(Amersham Biosciences) and Superdex 200 Hiload 26/60 gel
filtration columns (Amersham Biosciences). The samples were
injected in 5 cm3 volume of 100 mM Tris-HCl buffer pH 7.5 used
for the run, and 10 cm3 fractions were collected for subsequent
steps. CentriPrep YM-10 centrifugal filter devices (Biorad) were
used for fraction concentration.
and trifluoroethyl butanoate (0.100 M) in TBME. The reactions
were analyzed by GC equipped with a chiral a column (Varian
permethylated b-cyclodextrin).
Acknowledgements
Removal of glycosyl groups. Removal of potential N-linked
sugars was accomplished using commercial glycosidase PNGase
F (New England Biolabs) which cleaves between the innermost
GlNAc and asparagine residues of high mannose, hybrid and com-
plex oligosaccharides from N-linked glycoproteins. The procedure
was carried out as instructed by the manufacturer.
The authors thank the Technology Development Center of
Finland (TEKES) for financial support and ViaZym for ViaKit
containing immobilized CAL-A preparations.
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894 | Org. Biomol. Chem., 2010, 8, 886–895
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