Takahashi N, Matsuda T, Shikami K, Shimada I, Arata Y, Nakamura R
GlycoLab, Nakano Central Research Institute, Nakano Vinegar Co. Ltd, Handa-City, Japan.
Glycoconj J. 1993 Dec;10(6):425-34. doi: 10.1007/BF00737962.
Asparagine-linked oligosaccharides of duck ovomucoid were released quantitatively from the protein by digestion with glycoamidase A (from almond), the reducing ends of the oligosaccharide chains thus obtained were aminated with a fluorescent reagent, 2-aminopyridine, and the mixture of pyridylamino derivatives of the oligosaccharides was separated using two different types of high performance liquid chromatography (HPLC) on a reversed phase column and an amide adsorption column. More than sixteen different oligosaccharides were separated and the structures were characterized by a combination of the 2-dimensional sugar mapping technique using HPLC, exoglycosidase digestion, and proton nuclear magnetic resonance measurements (1- and 2-dimensional). Furthermore, the HPLC profile of duck ovomucoid oligosaccharides was compared with previously reported profiles obtained from quail and chicken ovomucoids.
用杏仁中的糖酰胺酶A将鸭卵类粘蛋白的天冬酰胺连接的寡糖从蛋白质中定量释放出来,用荧光试剂2-氨基吡啶对如此得到的寡糖链的还原端进行胺化,并用两种不同类型的高效液相色谱(HPLC)在反相柱和酰胺吸附柱上分离寡糖的吡啶基氨基衍生物混合物。分离出了十六种以上不同的寡糖,并用高效液相色谱二维糖图谱技术、外切糖苷酶消化和质子核磁共振测量(一维和二维)相结合的方法对其结构进行了表征。此外,还将鸭卵类粘蛋白寡糖的高效液相色谱图谱与先前报道的鹌鹑和鸡卵类粘蛋白的图谱进行了比较。