   Recombinant enzyme with no detectable endoglycosidase or other exoglycosidase contaminating

      activities

   Glycerol -free for optimal performance in HPLC and mass spectrometry analysis

   ≥95% purity, as determined by SDS-PAGE and intact ESI-MS

α1-2,4,6 Fucosidase O is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal

α1-2, α1-4 and α1-6 linked fucose residues from oligosaccharides. α1-2,4,6 Fucosidase O cleaves

α1-6 fucose residues more efficiently than other linkages.

Cloned from Omnitrophica bacterium and expressed in E. coli ⁽¹⁾.

1X GlycoBuffer 1.  Incubate at 37°C.

Heat Inactivation: 65°C for 10 min

10X GlycoBuffer 1 

1X GlycoBuffer 1
5 mM CaCl2
50 mM sodium acetate
(pH 5.5 @ 25°C)

Apparent: 49 kDa

One unit is defined as the amount of enzyme required to cleave > 95% of the fucose from 1 nmol of G0F

from human IgG [GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc(Fucα1-6)-

AMAC], in 1 hour at 37°C in a total reaction volume of 10 µl. 

4°C

1.   Reactions may be scaled-up linearly to accommodate larger reaction volumes.
2.   The enzymatic unit definition is calculated using a 37°C reaction temperature, to accommodate for

      simultaneous glycosidase digestions and N-glycan sequencing protocols. However, the optimal

      reaction temperature for α1-2,4,6 Fucosidase O is 50°C. A 37°C reaction temperature results in

      ~35% lower activity.
3.   In order to achieve complete removal of core α1-6 fucose residues from glycans with instant labels,

      it may be necessary to use longer incubation times (18 hours).
4.   α1-2,4,6 Fucosidase O is only active on an intact antibody if it is trimmed to the trimannosyl core.

1.   Vainauskas, S., New England Biolabs, Inc. Unpublished observation
2.   Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.