Preparation of GM1 ganglioside with sialidase-producing marine bacteria as a microbial biocatalyst.
This paper describes the preparation of monosialoganglioside GM1 with sialidase-producing marine bacteria as a microbial biocatalyst. A new sialidase-producing bacterium, identified tentatively as Pseudomonas sp. strain YF-2, was isolated from seawater by enrichment culture with ganglioside as the sole source of carbon. When YF-2 was cultured in a synthetic medium containing crude bovine brain gangliosides at 25 degrees C for 3 days, 80 to 90% of the gangliosides were converted to GM1. GM1 was then purified from the supernatant of YF-2 culture by C18 reverse-phased chromatography, followed by DEAE-Sephadex A25 anion-exchange chromatography. In a typical experiment, 178 mg of highly purified GM1 was obtained from 500 mg of the crude ganglioside fraction. The GM1 induced neurite outgrowth of neuroblastoma Neuro2a cells at a concentration of 33 to 100 microM in the presence of fetal calf serum. Sialidase was purified 33-fold with 13.3% recovery from the culture supernatant of YF-2. The purified enzyme hydrolyzed polysialogangliosides to produce GM1 but did not act on GM1. It was therefore concluded that polysialogangliosides in the culture of strain YF-2 were converted to GM1 by this sialidase.
Fukano
Y
Y
Laboratory of Marine Biochemistry, Faculty of Agriculture Kyushu University, Fukuoka, Japan.
Ito
M
M
D000818:Animals; D001681:Biological Assay; D002417:Cattle; D002852:Chromatography, Ion Exchange; D005732:Gangliosides / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D013058:Mass Spectrometry; D009439:Neuraminidase / Q000302:isolation & purification / Q000378:metabolism; D016501:Neurites / Q000187:drug effects; D011549:Pseudomonas / Q000302:isolation & purification / Q000378:metabolism*; D012623:Seawater; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014871:Water Microbiology
D005732:Gangliosides; C025447:sialogangliosides; C016481:trisialoganglioside GT1; D009439:Neuraminidase