Modification of di- and tetrasaccharides from shark cartilage keratan sulphate by refined anhydromethanolic hydrochloric acid-treatments and evaluation of their specific desulphation.
Highly sulphated keratan di- and tetrasaccharides were prepared from keratan sulphate (KS) of shark cartilage by enzymatic digestion with keratanase II and subsequent chromatography. The tetrasaccharide fraction carrying four sulphate groups was completely desulphated by 100 mM anhydromethanolic hydrochloric acid (MeOH-HCl) treatment at room temperature for 16 h. The conditions for the desulphation reaction by MeOH-HCl treatment were examined using sulphated keratan di- and tetrasaccharides as substrates by means of reversed phase high performance liquid chromatography (HPLC) and/or capillary electrophoresis, followed by the preparation of partially desulphated keratan oligosaccharides. Sulphate substitution patterns of monosulphated keratan disaccharide and trisulphated keratan tetrasaccharide were evaluated by methylation analysis. The results suggested that 6-O-sulphate groups of Gal moieties are cleaved faster than those of GlcNAc moieties under the present conditions adopted for the MeOH-HCl treatment of KS-derived oligosaccharides.
Kariya
Yutaka
Y
Central Research Laboratories, Seikagaku Corporation, 3-1253 Tateno, Higashiyamato, Tokyo 207-0021, Japan. kariya@seikagaku.co.jp
Watabe
Shugo
S
Mochizuki
Hideo
H
Imai
Kyoko
K
Kikuchi
Hiroshi
H
Suzuki
Kiyoshi
K
Kyogashima
Mamoru
M
Ishii
Tadashi
T
D000118:Acetylglucosaminidase / Q000378:metabolism; D000818:Animals; D002356:Cartilage / Q000737:chemistry* / Q000201:enzymology; D006851:Hydrochloric Acid / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007632:Keratan Sulfate / Q000737:chemistry* / Q000378:metabolism; D000432:Methanol / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D012754:Sharks; D013431:Sulfates / Q000378:metabolism*; D013997:Time Factors
D009844:Oligosaccharides; D013431:Sulfates; D007632:Keratan Sulfate; C086950:keratanase II; D000118:Acetylglucosaminidase; D006851:Hydrochloric Acid; D000432:Methanol