Enzyme actions on the blood group A- and B-active oligosaccharides isolated from A and B substances.

Rege et al.l)'2) and Painter et al.3~ isolated blood group-active f ucose-containing oligosaccharides by treating blood group substances in human ovarian cyst fluids with triethylamine. Lloid et al.4~ isolated f ucose-containing oligosaccharides by decomposing A and H substances from hog stomac mucin and A, B and H substances from human ovarian cyst fluids with alkaline borohydride. The present report concerns the actions of blood group substancedecomposing enzymes on the Aand B-active oligosaccharides isolated from A and B substances. Materials and methods. 1) Blood group substances. These were obtained from human stomach linings by the pepsin digestion method and purified by the phenol extraction method. 2) Enzyme samples. A-decomposing enzyme from Clostridium tertium A was prepared by the method of Yamamoto and Iseki,5~ A-decomposing enzyme from hog liver by the method of Iseki and Yamamoto,°~ Bdecomposing enzyme from Clostridium sporo genes Maebashi by the method of Iseki et a1.,7~ and H-decomposing enzyme from Bacillus f ulminans and Bacillus cereus H by the method of Iseki et al.8 Besides, a-galactosidase9~ from coffee bean was used. 3) Antisera. Eel serum was used as anti-H serum. In addition anti-Shigella dysenteriae chicken serum and anti-Diplococcus pneumoniae Type XIV chicken serum (anti-Pn XIV serum) were used. 4) Isolation and purification of oligosaccharides. Blood group substances were treated with triethylamine according to the method of Rege et al.' Oligosaccharides were isolated by Sephadex G-10 or G-15 column chromatography, paper chromatography and paper electrophoresis. Paper chromatography was performed by the descending method with the following solvents : Solvent I, n-butanol : pyridine: water (6: 4 : 3) ; solvent II, n-butanol : acetic acid; water (4: 1: 5) ; solvent III, ethylacetate : pyridine: water (10: 4 : 3) ; solvent IV, ethylacetate : acetic acid: formic acid: water (18: 4: 1: 3). Paper electro-