Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 May;63(5):1861–1865. doi: 10.1128/aem.63.5.1861-1865.1997

Preparation of GM1 ganglioside with sialidase-producing marine bacteria as a microbial biocatalyst.

Y Fukano 1, M Ito 1
PMCID: PMC168478  PMID: 9143118

Abstract

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.

Full Text

The Full Text of this article is available as a PDF (720.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ADA G. L., FRENCH E. L., LIND P. E. Purification and properties of neuraminidase from Vibrio cholerae. J Gen Microbiol. 1961 Mar;24:409–425. doi: 10.1099/00221287-24-3-409. [DOI] [PubMed] [Google Scholar]
  2. Argentino C., Sacchetti M. L., Toni D., Savoini G., D'Arcangelo E., Erminio F., Federico F., Milone F. F., Gallai V., Gambi D. GM1 ganglioside therapy in acute ischemic stroke. Italian Acute Stroke Study--Hemodilution + Drug. Stroke. 1989 Sep;20(9):1143–1149. doi: 10.1161/01.str.20.9.1143. [DOI] [PubMed] [Google Scholar]
  3. Arita M., Iwamori M., Higuchi T., Nagai Y. Negative ion fast atom bombardment mass spectrometry of gangliosides and asialo gangliosides: a useful method for the structural elucidation of gangliosides and related neutral glycosphingolipids. J Biochem. 1983 Jul;94(1):249–256. doi: 10.1093/oxfordjournals.jbchem.a134336. [DOI] [PubMed] [Google Scholar]
  4. Bremer E. G., Hakomori S., Bowen-Pope D. F., Raines E., Ross R. Ganglioside-mediated modulation of cell growth, growth factor binding, and receptor phosphorylation. J Biol Chem. 1984 Jun 10;259(11):6818–6825. [PubMed] [Google Scholar]
  5. Bremer E. G., Schlessinger J., Hakomori S. Ganglioside-mediated modulation of cell growth. Specific effects of GM3 on tyrosine phosphorylation of the epidermal growth factor receptor. J Biol Chem. 1986 Feb 15;261(5):2434–2440. [PubMed] [Google Scholar]
  6. Cassidy J. T., Jourdian G. W., Roseman S. The sialic acids. VI. Purification and properties of sialidase from Clostridium perfringens. J Biol Chem. 1965 Sep;240(9):3501–3506. [PubMed] [Google Scholar]
  7. Frieder B., Rapport M. M. The effect of antibodies to gangliosides on Ca2+ channel-linked release of gamma-aminobutyric acid in rat brain slices. J Neurochem. 1987 Apr;48(4):1048–1052. doi: 10.1111/j.1471-4159.1987.tb05625.x. [DOI] [PubMed] [Google Scholar]
  8. Geisler F. H., Dorsey F. C., Coleman W. P. Recovery of motor function after spinal-cord injury--a randomized, placebo-controlled trial with GM-1 ganglioside. N Engl J Med. 1991 Jun 27;324(26):1829–1838. doi: 10.1056/NEJM199106273242601. [DOI] [PubMed] [Google Scholar]
  9. HUGH R., LEIFSON E. The taxonomic significance of fermentative versus oxidative metabolism of carbohydrates by various gram negative bacteria. J Bacteriol. 1953 Jul;66(1):24–26. doi: 10.1128/jb.66.1.24-26.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ito M., Kurita T., Kita K. A novel enzyme that cleaves the N-acyl linkage of ceramides in various glycosphingolipids as well as sphingomyelin to produce their lyso forms. J Biol Chem. 1995 Oct 13;270(41):24370–24374. doi: 10.1074/jbc.270.41.24370. [DOI] [PubMed] [Google Scholar]
  11. KOVACS N. Identification of Pseudomonas pyocyanea by the oxidase reaction. Nature. 1956 Sep 29;178(4535):703–703. doi: 10.1038/178703a0. [DOI] [PubMed] [Google Scholar]
  12. Ledeen R. W. Biology of gangliosides: neuritogenic and neuronotrophic properties. J Neurosci Res. 1984;12(2-3):147–159. doi: 10.1002/jnr.490120204. [DOI] [PubMed] [Google Scholar]
  13. Nakamura K., Handa S. Coomassie brilliant blue staining of lipids on thin-layer plates. Anal Biochem. 1984 Nov 1;142(2):406–410. doi: 10.1016/0003-2697(84)90484-6. [DOI] [PubMed] [Google Scholar]
  14. Nojiri H., Stroud M., Hakomori S. A specific type of ganglioside as a modulator of insulin-dependent cell growth and insulin receptor tyrosine kinase activity. Possible association of ganglioside-induced inhibition of insulin receptor function and monocytic differentiation induction in HL-60 cells. J Biol Chem. 1991 Mar 5;266(7):4531–4537. [PubMed] [Google Scholar]
  15. Partington C. R., Daly J. W. Effect of gangliosides on adenylate cyclase activity in rat cerebral cortical membranes. Mol Pharmacol. 1979 May;15(3):484–491. [PubMed] [Google Scholar]
  16. Paulson J. C., Weinstein J., Dorland L., van Halbeek H., Vliegenthart J. F. Newcastle disease virus contains a linkage-specific glycoprotein sialidase. Application to the localization of sialic acid residues in N-linked oligosaccharides of alpha 1-acid glycoprotein. J Biol Chem. 1982 Nov 10;257(21):12734–12738. [PubMed] [Google Scholar]
  17. SVENNERHOLM L. CHROMATOGRAPHIC SEPARATION OF HUMAN BRAIN GANGLIOSIDES. J Neurochem. 1963 Sep;10:613–623. doi: 10.1111/j.1471-4159.1963.tb08933.x. [DOI] [PubMed] [Google Scholar]
  18. Schneider J. S., Roeltgen D. P., Rothblat D. S., Chapas-Crilly J., Seraydarian L., Rao J. GM1 ganglioside treatment of Parkinson's disease: an open pilot study of safety and efficacy. Neurology. 1995 Jun;45(6):1149–1154. doi: 10.1212/wnl.45.6.1149. [DOI] [PubMed] [Google Scholar]
  19. Svennerholm L. Gangliosides--a new therapeutic agent against stroke and Alzheimer's disease. Life Sci. 1994;55(25-26):2125–2134. doi: 10.1016/0024-3205(94)00393-9. [DOI] [PubMed] [Google Scholar]
  20. Uchida Y., Tsukada Y., Sugimori T. Enzymatic properties of neuraminidases from Arthrobacter ureafaciens. J Biochem. 1979 Nov;86(5):1573–1585. doi: 10.1093/oxfordjournals.jbchem.a132675. [DOI] [PubMed] [Google Scholar]
  21. Waki H., Murata A., Kon K., Maruyama K., Kimura S., Ogura H., Ando S. Isolation and characterization of a trisialyllactosylceramide, GT3, containing an O-acetylated sialic acid in cod fish brain. J Biochem. 1993 Apr;113(4):502–507. doi: 10.1093/oxfordjournals.jbchem.a124073. [DOI] [PubMed] [Google Scholar]
  22. Wu G., Ledeen R. W. Stimulation of neurite outgrowth in neuroblastoma cells by neuraminidase: putative role of GM1 ganglioside in differentiation. J Neurochem. 1991 Jan;56(1):95–104. doi: 10.1111/j.1471-4159.1991.tb02567.x. [DOI] [PubMed] [Google Scholar]
  23. Zheng M., Fang H., Tsuruoka T., Tsuji T., Sasaki T., Hakomori S. Regulatory role of GM3 ganglioside in alpha 5 beta 1 integrin receptor for fibronectin-mediated adhesion of FUA169 cells. J Biol Chem. 1993 Jan 25;268(3):2217–2222. [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES