title: Fucosyl-GM1a, an endoglycoceramidase-resistant ganglioside of porcine brain.
abstract: The use of bovine brain has been prohibited in many countries because of the world-wide prevalence of mad cow disease, and thus porcine brain is expected to be a new source for the preparation of gangliosides. Here, we report the presence of a ganglioside in porcine brain which is strongly resistant to hydrolysis by endoglycoceramidase, an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides of various glycosphingolipids. Five major gangliosides (designated PBG-1, 2, 3, 4, 5) were extracted from porcine brain by Folch's partition, followed by mild alkaline hydrolysis and PBA column chromatography. We found that PBG-2, but not the others, was strongly resistant to hydrolysis by the enzyme. After the purification of PBG-2 with Q-Sepharose, Silica gel 60 and Prosep-PB chromatographies, the structure of PBG-2 was determined by GC, GC-MS, FAB-MS and NMR spectroscopy as Fucalpha1-2Galbeta1-3GalNAcbeta1-4(NeuAcalpha2-3)Galbeta1-4Glcbeta1-1'Cer (fucosyl-GM1a). The ceramide was mainly composed of C18:0 and C20:0 fatty acids and d18:1 and d20:1 sphingoid bases. The apparent kcat/Km for fucosyl-GM1a was found to be 30 times lower than that for GM1a, indicating that terminal fucosylation makes GM1a resistant to hydrolysis by the enzyme. This report indicates the usefulness of endoglycoceramidase to prepare fucosyl-GM1a from porcine brain.
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan.
authors.forename: Xu
authors.initials: X
authors.lastname: Xu
authors.forename: Hatsumi
authors.initials: H
authors.lastname: Monjusho
authors.forename: Masanori
authors.initials: M
authors.lastname: Inagaki
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Hama
authors.forename: Kuniko
authors.initials: K
authors.lastname: Yamaguchi
authors.forename: Keishi
authors.initials: K
authors.lastname: Sakaguchi
authors.forename: Masao
authors.initials: M
authors.lastname: Iwamori
authors.forename: Nozomu
authors.initials: N
authors.lastname: Okino
authors.forename: Makoto
authors.initials: M
authors.lastname: Ito
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D005227:Fatty Acids / Q000032:analysis; D005677:G(M1) Ganglioside / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D007700:Kinetics; D009439:Neuraminidase / Q000378:metabolism; D013552:Swine
chemical_list: D005227:Fatty Acids; D005677:G(M1) Ganglioside; C039123:fucosyl GM1 ganglioside; D006026:Glycoside Hydrolases; C050427:endoglycoceramidase; D009439:Neuraminidase