Preparative method of functional oligasaccharides from bovine colostrum

初乳取自一头分娩后6小时的荷斯坦-弗里西亚奶牛。采用Schneir和Rafelson法对脱脂初乳进行透析分离。通过我们之前的方法，用Sp-Sephadex C-50离子交换层析实现了从透析液中完全去除多肽。采用凝胶过滤层析法（Bio-gel P-2），以蒸馏水为洗脱液对乳糖结晶去除后剩余的糖进行分离。另一方面，用NaB <H_4>的NaOH在<N_2>的黑暗条件下，50℃下处理36h，然后用Dowex 50W-X4树脂去除低分子肽和盐。最后以蒸馏水为洗脱液，用凝胶过滤层析（Biogel P-2）纯化所得低聚糖醇。采用Farrar等人的离子交换色谱法，在0.05 ~ 1.0 m的醋酸吡啶缓冲液线性浓度梯度下对纯化的低聚糖醇进行分离。

Colostrum was taken from a Holstein-Friesian cow 6h after parturition. Isolation of milk sugars from defatted colostrum by dialysis was fermormed by the method of Schneir and Rafelson. The complete removal of peptides from the dialyzate was achieved by ion-exchange chromatography with Sp-Sephadex C-50 by our earlier method. The sugars remaining after removal of lactose by crystallization were fractionated by gel filtration chromatography(Bio-gel P-2) with distilled water as eluent. On the otherhand, the colostrum glycopeptide was treated with NaOH of NaB <H_4> for 36h at 50 C in the dark under <N_2> , followed by removal of low-molecular peptides and salts using Dowex 50W-X4 resin. The resulting oligosacchariditols were finally purified by gel filtration chromatography(Biogel P-2) with distilled water as eluent. The purified oligosaccharditols were fractionated by ion-exchange chromatography using the method of Farrar et al in a linear concentration gradient of 0.05-1.0M of pyridine-acetate buffer.