Microscale preparation of even- and odd-numbered N-acetylheparosan oligosaccharides.
In order to prepare a series of N-acetylheparosan (NAH)-related oligosaccharides, bacterial NAH produced in Escherichia coli strain K5 was partially depolymerized with heparitinase I into a mixture of even-numbered NAH oligosaccharides, having an unsaturated uronic acid (DeltaUA) at the non-reducing end. A mixture of odd-numbered oligosaccharides was derived by removing this DeltaUA in the aforementioned mixture by a 'trimming' reaction using mercury(II) acetate. Each oligosaccharide mixture was subjected to gel-filtration chromatography to generate a series of size-uniform NAH oligosaccharides of satisfactory purity (assessed by analytical anion-exchange HPLC), and their structures were identified by MALDITOF-MS, ESIMS, and 1H NMR analysis. As a result, a microscale preparation of a series of both even- and odd-numbered NAH oligosaccharides was achieved for the first time. The developed procedure is simple and systematic, and thus, should be valuable for providing not only research tools for heparin/heparan sulfate-specific enzymes and their binding proteins, but also precursor substrates with medical applications.
Minamisawa
Toshikazu
T
Glycostructure Analysis Team, Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), Central-2, 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan.
Suzuki
Kiyoshi
K
Kajimoto
Naoko
N
Iida
Masami
M
Maeda
Hiroshi
H
Hirabayashi
Jun
J
D000117:Acetylglucosamine / Q000737:chemistry*; D020723:Glucuronic Acid / Q000737:chemistry; D006025:Glycosaminoglycans / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification
D006025:Glycosaminoglycans; C527794:N-acetylheparosan; D009844:Oligosaccharides; D020723:Glucuronic Acid; D000117:Acetylglucosamine