A new alpha-arabinofuranosidase gene (Hiabf43) was cloned from Humicola insolens Y1 and successfully expressed in Pichia pastoris GS115. Deduced HiAbf43 contained a putative signal peptide and a catalytic domain of glycoside hydrolase (GH) family 43. Purified recombinant HiAbf43 showed optimal activity at pH 5.0 and 50 degrees C, and was active over a broad pH range. The enzyme was specific for the cleavage of alpha-1,3-linkage and showed high activity against 4-nitrophenyl alpha-L-arabinofuranoside, debranched arabinan, and sugar beet arabinan. Sequential addition of HiAbf43 followed by Xyn11A increased the degradation efficiency of birchwood and beechwood xylans but not wheat arabinoxylan. The synergy degree was high up to 1.21-fold.