and other genes previously described to be involved in the metabolism of arabinose-containing glycans can in the majority cases explain the (in)ability of individual
B. longum
subsp.
longum
strains to grow on cereal-derived arabinoxylans and arabinan.