PURIFICATION AND CHARACTERIZATION OF A COFFEA-CANEPHORA ALPHA-D-GALACTOSIDASE ISOZYME

被引:23
作者
HAIBACH, F [1 ]
HATA, J [1 ]
MITRA, M [1 ]
DHAR, M [1 ]
HARMATA, M [1 ]
SUN, P [1 ]
SMITH, D [1 ]
机构
[1] UNIV MISSOURI,DEPT CHEM,COLUMBIA,MO 65201
关键词
D O I
10.1016/0006-291X(91)92117-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Exoglycosidases modify carbohydrate epitopes on glycoproteins and glycolipids. The α-D-galactosidase from Coffea canephora is an important exoglycosidase which degrades the human blood group B epitope. Although multiple isozymes have been described, they have never been demonstrably purified and thoroughly characterized. We have developed a technique to purify an isozyme to homogeneity. The isolated enzyme has a molecular weight of 36.7 kDa by SDS PAGE and 34.0 kDa by gel filtration. The isozyme is highly selective for α-D-galactosides and inactive against other low molecular weight substrates. It hydrolyzes the the terminal α-D-galactosyl residue from the blood group B epitope. Protease activity is below detectable limits. The isozyme has a broad pH optima at 6.3, a pl of 7.03, is unaffected by ionic strength, and is stable at 4° C. © 1991.
引用
收藏
页码:1564 / 1571
页数:8
相关论文
共 18 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]  
BRYANT N, 1982, IMMUNOHEMATOLOGY, P280
[3]   ALPHA-GALACTOSIDASE AND BETA-GALACTOSIDASE ACTIVITIES IN PROTEIN BODIES AND CELL-WALLS OF LENTIL SEED COTYLEDONS [J].
CORCHETE, MP ;
GUERRA, H .
PHYTOCHEMISTRY, 1987, 26 (04) :927-932
[4]  
Courtois J.E., 1966, METHODS ENZYMOLOGY, P565
[5]  
DEAN K, 1977, PRACTICAL ENZYMOLOGY, P202
[6]  
Flowers H M, 1979, Adv Enzymol Relat Areas Mol Biol, V48, P29
[7]  
GOLDSTEIN JUDITH L., 1965, PHYTOCHEMISTRY, V4, P185, DOI 10.1016/S0031-9422(00)86162-2
[8]   THE EFFECT OF GLYCOSIDASES ON THE SURVIVAL OF RAT ERYTHROCYTES IN CIRCULATION [J].
GUTOWSKI, KA ;
LINSEMAN, DA ;
AMINOFF, D .
CARBOHYDRATE RESEARCH, 1988, 178 :307-313
[9]  
Hagerman A. E., 1989, CHEM SIGNIFICANCE CO, P323, DOI DOI 10.1007/978-1-4684-7511-1_20
[10]   STUDIES ON B-ANTIGENIC SITES OF HUMAN ERYTHROCYTES BY USE OF COFFEE BEAN ALPHA-GALACTOSIDASE [J].
HARPAZ, N ;
FLOWERS, HM ;
SHARON, N .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1975, 170 (02) :676-683