Substrate specificity and transfucosylation activity of GH29 α-L-fucosidases for enzymatic production of human milk oligosaccharides.

•Seven GH29 α-l-fucosidases all possess transfucosylation activity.•α-l-Fucosidase from F. graminearum catalyses formation of 2′FL from xyloglucan.•α-l-Fucosidase CpAfc2 from C. perfringens efficiently catalyses formation of LNFP II.•Systematic characterization of substrate specificity for seven GH29 α-l-fucosidases.