A new endo-beta-galactosidase acting on the Gal beta 1-3Gal linkage of the proteoglycan linkage region.
A new type of endo-beta-galactosidase acting on the linkage region of peptidochondroitin sulfate was isolated from the mid-gut gland of the mollusk Patinopecten. The purification procedure included ammonium sulfate precipitation, Sephacryl S-200HR gel filtration, DEAE-Sephacel chromatography, and TSKgel Phenyl-5PW RP high performance liquid chromatography. The purified enzyme was free from exoglycosidases, sulfatases, and phosphatases. The specificity of the enzyme was as follows. 1) It acted on the internal galactoside linkage of sugar chains; 2) it specifically hydrolyzed the galactosylgalactose (Gal beta 1-3Gal) linkage, but not the galactosylxylose (Gal beta 1-4Xyl) linkage in the linkage region of peptidoglycans; 3) the enzyme activity was unaffected by the type of glycosaminoglycan, chondroitin sulfate, dermatan sulfate or heparan sulfate used as a substrate; 4) keratan sulfate and some oligosaccharides from glycolipid were not degraded by the enzyme. These properties of the endo-beta-galactosidase characterize it as a new endo-beta-galactosidase with unique specificity.
Takagaki
K
K
Department of Biochemistry, Hirosaki University School of Medicine, Japan.
Nakamura
T
T
Takeda
Y
Y
Daidouji
K
K
Endo
M
M
D000818:Animals; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002854:Chromatography, Paper; D004064:Digestive System / Q000201:enzymology*; D004187:Disaccharides / Q000378:metabolism*; D006026:Glycoside Hydrolases; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008974:Mollusca; D011509:Proteoglycans / Q000378:metabolism*; D013379:Substrate Specificity; D001616:beta-Galactosidase / Q000302:isolation & purification / Q000378:metabolism*
D004187:Disaccharides; D011509:Proteoglycans; C055075:galactosyl-(1-3)galactose; D006026:Glycoside Hydrolases; C026093:keratan-sulfate endo-1,4-beta-galactosidase; D001616:beta-Galactosidase