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Purification and Substrate-Specificity of β-Xylosidase from Sycamore Cell (Acer-Pseudoplatanus L.): Application for Structural Analysis of Xylose-Containing N-Linked Oligosaccharides

✍ Scribed by K. Tezuka; M. Hayashi; H. Ishihara; M. Nishimura; K. Onozaki; N. Takahashi


Publisher
Elsevier Science
Year
1993
Tongue
English
Weight
340 KB
Volume
211
Category
Article
ISSN
0003-2697

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✦ Synopsis


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.