Purification and substrate specificity of beta-xylosidase from sycamore cell (Acer pseudoplatanus L.): application for structural analysis of xylose-containing N-linked oligosaccharides.
A beta-xylosidase was purified 51-fold from culture medium of sycamore (Acer pseudoplatanus L.) cells using p-nitrophenyl beta-D-xylopyranoside as a substrate. This enzyme can remove a xylose residue from asparagine-linked oligosaccharides, derivatized with 2-aminopyridine. A pentasaccharide, Xy1 beta 2Man beta 4GlcNAc beta 4(Fuc-alpha 3)GlcNAc was the favorite substrate in N-linked oligosaccharides, but a xylose residue in Xy1 beta 2(Man-alpha 3)Man beta sequence could not be removed by the enzyme. We also propose an efficient method for detection of xylose residue in N-linked oligosaccharides by a combination of the two-dimensional sugar mapping technique and the xylosidase digestion.
Tezuka
K
K
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Japan.
Hayashi
M
M
Ishihara
H
H
Nishimura
M
M
Onozaki
K
K
Takahashi
N
N
D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D059828:Plant Cells; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D010944:Plants / Q000201:enzymology*; D013379:Substrate Specificity; D014994:Xylose / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D014995:Xylosidases / Q000302:isolation & purification* / Q000378:metabolism*
D009844:Oligosaccharides; D010940:Plant Proteins; D014994:Xylose; D014995:Xylosidases; C026579:exo-1,4-beta-D-xylosidase